(12) Patent Application Publication (48) Pub

Home , Ecadotril, Isovaline, Kyotorphin, Trimebutine

US 20120283411A9 (19) United States (10) Pub. No.: US 2012/0283411 A9 (12) Patent Application Publication (48) Pub. Date: Nov. 8, 2012 Currie et al. CORRECTED PUBLICATION

(54) METHODS AND COMPOSITIONS FOR THE Related U.S. Application Data TREATMENT OF GASTROINTESTINAL (60) Provisional application No. 60/817.843, filed on Jun. DISORDERS 29, 2006. (75) Inventors: Mark G. Currie, Sterling, MA Publication Classification (US); Daniel P. Zimmer, (51) Int. Cl. Somerville, MA (US) C07K 14/00 (2006.01) C07K 7/08 (2006.01) (73) Assignee: IRONWOOD C07K 7/06 (2006.01) PHARMACEUTICALS, INC., (52) U.S. Cl...... 530/324; 530/328; 530/327: 530/326; Cambridge, MA (US) 530/325 (57) ABSTRACT (21) Appl. No.: 12/306,788 Compositions and related methods for treating IBS and other gastrointestinal disorders and conditions (e.g., gastrointesti (22) PCT Filed: Jun. 27, 2007 nal motility disorders, functional gastrointestinal disorders, gastroesophageal reflux disease (GERD), duodenogastric reflux, Crohn's disease, ulcerative colitis, inflammatory (86) PCT No.: PCT/US07/72223 bowel disease, functional heartburn, dyspepsia (including functional dyspepsia or nonulcer dyspepsia), gastroparesis, S371 (c)(1), chronic intestinal pseudo-obstruction (or colonic pseudoob (2), (4) Date: Nov. 9, 2009 struction), disorders and conditions associated with constipa tion, e.g., constipation associated with use of opiate pain Prior Publication Data killers, post-surgical constipation, and constipation associ ated with neuropathic disorders and disorders and conditions (15) Correction of US 2012/0088902 A1 Apr. 12, 2012 associated with excess fluid and/or salt retention as well as See Claims 1, 5, 7, and 12. other conditions and disorders are described. The composi tions feature polypeptides that activate the guanylate cyclase (65) US 2012/0088902 A1 Apr. 12, 2012 C (GC-C) receptor.

: - 3×3 espp.

::::::::: : 43.84.82&3.

Patent Application Publication Nov. 8, 2012 Sheet 2 of 5 US 2012/0283411 A9

YYYYYYYYYYYYYYYaaalaaaaaaaaaa %% &########

----

§############3

{ || ––– ------| Siz Patent Application Publication Nov. 8, 2012 Sheet 3 of 5 US 2012/0283411 A9

:::::::::::"?????????

?,ø38843

------~--~~~~~~~~~--~~~~~~~~~~~~~~~~~~~~~~$2~~~~~~~~~~~~.~~~~~~--~~~~--~~~~------3---

SS Patent Application Publication Nov. 8, 2012 Sheet 4 of 5 US 2012/0283411 A9

}

~~~~-4------~--~--~~~~

gy Patent Application Publication Nov. 8, 2012 Sheet 5 of 5 US 2012/0283411 A9

¿???????%

------•••••••••••••••••••••••••••••••••••••••••••+~~~~~~#~~~~·······

------.-.-.-………>.>--~~------~--~~~~…………………………………~~~~},~~~~~

G/9 US 2012/0283411 A9 Nov. 8, 2012

METHODS AND COMPOSITIONS FOR THE three interacting mechanisms: altered bowel motility, an TREATMENT OF GASTRONTESTINAL increased sensitivity of the intestine or colon to pain stimuli DISORDERS (visceral sensitivity) and psychosocial factors (Camilleri 2001, Gastroenterology 120:652-668). Recently, there has TECHNICAL FIELD been increasing evidence for a role of inflammation in etiol ogy of IBS. Reports indicate that subsets of IBS patients have 0001. This invention relates to methods and compositions Small but significant increases in colonic inflammatory and for treating gastrointestinal disorders, obesity, congestive mast cells, increased inducible nitric oxide (NO)and synthase heart failure, benign prostatic hyperplasia (BPH) and other (iNOS) and altered expression of inflammatory cytokines disorders. (reviewed by Talley 2000, Medscape Coverage of DDW week). BACKGROUND 0005. The guanylate cyclase-C (GC-C) receptor (re 0002 Irritable bowel syndrome (IBS) is a common viewed by Lucas et al. 2000 Pharmacol. Rev 52:375-414 and chronic disorder of the intestine that affects 20 to 60 million Vaandrager et al. 2002 Molecular and Cellular Biochemistry individuals in the US alone (Lehman Brothers, Global 230:73-83) is a key regulator in mammals of intestinal func Healthcare-Irritable Bowel Syndrome Industry Update, Sep tion (although low levels of GC-C have been detected in other tember 1999). IBS is the most common disorder diagnosed by tissues). GC-C responds to the endogenous hormones, gua gastroenterologists (28% of patients examined) and accounts nylin and uroguanylin, and to enteric bacterial polypeptides for 12% of visits to primary care physicians (Camilleri 2001 from the heat stable enterotoxin family (ST polypeptides). Gastroenterology 120:652-668). In the US, the economic When an agonist binds to GC-C, there is an elevation of the impact of IBS is estimated at $25 billion annually, through second messenger, cyclic GMP, and an increase in chloride direct costs of health care use and indirect costs of absentee and bicarbonate secretion, resulting in an increase in intesti ism from work (Talley 1995 Gastroenterology 109:1736 nal fluid secretion. The Genbank protein GI accession num 1741). Patients with IBS have three times more absenteeism ber for guanylyl cyclase C homologs from multiple organ from work and report a reduced quality of life. Sufferers may isms are: be unable or unwilling to attend social events, maintain employment, or travel even short distances (Drossman 1993 Dig Dis Sci 38:1569-1580). There is a tremendous unmet Genbank GI medical need in this population since few prescription options number organism exist to treat IBS. 27806993 Cattle 0003 Patients with IBS suffer from abdominal pain and a 16SSS439 Eel disturbed bowel pattern. Three subgroups of IBS patients 16555437 Eel have been defined based on the predominant bowel habit: 45211 69 Fish constipation-predominant (c-IBS), diarrhea-predominant 1850774 Frog 149S352 Guinea pig (d-IBS) or alternating between the two (a-IBS). Estimates of 2494.861 Guinea pig individuals who suffer from c-IBS range from 20-50% of the 4826,752 human IBS patients with 30% frequently cited. In contrast to the 4SOS441 human 1184046 human other two subgroups that have a similar gender ratio, c-IBS is 12306.17 OUSE more common in women (ratio of 3:1) (Talley et al. 1995 Am 2708786 OUSE J Epidemiol 142:76-83). 71894985 OUSE 47S23018 Pig 0004. The definition and diagnostic criteria for IBS have 593OO67 rabbit been formalized in the “Rome Criteria' (Drossman et al. 6981OOO Rat 1999, Gut 45:Suppl II: 1-81), which are well accepted in 40445437 Worm clinical practice. Briefly, the criteria specify that for at least 12 weeks (consecutive or non-consecutive in the preceding 12 months of abdominal discomfort or pain at least two of the following three features must occur: (1) relieved with defeca SUMMARY DESCRIBED HEREIN tion, (2) onset associated with a change in frequency of stool, 0006. Described herein are compositions and related and (3) onset associated with a change in form (appearance) methods for treating a variety of disorders, including IBS and of stool. The Rome II criteria also state that the symptoms that other gastrointestinal disorders and conditions (e.g., gas cumulatively support the diagnosis of irritable bowel syn trointestinal motility disorders, inflammatory bowel disease drome include: abnormal stool frequency ("abnormal” may (IBD), chronic intestinal pseudo-obstruction, colonic be defined as greater than 3 bowel movements per day and pseudo-obstruction, Crohn's disease, duodenogastric reflux, less than 3 bowel movements per week), abnormal stool form dyspepsia, functional dyspepsia, nonulcer dyspepsia, a func (lumpy/hard or loose/watery stool), abnormal stool passage tional gastrointestinal disorder, functional heartburn, gastroe (straining, urgency, or feeling of incomplete evacuation), pas sophageal reflux disease (GERD), gastroparesis, irritable sage of mucus, and bloating or feeling of abdominal disten bowel syndrome, post-operative ileus, ulcerative colitis, Sion. However, the complexity of symptoms has not been chronic constipation, and disorders and conditions associated explained by anatomical abnormalities or metabolic changes. with constipation (e.g. constipation associated with use of This has led to the classification of IBS as a functional GI opiate pain killers, post-Surgical constipation, and constipa disorder, which is diagnosed on the basis of the Rome criteria tion associated with neuropathic disorders as well as other and limited evaluation to exclude organic disease (Ringel et conditions and disorders are described herein. al. 2001, Annu Rev MedS2:319-338). IBS is considered to be 0007. The therapeutic compositions and methods a “biopsychosocial disorder resulting from a combination of described herein employ polypeptides that include at least a US 2012/0283411 A9 Nov. 8, 2012 portion of the pro sequence of guanylin or uroguanylin or tion of the pro sequence of human uroguanylin together with variants of such polypeptides. Without being bound by any all or a portion of the sequence of mature human uroguanylin. particular theory, polypeptides that include portions of the pro Thus, certain useful polypeptides include all or a portion of sequence may bind to and activate the GC-C receptor and/or the sequence: another target. 0008 Also described herein are compositions and related methods for treating obesity, congestive heart failure (includ (SEQ ID NO: ZZ-human uroguanylin; mature ing congestive heart failure at any of stages I-IV according to uroguanylin portion underlined) . New York Heart Association (NYHA) Functional Classifica WYIQYOGFRVOLESMKKLSDLEAOWAPSPRLOAQSLLPAVCHHPALPODL tion) and benign prostatic hyperplasia (BPH). QPWCASQEASSIFKTLRTIANDDCELCWNWACTGCL 0009. Without being bound by any particular theory, in the case of IBS and other gastrointestinal disorders the polypep 0016 Other useful polypeptides include all or a portion of tides are useful, in part, because they can increase gastrointes a polypeptide, SEQ ID NO:X1, that is related to human tinal motility. proguanylin. Thus, useful polypeptides include a polypeptide 0010 Without being bound by any particular theory, in the comprising (or consisting of or consisting essentially of) at case of IBS and other gastrointestinal disorders the polypep least 10 contiguous amino acid of a polypeptide having the tides are useful, in part, because they can decrease inflamma Sequence: XXXX XXX, XXXXL EXVK tion. X17 L X19 X20 L X22 X2s X24X2s X26 X27X28X29 Xso Xs 0011. Without being bound by any particular theory, in the X32Xss X34Xss X36 X37 Xss X39 Xao X4 X42X4s X44X4s case of IBS and other gastrointestinal disorders the polypep X46X47X4s X49 Xso CXs2 Xss XS4Xss Xs, Xs7 PXso Xso tides are also useful because they may decrease gastrointes X61 X62 PX64 CX66 X67 X68 X69X7o X7 X72X-7s X74X-7s R tinal pain, visceral pain, chronic visceral hypersensitivity, or LX-7s XXso Xs, Xs, Xs PG TC EICAYAACTG CXoo hypersensitivity to colorectal distension. (SEQID NO:X1-guanylin consensus 1) 0012. Without being bound by any particular theory, in the wherein: case of salt retention, fluid retention disorders and combina tions thereof the polypeptides are also useful because they X is V or S; may elicit one or more of diuresis, naturesis and/or kaliuresis. X is T. L., I, Y or E: Thus the peptides described herein may be diuretics. 0013 Described are pharmaceutical compositions com X is V or F: prising a polypeptide described hereinas well as combination compositions comprising a polypeptide described herein and X is Q or K. one or more additional therapeutic agents including, without Xs is D or E: limitation, the agents described herein. The other agents can be administered with the polypeptides described herein (si X is G or N: multaneously or sequentially). They can also be linked to a X, is D, N, E or G: polypeptide described herein to create therapeutic conju gates. X is For L: 0014 Described herein are various useful polypeptides X is S. T or K: that include all or a portion of the sequence of the pro sequence of human guanylin. Thus, certain useful polypep X is ForY: tides include all or a portion of the sequence: X is S or P; VTVQDGNFSFSLES VKKLKDLOEPQEPRVGKLRN X is S or A: FAPIPGEPVVPILCSNPNFPEELKPLC KEPNAQEILQR X, is K, Q or R: LEEIAED (SEQ ID NO:X'; human guanylin pro sequence). Other useful polypeptides include all or portion of the pro X is K or H: sequence of human guanylin together with all or a portion of the sequence of mature human guanylin. Thus, certain useful X is D, E, A, H or G; polypeptides include all or a portion of the sequence: X is Q, R. G. M. A.

(SEQ ID NO: X; human proguanylin; mature guanylin portion underlined). VTVODGNFSFSLESVKKLKDLOEPOEPRVGKLRNFAPIPGEPVWPILCSNPNFPEELKPLC

KEPNAQEILQRLEEIAEDPGTCEICAYAACTGC

00.15 Described herein are various useful polypeptides X is E. Q or D; that include all or a portion of the sequence of the pro sequence of human uroguanylin. Thus, certain useful X is A. S. E. V. L or P; polypeptides include all or a portion of the sequence: Xs is Q. N. P. G or S; VYIQYOGFRVQLESMKKLSDLE X is E. K. MorV: AQWAPSPRLQAQSLLPAVCHHPALPQDLQPVCASQE ASSIFKTLRTIA (SEQ ID NO:ZZ-human uroguanylin 0017 X, is G, L or is missing: prosequence). Other useful polypeptides include all or por Xs is Q, S, R, A or is missing:

US 2012/0283411 A9 Nov. 8, 2012

X is T or S. and the examples of a purified polypeptide consisting of a polypeptide fragment of SEQID NO:X comprising at least 10 Xzo is I or M: contiguous amino acids of SEQID NO:X are: X, is A, S or D; 0.038 a) a polypeptide comprising amino acids 1-16 of SEQ ID NO:X; X, is N. T. G or Q; 0.039 b) a polypeptide comprising amino acids 1-47 of X7 is D or E; and SEQ ID NO:X; 0040 c) a polypeptide comprising amino acids 1-61 of X7 is V or I. SEQ ID NO:X; 0034. Thus, described herein are 1) purified polypeptides 0041 d) a polypeptide comprising amino acids 1-72 of comprising (or consisting of or consisting essentially of) at SEQ ID NO:X; least 10 contiguous amino acids of SEQ ID NO:X' and 2) 0.042 e) a polypeptide comprising amino acids 17-47 of purified polypeptides consisting of a polypeptide fragment of SEQ ID NO:X; SEQ ID NO:X'. comprising (or consisting of or consisting 0.043 f) a polypeptide comprising amino acids 17-61 of essentially of) at least 10 contiguous amino acids of SEQID SEQ ID NO:X; NO:X VTVQDGNFSFSLES 0044 g) a polypeptide comprisingamino acids 17-72 of VKKLKDLOEPQEPRVGKLRN SEQ ID NO:X; FAPIPGEPVVPILCSNPNFPEELKPLC KEPNAQEILQR 0.045 h) a polypeptide comprisingamino acids 17-94 of LEEIAED (SEQ ID NO:X'; human guanylin prosequence). SEQ ID NO:X; In various embodiments the purified polypeptide comprises 0046 i) a polypeptide comprising amino acids 48-61 of at least 15 amino acids, at least 20 amino acids, at least 25 SEQ ID NO:X; amino acids, at least 30 amino acids, at least 40 amino acids, 0047 j) a polypeptide comprising amino acids 48-72 of at least 45 amino acids, at least 50 amino acids, at least 55 SEQ ID NO:X; amino acids, at least 60 amino acids, at least 65 amino acids, 0.048 k) a polypeptide comprisingamino acids 48-94 of at least 70 amino acids, at least 75 amino acids, or at least 80 SEQ ID NO:X; amino acids. In various embodiments the purified polypep 0049. 1) a polypeptide comprising amino acids 62-72 of tide comprises fewerthan 80, 75,7065, 60,55, 50, 45, 40,35, SEQ ID NO:X; 30, 25 or 20 contiguous amino acids of SEQID NO:X', and 0050 m) a polypeptide comprising amino acids 62-94 has the ability to bind and/or activate the GC-C receptor of SEQID NO:X; and/or elicit diuresis when administered to a subject. 0051 n) a polypeptide comprisingamino acids 73-94 of 0035. Thus, described herein are 1) purified polypeptides SEQ ID NO:X; comprising (or consisting of or consisting essentially of) at 0.052 o) a polypeptide comprising amino acids 1-23 of least 10 contiguous amino acids of SEQ ID NO:X and 2) purified polypeptides consisting of a polypeptide fragment of SEQ ID NO:X; SEQ ID NO:X comprising (or consisting of or consisting 0.053 p) a polypeptide comprisingamino acids 48-66 of essentially of) at least 10 contiguous amino acids of SEQID SEQ ID NO:X; NOX 0.054 q) a polypeptide comprisingamino acids 48-71 of VTVQDGNFSFSLES SEQ ID NO:X; VKKLKDLOEPQEPRVGKLRN 0.055 r) a polypeptide comprising amino acids 48-76 of FAPIPGEPVVPILCSNPNFPEELKPLC KEPNAQEILQR SEQ ID NO:X; LEEIAEDPGTCEICAYAACTGC (SEQ ID NO:X; human 005.6 s) a polypeptide comprising amino acids 43-61 of proguanylin). In various embodiments: the purified polypep SEQ ID NO:X; tide comprises at least 15 amino acids, at least 20 amino acids, 0057 t) a polypeptide comprising amino acids 38-61 of at least 25 amino acids, at least 30 amino acids, at least 40 SEQ ID NO:X; amino acids, at least 45 amino acids, at least 50 amino acids, 0.058 u) a polypeptide comprisingamino acids 33-61 of at least 55 amino acids, at least 60 amino acids, at least 65 SEQ ID NO:X; amino acids, at least 70 amino acids, at least 75 amino acids, 0059 v) a polypeptide comprisingamino acids 43-66 of or at least 80 amino acids. In various embodiments the puri SEQ ID NO:X; fied polypeptide comprises fewer than 80, 75, 7065, 60, 55, 0060 w) a polypeptide comprising amino acids 38-66 50, 45,40,35, 30, 25 or 20 contiguous amino acids of SEQID of SEQID NO:X; NO:X and has the ability to bind and/or activate the GC-C 0061 x) a polypeptide comprisingamino acids 33-66 of receptor and/or elicit diuresis when administered to a subject. SEQ ID NO:X; 0.036 Compositions, including pharmaceutical composi 0062 y) a polypeptide comprisingamino acids 43-71 of tions, can include at least one such polypeptide or can include SEQ ID NO:X; at least two (three, four or more) such polypeptides which are 0.063 Z) a polypeptide comprising amino acids 38-71 of different. In the compositions containing two or more Such SEQ ID NO:X; polypeptides the polypeptides can be separate or they can be 0.064 aa) a polypeptide comprising amino acids 33-71 covalently direct linked, e.g., by a peptide bond or a linker or of SEQID NO:X; they can be indirectly linked. For example, two Such polypep 0065 ab) a polypeptide comprising amino acids 43-76 tide sequences can be contained within a larger polypeptide of SEQID NO:X; and the two polypeptide sequences can be separated by other 0.066 ac) a polypeptide comprising amino acids 38-76 polypeptide sequences. of SEQID NO:X; and 0037 Among the examples of a purified polypeptide com 0067 ad) a polypeptide comprising amino acids 33-76 prising at least 10 contiguous amino acids of SEQID NO:X of SEQID NO:X. US 2012/0283411 A9 Nov. 8, 2012

0068. The contemplated purified polypeptides include any 0.098 ac) a polypeptide consisting of amino acids 38-76 Subset of the aforementioned polypeptides as well as each one of SEQID NO:X; and of the forgoing polypeptides. 0099 ad) a polypeptide consisting of amino acids 33-76 0069. Among the examples of a purified polypeptide com of SEQID NO:X. prising at least 10 contiguous amino acids of SEQID NO:X 0100. The contemplated purified polypeptides include any and the examples of a purified polypeptide consisting of a Subset of the aforementioned polypeptides as well as each one polypeptide fragment of SEQID NO:X comprising at least 10 of the forgoing polypeptides. contiguous amino acids of SEQID NO:X are: 0101. Among the examples of a purified polypeptide com 0070 a) a polypeptide consisting of amino acids 1-16 of prising at least 10 contiguous amino acids of SEQID NO:X SEQID NO:X; and the examples of a purified polypeptide consisting of a 0071 b) a polypeptide consisting of amino acids 1-47 of polypeptide fragment of SEQID NO:X comprising at least 10 SEQID NO:X; contiguous amino acids of SEQID NO:X are: 0072 c) a polypeptide consisting of amino acids 1-61 of 0102 a) a polypeptide consisting essentially of amino SEQID NO:X; acids 1-16 of SEQID NO:X: 0073 d) a polypeptide consisting of amino acids 1-72 of 0.103 b) a polypeptide consisting essentially of amino SEQID NO:X; acids 1-47 of SEQID NO:X; 0074 e) a polypeptide consisting of amino acids 17-47 0104 c) a polypeptide consisting essentially of amino of SEQID NO:X; acids 1-61 of SEQID NO:X: 0075 f) a polypeptide consisting of amino acids 17-61 0105 d) a polypeptide consisting essentially of amino of SEQID NO:X; acids 1-72 of SEQID NO:X; 0076 g) a polypeptide consisting of amino acids 17-72 0106 e) a polypeptide consisting essentially of amino of SEQID NO:X; acids 17-47 of SEQID NO:X; 0077 h) a polypeptide consisting of amino acids 17-94 0.107 f) a polypeptide consisting essentially of amino of SEQID NO:X; acids 17-61 of SEQID NO:X; 0078 i) a polypeptide consisting of amino acids 48-61 0.108 g) a polypeptide consisting essentially of amino of SEQID NO:X; acids 17-72 of SEQID NO:X; 0079 j) a polypeptide consisting of amino acids 48-72 0.109 h) a polypeptide consisting essentially of amino of SEQID NO:X; acids 17-94 of SEQID NO:X; 0080 k) a polypeptide consisting of amino acids 48-94 0110 i) a polypeptide consisting essentially of amino of SEQID NO:X; acids 48-61 of SEQID NO:X; I0081 l) a polypeptide consisting of amino acids 62-72 0111 ) a polypeptide consisting essentially of amino of SEQID NO:X; acids 48-72 of SEQID NO:X; I0082 m) a polypeptide consisting of amino acids 62-94 0112 k) a polypeptide consisting essentially of amino of SEQID NO:X; acids 48-94 of SEQID NO:X; I0083 n) a polypeptide consisting of amino acids 73-94 0113. 1) a polypeptide consisting essentially of amino of SEQID NO:X; acids 62-72 of SEQID NO:X; I0084 o) a polypeptide consisting of amino acids 1-23 of 0114 m) a polypeptide consisting essentially of amino SEQID NO:X; acids 62-94 of SEQID NO:X; I0085 p) a polypeptide consisting of amino acids 48-66 0115 n) a polypeptide consisting essentially of amino of SEQID NO:X; acids 73-94 of SEQID NO:X; I0086 q) a polypeptide consisting of amino acids 48-71 0116 o) a polypeptide consisting essentially of amino of SEQID NO:X; acids 1-23 of SEQID NO:X; I0087 r) a polypeptide consisting of amino acids 48-76 0117 p) a polypeptide consisting essentially of amino of SEQID NO:X; acids 48-66 of SEQID NO:X; I0088 s) a polypeptide consisting of amino acids 43-61 0118 q) a polypeptide consisting essentially of amino of SEQID NO:X; acids 48-71 of SEQID NO:X; I0089 t) a polypeptide consisting of amino acids 38-61 0119 r) a polypeptide consisting essentially of amino of SEQID NO:X; acids 48-76 of SEQID NO:X; 0090 u) a polypeptide consisting of amino acids 33-61 0120 s) a polypeptide consisting essentially of amino of SEQID NO:X; acids 43-61 of SEQID NO:X; 0091 v) a polypeptide consisting of amino acids 43-66 0121 t) a polypeptide consisting essentially of amino of SEQID NO:X; acids 38-61 of SEQID NO:X; 0092 w) a polypeptide consisting of amino acids 38-66 0.122 u) a polypeptide consisting essentially of amino of SEQID NO:X; acids 33-61 of SEQID NO:X; 0093 x) a polypeptide consisting of amino acids 33-66 0123 V) a polypeptide consisting essentially of amino of SEQID NO:X; acids 43-66 of SEQID NO:X; 0094 y) a polypeptide consisting of amino acids 43-71 0.124 w) a polypeptide consisting essentially of amino of SEQID NO:X; acids 38-66 of SEQID NO:X; 0.095 Z) a polypeptide consisting of amino acids 38-71 0.125 X) a polypeptide consisting essentially of amino of SEQID NO:X; acids 33-66 of SEQID NO:X; 0096 aa) a polypeptide consisting of amino acids 33-71 0.126 y) a polypeptide consisting essentially of amino of SEQID NO:X; acids 43-71 of SEQID NO:X; 0097 ab) a polypeptide consisting of amino acids 43-76 0127 Z) a polypeptide consisting essentially of amino of SEQID NO:X; acids 38-71 of SEQID NO:X; US 2012/0283411 A9 Nov. 8, 2012

0128 aa) a polypeptide consisting essentially of amino 0.137 polypeptide (c) and at least one of polypeptides acids 33-71 of SEQID NO:X; (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (p), (q). I0129 ab) a polypeptide consisting essentially of amino (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); acids 43-76 of SEQID NO:X; 0.138 polypeptide (d) and at least one of polypeptides 0.130 ac) a polypeptide consisting essentially of amino (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r). acids 38-76 of SEQID NO:X; and (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0131 ad) a polypeptide consisting essentially of amino 0.139 polypeptide (e) and at least one of polypeptides acids 33-76 of SEQID NO:X. 0132) The contemplated purified polypeptides include any (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (S), Subset of the aforementioned polypeptides as well as each one (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); of the forgoing polypeptides. 0140 polypeptide (f) and at least one of polypeptides 0133) For SEQ ID NO:X useful compositions, including (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), (s), (t), pharmaceutical compositions, that include, for example, (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); more than one of polypeptides (a)-(ad) (or polypeptides com 0141 polypeptide (g) and at least one of polypeptides prising, consisting of or consisting essentially of one or more (h), (i), (), (k), (1), (m), (n), (o), (p), (q). (r). (S), (t), (u), of (a)-(ad) or a consisting of a polypeptide fragment of SEQ (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); ID NO:X comprising one or more of (a)-(ad)) include com 0.142 polypeptide (h) and at least one of polypeptides binations 1-38 below: (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0.143 polypeptide (i) and at least one of polypeptides (), (k), (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), 1 (a), (e) 2 (a), (e), (i) (X), (y), (Z), (aa), (ab), (ac), (ad); 3 (a), (e), (ii), (I) 0.144 polypeptide () and at least one of polypeptides 4 (a), (e), (i), (), (n) (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X). 5 (a), (e), (i), (m) (y), (Z), (aa), (ab), (ac), (ad); 6 (a), (e), (k) 0145 polypeptide (k) and at least one of polypeptides 7 (a), (f) 8 (a), (f), (I) (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y). 9 (a), (f), (), (n) (Z), (aa), (ab), (ac), (ad); 10 (a), (f), (m) 0146 polypeptide (1) and at least one of polypeptides 11 (a), (g) 12 (a), (g), (n) (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z). 13 (a), (h) (aa), (ab), (ac), (ad); 14 (b), (i) 0147 polypeptide (m) and at least one of polypeptides 15 (b), (i), (I) (n), (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa). 16 (b), (i), (), (n) 17 (b), (i), (m) (ab), (ac), (ad); 18 (b), () 0.148 polypeptide (n) and at least one of polypeptides 19 (b), (), (n) (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa). 2O (b), (k) (ab), (ac), (ad); 21 (c), (I) 0.149 polypeptide (o) and at least one of polypeptides 22 (c), (), (n) 23 (d), (n) (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab). 24 (e), (i) (ac), (ad); 25 (e), (i), (I) 0.150 polypeptide (p) and at least one of polypeptides 26 (e), (i), (), (n) 27 (e), (i), (m) s'). (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac). 28 (e), () aCl), 29 (e), (), (n) 0151 polypeptide (q) and at least one of polypeptides 30 (e), (k) (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 31 (f), (I) 32 (f), (), (n) 0152 polypeptide (r) and at least one of polypeptides 33 (f), (m) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 34 (g), (n) 0.153 polypeptide (s) and at least one of polypeptides 35 (i), (), (n) 36 (i), (m) (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 37 (), (n) 0154 polypeptide (t) and at least one of polypeptides 38 (l), (n) (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0.155 polypeptide (u) and at least one of polypeptides 0134) For SEQ ID NO:X, useful pharmaceutical compo (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); sitions include: a pharmaceutical composition comprising at 0156 polypeptide (V) and at least one of polypeptides least two (three, four or more) of polypeptides (a)-(ad) and a (W), (X), (y), (Z), (aa), (ab), (ac), (ad); pharmaceutically acceptable carrier. In various embodiments 0157 polypeptide (w) and at least one of polypeptides the pharmaceutical composition comprises: (X), (y), (Z), (aa), (ab), (ac), (ad); 0.135 polypeptide (a) and at least one of: polypeptides 0158 polypeptide (x) and at least one of polypeptides (b), (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (y), (Z), (aa), (ab), (ac), (ad); (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), 0159 polypeptide (y) and at least one of polypeptides (ac), (ad); (Z), (aa), (ab), (ac), (ad); 0.136 polypeptide (b) and at least one of polypeptides 0.160 polypeptide (Z) and at least one of polypeptides (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (p), (aa), (ab), (ac), (ad); S.) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), 0.161 polypeptide (aa) and at least one of polypeptides aCl), (ab), (ac), (ad);

US 2012/0283411 A9 Nov. 8, 2012 10

X-7s is E, A, G or C. Xs is Q or P; X7 is E. A. V. S. L or M: X is E or K. Xso is I or V: 0.174 X, is missing: X2s is missing: Xs is A or P; X2 is missing: Xs is E. Q. A or S. X is Por V: Xs is D or E; and X is R 0170 X is For is missing. 0171 In various embodiments: the purified polypeptide X is L or V. comprises at least 15 amino acids, at least 20 amino acids, at X is P or G; least 25 amino acids, at least 30 amino acids, at least 40 amino acids, at least 45 amino acids, at least 50 amino acids, at least X is S. R or K. 55 amino acids, at least 60 amino acids, at least 65 amino Xs is H or L. acids, at least 70 amino acids, at least 75 amino acids, or at least 80 amino acids. In various embodiments the purified 0.175 X is R. K or is missing: polypeptide comprises fewer than 80, 75, 7065, 60, 55, 50, X, is N. K or is missing: 45, 40, 35, 30, 25 or 20 contiguous amino acids of SEQ ID Xss is For is missing: NO:X1 and has the ability to bind and/or activate the GC-C X is A or is missing: receptor and/or elicit diuresis when administered to a subject. Xao is P or is missing: 0172 Compositions, including pharmaceutical composi X is I, R or is missing: tions, can include at least one such polypeptide or can include at least two (three, four or more) such polypeptides which are X is L or P; different. In the compositions containing two or more Such X is G or L. polypeptides the polypeptides can be separate or they can be covalently direct linked, e.g., by a peptide bond or a linker or X is G. E. or K, they can be indirectly linked. For example, two Such polypep Xas is P or S; tide sequences can be contained within a larger polypeptide and the two polypeptide sequences can be separated by other X is V or A: polypeptide sequences. Xa, is A or V. 0173. In certain embodiments of the purified polypeptide comprising at least 10 contiguous amino acids of SEQ ID 0176 X is P or is missing: NO:X1 and in certain embodiments of the polypeptide con sisting of a polypeptide fragment of SEQ ID NO:X1 com Xao is I or Q. prising at least 10 contiguous amino acids of SEQID NO:X1: Xso is L. X is V: Xs is S. X is T or L; 0177 Xs is missing: X is V or F: Xs is H, N, or D; X is Q or K. Xss is P or S; Xs is D or E: Xs is A, K or N: X is G or N: Xs, is For L. X, is D or N: Xso is E. X is For L: X is E or A: X is S; X61 is L. X is ForY: X is K or R: X is S or P; X is L. I or V. X is S. X is K, E, Q, Tor R: X, is K. X, is E or K. Xo is K. Xs is P. X is D or E: X is N: X is Q or R; Xzo is A. X2 is E. X, is E or Q. X is V. L or P; X72 is E.

US 2012/0283411 A9 Nov. 8, 2012 12

X is N, D or G; Xs is For L; Xzo is A or S; X is S; X, is E, Q, P, A or S; X is ForY: X, is E. D. Q. Mor A: X is S or P; X-7 is I, A, or S; X is S; X7 is L. For V: X, is K. X-7s is Q, E, D, N. G or A: Xo is K. X-7s is E, A, G or C. X is D or E: X7 is E. A. V. S. L or M: X is Q or R: Xso is I or V: X2 is E. Xs is A or P; X is V. L or P; Xs is E. Q. A or S. Xs is Q or P; Xs is D or E: X is E or K. (0189 X, is missing: Xss is G, S, R, or N: X2s is missing: Xs is S or T. X2 is missing: X is Y or F: X is Por V: X is T or A.; and X is R 0185. X is For is missing. X is L or V. 0186. In various embodiments: the purified polypeptide comprises at least 15 amino acids, at least 20 amino acids, at X is P or G; least 25 amino acids, at least 30 amino acids, at least 40 amino X is S. R or K. acids, at least 45 amino acids, at least 50 amino acids, at least 55 amino acids, at least 60 amino acids, at least 65 amino Xs is H or L. acids, at least 70 amino acids, at least 75 amino acids, or at 0.190 X is R. K or is missing: least 80 amino acids. In various embodiments the purified X, is N. K or is missing: polypeptide comprises fewer than 80, 75, 7065, 60, 55, 50, Xss is For is missing: 45, 40, 35, 30, 25 or 20 contiguous amino acids of SEQ ID X is A or is missing: NO:X2 and has the ability to bind and/or activate the GC-C Xao is P or is missing: receptor and/or elicit diuresis when administered to a subject. 0187 Compositions, including pharmaceutical composi X is I, R or is missing: tions, can include at least one such polypeptide or can include X is L or P; at least two (three, four or more) such polypeptides which are different. In the compositions containing two or more Such X is G or L. polypeptides the polypeptides can be separate or they can be covalently direct linked, e.g., by a peptide bond or a linker or X is G, E or K, they can be indirectly linked. For example, two Such polypep X is P or S; tide sequences can be contained within a larger polypeptide and the two polypeptide sequences can be separated by other X is V or A: polypeptide sequences. X, is A or V. 0188 In certain embodiments of the purified polypeptide comprising at least 10 contiguous amino acids of SEQ ID 0191 X is P or is missing: NO:X2 and in certain embodiments of the polypeptide con sisting of a polypeptide fragment of SEQ ID NO:X2 com Xao is I or Q. prising at least 10 contiguous amino acids of SEQID NO:X2: Xso is L. X is V: Xs is S. X is T or L; 0.192 Xs is missing: X is V or F: Xs is H, N, or D; X is Q or K. Xss is P or S; Xs is D or E: Xs is A, K or N: X is G or N: Xs, is For L. X, is D or N: Xso is E. US 2012/0283411 A9 Nov. 8, 2012

Xo is E or A: 0205 k) a polypeptide comprisingamino acids 51-99 of SEQ ID NO:X1 OR SEQ ID NO:X2. X61 is L. 0206 1) a polypeptide comprising amino acids 66-76 of X is K or R: SEQ ID NO:X1 OR SEQ ID NO:X2: 0207 m) a polypeptide comprising amino acids 66-99 X is L. I or V. of SEQID NO:X1 OR SEQID NO:X2. X is K, E, Q, Tor R: 0208 n) a polypeptide comprisingamino acids 77-99 of SEQ ID NO:X1 OR SEQ ID NO:X2: X, is E or K. 0209 o) a polypeptide comprising amino acids 1-23 of Xs is P. SEQ ID NO:X1 OR SEQ ID NO:X2: 0210 p) a polypeptide comprisingamino acids 51-70 of X is N: SEQ ID NO:X1 OR SEQ ID NO:X2: Xzo is A. 0211 q) a polypeptide comprisingamino acids 51-75 of SEQ ID NO:X1 OR SEQ ID NO:X2: X, is E or Q; 0212 r) a polypeptide comprising amino acids 51-80 of X72 is E. SEQ ID NO:X1 OR SEQ ID NO:X2: 0213 s) a polypeptide comprising amino acids 46-65 of X-7 is I; SEQ ID NO:X1 OR SEQ ID NO:X2: X74 is L. 0214 t) a polypeptide comprising amino acids 41-65 of SEQ ID NO:X1 OR SEQ ID NO:X2: X-7s is Q or E: 0215 u) a polypeptide comprisingamino acids 36-65 of X-7s is E or A: SEQ ID NO:X1 OR SEQ ID NO:X2: 0216 V) a polypeptide comprisingamino acids 46-70 of X-7 is E or A: SEQ ID NO:X1 OR SEQ ID NO:X2: Xso is I; 0217 w) a polypeptide comprising amino acids 41-70 of SEQID NO:X1 OR SEQID NO:X2: Xs is A. 0218. X) a polypeptide comprisingamino acids 36-70 of Xs is E or Q; SEQ ID NO:X1 OR SEQ ID NO:X2: 0219 y) a polypeptide comprisingamino acids 46-75 of Xss is D; SEQ ID NO:X1 OR SEQ ID NO:X2: Xss is G, or S. 0220 Z) a polypeptide comprising amino acids 41-75 of SEQ ID NO:X1 OR SEQ ID NO:X2: Xs is T. 0221 aa) a polypeptide comprising amino acids 36-75 X is Y: of SEQID NO:X1 OR SEQID NO:X2: 0222 ab) a polypeptide comprising amino acids 46-80 X is T. and of SEQID NO:X1 OR SEQID NO:X2: 0193 X is missing. 0223 ac) a polypeptide comprising amino acids 41-80 0194 Among the examples of a purified polypeptide com of SEQID NO:X1 OR SEQID NO:X2; and prising at least 10 contiguous amino acids of SEQID NO:X1 0224 ad) a polypeptide comprising amino acids 36-80 or X2 and the examples of a purified polypeptide consisting of of SEQID NO:X1 OR SEQID NO:X2. a polypeptide fragment of SEQID NO:X1 or X2 comprising 0225. The contemplated purified polypeptides include any at least 10 contiguous amino acids of SEQID NO:X1 or X2 Subset of the aforementioned polypeptides as well as each one a. of the forgoing polypeptides. 0.195 a) a polypeptide comprising amino acids 1-16 of 0226. Among the examples of a purified polypeptide com SEQID NO:X1 OR SEQ ID NO:X2: prising at least 10 contiguous amino acids of SEQID NO:X1 0196) b) a polypeptide comprising amino acids 1-50 of or X2 and the examples of a purified polypeptide consisting of SEQID NO:X1 OR SEQ ID NO:X2: a polypeptide fragment of SEQID NO:X1 or X2 comprising 0.197 c) a polypeptide comprising amino acids 1-65 of at least 10 contiguous amino acids of SEQID NOX1 or X2 SEQID NO:X1 OR SEQ ID NO:X2: a. 0198 d) a polypeptide comprising amino acids 1-76 of 0227 a) a polypeptide consisting of amino acids 1-16 of SEQID NO:X1 OR SEQ ID NO:X2: SEQ ID NO:X1 OR SEQ ID NO:X2: 0199 e) a polypeptide comprising amino acids 17-50 of 0228 b) a polypeptide consisting of amino acids 1-50 of SEQID NO:X1 OR SEQ ID NO:X2: SEQ ID NO:X1 OR SEQ ID NO:X2: 0200 f) a polypeptide comprising amino acids 17-65 of 0229 c) a polypeptide consisting of amino acids 1-65 of SEQID NO:X1 OR SEQ ID NO:X2: SEQ ID NO:X1 OR SEQ ID NO:X2: 0201 g) a polypeptide comprising amino acids 17-76 of 0230 d) a polypeptide consisting of amino acids 1-76 of SEQID NO:X1 OR SEQ ID NO:X2: SEQ ID NO:X1 OR SEQ ID NO:X2: 0202 h) a polypeptide comprising amino acids 17-99 of 0231 e) a polypeptide consisting of amino acids 17-50 SEQID NO:X1 OR SEQ ID NO:X2. of SEQID NO:X1 OR SEQID NO:X2: 0203 i) a polypeptide comprising amino acids 51-65 of 0232 f) a polypeptide consisting of amino acids 17-65 SEQID NO:X1 OR SEQ ID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0204 j) a polypeptide comprising amino acids 51-76 of 0233 g) a polypeptide consisting of amino acids 17-76 SEQID NO:X1 OR SEQ ID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: US 2012/0283411 A9 Nov. 8, 2012

0234 h) a polypeptide consisting of amino acids 17-99 0263 e) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2. acids 17-50 of SEQID NO:X1 OR SEQID NO:X2: 0235 i) a polypeptide consisting of amino acids 51-65 0264 f) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 17-65 of SEQID NO:X1 OR SEQID NO:X2: 0236 ) a polypeptide consisting of amino acids 51-76 0265 g) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 17-76 of SEQID NO:X1 OR SEQID NO:X2: 0237) k) a polypeptide consisting of amino acids 51-99 0266 h) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2. acids 17-99 of SEQID NO:X1 OR SEQID NO:X2. 0238 1) a polypeptide consisting of amino acids 66-76 0267 i) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 51-65 of SEQID NO:X1 OR SEQID NO:X2: 0239) m) a polypeptide consisting of amino acids 66-99 0268 ) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2. acids 51-76 of SEQID NO:X1 OR SEQID NO:X2: 0240 n) a polypeptide consisting of amino acids 77-99 0269 k) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 51-99 of SEQID NO:X1 OR SEQID NO:X2. 0241 o) a polypeptide consisting of amino acids 1-23 of 0270 l) a polypeptide consisting essentially of amino SEQID NO:X1 OR SEQ ID NO:X2: acids 66-76 of SEQID NO:X1 OR SEQID NO:X2: 0242 p) a polypeptide consisting of amino acids 51-70 0271 m) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 66-99 of SEQID NO:X1 OR SEQID NO:X2. 0243 q) a polypeptide consisting of amino acids 51-75 0272 n) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 77-99 of SEQID NO:X1 OR SEQID NO:X2: 0244 r) a polypeptide consisting of amino acids 51-80 0273 o) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 1-23 of SEQID NO:X1 OR SEQID NO:X2: 0245 S) a polypeptide consisting of amino acids 46-65 0274 p) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: acids 51-70 of SEQID NO:X1 OR SEQID NO:X2: 0246 t) a polypeptide consisting of amino acids 41-65 0275 q) a polypeptide consisting essentially of amino of SEQID NO:X1 OR SEQID NO:X2: 0247 u) a polypeptide consisting of amino acids 36-65 acids 51-75 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0276 r) a polypeptide consisting essentially of amino 0248 V) a polypeptide consisting of amino acids 46-70 acids 51-80 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0277 s) a polypeptide consisting essentially of amino 0249 w) a polypeptide consisting of amino acids 41-70 acids 46-65 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0278 t) a polypeptide consisting essentially of amino 0250 X) a polypeptide consisting of amino acids 36-70 acids 41-65 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0279 u) a polypeptide consisting essentially of amino 0251 y) a polypeptide consisting of amino acids 46-75 acids 36-65 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0280 v) a polypeptide consisting essentially of amino 0252) Z) a polypeptide consisting of amino acids 41-75 acids 46-70 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0281 w) a polypeptide consisting essentially of amino 0253 aa) a polypeptide consisting of amino acids 36-75 acids 41-70 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0282 X) a polypeptide consisting essentially of amino 0254 ab) a polypeptide consisting of amino acids 46-80 acids 36-70 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2: 0283 y) a polypeptide consisting essentially of amino 0255 ac) a polypeptide consisting of amino acids 41-80 acids 46-75 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2; and 0284 Z) a polypeptide consisting essentially of amino 0256 ad) a polypeptide consisting of amino acids 36-80 acids 41-75 of SEQID NO:X1 OR SEQID NO:X2: of SEQID NO:X1 OR SEQID NO:X2. 0285 aa) a polypeptide consisting essentially of amino 0257 The contemplated purified polypeptides include any acids 36-75 of SEQID NO:X1 OR SEQID NO:X2: Subset of the aforementioned polypeptides as well as each one 0286 ab) a polypeptide consisting essentially of amino of the forgoing polypeptides. acids 46-80 of SEQID NO:X1 OR SEQID NO:X2: 0258 Among the examples of a purified polypeptide com 0287 ac) a polypeptide consisting essentially of amino prising at least 10 contiguous amino acids of SEQID NO:X1 acids 41-80 of SEQID NO:X1 OR SEQID NO:X2; and or X and the examples of a purified polypeptide consisting of 0288 ad) a polypeptide consisting essentially of amino a polypeptide fragment of SEQID NO:X1 or X2 comprising acids 36-80 of SEQID NO:X1 OR SEQID NO:X2. at least 10 contiguous amino acids of SEQID NOX1 or X2 0289. The contemplated purified polypeptides include any a. Subset of the aforementioned polypeptides as well as each one 0259 a) a polypeptide consisting essentially of amino of the forgoing polypeptides. acids 1-16 of SEQID NO:X1 OR SEQID NO:X2: 0290 For SEQ ID NO:X1 and SEQ ID NO:X2 useful 0260 b) a polypeptide consisting essentially of amino compositions, including pharmaceutical compositions, that acids 1-50 of SEQID NO:X1 OR SEQID NO:X2: include, for example, more than one of polypeptides (a)-(ad) 0261 c) a polypeptide consisting essentially of amino (or polypeptides comprising, consisting of or consisting acids 1-65 of SEQID NO:X1 OR SEQID NO:X2: essentially of one or more of (a)-(ad) or a consisting of a 0262 d) a polypeptide consisting essentially of amino polypeptide fragment of SEQID NO:X1 or X2 comprising acids 1-76 of SEQID NO:X1 OR SEQID NO:X2: one or more of (a)-(ad)) include combinations 1-38 below: US 2012/0283411 A9 Nov. 8, 2012 15

0299 polypeptide (h) and at least one of polypeptides (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 1 (a), (e) 2 (a), (e), (i) 0300 polypeptide (i) and at least one of polypeptides 3 (a), (e), (ii), (I) (), (k), (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), 4 (a), (e), (i), (), (n) (X), (y), (Z), (aa), (ab), (ac), (ad); 5 (a), (e), (i), (m) 0301 polypeptide () and at least one of polypeptides 6 (a), (e), (k) 7 (a), (f) (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X). 8 (a), (f), (I) (y), (Z), (aa), (ab), (ac), (ad); 9 (a), (f), (), (n) 0302 polypeptide (k) and at least one of polypeptides 10 (a), (f), (m) 11 (a), (g) (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y). 12 (a), (g), (n) (Z), (aa), (ab), (ac), (ad); 13 (a), (h) 0303 polypeptide (1) and at least one of polypeptides 14 (b), (i) (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z). 15 (b), (i), (I) 16 (b), (i), (), (n) (aa), (ab), (ac), (ad); 17 (b), (i), (m) 0304 polypeptide (m) and at least one of polypeptides 18 (b), () (n), (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa). 19 (b), (), (n) (ab), (ac), (ad); 2O (b), (k) 0305 polypeptide (n) and at least one of polypeptides 21 (c), (I) 22 (c), (), (n) (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa). 23 (d), (n) (ab), (ac), (ad); 24 (e), (i) 0306 polypeptide (o) and at least one of polypeptides 25 (e), (i), (I) (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab). 26 (e), (i), (), (n) 27 (e), (i), (m) (ac), (ad); 28 (e), () 0307 polypeptide (p) and at least one of polypeptides 29 (e), (), (n) S.) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac). 30 (e), (k) aCl), 31 (f), (I) 32 (f), (), (n) 0308 polypeptide (q) and at least one of polypeptides 33 (f), (m) (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 34 (g), (n) 0309 polypeptide (r) and at least one of polypeptides 35 (i), (), (n) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 36 (i), (m) 37 (), (n) 0310 polypeptide (s) and at least one of polypeptides 38 (l), (n) (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0311 polypeptide (t) and at least one of polypeptides (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0291 For SEQ ID NO:X1 and SEQ ID NO:X2, useful pharmaceutical compositions include: a pharmaceutical 0312 polypeptide (u) and at least one of polypeptides composition comprising at least two (three, four or more) of (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); polypeptides (a)-(ad) and a pharmaceutically acceptable car 0313 polypeptide (V) and at least one of polypeptides rier. In various embodiments the pharmaceutical composition (W), (X), (y), (Z), (aa), (ab), (ac), (ad); comprises: 0314 polypeptide (w) and at least one of polypeptides (X), (y), (Z), (aa), (ab), (ac), (ad); 0292 polypeptide (a) and at least one of: polypeptides 0315 polypeptide (x) and at least one of polypeptides (b), (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (y), (Z), (aa), (ab), (ac), (ad); (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), 0316 polypeptide (y) and at least one of polypeptides (ac), (ad); (Z), (aa), (ab), (ac), (ad); 0293 polypeptide (b) and at least one of polypeptides 0317 polypeptide (Z) and at least one of polypeptides (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (p), (aa), (ab), (ac), (ad); S.) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), 0318 polypeptide (aa) and at least one of polypeptides aCl), (ab), (ac), (ad); 0294 polypeptide (c) and at least one of polypeptides 0319 polypeptide (ab) and at least one of polypeptides (d), (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (ac), (ad); and (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0320 polypeptide (ac) and at least polypeptide (ad). 0295 polypeptide (d) and at least one of polypeptides 0321) Described herein are purified polypeptides compris (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), ing (or consisting of or consisting essentially of) at least 10 (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); contiguous amino acids of SEQ ID NO:ZZ and purified 0296 polypeptide (e) and at least one of polypeptides polypeptides consisting of a polypeptide fragment of SEQID (f), (g), (h), (i), (), (k), (l), (m), (n), (o), (p), (q). (r), (S), NO:ZZ comprising (or consisting of or consisting essentially (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); of) at least 10 contiguous amino acids of SEQ ID NO:ZZ 0297 polypeptide (f) and at least one of polypeptides VYIQYOGFRVQLESMKKLSDLE (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), (S), (t), AQWAPSPRLQAQSLLPAVCHHPALPQDLQPVCASQE (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); ASSIFKTLRTIA (SEQ ID NO:ZZ'; human uroguanylin 0298 polypeptide (g) and at least one of polypeptides prosequence). (h), (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (S), (t), (u), 0322. In various embodiments: the purified polypeptide (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); comprises at least 15 amino acids, at least 20 amino acids, at US 2012/0283411 A9 Nov. 8, 2012

least 25 amino acids, at least 30 amino acids, at least 40 amino 0332 e) a polypeptide comprising amino acids 17-40 of acids, at least 45 amino acids, at least 50 amino acids, at least SEQ ID NO:ZZ: 55 amino acids, at least 60 amino acids, at least 65 amino 0333 f) a polypeptide comprising amino acids 17-54 of acids, at least 70 amino acids, at least 75 amino acids, or at SEQ ID NO:ZZ: least 80 amino acids. In various embodiments the purified 0334 g) a polypeptide comprisingamino acids 17-66 of polypeptide comprises fewer than 80, 75, 7065, 60, 55, 50, SEQ ID NO:ZZ: 45, 40, 35, 30, 25 or 20 contiguous amino acids of SEQ ID 0335 h) a polypeptide comprisingamino acids 17-86 of NO:ZZ and has the ability to bind and/or activate the GC-C SEQ ID NO:ZZ: receptor and/or elicit diuresis when administered to a subject. 0336 i) a polypeptide comprising amino acids 41-54 of 0323 Compositions, including pharmaceutical composi SEQ ID NO:ZZ: tions, can include at least one such polypeptide or can include 0337 j) a polypeptide comprising amino acids 41-66 of at least two (three, four or more) such polypeptides which are SEQ ID NO:ZZ: different. In the compositions containing two or more Such 0338 k) a polypeptide comprisingamino acids 41-86 of polypeptides the polypeptides can be separate or they can be SEQ ID NO:ZZ: covalently direct linked, e.g., by a peptide bond or a linker or 0339) 1) a polypeptide comprising amino acids 55-66 of they can be indirectly linked. For example, two Such polypep SEQ ID NO:ZZ: tide sequences can be contained within a larger polypeptide 0340 m) a polypeptide comprising amino acids 55-86 and the two polypeptide sequences can be separated by other of SEQID NO:ZZ: polypeptide sequences. 0341 n) a polypeptide comprisingamino acids 67-86 of 0324 Described herein are purified polypeptides compris SEQ ID NO:ZZ: ing (or consisting of or consisting essentially of) at least 10 0342 o) a polypeptide comprising amino acids 1-21 of contiguous amino acids of SEQ ID NO:ZZ and purified SEQ ID NO:ZZ: polypeptides consisting of a polypeptide fragment of SEQID 0343 p) a polypeptide comprisingamino acids 41-59 of NO:ZZ comprising (or consisting of or consisting essentially SEQ ID NO:ZZ: of) at least 10 contiguous amino acids of SEQ ID NO:ZZ 0344 q) a polypeptide comprisingamino acids 41-64 of VYIQYOGFRVQLESMKKLSDLE SEQ ID NO:ZZ: AQWAPSPRLQAQSLLPAVCHHPALPQDLQPVCASQE 0345 r) a polypeptide comprising amino acids 41-69 of ASSIFKTLRTIANDDCELCVNVACTGCL (SEQ ID SEQ ID NO:ZZ: NO:ZZ; human prouroguanylin). 0346 s) a polypeptide comprising amino acids 36-54 of 0325 In various embodiments: the purified polypeptide SEQ ID NO:ZZ: comprises at least 15 amino acids, at least 20 amino acids, at 0347 t) a polypeptide comprising amino acids 31-54 of least 25 amino acids, at least 30 amino acids, at least 40 amino SEQ ID NO:ZZ: acids, at least 45 amino acids, at least 50 amino acids, at least 0348 u) a polypeptide comprisingamino acids 26-54 of 55 amino acids, at least 60 amino acids, at least 65 amino SEQ ID NO:ZZ: acids, at least 70 amino acids, at least 75 amino acids, or at 0349 v) a polypeptide comprisingamino acids 36-59 of least 80 amino acids. In various embodiments the purified SEQ ID NO:ZZ: polypeptide comprises fewer than 80, 75, 7065, 60, 55, 50, 0350 w) a polypeptide comprising amino acids 31-59 45, 40, 35, 30, 25 or 20 contiguous amino acids of SEQ ID of SEQID NO:ZZ: NO:ZZ and has the ability to bind and/or activate the GC-C 0351 x) a polypeptide comprisingamino acids 26-59 of receptor and/or elicit diuresis when administered to a subject. SEQ ID NO:ZZ: 0326 Compositions, including pharmaceutical composi 0352 y) a polypeptide comprisingamino acids 36-64 of tions, can include at least one such polypeptide or can include SEQ ID NO:ZZ: at least two (three, four or more) such polypeptides which are 0353 Z) a polypeptide comprising amino acids 31-64 of different. In the compositions containing two or more Such SEQ ID NO:ZZ: polypeptides the polypeptides can be separate or they can be 0354 aa) a polypeptide comprising amino acids 26-64 covalently direct linked, e.g., by a peptide bond or a linker or of SEQID NO:ZZ: they can be indirectly linked. For example, two Such polypep 0355 ab) a polypeptide comprising amino acids 36-69 tide sequences can be contained within a larger polypeptide of SEQID NO:ZZ: and the two polypeptide sequences can be separated by other 0356 ac) a polypeptide comprising amino acids 31-69 polypeptide sequences. of SEQID NO:ZZ; and 0327. Among the examples of a purified polypeptide com 0357 ad) a polypeptide comprising amino acids 26-69 prising at least 10 contiguous amino acids of SEQID NO:ZZ of SEQID NO:ZZ. and the examples of a polypeptide consisting of a polypeptide 0358. The contemplated purified polypeptides include any Subset of the aforementioned polypeptides as well as each one fragment of SEQID NO:ZZ comprising at least 10 contigu of the forgoing polypeptides. ous amino acids of SEQID NO:ZZ are: 0359 Among the examples of a purified polypeptide com 0328 a) a polypeptide comprising amino acids 1-16 of prising at least 10 contiguous amino acids of SEQID NO:ZZ SEQID NO:ZZ: and the examples of a polypeptide consisting of a polypeptide 0329 b) a polypeptide comprising amino acids 1-40 of fragment of SEQID NO:ZZ comprising at least 10 contigu SEQID NO:ZZ: ous amino acids of SEQID NO:ZZ are: 0330 c) a polypeptide comprising amino acids 1-54 of 0360 a) a polypeptide consisting of amino acids 1-16 of SEQID NO:ZZ: SEQ ID NO:ZZ: 0331 d) a polypeptide comprising amino acids 1-66 of 0361 b) a polypeptide consisting of amino acids 1-40 of SEQID NO:ZZ: SEQ ID NO:ZZ: US 2012/0283411 A9 Nov. 8, 2012 17

0362 c) a polypeptide consisting of amino acids 1-54 of 0392 a) a polypeptide consisting essentially of amino SEQID NO:ZZ: acids 1-16 of SEQID NO:ZZ: 0363 d) a polypeptide consisting of amino acids 1-66 of 0393 b) a polypeptide consisting essentially of amino SEQID NO:ZZ: acids 1-40 of SEQID NO:ZZ: 0364 e) a polypeptide consisting of amino acids 17-40 0394 c) a polypeptide consisting of amino acids 1-54 of of SEQID NO:ZZ: SEQ ID NO:ZZ: 0365 f) a polypeptide consisting of amino acids 17-54 0395 d) a polypeptide consisting of amino acids 1-66 of of SEQID NO:ZZ: SEQ ID NO:ZZ: 0366 g) a polypeptide consisting of amino acids 17-66 0396 e) a polypeptide consisting of amino acids 17-40 of SEQID NO:ZZ: of SEQID NO:ZZ: 0367 h) a polypeptide consisting of amino acids 17-86 0397) f) a polypeptide consisting of amino acids 17-54 of SEQID NO:ZZ: of SEQID NO:ZZ: 0368 i) a polypeptide consisting of amino acids 41-54 0398 g) a polypeptide consisting of amino acids 17-66 of SEQID NO:ZZ: of SEQID NO:ZZ: 0369 ) a polypeptide consisting of amino acids 41-66 0399 h) a polypeptide consisting of amino acids 17-86 of SEQID NO:ZZ: of SEQID NO:ZZ: 0370 k) a polypeptide consisting of amino acids 41-86 04.00 i) a polypeptide consisting of amino acids 41-54 of SEQID NO:ZZ: of SEQID NO:ZZ: 0371 l) a polypeptide consisting of amino acids 55-66 0401 ) a polypeptide consisting of amino acids 41-66 of SEQID NO:ZZ: 0372 m) a polypeptide consisting of amino acids 55-86 of SEQID NO:ZZ: of SEQID NO:ZZ: 0402 k) a polypeptide consisting of amino acids 41-86 0373) n) a polypeptide consisting of amino acids 67-86 of SEQID NO:ZZ: of SEQID NO:ZZ: 0403 1) a polypeptide consisting of amino acids 55-66 0374 o) a polypeptide consisting of amino acids 1-21 of of SEQID NO:ZZ: SEQID NO:ZZ: 04.04 m) ap olypeptide consisting of amino acids 55-86 0375 p) a polypeptide consisting of amino acids 41-59 of SEQID NO:ZZ: of SEQID NO:ZZ: 0405 n) a polypeptide consisting of amino acids 67-86 0376 q) a polypeptide consisting of amino acids 41-64 of SEQID NO:ZZ: of SEQID NO:ZZ: 0406 o) a polypeptide consisting of amino acids 1-21 of 0377 r) a polypeptide consisting of amino acids 41-69 SEQ ID NO:ZZ: of SEQID NO:ZZ: 0407 p) a polypeptide consisting of amino acids 41-59 0378 s) a polypeptide consisting of amino acids 36-54 of SEQID NO:ZZ: of SEQID NO:ZZ: 04.08 q) a polypeptide consisting of amino acids 41-64 0379 t) a polypeptide consisting of amino acids 31-54 of SEQID NO:ZZ: of SEQID NO:ZZ: 04.09 r) a polypeptide consisting of amino acids 41-69 0380 u) a polypeptide consisting of amino acids 26-54 of SEQID NO:ZZ: of SEQID NO:ZZ: 0410 S) a polypeptide consisting of amino acids 36-54 0381 v) a polypeptide consisting of amino acids 36-59 of SEQID NO:ZZ: of SEQID NO:ZZ: 0411 t) a polypeptide consisting of amino acids 31-54 0382 w) a polypeptide consisting of amino acids 31-59 of SEQID NO:ZZ: of SEQID NO:ZZ: 0412 u) a polypeptide consisting of amino acids 26-54 0383 x) a polypeptide consisting of amino acids 26-59 of SEQID NO:ZZ: of SEQID NO:ZZ: 0413 V) a polypeptide consisting of amino acids 36-59 0384 y) a polypeptide consisting of amino acids 36-64 of SEQID NO:ZZ: of SEQID NO:ZZ: 0414 w) ap olypeptide consisting of amino acids 31-59 0385 Z) a polypeptide consisting of amino acids 31-64 of SEQID NO:ZZ: of SEQID NO:ZZ: 0415 X) a polypeptide consisting of amino acids 26-59 0386 aa) a polypeptide consisting of amino acids 26-64 of SEQID NO:ZZ: of SEQID NO:ZZ: 0416 y) a polypeptide consisting of amino acids 36-64 0387 ab) a polypeptide consisting of amino acids 36-69 of SEQID NO:ZZ: of SEQID NO:ZZ: 0417 Z) a polypeptide consisting of amino acids 31-64 0388 ac) a polypeptide consisting of amino acids 31-69 of SEQID NO:ZZ: of SEQID NO:ZZ; and 0418 aa) a polypeptide consisting of amino acids 26-64 0389 ad) a polypeptide consisting of amino acids 26-69 of SEQID NO:ZZ: of SEQID NO:ZZ. 0419 polypeptide consisting of amino acids 36-69 0390 The contemplated purified polypeptides include any of SEQID NO:ZZ: Subset of the aforementioned polypeptides as well as each one 0420 ac)a polypeptide consisting of amino acids 31-69 of the forgoing polypeptides. of SEQID NO:ZZ; and 0391 Among the examples of a purified polypeptide com 0421 ad) a polypeptide consisting of amino acids 26-69 prising at least 10 contiguous amino acids of SEQID NO:ZZ of SEQID NO:ZZ. and the examples of a polypeptide consisting of a polypeptide 0422 The contemplated purified polypeptides include any fragment of SEQID NO:ZZ comprising at least 10 contigu Subset of the aforementioned polypeptides as well as each one ous amino acids of SEQID NO:ZZ are: of the forgoing polypeptides. US 2012/0283411 A9 Nov. 8, 2012

0423 For SEQID NO:ZZ useful compositions, including 0430 polypeptide (f) and at least one of polypeptides pharmaceutical compositions, that include, for example, (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), (s), (t), more than one of polypeptides (a)-(ad) (or polypeptides com (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); prising, consisting of or consisting essentially of one or more 0431 polypeptide (g) and at least one of polypeptides of (a)-(ad) or a consisting of a polypeptide fragment of SEQ (h), (i), (), (k), (1), (m), (n), (o), (p), (q). (r). (S), (t), (u), ID NO:ZZ comprising one or more of (a)-(ad)) include com (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); binations 1-38 below: 0432 polypeptide (h) and at least one of polypeptides (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0433 polypeptide (i) and at least one of polypeptides 1 (a), (e) 2 (a), (e), (i) (), (k), (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), 3 (a), (e), (ii), (I) (X), (y), (Z), (aa), (ab), (ac), (ad); 4 (a), (e), (i), (), (n) 0434 polypeptide () and at least one of polypeptides 5 (a), (e), (i), (m) (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X). 6 (a), (e), (k) 7 (a), (f) (y), (Z), (aa), (ab), (ac), (ad); 8 (a), (f), (I) 0435 polypeptide (k) and at least one of polypeptides 9 (a), (f), (), (n) (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y). 10 (a), (f), (m) 11 (a), (g) (Z), (aa), (ab), (ac), (ad); 12 (a), (g), (n) 0436 polypeptide (1) and at least one of polypeptides 13 (a), (h) (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z). 14 (b), (i) (aa), (ab), (ac), (ad); 15 (b), (i), (I) 0437 polypeptide (m) and at least one of polypeptides 16 (b), (i), (), (n) 17 (b), (i), (m) (n), (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa). 18 (b), () (ab), (ac), (ad); 19 (b), (), (n) 0438 polypeptide (n) and at least one of polypeptides 2O (b), (k) (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa). 21 (c), (I) 22 (c), (), (n) (ab), (ac), (ad); 23 (d), (n) 0439 polypeptide (o) and at least one of polypeptides 24 (e), (i) (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab). 25 (e), (i), (I) 26 (e), (i), (), (n) (ac), (ad); 27 (e), (i), (m) 0440 polypeptide (p) and at least one of polypeptides 28 (e), () S.) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac). 29 (e), (), (n) aCl), 30 (e), (k) 0441 polypeptide (q) and at least one of polypeptides 31 (f), (I) 32 (f), (), (n) (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 33 (f), (m) 0442 polypeptide (r) and at least one of polypeptides 34 (g), (n) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 35 (i), (), (n) 0443 polypeptide (s) and at least one of polypeptides 36 (i), (m) 37 (), (n) (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 38 (l), (n) 0444 polypeptide (t) and at least one of polypeptides (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0424 For SEQID NO:ZZ, useful pharmaceutical compo 0445 polypeptide (u) and at least one of polypeptides sitions include: a pharmaceutical composition comprising at (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); least two (three, four or more) of polypeptides (a)-(ad) and a 0446 polypeptide (V) and at least one of polypeptides pharmaceutically acceptable carrier. In various embodiments (W), (X), (y), (Z), (aa), (ab), (ac), (ad); the pharmaceutical composition comprises: 0447 polypeptide (w) and at least one of polypeptides (X), (y), (Z), (aa), (ab), (ac), (ad); 0425 polypeptide (a) and at least one of: polypeptides 0448 polypeptide (x) and at least one of polypeptides (b), (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (y), (Z), (aa), (ab), (ac), (ad); (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), 0449 polypeptide (y) and at least one of polypeptides (ac), (ad); (Z), (aa), (ab), (ac), (ad); 0426 polypeptide (b) and at least one of polypeptides 0450 polypeptide (Z) and at least one of polypeptides (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (p), (aa), (ab), (ac), (ad); S.) (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), 0451 polypeptide (aa) and at least one of polypeptides aCl), (ab), (ac), (ad); 0427 polypeptide (c) and at least one of polypeptides 0452 polypeptide (ab) and at least one of polypeptides (d), (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (ac), (ad); and (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 0453 polypeptide (ac) and at least polypeptide (ad). 0428 polypeptide (d) and at least one of polypeptides 0454. Also described is: 1) a purified polypeptide com (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), prising (or consisting of or consisting essentially of) at least (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 10 contiguous amino acids of SEQ ID NO:ZZ1; and 2) a 0429 polypeptide (e) and at least one of polypeptides purified polypeptide consisting of a polypeptide fragment of (f), (g), (h), (i), (), (k), (l), (m), (n), (o), (p), (q). (r), (S), SEQID NO:ZZ1 comprising (or consisting of or consisting (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); essentially of) at least 10 contiguous amino acids of SEQID US 2012/0283411 A9 Nov. 8, 2012

NO:ZZ1 VX, X YXGXXVX LX SX's KX, L Xs, is S or N: X19 X20 L X22 X23 X24 X2s X26 X27 X28 X29 Xso Xs X32 Xss X34Xss X36 X37Xss X39X40X4 CX4s X44X4s ALPX49D Xss is Q., E. K or S. L Xs2 P Xsa C Xso X57 Xss Xso Xso X61 X62 Xos X64 X6's X66 Xs is E. D or Q; L RXXX, ND D C E L CV NVA (SEQID NO:ZZ1 uroguanylin consensus 1), wherein: Xo is A or V. X is S or A: X, is Y or D: X is S or N: X is Q or K. X is I or T. X is Q, H or E: X is For L.; X is For Y: X is R or Q; Xs is K, Q or L.; X is Q or K. X is T or A: X is E. K or D; X is T or S. Xs is MorV: Xzo is I or M; and X, is K or Q. X, is A, S or D. 0456. In various embodiments: the purified polypeptide X is S, N, K or D: comprises at least 15 amino acids, at least 20 amino acids, at X is D, E or A: least 25 amino acids, at least 30 amino acids, at least 40 amino acids, at least 45 amino acids, at least 50 amino acids, at least X is E. V or L. 55 amino acids, at least 60 amino acids, at least 65 amino X is A, E or G; acids, at least 70 amino acids, at least 75 amino acids, or at least 80 amino acids. In various embodiments the purified X is Q or K. polypeptide comprises fewer than 80, 75, 7065, 60, 55, 50, X is W. Q., E or P; 45, 40, 35, 30, 25 or 20 contiguous amino acids of SEQID NO:ZZ1 and has the ability to bind and/or activate the GC-C X is A, M.V or R: receptor and/or elicit diuresis when administered to a Subject. X, is P or S; 0457 Compositions, including pharmaceutical composi Xs is S, N, D or F: tions, can include at least one such polypeptide or can include at least two (three, four or more) such polypeptides which are X is P or R: different. In the compositions containing two or more Such polypeptides the polypeptides can be separate or they can be X is R. Q. G or H: covalently direct linked, e.g., by a peptide bond or a linker or X is L., P, Q or R: they can be indirectly linked. For example, two Such polypep tide sequences can be contained within a larger polypeptide X is Q, R or M: and the two polypeptide sequences can be separated by other X is A, K, R., D or G; polypeptide sequences. 0458 In certain embodiments of the purified polypeptide X is Q, S or T. comprising at least 10 contiguous amino acids of SEQ ID X is S. G. D or Q; NO:ZZ1 and in certain embodiments of the polypeptide con sisting of a polypeptide fragment of SEQID NO:ZZ1 com 0455 X is L., R or is missing: prising at least 10 contiguous amino acids of SEQ ID X, is L., P or D; NO:ZZ1: Xs is L. Q or P; X is Y: X is P or S; X is Q or K. X is A, S, D or V: X is Q, or H: X is V or L.; Xs is F: X is H. Y or S; X is R or Q; X is H. N or D: X is Q; Xas is P or S; X is E: X is Q, L, Por S. X is Mor V: Xs is Q or R; X, is K. Xs is V or I; X is S. Xs is A, Q, T or E: X20 is D;

US 2012/0283411 A9 Nov. 8, 2012 21

Xs is A, Q, T or E: Xs is Mor V: Xs, is S or N: X, is K. Xss is Q., E. K or S. X is S. Xso is E. D or Q; X20 is D; X is A or V. X2 is E. X is S or A: X is A or E: X is S or N: X is Q or K. X is I or T. Xs is W: X is For L.; X is A or M: Xs is K, Q or L.; X, is P or S; X is T or A: Xs is S. X is T or S. X is P. Xzo is I or M: X is R or Q; X, is A, S or D: X is L.; X, is N. T. G or Q; X is Q or R; X is A or K. X-7 is D or E; and X is Q or S. X is V or I. Xs is S or G; 0462. In various embodiments: the purified polypeptide comprises at least 15 amino acids, at least 20 amino acids, at 0465 X is missing: least 25 amino acids, at least 30 amino acids, at least 40 amino acids, at least 45 amino acids, at least 50 amino acids, at least X, is L or P; 55 amino acids, at least 60 amino acids, at least 65 amino Xss is L or Q. acids, at least 70 amino acids, at least 75 amino acids, or at least 80 amino acids. In various embodiments the purified X is P. polypeptide comprises fewer than 80, 75, 7065, 60, 55, 50, Xao is A. 45, 40, 35, 30, 25 or 20 contiguous amino acids of SEQ ID NO:ZZ2 and has the ability to bind and/or activate the GC-C X is V: receptor and/or elicit diuresis when administered to a subject. Xas is H. 0463 Compositions, including pharmaceutical composi tions, can include at least one such polypeptide or can include X is Hor N: at least two (three, four or more) such polypeptides which are Xs is P. different. In the compositions containing two or more Such polypeptides the polypeptides can be separate or they can be Xao is Q or L. covalently direct linked, e.g., by a peptide bond or a linker or Xs is Q. they can be indirectly linked. For example, two Such polypep tide sequences can be contained within a larger polypeptide Xs is V or I; and the two polypeptide sequences can be separated by other Xs is A. polypeptide sequences. 0464. In certain embodiments of the purified polypeptide Xs, is S. comprising at least 10 contiguous amino acids of SEQ ID Xss is Q; NO:ZZ2 and in certain embodiments of the polypeptide con sisting of a polypeptide fragment of SEQID NO:ZZ2 com Xso is E. prising at least 10 contiguous amino acids of SEQ ID Xgo is A. NO:ZZ2: X is S or A: X is Y: X is S. X is Q or K. X is I or T. X is Q, or H: X is F: X is F: Xes is K. X is R or Q; X is T or A: X is Q; X is T. X is E: Xzo is I; US 2012/0283411 A9 Nov. 8, 2012 22

X-7 is A: 0493 aa) a polypeptide comprising amino acids 27-65 of SEQID NO:ZZ1 or SEQID NO:ZZ2: X, is N or T. 0494 ab) a polypeptide comprising amino acids 37-70 X7 is D or E; and of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0495 ac) a polypeptide comprising amino acids 32-70 X7 is V or I. of SEQID NO:ZZ1 or SEQID NO:ZZ2; and 0466 Among the examples of a purified polypeptide com 0496 ad) a polypeptide comprising amino acids 27-70 prising at least 10 contiguous amino acids of SEQIDNO:ZZ1 of SEQID NO:ZZ1 or SEQID NO:ZZ2. or ZZ2 and the examples of a purified polypeptide consisting 0497. Among the examples of a purified polypeptide com of a polypeptide fragment of SEQID NO:ZZ1 or ZZ2 com prising at least 10 contiguous amino acids of SEQIDNO:ZZ1 prising at least 10 contiguous amino acids of SEQIDNO:ZZ1 or ZZ2 and the examples of a polypeptide consisting of a or ZZ2 are: polypeptide fragment of SEQID NO:ZZ1 or ZZ2 comprising 0467 a) a polypeptide consisting of amino acids 1-16 of at least 10 contiguous amino acids of SEQID NO:ZZ1 or ZZ2 SEQID NO:ZZ1 or SEQID NO:ZZ2: a. 0468 b) a polypeptide consisting of amino acids 1-41 of 0498 a) a polypeptide consisting of amino acids 1-16 of SEQID NO:ZZ1 or SEQID NO:ZZ2: SEQ ID NO:ZZ1 or SEQID NO:ZZ2; 0469 c) a polypeptide consisting of amino acids 1-55 of 0499 b) a polypeptide consisting of amino acids 1-41 of SEQID NO:ZZ1 or SEQID NO:ZZ2: SEQ ID NO:ZZ1 or SEQID NO:ZZ2; 0470 d) a polypeptide consisting of amino acids 1-67 of 0500 c) a polypeptide consisting of amino acids 1-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: SEQ ID NO:ZZ1 or SEQID NO:ZZ2; 0471 e) a polypeptide consisting of amino acids 17-41 0501 d) a polypeptide consisting of amino acids 1-67 of of SEQID NO:ZZ1 or SEQID NO:ZZ2: SEQ ID NO:ZZ1 or SEQID NO:ZZ2; 0472 f) a polypeptide consisting of amino acids 17-55 0502 e) a polypeptide consisting of amino acids 17-41 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0473 g) a polypeptide consisting of amino acids 17-67 0503 f) a polypeptide consisting of amino acids 17-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0474 h) a polypeptide consisting of amino acids 17-82 0504 g) a polypeptide consisting of amino acids 17-67 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0475 i) a polypeptide consisting of amino acids 42-55 0505 h) a polypeptide consisting of amino acids 17-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2. 0476 j) a polypeptide consisting of amino acids 42-67 0506 i) a polypeptide consisting of amino acids 42-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0477 k) a polypeptide consisting of amino acids 42-82 0507 j) a polypeptide consisting of amino acids 42-67 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0478 1) a polypeptide consisting of amino acids 56-67 05.08 k) a polypeptide consisting of amino acids 42-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2. 0479 m) a polypeptide consisting of amino acids 56-82 0509 1) a polypeptide consisting of amino acids 56-67 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0480 n) a polypeptide consisting of amino acids 68-82 0510 m) a polypeptide consisting of amino acids 56-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2. 0481 o) a polypeptide consisting of amino acids 1-21 of 0511 n) a polypeptide consisting of amino acids 68-82 SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0482 p) a polypeptide consisting of amino acids 41-60 0512 o) a polypeptide consisting of amino acids 1-21 of of SEQID NO:ZZ1 or SEQID NO:ZZ2: SEQ ID NO:ZZ1 or SEQID NO:ZZ2; 0483 q) a polypeptide consisting of amino acids 41-65 0513 p) a polypeptide consisting of amino acids 41-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0484 r) a polypeptide consisting of amino acids 41-70 0514 q) a polypeptide consisting of amino acids 41-65 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0485 s) a polypeptide consisting of amino acids 37-55 0515 r) a polypeptide consisting of amino acids 41-70 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0486 t) a polypeptide consisting of amino acids 32-55 0516 s) a polypeptide consisting of amino acids 37-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0487 u) a polypeptide consisting of amino acids 27-55 0517 t) a polypeptide consisting of amino acids 32-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0488 V) a polypeptide consisting of amino acids 37-60 0518 u) a polypeptide consisting of amino acids 27-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0489 w) a polypeptide consisting of amino acids 32-60 0519 v) a polypeptide consisting of amino acids 37-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0490 x) a polypeptide consisting of amino acids 27-60 0520 w) a polypeptide consisting of amino acids 32-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0491 y) a polypeptide consisting of amino acids 37-65 0521 x) a polypeptide consisting of amino acids 27-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0492 Z) a polypeptide consisting of amino acids 32-65 0522 y) a polypeptide consisting of amino acids 37-65 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of SEQID NO:ZZ1 or SEQID NO:ZZ2: US 2012/0283411 A9 Nov. 8, 2012 23

0523 Z) a polypeptide consisting of amino acids 32-65 0553 x) a polypeptide consisting essentially of amino of SEQID NO:ZZ1 or SEQID NO:ZZ2: acids 27-60 of SEQID NO:ZZ1 or SEQ ID NO:ZZ2: 0524 aa) a polypeptide consisting of amino acids 27-65 0554 y) a polypeptide consisting essentially of amino of SEQID NO:ZZ1 or SEQID NO:ZZ2: acids 37-65 of SEQID NO:ZZ1 or SEQ ID NO:ZZ2: 0525) ab) a polypeptide consisting of amino acids 37-70 0555 Z) a polypeptide consisting essentially of amino of SEQID NO:ZZ1 or SEQID NO:ZZ2: acids 32-65 of SEQID NO:ZZ1 or SEQ ID NO:ZZ2: 0526 ac) a polypeptide consisting of amino acids 32-70 0556 aa) a polypeptide consisting essentially of amino of SEQID NO:ZZ1 or SEQID NO:ZZ2; and acids 27-65 of SEQID NO:ZZ1 or SEQ ID NO:ZZ2: 0527 ad) a polypeptide consisting of amino acids 27-70 0557 ab) a polypeptide consisting essentially of amino of SEQID NO:ZZ1 or SEQID NO:ZZ2. acids 37-70 of SEQID NO:ZZ1 or SEQ ID NO:ZZ2: 0528 Among the examples of a purified polypeptide com 0558 ac) a polypeptide consisting essentially of amino prising at least 10 contiguous amino acids of SEQIDNO:ZZ1 acids 32-70 of SEQ ID NO:ZZ1 or SEQ ID NO:ZZ2: or ZZ2 and the examples of a polypeptide consisting of a and polypeptide fragment of SEQID NO:ZZ1 or ZZ2 comprising 0559 ad) a polypeptide consisting essentially of amino at least 10 contiguous amino acids of SEQID NO:ZZ1 or ZZ2 acids 27-70 of SEQID NO:ZZ1 or SEQ ID NO:ZZ2. a. 0560. The contemplated purified polypeptides include any 0529 a) a polypeptide consisting essentially of amino Subset of the aforementioned polypeptides as well as each one acids 1-16 of SEQID NO:ZZ1 or SEQID NO:ZZ2: of the forgoing polypeptides. b) a polypeptide consisting essentially of amino 0530 0561. For SEQ ID NO:ZZ1 and SEQID NO:ZZ2 useful acids 1-41 of SEQID NO:ZZ1 or SEQID NO:ZZ2: compositions, including useful pharmaceutical composi 0531 c) a polypeptide consisting essentially of amino tions, that include, for example, more than one of polypep acids 1-55 of SEQID NO:ZZ1 or SEQ tides (a)-(ad) (or polypeptides comprising, consisting of or 0532 ID NO:ZZ2: consisting essentially of one or more of (a)-(ad) or a consist 0533 d) a polypeptide consisting essentially of amino ing of a polypeptide fragment of SEQ ID NO:ZZ1 or ZZ2) acids 1-67 of SEQID NO:ZZ1 or SEQIDN comprising one or more of (a)-(ad)) include combinations 0534 e) a polypeptide consisting essentially of amino 1-38 below: acids 17-41 of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0535 f) a polypeptide consisting essentially of amino acids 17-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2; 0536 g) a polypeptide consisting essentially of amino (a), (e) (a), (e), (i) acids 17-67 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (a), (e), (i), (I) 0537 h) a polypeptide consisting essentially of amino (a), (e), (i), (), (n) acids 17-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2. (a), (e), (i), (m) i) a polypeptide consisting essentially of amino (a), (e), (k) 0538 (a), (f) acids 42-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (a), (f), (I) 0539 j) a polypeptide consisting essentially of amino (a), (f), (), (n) 1 (a), (f), (m) acids 42-67 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (a), (g) (0540 k) a polypeptide consisting essentially of amino (a), (g), (n) acids 42-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2. (a), (h) (0541 1) a polypeptide consisting essentially of amino (b), (i) (b), (i), (I) acids 56-67 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (b), (i), (), (n) (0542 m) a polypeptide consisting essentially of amino (b), (i), (m) acids 56-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2. (b), () (0543 n) a polypeptide consisting essentially of amino (b), (), (n) (b), (k) acids 68-82 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (c), (I) (0544 o) a polypeptide consisting essentially of amino (c), (), (n) acids 1-21 of SEQID NO:ZZ1 or SEQIDN (d), (n) p) a polypeptide consisting essentially of amino (e), (i) 0545 (e), (i), (I) acids 41-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (e), (i), (), (n) (0546 q) a polypeptide consisting essentially of amino (e), (i), (m) acids 41-65 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (e), () consisting essentially of amino (e), (), (n) 0547 r) a polypeptide (e), (k) acids 41-70 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (f), (I) (0548 S) a polypeptide consisting essentially of amino (f), (), (n) (f), (m) acids 37-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (g), (n) 0549 t) a polypeptide consisting essentially of amino (i), (), (n) acids 32-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (i), (m) 0550 u) a polypeptide consisting essentially of amino (), (n) acids 27-55 of SEQID NO:ZZ1 or SEQID NO:ZZ2: (l), (n) 0551 V) a polypeptide consisting essentially of amino acids 37-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: 0562 For SEQID NO:ZZ1 and SEQID NO:ZZ2, useful 0552) w) a polypeptide consisting essentially of amino pharmaceutical compositions include: a pharmaceutical acids 32-60 of SEQID NO:ZZ1 or SEQID NO:ZZ2: composition comprising at least two (three, four or more) of US 2012/0283411 A9 Nov. 8, 2012 24 polypeptides (a)-(ad) and a pharmaceutically acceptable car 0584) polypeptide (V) and at least one of polypeptides rier. In various embodiments the pharmaceutical composition (W), (X), (y), (Z), (aa), (ab), (ac), (ad); comprises: 0585 polypeptide (w) and at least one of polypeptides 0563 polypeptide (a) and at least one of: polypeptides (X), (y), (Z), (aa), (ab), (ac), (ad); (b), (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), 0586 polypeptide (x) and at least one of polypeptides (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (y), (Z), (aa), (ab), (ac), (ad); (ac), (ad); 0587 polypeptide (y) and at least one of polypeptides 0564) polypeptide (b) and at least one of polypeptides (Z), (aa), (ab), (ac), (ad); (c), (d), (e), (f), (g), (h), (i), (), (k), (1), (m), (n), (o), (p), 0588 polypeptide (Z) and at least one of polypeptides s'). (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (aa), (ab), (ac), (ad); aCl), 0589 polypeptide (aa) and at least one of polypeptides 0565 polypeptide (c) and at least one of polypeptides (ab), (ac), (ad); (d), (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). 0590 polypeptide (ab) and at least one of polypeptides (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); (ac), (ad); and 0566 polypeptide (d) and at least one of polypeptides 0591 polypeptide (ac) and at least polypeptide (ad). (e), (f), (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), 0592 Useful polypeptides include those comprising, con (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); sisting of, or consisting essentially of amino acids 1-23 of 0567 polypeptide (e) and at least one of polypeptides SEQ ID NO:X; amino acids 48-66 of SEQID NO:X; amino (f), (g), (h), (i), (), (k), (l), (m), (n), (o), (p), (q). (r), (S), acids 48-71 of SEQID NO:X; amino acids 48-76 of SEQ ID (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); NO:X; amino acids 43-61 of SEQ ID NO:X; amino acids 0568 polypeptide (f) and at least one of polypeptides 38-61 of SEQID NO:X; amino acids 33-61 of SEQID NO:X; (g), (h), (i), (), (k), (1). (m), (n), (o), (p), (q). (r), (S), (t), amino acids 43-66 of SEQID NO:X; amino acids 38-66 of (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); SEQ ID NO:X; amino acids 33-66 of SEQID NO:X; amino 0569 polypeptide (g) and at least one of polypeptides acids 43-71 of SEQID NO:X; amino acids 38-71 of SEQ ID (h), (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (S), (t), (u), NO:X; amino acids 33-71 of SEQ ID NO:X; amino acids (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); 43-76 of SEQID NO:X; amino acids 38-76 of SEQID NO:X; 0570 polypeptide (h) and at least one of polypeptides amino acids 33-76 of SEQ ID NO:X; amino acids 1-23 of (i), (), (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), SEQ ID NO:X1; amino acids 51-70 of SEQ ID NO:X1; (W), (X), (y), (Z), (aa), (ab), (ac), (ad); amino acids 51-75 of SEQID NO:X1; amino acids 51-80 of 0571 polypeptide (i) and at least one of polypeptides SEQ ID NO:X1; amino acids 46-65 of SEQ ID NO:X1; (), (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), amino acids 41-65 of SEQID NO:X1; amino acids 36-65 of (X), (y), (Z), (aa), (ab), (ac), (ad); SEQ ID NO:X1; amino acids 46-70 of SEQ ID NO:X1; 0572 polypeptide () and at least one of polypeptides amino acids 41-70 of SEQID NO:X1; amino acids 36-70 of (k), (1), (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), SEQ ID NO:X1; amino acids 46-75 of SEQ ID NO:X1; (y), (Z), (aa), (ab), (ac), (ad); amino acids 41-75 of SEQID NO:X1; amino acids 36-75 of 0573 polypeptide (k) and at least one of polypeptides SEQ ID NO:X1; amino acids 46-80 of SEQ ID NO:X1; (1). (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y). amino acids 41-80 of SEQID NO:X1; amino acids 36-80 of (Z), (aa), (ab), (ac), (ad); SEQIDNO:X1; amino acids 1-23 of SEQID NO: X2:amino 0574 polypeptide (1) and at least one of polypeptides acids 51-70 of SEQID NO:X2:amino acids 51-75 of SEQID (m), (n), (o), (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), NO: X2: amino acids 51-80 of SEQID NO: X2: amino acids (aa), (ab), (ac), (ad); 46-65 of SEQID NO: X2: amino acids 41-65 of SEQID NO: 0575 polypeptide (m) and at least one of polypeptides X2:amino acids 36-65 of SEQID NO: X2:amino acids 46-70 (n), (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), of SEQID NO: X2: amino acids 41-70 of SEQID NO: X2: (ab), (ac), (ad); amino acids 36-70 of SEQID NO: X2: amino acids 46-75 of 0576 polypeptide (n) and at least one of polypeptides SEQ ID NO: X2: amino acids 41-75 of SEQ ID NO: X2: (o), (p), (q). (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), amino acids 36-75 of SEQID NO: X2: amino acids 46-80 of SEQID NO: X2: amino acids 41-80 of SEQID NO: X2; and (ab), (ac), (ad); amino acids 36-80 of SEQ ID NO: X2. The contemplated 0577 polypeptide (o) and at least one of polypeptides compositions include any Subset of the aforementioned (p), (q). (r), (s), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), polypeptides as well as each one of the forgoing polypeptides. (ac), (ad); 0593] Useful polypeptides include those comprising, con 0578 polypeptide (p) and at least one of polypeptides sisting of, or consisting essentially of amino acids 1-21 of s'). (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), SEQ ID NO:ZZ; amino acids 41-59 of SEQ ID NO:ZZ: aCl), amino acids 41-64 of SEQID NO:ZZ; amino acids 41-69 of 0579 polypeptide (q) and at least one of polypeptides SEQ ID NO:ZZ; amino acids 36-54 of SEQ ID NO:ZZ: (r), (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); amino acids 31-54 of SEQID NO:ZZ; amino acids 26-54 of 0580 polypeptide (r) and at least one of polypeptides SEQ ID NO:ZZ; amino acids 36-59 of SEQ ID NO:ZZ: (S), (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); amino acids 31-59 of SEQID NO:ZZ: amino acids 26-59 of 0581 polypeptide (s) and at least one of polypeptides SEQ ID NO:ZZ: amino acids 36-64 of SEQ ID NO:ZZ: (t), (u). (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); amino acids 31-64 of SEQID NO:ZZ; amino acids 26-64 of 0582 polypeptide (t) and at least one of polypeptides SEQ ID NO:ZZ; amino acids 36-69 of SEQ ID NO:ZZ: (u), (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); amino acids 31-69 of SEQID NO:ZZ: amino acids 26-69 of 0583 polypeptide (u) and at least one of polypeptides SEQ ID NO:ZZ: amino acids 1-21 of SEQ ID NO:ZZ1; (V), (W), (X), (y), (Z), (aa), (ab), (ac), (ad); amino acids 41-60 of SEQID NO:ZZ1; amino acids 41-65 of US 2012/0283411 A9 Nov. 8, 2012

SEQ ID NO:ZZ1; amino acids 41-70 of SEQ ID NO:ZZ1; composition comprising an aforementioned polypeptide to a amino acids 37-55 of SEQID NO:ZZ1; amino acids 32-55 of patient in need thereof. In various embodiments the gas SEQ ID NO:ZZ1; amino acids 27-55 of SEQ ID NO:ZZ1; trointestinal disorder is: a gastrointestinal motility disorder, amino acids 37-60 of SEQID NO:ZZ1; amino acids 32-60 of irritable bowel syndrome, a functional gastrointestinal disor SEQ ID NO:ZZ1; amino acids 27-60 of SEQ ID NO:ZZ1; der, gastroesophageal reflux disease, duodenogastric reflux, amino acids 37-65 of SEQID NO:ZZ1; amino acids 32-65 of functional heartburn, dyspepsia, functional dyspepsia, non SEQ ID NO:ZZ1; amino acids 27-65 of SEQ ID NO:ZZ1; ulcer dyspepsia, gastroparesis, chronic intestinal pseudo-ob amino acids 37-70 of SEQID NO:ZZ1; amino acids 32-70 of struction, or colonic pseudo-obstruction. SEQ ID NO:ZZ1; amino acids 27-70 of SEQ ID NO:ZZ1; 0600 Also described is a method of treating obesity com amino acids 1-21 of SEQID NO: ZZ2; amino acids 41-60 of prising administering a pharmaceutical composition com SEQ ID NO: ZZ2; amino acids 41-65 of SEQID NO: ZZ2: prising an aforementioned polypeptide to a patient in need amino acids 41-70 of SEQID NO: ZZ2; amino acids 37-55 of thereof. SEQID NO: ZZ2; amino acids 32-55 of SEQID NO: ZZ2: 0601 Also described is a method of treating congestive amino acids 27-55 of SEQID NO: ZZ2; amino acids 37-60 of heart failure comprising administering a pharmaceutical SEQID NO: ZZ2; amino acids 32-60 of SEQID NO: ZZ2: composition of any comprising an aforementioned polypep amino acids 27-60 of SEQID NO: ZZ2; amino acids 37-65 of tide to a patient in need thereof. In various embodiments the SEQID NO: ZZ2; amino acids 32-65 of SEQID NO: ZZ2: congestive heart failure is categorized as Class II congestive amino acids 27-65 of SEQID NO: ZZ2; amino acids 37-70 of heart failure; the congestive heart failure is categorized as SEQID NO: ZZ2; amino acids 32-70 of SEQID NO: ZZ2: Class III congestive heart failure; and the congestive heart and amino acids 27-70 of SEQ ID NO: ZZ2. The contem failure is categorized as Class IV congestive heart failure. The plated compositions include any Subset of the aforemen New York Heart Association (NYHA) functional classifica tioned polypeptides as well as each one of the forgoing tion system relates congestive heart failure symptoms to polypeptides. everyday activities and the patient’s quality of life. The 0594. In various embodiments of the aforementioned puri NYHA defines the classes of patient symptoms relating to fied polypeptides: the polypeptide binds to the GC-C recep congestive heart failure as: Class II-slight limitation of physi tor; the polypeptide increases cGMP levels when adminis cal activity, comfortable at rest, but ordinary physical activity tered to a subject; the polypeptide increases cGMP levels in results in fatigue, palpitation, or dyspnea; Class III—marked the T84 assay; the polypeptide increases intestinal transit limitation of physical activity, comfortable at rest, but less when administered to a subject; the polypeptide decreases than ordinary activity causes fatigue, palpitation, or dyspnea intestinal transit when administered to a subject; the polypep and Class IV unable to carry out any physical activity with tide decreases stool firmness when administered to a subject; out discomfort, symptoms of cardiac insufficiency at rest, if the polypeptide increases stool frequency when administered any physical activity is undertaken, discomfort is increased. to a Subject; the polypeptide decreases visceral pain when Heart failure treatment using the polypeptides and methods administered to a Subject; the polypeptide modulates activity described herein can also be classified according to the ACC/ of the GC-C receptor; the polypeptide increases the activity of AHA guidelines (Stage A: At risk for developing heart failure the GC-C receptor and the polypeptide decreases the activity without evidence of cardiac dysfunction; Stage B: Evidence of the GC-C receptor, the polypeptide increases urine output of cardiac dysfunction without symptoms; Stage C. Evidence in a rodent diuresis assay, the polypeptide elicits diuresis of cardiac dysfunction with symptoms; and Stage D: Symp when administered to a Subject, the polypeptide elicits nature toms of heart failure despite maximal therapy). sis when administered to a subject, the polypeptide elicits 0602 Also described is a method of treating benign pros kaluresis when administered to a subject tatic hyperplasia comprising administering a pharmaceutical 0595 Also described is a method of treating a disorder composition comprising an aforementioned polypeptide to a associated with reduced gastrointestinal transit rate or reduced gastrointestinal motility comprising administering a patient in need thereof. pharmaceutical composition comprising an aforementioned 0603 Also described is a method of treating constipation polypeptide to a patient in need thereof. comprising administering the pharmaceutical composition 0596. Also described is a method of treating a gastrointes comprising an aforementioned polypeptide to a patient in tinal hypomotility disorder comprising administering a phar need thereof. maceutical composition comprising an aforementioned 0604. Also described is a method of treating a gastrointes polypeptide to a patient in need thereof. In various embodi tinal motility disorder in a patient, the method comprising ments: the disorder is selected from the group consisting of administering a pharmaceutical composition comprising an constipation, constipation dominant irritable bowel syn aforementioned polypeptide to a patient in need thereof. drome and pelvic floor dyssynergia. 0605 Also described is a method of treating gastrointes 0597 Also described is a method of treating a non-inflam tinal pain or visceral pain in a patient, the method comprising matory gastrointestinal disorder comprising administering a administering a pharmaceutical composition comprising an pharmaceutical composition comprising an aforementioned aforementioned polypeptide to a patient in need thereof. polypeptide to a patient in need thereof. 0606. Also described is a method for increasing the activ 0598. Also described is a method of treating a gastrointes ity of an intestinal guanylate cyclase (GC-C) receptor in a tinal disorder other than Crohn's disease and ulcerative colitis patient, the method comprising administering a pharmaceu comprising administering a pharmaceutical composition tical composition comprising an aforementioned polypeptide comprising an aforementioned polypeptide to a patient in to a patient in need thereof. need thereof. 0607 Also described is a method of treating a disorder 0599 Also described is a method of treating a gastrointes characterized by salt retention, fluid retention, and combina tinal disorder comprising administering a pharmaceutical tions thereof, in a patient in need thereof, the method com US 2012/0283411 A9 Nov. 8, 2012 26 prising administering a pharmaceutical composition com 0613. In certain embodiments the patient has been diag prising an aforementioned polypeptide to a patient in need nosed as suffering from IBS according to the Rome criteria. In thereof. certain embodiments the patient is female. 0608 Unless otherwise specified, the term “agonist used 0614 The polypeptide can contain additional carboxy ter herein refers to an agonist of the GC-C receptor. The polypep minal or amino terminal amino acids or both. For example, tide can contain up to four cysteines that form one or two the polypeptide can include an amino terminal sequence that disulfide bonds or do not form a disulfide bond. In certain facilitates a recombinant production of the polypeptide and is embodiments the disulfide bonds are replaced by other cova cleaved prior to administration of the polypeptide to a patient. The polypeptide can also include other amino terminal or lent cross-links and in Some cases the cysteines are Substi carboxy terminal amino acids. In some cases the additional tuted by other residues to provide for alternative covalent amino acids protect the polypeptide, stabilize the polypeptide cross-links. The polypeptides may also include at least one or alter the activity of the polypeptide. In some cases some or trypsin or chymotrypsin cleavage site and/or a carboxy-ter all of these additional amino acids are removed prior to minal analgesic polypeptide or Small molecule, e.g., AspPhe administration of the polypeptide to a patient. The polypep or some other analgesic polypeptide. When present within the tide can include 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70 polypeptide, the analgesic polypeptide or Small molecule 80, 90, 100 or more amino acids at its amino terminus or may be preceded by a chymotrypsin or trypsin cleavage site carboxy terminus or both. The number of flanking amino that allows release of the analgesic polypeptide or Small mol acids need not be the same. For example, there can be 10 ecule. The polypeptides and methods are also useful for treat additional amino acids at the amino terminus of the polypep ing pain and inflammation associated with various disorders, tide and none at the carboxy terminus. including gastrointestinal disorders. Certain polypeptides 0615. The polypeptides described herein can be adminis include a functional chymotrypsin or trypsin cleavage site tered together with: mature human guanylin or a polypeptide located so as to allow inactivation of the polypeptide upon comprising the amino acid sequence of mature human gua cleavage. Certain polypeptides having a functional cleavage nylin, mature human uroguanylin or a polypeptide compris site undergo cleavage and gradual inactivation in the digestive ing the amino acid sequence of mature human uroguanylin, or tract, and this is desirable in some circumstances. In certain certain other polypeptides that act as GC-C receptoragonists. polypeptides, a functional chymotrypsin site is altered, 0616. Useful variants of PGTCEICAYAACTGC (human increasing the stability of the polypeptide in vivo (e.g., gua guanylin) (SEQ ID NO:) that can be combined with the nylin). polypeptide described herein include: 0609 Among the methods described herein are: a method for increasing intestinal motility comprising administering a GC-C receptor agonist, e.g., a polypeptide described herein, PGTCECATAACTGC (SEQ ID NO: ) to a patient in need thereof: a method for treating a disorder PGTCEICANAACTGC (SEQ ID NO: ) associated with reduced gastrointestinal transit rates or reduced gastrointestinal motility comprising administering a PGTCEICAOAACTGC (SEQ ID NO: ) GC-C receptor agonist, e.g., a polypeptide described herein, PGTCEICARAACTGC (SEQ ID NO: ) to a patient in need thereof a method for treating a gas trointestinal hypomotility disorder comprising administering PGTCEICAEAACTGC (SEQ ID NO: ) a GC-C receptoragonist, e.g., a polypeptide described herein, to a patient in need thereof a method treating a non-inflam PGTCEICADAACTGC (SEQ ID NO: ) matory gastrointestinal disorder comprising administering a PGTCEICAGAACTGC (SEQ ID NO: ) GC-C receptor agonist, e.g., a polypeptide described herein, to a patient in need thereof...; a method treating a gastrointes PGTCEICAAAACTGC (SEQ ID NO: ) tinal disorder other than Crohn's disease and ulcerative colitis PGTCEICAMAACTGC (SEQ ID NO: ) comprising administering a GC-C receptor agonist, e.g., a polypeptide described herein, to a patient in need thereof. PGTCEICAIAACTGC (SEQ ID NO: ) 0610 The disorders which can be treated by administering PGTCEICALAACTGC (SEQ ID NO: ) a GC-C receptoragonist, e.g., a polypeptide described herein, include constipation, constipation dominant irritable bowel PGTCEICAWAACTGC (SEQ ID NO: ) syndrome and pelvic floor dyssynergia. PGTCEICAHAACTGC (SEQ ID NO: ) 0611. Also described are methods for treating other disor ders such as congestive heart failure and benign prostatic PGTCEGICAYAACTGC (SEQ ID NO: ) hyperplasia by administering a polypeptide or Small molecule (parenterally or orally) that acts as an agonist of the GC-C PGTCEIGCAYAACTGC (SEQ ID NO: ) receptor. Such agents can be used in combination with natri PGTCEICGAYAACTGC (SEQ ID NO: ) uretic polypeptides (e.g., atrial natriuretic polypeptide, brain natriuretic polypeptide or C-type natriuretic polypeptide), a PGTCEICAGYAACTGC (SEQ ID NO: ) diuretic, or an inhibitor of angiotensin converting enzyme. PGTCEICAYGAACTGC (SEQ ID NO: ) 0612 Also described are methods and compositions for increasing intestinal motility. Intestinal motility involves PGTCEICAYAGACTGC (SEQ ID NO: ) spontaneous coordinated distentions and contractions of the PGTCEICAYAAGCTGC (SEQ ID NO: ) stomach, intestines, colon and rectum to move food through the gastrointestinal tract during the digestive process. US 2012/0283411 A9 Nov. 8, 2012 27

Chem 21:117) to form alternative internal cross-links at the - Continued positions of the normal disulfide bonds. PGTCEICAYAACGTGC (SEQ ID NO: ) 0622. In addition, one or more disulfide bonds can be PGTCEICAYAACTGGC (SEQ ID NO: ) replaced by alternative covalent cross-links, e.g., an amide linkage (-CHCH(O)NHCH- or -CH-NHCH(O) PGTCAEICAYAACTGC (SEQ ID NO: ) CH2—), an ester linkage, a thioester linkage, a lactambridge, a carbamoyl linkage, a urea linkage, a thiourea linkage, a PGTCEAICAYAACTGC (SEQ ID NO: ) phosphonate ester linkage, an alkyl linkage PGTCEIACAYAACTGC (SEQ ID NO: ) (—CHCHCHCH ), al alkenyl linkage ( CH-CH=CHCH ), al ether linkage PGTCEICAAYAACTGC (SEQ ID NO: ) (—CH2CHOCH2—, or —CHOCH2CH2—), a thioether PGTCEICAYAAACTGC (SEQ ID NO: ) linkage ( CHCH-SCH or —CH-SCH-CH ), an amine linkage ( CHCH-NHCH- O PGTCEICAYAACATGC (SEQ ID NO: ) —CH-NHCHCH ) or a thioamide linkage ( CHCH(S) HNHCH-or-CHNHCH(S)CH, ). For example, Ledu PGTCEICAYAACTAGC (SEQ ID NO: ) et al. (Proc Natl Acad. Sci. 100:11263-78, 2003) describe PGTCEICAYAACTGAC (SEQ ID NO: ) methods for preparing lactam and amide cross-links. Schafmeister et al. (J. Am. Chem. Soc. 122:5891, 2000) PGTCAEICAAYAACTGC (SEQ ID NO: ) describe stable, hydrocarbon cross-links. Hydrocarbon cross PGTCEAICAAYAACTGC (SEQ ID NO: ) links can be produced via metathesis (or methathesis fol lowed by hydrogenation in the case of saturated hydrocarbons PGTCEIACAAYAACTGC (SEQ ID NO: ) cross-links) using one or another of the Grubbs catalysts 0617 The polypeptides described herein can be attached (available from Materia, Inc. and Sigma-Aldrich and to one, two or more different moieties each providing the described, for example, in U.S. Pat. Nos. 5,831,108 and same or different functions. For example, the polypeptide can 6,111,121). In some cases, the generation of Such alternative be linked to a molecule that is an analgesic and to a polypep cross-links requires replacing the Cys residues with other tide that is used to treat obesity. The polypeptide and various residues such as Lys or Glu or non-naturally occurring amino moieties can be ordered in various ways. For example, a acids. In addition the lactam, amide and hydrocarbon cross polypeptide described herein can have an analgesic polypep links can be used to stabilize the polypeptide even if they link tide linked to its amino terminus and an anti-obesity polypep amino acids at positions other than those occupied by Cys. tide linked to its carboxy terminus. The additional moieties Such cross-links can occur between two amino acids that are can be directly covalently bonded to the polypeptide or can be separated by two amino acids or between two amino acids bonded via linkers. that are separated by six amino acids (see, e.g., Schafmeister 0618. The polypeptides described herein can be a cyclic et al. (J. Am. Chem. Soc. 122:5891, 2000)). polypeptide or a linear polypeptide. In addition, multiple 0623. In certain embodiments, one or more amino acids copies of the same polypeptide can be incorporated into a can be replaced by a non-naturally occurring amino acid or a single cyclic or linear polypeptide. naturally or non-naturally occurring amino acid analog. 0619. The polypeptides can include the amino acid There are many amino acids beyond the standard 20 (Ala, sequence of a polypeptide that occurs naturally in a vertebrate Arg, ASn, Asp, Cys, Gln, Glu, Gly, H is, Ile, Leu, Lys, Met, (e.g., mammalian) species or in a bacterial species. In addi Phe, Pro, Ser. Thr, Tip, Tyr, and Val). Some are naturally tion, the polypeptides can be partially or completely non occurring others are not (see, for example, Hunt. The Non naturally occurring polypeptides. Also within the invention Protein Amino Acids: In Chemistry and Biochemistry of the are peptidomimetics corresponding to the polypeptides Amino Acids, Barrett, Chapman and Hall, 1985). For described herein. example, an aromatic amino acid can be replaced by 3.4- 0620. In the case of mature uroguanylin and mature gua dihydroxy-L-phenylalanine, 3-iodo-L-tyrosine, triiodothyro nylin, when fully folded, disulfide bonds are present between nine, L-thyroxine, phenylglycine (Phg) or nor-tyrosine the first and third cysteines and between the second and fourth (norTyr). Phg and norTyrand otheramino acids including Phe cysteines in the mature protein, e.g., in mature uroguanylin and Tyrcan be substituted by, e.g., a halogen, —CH-OH, there is a disulfide bond between Cys and Cys and a dis —CH-NH. —C(O)H, —CHCH. —CN, —CH2CHCH ulfide bond between Cys, and Cysis. In some embodiments, —SH, or another group. Anyamino acid can be substituted by the polypeptide has only one disulfide bond, e.g., between the the D-form of the amino acid. first and third cysteines. In certain embodiments one or more 0624 With regard to non-naturally occurring amino acids Cys can be replaced by Mpt (mercaptoproline) or Pen (peni or a naturally and non-naturally occurring amino acid ana cillamine) or Dpr (diaminopropionic acid) or some other logs, a number of Substitutions in the polypeptide and ago amino acid that can covalently link to another amino acid nists described herein are possible alone or in combination. (e.g., Cys, Mpt, Pen or Dpr). In other embodiments, the 0625 For example, glutamine residues can be substituted polypeptide is a reduced polypeptide having no disulfide with gamma-Hydroxy-Glu or gamma-Carboxy-Glu. bonds. Tyrosine residues can be substituted with an alpha substituted 0621. In some embodiments, one or both members of a amino acid such as L-alpha-methylphenylalanine or by ana pair of Cys residues which normally form a disulfide bond can logues such as: 3-Amino-Tyr; Tyr(CH3); Tyr(PO(CH)); be replaced by homocysteine, penicillamine, 3-mercaptopro Tyr(SOH); beta-Cyclohexyl-Ala; beta-(1-Cyclopentenyl)- line (Kolodzie et al. 1996 Int J Pept Protein Res 48:274): Ala; beta-Cyclopentyl-Ala; beta-Cyclopropyl-Ala; beta BB-dimethylcysteine (Hunt et al. 1993 IntJ Pept Protein Res Quinolyl-Ala; beta-(2-Thiazolyl)-Ala; beta-(Triazole-1-yl)- 42:249) or diaminopropionic acid (Smith et al. 1978 J Med Ala; beta-(2-Pyridyl)-Ala; beta-(3-Pyridyl)-Ala; Amino-Phe: US 2012/0283411 A9 Nov. 8, 2012 28

Fluoro-Phe: Cyclohexyl-Gly; tBul-Gly; beta-(3-benzothie other than proline; an O-methyl-L-tyrosine; an L-3-(2-naph nyl)-Ala; beta-(2-thienyl)-Ala; 5-Methyl-Trp.; and 4-Methyl thyl)alanine; a 3-methyl-phenylalanine; a p-acetyl-L-pheny Trp. Proline residues can be substituted with homopro lalanine; an 0-4-allyl-L-tyrosine; a 4-propyl-L-tyrosine; a tri (L-pipecolic acid); hydroxy-Pro; 3,4-Dehydro-Pro; 4-fluoro O-acetyl-GlcNAc-serine; an L-Dopa; a fluorinated Pro; or alpha-methyl-Pro or an N(alpha)-C(alpha) cyclized phenylalanine; an isopropyl-L-phenylalanine; a p-azido-L- amino acid analogues with the structure: phenylalanine; a p-acyl-L-phenylalanine; a p-benzoyl-L- phenylalanine; an L-phosphoserine; a phosphonoSerine; a

phosphonotyrosine; a p-iodo-phenylalanine; a 4-fluorophe nylglycine; a p-bromophenylalanine; a p-amino-L-phenyla lanine; an isopropyl-L-phenylalanine; L-3-(2-naphthyl)ala nine; an amino-, isopropyl-, or O-allyl-containing phenylalanine analogue; a dopa, O-methyl-L-tyrosine; a gly cosylated amino acid; a p-(propargyloxy)phenylalanine; dimethyl-Lysine; hydroxy-proline; mercaptopropionic acid; methyl-lysine; 3-nitro-tyrosine; norleucine; gyro-glutamic acid; Z (Carbobenzoxyl); e-Acetyl-Lysine; B-alanine; ami 0626. Alanine residues can be substituted with alpha-sub nobenzoyl derivative; aminobutyric acid (Abu); citrulline; stituted or N-methylated amino acid Such as alpha-amino aminohexanoic acid; aminoisobutyric acid; cyclohexylala isobutyric acid (aib), L/D-alpha-ethylalanine (L/D-isova nine; d-cyclohexylalanine, hydroxyproline; nitro-arginine; line), L/D-methylvaline, or L/D-alpha-methyleucine or a nitro-phenylalanine; nitro-tyrosine; norvaline; octahydroin non-natural amino acid Such as beta-fluoro-Ala. Alanine can dole carboxylate; ornithine; penicillamine; tetrahydroiso also substituted with: quinoline; acetamidomethyl protected amino acids and pegy lated amino acids. Further examples of unnatural amino acids and amino acid analogs can be found in U.S. 20030108885, U.S. 20030082575, US20060019347 (paragraphs 410-418) and the references cited therein. The polypeptides of the R invention can include further modifications including those described in US20060019347, paragraph 589. 0629. In some embodiments, an amino acid can be replaced by a naturally-occurring, non-essential amino acid, e.g., taurine. 0630 Methods to manufacture polypeptides containing 0627 Glycine residues can be substituted with alpha unnatural amino acids can be found in, for example, U.S. amino isobutyric acid (aib) or L/D-alpha-ethylalanine (L/D- 20030108885, U.S. 20030082575, US20060019347, Deiters isovaline). et al., J. Am Chem. Soc. (2003) 125:11782-3, Chin et al., 0628. Further examples of unnatural amino acids include: Science (2003)301:964-7, and the references cited therein. an unnatural analogue of tyrosine; an unnatural analogue of 0631 Peptides that include non-natural amino acids can glutamine; an unnatural analogue of phenylalanine; an also be prepared using the methods described in unnatural analogue of serine; an unnatural analogue of threo WOO2O86O75. nine; an alkyl, aryl, acyl, azido, cyano, halo, hydrazine, 0632. The polypeptides can have one or more conven hydrazide, hydroxyl, alkenyl, alkynl, ether, thiol, Sulfonyl, tional polypeptide bonds replaced by an alternative bond. Seleno, ester, thioacid, borate, boronate, phospho, Such replacements can increase the stability of the polypep phosphono, phosphine, heterocyclic, enone, imine, aldehyde, tide. For example, replacement of the polypeptide bond hydroxylamine, keto, or amino Substituted amino acid, or any between a residue amino terminal to an aromatic residue (e.g. combination thereof, an amino acid with a photoactivatable Tyr, Phe, Trp) with an alternative bond can reduce cleavage by cross-linker, a spin-labeled amino acid; a fluorescent amino carboxypeptidases and may increase half-life in the digestive acid; an amino acid with a novel functional group; an amino tract. Bonds that can replace polypeptide bonds include: a acid that covalently or noncovalently interacts with another retro-inverso bond (C(O) NH instead of NH C(O); a molecule; a metal binding amino acid; an amino acid that is reduced amide bond (NH-CH2); a thiomethylene bond amidated at a site that is not naturally amidated, a metal (S CH or CH S); an oxomethylene bond (O CH or containing amino acid; a radioactive amino acid; a photo CH-O); an ethylene bond (CH, CH); a thioamide bond caged and/or photoisomerizable amino acid; a biotin or (C(S) NH); a trans-olefine bond (CH=CH); a fluoro sub biotin-analogue containing amino acid; a glycosylated or car stituted trans-olefine bond (CF=CH); a ketomethylene bond bohydrate modified amino acid; a keto containing amino (C(O) CHR or CHR C(O) wherein R is H or CH; and a acid; amino acids comprising polyethylene glycol or poly fluoro-ketomethylene bond (C(O) CFR or CFR C(O) ether; a heavy atom Substituted amino acid (e.g., an amino wherein R is H or For CH. acid containing deuterium, tritium, C, Nor'O); a chemi 0633. The polypeptides can be modified using standard cally cleavable or photocleavable amino acid; an amino acid modifications. Modifications may occur at the amino (N ). with an elongated side chain; an amino acid containing a toxic carboxy (C ) terminus, internally or a combination of any of group; a Sugar Substituted amino acid, e.g., a Sugar Substituted the preceeding. In one aspect described herein, there may be serine or the like; a carbon-linked Sugar-containing amino more than one type of modification on the polypeptide. Modi acid; a redox-active amino acid; an O.-hydroxy containing fications include but are not limited to: acetylation, amida acid; an amino thio acid containing amino acid; an O.C. dis tion, biotinylation, cinnamoylation, farnesylation, formyla ubstituted amino acid; a B-amino acid; a cyclic amino acid tion, myristoylation, palmitoylation, phosphorylation (Ser, US 2012/0283411 A9 Nov. 8, 2012 29

Tyr or Thr), Stearoylation, Succinylation, Sulfurylation and diisopropylammonium salts, cyclohexyl- or dicyclohexylam cyclisation (via disulfide bridges or amide cyclisation), and monium salts and dibenzylethylenediammonium salts). Acid modification by Cy3 or Cy5. The polypeptides described addition salts include, but are note limited to: hydrochloride, herein may also be modified by 2,4-dinitrophenyl (DNP), acetate and trifluoroacetate salts. Inorganic acids which can DNP-lysin, modification by 7-Amino-4-methyl-coumarin be used to form acid addition salts include, but are not limited (AMC), flourescein, NBD (7-Nitrobenz-2-Oxa-1,3-Diazole), to: Sulfuric acid, nitric acid, hydrohalic acids such as hydro p-nitro-anilide, rhodamine B, EDANS (5-((2-aminoethyl) amino)naphthalene-1-sulfonic acid), dabcyl, dabsyl, dansyl, chloric acid or hydrobromic acid, phosphoric acids, and Sul texas red, FMOC, and Tamra (Tetramethylrhodamine). The famic acid. Organic acids which can be used to form acid polypeptides described herein may also be conjugated to, for addition salts include, but are not limited to: e.g. formic acid, example, polyethylene glycol (PEG); alkyl groups (e.g., acetic acid, propionic acid, pivalic acid, diethylacetic acid, C1-C20 straight or branched alkyl groups); fatty acid radi malonic acid. Succinic acid, pimelic acid, fumaric acid, cals; combinations of PEG, alkyl groups and fatty acid radi maleic acid, lactic acid, tartaric acid, malic acid, citric acid, cals (see U.S. Pat. No. 6,309,633; Soltero et al., 2001 Inno gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, vations in Pharmaceutical Technology 106-110); BSA and methane- or ethane-Sulfonic acid, ethanedisulfonic acid, KLH (Keyhole Limpet Hemocyanin). The addition of PEG 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-tolu and other polymers which can be used to modify polypeptides ene-Sulfonic acid, naphthalenemono- and -disulfonic acids, of the invention is described in US2006.019347 section IX. and laurylsulfuric acid. 0634. The polypeptides and agonists described herein can 0637. The various polypeptides can be present with a be chemically modified to increase therapeutic activity by counterion. Useful counterions include salts of acetate, ben synthetically adding sugar moieties (WO 88/02756; WO Zenesulfonate, benzoate, calcium edetate, camsylate, carbon 89/09786; DE 3910667 A1, EP 0374 089 A2; and U.S. Pat. ate, citrate, edetate (EDTA), edisylate, embonate, esylate, No. 4,861,755), adding cationic anchors (EP0363589), lipid fumarate, gluceptate, gluconate, glutamate, glycolylarsa moieties (WO91/09837; U.S. Pat. No. 4,837,303) or the sub nilate, hexylresorcinate, iodide, bromide, chloride, hydrox stituents described as compounds I, II, and III in U.S. Pat. No. ynaphthoate, isethionate, lactate, lactobionate, estolate, 5,552,520. maleate, malate, mandelate, mesylate, mucate, napsylate, 0635 Two or more polypeptides described herein can be nitrate, pantothenate, phosphate, Salicylate, Stearate. Succi joined by a peptide bond or by a polypeptide sequence (e.g., nate, Sulfate, tartarate, theoclate, acetamidobenzoate, adi a sequence of 1 or more amino acids) or they can be joined by pate, alginate, aminosalicylate, anhydromethylenecitrate, a linker. Many methods for protein cross-linking are known. ascorbate, aspartate, camphorate, caprate, caproate, capry Many cross-linking methods employ a thiol group and vari late, cinnamate, cyclamate, dichloroacetate, formate, genti ous methods can be used to introduce a thiol group, including sate, glucuronate, glycerophosphate, glycolate, hippurate, the reduction of intrinsic disulfides, as well as the conversion fluoride, malonate, napadisylate, nicotinate, oleate, orotate, of amine or carboxylic acid groups to thiol groups. In the oxalate, oxoglutarate, palmitate, pectinate, pectinate poly present polypeptide it is often desirable to preserve disulfide mer, phenylethylbarbiturate, picrate, propionate, pidolate, bonds. Thus, it may be desirable to introduce a thiol group. sebacate, rhodanide, tosylate, and tannate. Amines can be indirectly thiolated by reaction with succin 0638 For the treatment of gastrointestinal disorders, the imidyl 3-(2-pyridyldithio)propionate 4 (SPDP), followed by polypeptide can be administered orally, by rectal Suppository reduction of the 3-(2-pyridyldithio)propionyl conjugate with or parenterally. DTT or TCEP. Amines can be indirectly thiolated by reaction 0639. In various embodiments of the treatment methods: with succinimidyl acetylthioacetate 5 (SATA), followed by the patient is suffering from a gastrointestinal disorder, the removal of the acetyl group. Thiols can be incorporated at patient is Suffering from a disorder selected from the group carboxylic acid groups by an EDAC-mediated reaction with consisting of gastrointestinal motility disorders, chronic cystamine, followed by reduction of the disulfide. Polypep intestinal pseudo-obstruction, colonic pseudo-obstruction, tides can also be crosslinked through amines. An amine on a Crohn's disease, duodenogastric reflux, dyspepsia, functional first polypeptide can be thiolated and an amines on the second dyspepsia, nonulcer dyspepsia, a functional gastrointestinal polypeptide can be converted to a thiol-reactive functional disorder, functional heartburn, gastroesophageal reflux dis group Such as a maleimide or iodoacetamide. Indirect ease (GERD), gastroparesis, irritable bowel syndrome, post crosslinking of the amines in a first polypeptide to the thiols in operative ileus, ulcerative colitis, chronic constipation, and a second polypeptide is useful for forming a heteroconjugate. disorders and conditions associated with constipation (e.g. Thiol-reactive groups such as maleimides are typically intro constipation associated with use of opiate pain killers, post duced into the second polypeptide by modifying an amine Surgical constipation, and constipation associated with neu with a heterobifunctional crosslinker containing both a suc ropathic disorders as well as other conditions and disorders cinimidyl ester and a maleimide. The maleimide-modified are described herein); the patient is Suffering from a gas polypeptide is then reacted with the thiol-containing biomol trointestinal motility disorder, chronic intestinal pseudo-ob ecule to form a stable thioether crosslink. struction, colonic pseudo-obstruction, Crohn's disease, 0636. The peptides described herein can be in the form or duodenogastric reflux, dyspepsia, functional dyspepsia, non a salt, e.g., a pharmaceutically acceptable salt. A salt of a ulcer dyspepsia, a functional gastrointestinal disorder, func peptide can be formed by acid or base addition depending on tional heartburn, gastroesophageal reflux disease (GERD), the nature of the peptide. Suitable salts include base salts such gastroparesis, inflammatory bowel disease, irritable bowel as alkali metal salts, e.g., Sodium, potassium, magnesium, and syndrome, post-operative ileus, ulcerative colitis, chronic calcium salts. Other suitable base salts include substituted constipation, and disorders and conditions associated with and unsubstituted ammonium salts (e.g., dimethyl-, diethyl constipation (e.g. constipation associated with use of opiate or diisopropylammonium salts, monoethanol-, diethanol- or pain killers, post-Surgical constipation, and constipation US 2012/0283411 A9 Nov. 8, 2012 30 associated with neuropathic disorders as well as other condi a fusion protein with a polypeptide described herein. Thus, tions and disorders are described herein); and the composi for example, PYY can be fused to the carboxy or amino tion is administered orally. terminus of a polypeptide described herein. Such a fusion 0640. As noted, the polypeptides can be used to treat a protein can include a chymostrypsin or trypsin cleavage site patient Suffering from constipation. Clinically accepted cri that can permit cleavage to separate the two polypeptides. teria that define constipation range from the frequency of 0644 As noted, the polypeptides can be used to treat a bowel movements, the consistency of feces and the ease of patient Suffering from congestive heart failure. The polypep bowel movement. One common definition of constipation is tide can be administered in combination with one or more less than three bowel movements per week. Other definitions agents for treatment of congestive heart failure, for example, include abnormally hard stools or defecation that requires a natriuretic polypeptide Such as atrial natriuretic polypep excessive straining (Schiller 2001 Aliment Pharmacol Ther tide, brain natriuretic polypeptide or C-type natriuretic 15:749-763). Constipation may be idiopathic (functional polypeptide), nesiritide (NatrecorR), a diuretic, or an inhibi constipation or slow transit constipation) or secondary to tor of angiotensin converting enzyme. other causes including neurologic, metabolic or endocrine 0645. As noted, the polypeptides can be used to treat con disorders. These disorders include diabetes mellitus, gestive heart failure. The polypeptide can act in the kidney hypothyroidism, hyperthyroidism, hypocalcaemia, Multiple and adrenal gland to control natriuresis, kaliuresis, and diure Sclerosis, Parkinson's disease, spinal cord lesions, Neurofi sis thereby reducing the build-up of fluid associated with bromatosis, autonomic neuropathy, Chagas disease, Hirschs congestive heart failure (Lorenz et al...J Clin Invest 112:1138, prung disease and cystic fibrosis. Constipation may also be 2003: Carrithers et al. Kidney Int 65:40, 2004). The polypep the result of Surgery or due to the use of drugs such as anal tide can be administered in combination with one or more gesics (like opioids), antihypertensives, anticonvulsants, agents for treatment of congestive heart failure, including but antidepressants, antispasmodics and antipsychotics. In vari not limited to the agents useful for combitherapy described ous embodiments, the constipation is associated with use of a herein. For example, the polypeptide can be administered in therapeutic agent; the constipation is associated with a neu combination with a natriuretic polypeptide such as atrial ropathic disorder; the constipation is post-Surgical constipa natriuretic polypeptide, brain natriuretic polypeptide or tion; the constipation is associated with a gastrointestinal C-type natriuretic polypeptide), a diuretic, or an inhibitor of disorder; the constipation is idiopathic (functional constipa angiotensin converting enzyme. tion or slow transit constipation); the constipation is associ 0646. The polypeptides can be used to treat, for example, ated with neuropathic, metabolic or endocrine disorder (e.g., constipation, decreased intestinal motility, slow digestion, diabetes mellitus, hypothyroidism, hyperthyroidism, hypoc slow stomach emptying. The polypeptides can be used to alcaemia, Multiple Sclerosis, Parkinson's disease, spinal cord relieve one or more symptoms of IBS (bloating, pain, consti lesions, neurofibromatosis, autonomic neuropathy, Chagas pation), GERD (acid reflux into the esophagus), duodenogas disease, Hirschsprung disease or cystic fibrosis). Constipa tric reflux, functional dyspepsia, or gastroparesis (nausea, tion may also be the result of Surgery or due to the use of drugs Vomiting, bloating, delayed gastric emptying) and other dis Such as analgesics (e.g., opioids), antihypertensives, anticon orders described herein. Vulsants, antidepressants, antispasmodics and antipsychot 0647 Clinically accepted criteria that define constipation 1CS range from the frequency of bowel movements, the consis 0641 As noted, the polypeptides can be used to treat a tency of feces and the ease of bowel movement. One common patient Suffering from a gastrointestinal disorder. In various definition of constipation is less than three bowel movements embodiments, the patient is suffering from a gastrointestinal per week. Other definitions include abnormally hard stools or disorder; the patient is suffering from a disorder selected from defecation that requires excessive straining (Schiller 2001, the group consisting of gastrointestinal motility disorders, Aliment Pharmacol Ther 15:749-763). Constipation may be chronic intestinal pseudo-obstruction, colonic pseudo-ob idiopathic (functional constipation or slow transit constipa struction, Crohn's disease, duodenogastric reflux, dyspepsia, tion) or secondary to other causes including neurologic, meta functional dyspepsia, nonulcer dyspepsia, a functional gas bolic or endocrine disorders. These disorders include diabetes trointestinal disorder, functional heartburn, gastroesophageal mellitus, hypothyroidism, hyperthyroidism, hypocalcaemia, reflux disease (GERD), gastroparesis, irritable bowel syn Multiple Sclerosis, Parkinson's disease, spinal cord lesions, drome, post-operative ileus, ulcerative colitis, chronic consti Neurofibromatosis, autonomic neuropathy, Chagas disease, pation, and disorders and conditions associated with consti Hirschsprung's disease and cystic fibrosis. Constipation may pation (e.g. constipation associated with use of opiate pain also be the result of Surgery or due to the use of drugs such as killers, post-Surgical constipation, and constipation associ analgesics (like opioids), antihypertensives, anticonvulsants, ated with neuropathic disorders as well as other conditions antidepressants, antispasmodics and antipsychotics. and disorders are described herein), obesity, congestive heart 0648. Also featured are isolated nucleic acid molecules failure, or benign prostatic hyperplasia comprising or consisting of a sequence encoding a polypep 0642 Also featured is an isolated nucleic acid molecule tide described herein. The invention also features vectors, comprising a nucleotide sequence encoding a polypeptide e.g., expression vectors that include Such nucleic acid mol above. ecules and can be used to express a polypeptide described 0643. As noted, the polypeptides can be used to treat a herein in a cultured cell (e.g., a eukaryotic cell or a prokary patient Suffering from obesity. The polypeptide can be admin otic cell). The vector can further include one or more regula istered in combination with one or more agents for treatment tory elements, e.g., a heterologous promoter or elements of obesity, including, without limitation, the anti-obesity required for translation operably linked to the sequence agents described herein. A polypeptide useful for treating encoding the polypeptide. In some cases the nucleic acid obesity can be administered as a co-therapy with a polypep molecule will encode an amino acid sequence that includes tide described herein either as a distinct molecule or as part of the amino acid sequence of a polypeptide described herein. US 2012/0283411 A9 Nov. 8, 2012

For example, the nucleic acid molecule can encode a prepro 0654 The invention features a method for preparing a tein or a preproprotein that can be processed to produce a polypeptide described herein by: providing a host cells (e.g., polypeptide described herein. In cases where unnatural a bacterial or mammalian or insect cell) harboring a nucleic amino acids are present in the polypeptides described herein, acid molecule encoding the polypeptide, culturing the cell selector codons can be utilized in the synthesis of such under conditions suitable for expression of the polypeptide, polypeptides similar to that described in US20060019347 and at least partially purifying the polypeptide from the cellor (for example, paragraphs 398-408, 457-499, and 576-588) the culture media in which the cell is cultured. herein incorporated by reference. 0655 The invention features a method for treating inflam mation, including inflammation of the gastrointestinal tract, 0649. A vector that includes a nucleotide sequence encod e.g., inflammation associated with a gastrointestinal disorder ing a polypeptide described herein or a polypeptide or or infection or some other disorder, the method comprising polypeptide comprising a polypeptide described herein may administering to a patient a pharmaceutical composition be either RNA or DNA, single- or double-stranded, prokary comprising a purified polypeptide described herein. In vari otic, eukaryotic, or viral. Vectors can include transposons, ous embodiments the inflammation is associated with a gas viral vectors, episomes, (e.g., plasmids), chromosomes trointestinal disorder, the inflammation is not associated with inserts, and artificial chromosomes (e.g. BACs or YACs). a gastrointestinal disorder. Suitable bacterial hosts for expression of the encode polypep tide or polypeptide include, but are not limited to, E. coli. 0656. The invention features a method for treating hyper Suitable eukaryotic hosts include yeast Such as S. cerevisiae, tension. The method comprises: administering to the patienta other fungi, Vertebrate cells, invertebrate cells (e.g., insect pharmaceutical composition comprising, consisting essen cells), plant cells, human cells, human tissue cells, and whole tially of, or consisting of a polypeptide described herein and eukaryotic organisms. (e.g., a transgenic plant or a transgenic a pharmaceutically acceptable carrier. The composition can animal). Further, the vector nucleic acid can be used to gen be administered in combination with another agent for treat erate a virus Such as vaccinia or baculovirus (for example ment of hypertension, for example, a diuretic, an ACE inhibi using the Bac-to-BacR. Baculovirus expression system (Invit tor, an angiotensin receptor blocker, a beta-blocker, or a cal rogen Life Technologies, Carlsbad, Calif.)). cium channel blocker. 0650. As noted above the invention includes vectors and 0657 The invention features a method for treating second genetic constructs Suitable for production of a polypeptide ary hyperglycemias in connection with pancreatic diseases described herein or a polypeptide or polypeptide comprising (chronic pancreatitis, pancreasectomy, hemochromatosis) or Such a polypeptide. Generally, the genetic construct also endocrine diseases (acromegaly, Cushing's syndrome, pheo includes, in addition to the encoding nucleic acid molecule, chromocytoma or hyperthyreosis), drug-induced hyperglyce elements that allow expression, such as a promoter and regu mias (benzothiadiazine Saluretics, diaZOxide or glucocorti latory sequences. The expression vectors may contain tran coids), pathologic glucose tolerance, hyperglycemias, Scriptional control sequences that control transcriptional ini dyslipoproteinemias, adiposity, hyperlipoproteinemias and/ tiation, Such as promoter, enhancer, operator, and repressor or hypotensions is described. The method comprises: admin sequences. A variety of transcriptional control sequences are istering to the patient a pharmaceutical composition compris well known to those in the art and may be functional in, but are ing, consisting essentially of, or consisting of a polypeptide not limited to, a bacterium, yeast, plant, or animal cell. The described herein and a pharmaceutically acceptable carrier. expression vector can also include a translation regulatory 0658 Also described are methods for producing any of the sequence (e.g., an untranslated 5' sequence, an untranslated 3' forgoing polypeptides comprising providing a cell harboring sequence, a poly Aaddition site, oran internal ribosome entry a nucleic acid molecule encoding the polypeptide, culturing site), a splicing sequence or splicing regulatory sequence, and the cell under conditions in which the polypeptide is a transcription termination sequence. The vector can be expressed, and isolating the expressed polypeptide. capable of autonomous replication or it can integrate into host 0659 Also described are methods for producing any of the DNA. forgoing polypeptides comprising chemically synthesizing 0651. The invention also includes isolated host cells har the polypeptide and then purifying the synthesized polypep boring one of the forgoing nucleic acid molecules and meth tide. ods for producing a polypeptide by culturing Such a cell and 0660 Also described are pharmaceutical compositions recovering the polypeptide or a precursor of the polypeptide. comprising the forgoing polypeptides. Recovery of the polypeptide or precursor may refer to col 0661. Also described are nucleic acid molecules encoding lecting the growth solution and need not involve additional any of the forgoing polypeptides, vectors (e.g., expression steps of purification. Proteins of the present invention, how vectors) containing such nucleic acid molecules and host ever, can be purified using standard purification techniques, cells harboring the nucleic acid molecules or vectors. Such as, but not limited to, affinity chromatography, therma 0662. The various polypeptides described herein can precipitation, immunoaffinity chromatography, ammonium include at least 10 contiguous amino acids of the pro Sulfate precipitation, ion exchange chromatography, filtra sequence, a pre sequence or both a pre sequence and a pro tion, electrophoresis and hydrophobic interaction chromatog sequence (a "prepro sequence') of guanylin or uroguanylin raphy. from humans or other species. Thus, useful polypeptides can 0652 The polypeptides can be purified. Purified polypep include a pre sequence, a pro sequence or a prepro sequence tides are polypeptides separated from other proteins, lipids, preceding (amino-terminal to) a GC-C receptor agonist and nucleic acids or from the compounds from which is it polypeptide described herein. FIG. 1 depicts the pre sequence synthesized. The polypeptide can constitute at least 10, 20, 50 (SEQID NOs: ), pro sequence (SEQID NOs: 70, 80 or 95% by dry weight of the purified preparation. ), prepro sequence (SEQ ID NOs: 0653. The invention features a method for preparing a ), and mature sequence for a number guanylin polypeptide described herein by: chemically synthesizing the and uroguanylin polypeptides as well a various combinations polypeptide and at least partially purifying the synthesized thereof (e.g., a polypeptide consisting of a pre sequence and a polypeptide. mature polypeptide). US 2012/0283411 A9 Nov. 8, 2012 32

0663 Among the useful GC-C receptor polypeptides that patient. In other cases the prepropolypeptide, propolypeptide can modified by the addition of a polypeptide described or the prepolypeptide is administered to the patient. The pre herein are: sequence and/or the pro sequence may stabilize the polypep tide or an active isomer thereof, facilitate efficient folding of the polypeptide or protect the polypeptide from degradation PGTCEICASAACTGC (SEQ ID NO: ) in the patient's body. Thus, pre sequences, pro sequences and/or preprosequences that do not significantly interfere PGTCEICATAACTGC (SEQ ID NO: ) with GC-C receptor agonist activity can be beneficial. In PGTCEICANAACTGC (SEQ ID NO: ) Some cases the pre sequence and/or the prosequence are removed by physiological processes after administration. PGTCEICAOAACTGC (SEQ ID NO: ) 0665 Pro sequences that include Cys residues that may PGTCEICARAACTGC (SEQ ID NO: ) form a disulfide bond. For example, many pro sequences include two Cys residues separated by 12 or 13 amino acids. PGTCEICAEAACTGC (SEQ ID NO: ) These Cys residues may form a disulfide bond. These Cys residues can be replaced by homocysteine, penicillamine, PGTCEICADAACTGC (SEQ ID NO: ) 3-mercaptoproline (Kolodzie et al. 1996 Int J Pept Protein PGTCEICAGAACTGC (SEQ ID NO: ) Res 48:274); B.B-dimethylcysteine (Hunt et al. 1993 Int J Pept Protein Res 42:249) or diaminopropionic acid (Smith et PGTCEICAAAACTGC (SEQ ID NO: ) al. 1978 J. Med Chem 21:117) to form alternative internal PGTCEICAMAACTGC (SEQ ID NO: ) cross-links at the positions of the normal disulfide bonds. PGTCEICAIAACTGC (SEQ ID NO: ) Metabolites of Asparagine PGTCEICALAACTGC (SEQ ID NO: ) 0666. In some cases an asparagine (ASn) within a polypep tide can be metabolized to have a different structure and the PGTCEICAWAACTGC (SEQ ID NO: ) GC receptoragonist containing Such a metabolite of ASn may PGTCEICAHAACTGC (SEQ ID NO: ) retain activity. Polypeptides where one or more ASn, e.g., one or more Asn within an embodiment of one or more of SEQID PGTCEGICAYAACTGC (SEQ ID NO: ) NO:X, SEQ ID NO:X, SEQ ID NO:X1, SEQ ID NO:X2, PGTCEIGCAYAACTGC (SEQ ID NO: ) SEQIDNO:ZZ, SEQIDNO:ZZ, SEQIDNO:ZZ1, and SEQ ID NO:ZZ2 described herein are replaced by a metabolite of PGTCEICGAYAACTGC (SEQ ID NO: ) Asn can be useful in the methods described herein and can be present in a pharmaceutical composition that optionally con PGTCEICAGYAACTGC (SEQ ID NO: ) tains one or more additional active ingredients. PGTCEICAYGAACTGC (SEQ ID NO: ) 0667 For example, one or more Asn within a polypeptide PGTCEICAYAGACTGC (SEQ ID NO: ) and the peptide bond carboxy terminal thereto having the Structure: PGTCEICAYAAGCTGC (SEQ ID NO: )

PGTCEICAYAACGTGC (SEQ ID NO: ) O PGTCEICAYAACTGGC (SEQ ID NO: ) HN PGTCAEICAYAACTGC (SEQ ID NO: ) NH PGTCEAICAYAACTGC (SEQ ID NO: ) HN

PGTCEIACAYAACTGC (SEQ ID NO: ) O PGTCEICAAYAACTGC (SEQ ID NO: ) PGTCEICAYAAACTGC (SEQ ID NO: ) can replaced by a group having a structure selected from: PGTCEICAYAACATGC (SEQ ID NO: ) PGTCEICAYAACTAGC (SEQ ID NO: ) PGTCEICAYAACTGAC (SEQ ID NO: ) PGTCAEICAAYAACTGC (SEQ ID NO: ) PGTCEAICAAYAACTGC (SEQ ID NO: ) and A ch 7, PGTCEIACAAYAACTG.C. (SEQ ID NO: ) 0664. In some cases a polypeptide will be produced, e.g., recombinantly, with a pre sequence and/or a pro sequence. In certain cases the pre sequence and/or pro sequence is removed prior to administration of the polypeptide to a US 2012/0283411 A9 Nov. 8, 2012 33

Thus, the Asn and the peptide bond carboxy terminal there to can be replaced by a cyclic imide: -continued (b) O yHO N H A ch O (c) HO O Asp: y 0668 H O

O provided that an ASn at the carboxy terminus is not replaced by structure (a) or structure (c). When the ASn is at the car boxy terminus of the peptide, structure (a) cannot form. Since N structure (c) is formed through structure (a), structure (c) cannot be formed when the ASn is at the carboxy terminus. x NH-- y O 0671 The formation of the various metabolites of Asp is depicted below. or isoASn: O R HO O Fragments O O s O OH A.C.X.H O

0669. The Asp can be L-Asp or D-Asp. The iso Asn can be D-isoASn or L-isoASn. O 0670 Considering the asparagine only, one or more Asn O O R having the structure: direct hydrolysis: O pH 1-2 N N 3 O HN O R. D. L. Aspartyl peptide

N N 3 O

maximum stability at cyclic imide pH3-4 formation: slow at acidic pH more prevalent at pH 7-11; Subject to buffer s catalysis is can be optionally replaced by a group having a structure D. L. cyclic imide selected from (a), (b) and (c):

R O

N H s OH N 3 A ch L-iso Aspartyl peptide US 2012/0283411 A9 Nov. 8, 2012 34

0672. The details of one or more embodiments described 1998 Clin Pharmacol Ther 63:580-93). In this manner such herein are set forth in the accompanying description. All of polypeptides can treat both pain and inflammation. Other the publications, patents and patent applications are hereby analgesic polypeptides can be present at the carboxy terminus incorporated by reference. of the polypeptide (following a cleavage site) including: endomorphin-1, endomorphin-2, nocistatin, dalargin, lupron, FIGURE Ziconotide, and Substance P. As described in greater detail below, various analgesic polypeptides and compounds can be 0673. The FIGURE depicts the pre sequence (SEQ ID covalently linked to or used in combination therapy with the NOS: ), pro sequence (SEQ ID NOs: therapeutic polypeptides described herein. ), prepro sequence (SEQ ID NOs: 0679. In the human body an inactive form of chymot ), and mature sequence for a number guanylin rypsin, chymotrypsinogen is produced in the pancreas. When and uroguanylin polypeptides as well a various combinations this inactive enzyme reaches the Small intestine it is converted thereof (e.g., a polypeptide consisting of a pre sequence and a to active chymotrypsin by the excision of two di-peptides. mature polypeptide). Active chymotrypsin will cleave polypeptides at the polypep tide bond on the carboxy-terminal side of Trp, Tyror Phe. The DETAILED DESCRIPTION presence of active chymotrypsin in the intestinal tract will 0674. The polypeptides described herein bind to the gua lead to cleavage of certain of the polypeptides described nylate cyclase (GC-C) receptor, a key regulator of fluid and herein having an appropriately positioned chymotrypsin electrolyte balance in the intestine and kidney. When stimu cleavage site. Certain of the polypeptides described herein lated, this receptor, which is located on the apical membrane include a Trp, Tyr or Phe immediately followed by a carboxy of the intestinal epithelial Surface, causes an increase in intes terminal analgesic polypeptide. It is expected that chymot tinal epithelial cyclic GMP (cGMP). This increase incCMP rypsin cleavage will release the analgesic polypeptide from is believed to cause a decrease in water and Sodium absorption polypeptide described herein having an appropriately posi and an increase in chloride and potassium ion secretion, lead tioned chymotrypsin cleavage site as the polypeptide passes ing to changes in intestinal fluid and electrolyte transport and through the intestinal tract. increased intestinal motility. The intestinal GC-C receptor 0680 Trypsinogen, like chymotrypsin, is a serine protease possesses an extracellular ligand binding region, a transmem that is produced in the pancreas and is present in the digestive brane region, an intracellular protein kinase-like region and a tract. The active form, trypsin, will cleave polypeptides hav cyclase catalytic domain. Proposed functions for the GC-C ing a Lys or Arg. The presence of active trypsin in the intes receptor are the fluid and electrolyte homeostasis, the regu tinal tract will lead to cleavage of certain of the polypeptides lation of epithelial cell proliferation and the induction of described herein having an appropriately positioned trypsin apoptosis (Shailhubhai 2002 Curr Opin Drug Dis Devel cleavage site. It is expected that chymotrypsin cleavage will 5:261-268). release the analgesic polypeptide from polypeptide described 0675. In addition to being expressed in gastrointestinal herein having an appropriately positioned trypsin cleavage epithelial cells, GC-C is expressed in extra-intestinal tissues site as the polypeptide passes through the intestinal tract. including kidney, lung, pancreas, pituitary, adrenal, develop 0681. In some cases, the polypeptides described herein are ing liver, heart and male and female reproductive tissues produced as a prepro protein. The prepro protein can include (reviewed in Vaandrager 2002 Mol Cell Biochem 230:73-83). any suitable prepro sequence, including but not limited to, for This suggests that the GC-C receptor agonists can be used in example, mnafillsalc llgawaalag gvtvqdgnifs fslesvkklk the treatment of disorders outside the GI tract, for example, dlqepqeprygklrnfapipgepVVpilcs npnfpeelkplckepnaqeildr congestive heart failure and benign prostatic hyperplasia. leeiaed (SEQID NO:), mgcraasgll pgvaVVlllllqstdSVyiqyag 0676 Ghrelin, a polypeptide hormone secreted by the frvgles mkklsdlead wapsprldaq sllpavchhp alpqdlqpvc asqe stomach, is a key regulator of appetite in humans. Ghrelin assifktlrtia (SEQ ID NO:), lrtia (SEQ ID NO.), minawllsvlc expression levels are regulated by fasting and by gastric emp lgalavlve gvtvqdgdls fplesvkalk hlrevdeptl mshkkfalrl tying. (Kim et al., 2003, Neuroreprt 14:1317-20; Gualillo et pkpvapelcs qsafpealrplcekpnaeeildrleaiaqd (SEQID NO:), al., 2003, FEBS Letts 552: 105-9). Thus, by increasing gas and msg.sqlwaav Illvilqsaq gVyikyhgfa Valesvkkln trointestinal motility, GC-C receptor agonists may also be eleekqmsdp qqqksgllpd Vcynpalpla ldpvcasqea astfkalrtia used to regulate obesity. (SEQID NO:) or a bacterial leader sequence such as: mkksil 0677. In humans, the GC-C receptor is activated by gua fiflsvlsfspfaqdakpVesskekitleskkcniakkSnksgpesmn. Where nylin (Gn) (U.S. Pat. No. 5.96,097), uroguanylin (Ugn) (U.S. the polypeptide is produced by a bacterial cell, e.g., E. coli, Pat. No. 5,140,102) and lymphoguanylin (Forte et al. 1999 the forgoing leader sequence will be cleaved and the mature Endocrinology 140:1800-1806). polypeptide will be efficiently secreted from the bacterial 0678. Many gastrointestinal disorders, including IBS, are cell. U.S. Pat. No. 5,395,490 describes vectors, expression associated with abdominal or visceral pain. Certain of the systems and methods for the efficient production of certain polypeptides described herein include the analgesic or anti mature polypeptides having disulfide bonds in bacterial cells nociceptive tags such as the carboxy-terminal sequence Asp and methods for achieving efficient Secretion of such mature Phe immediately following a Trp, Tyr or Phe (i.e., a chymot polypeptides. The vectors, expression systems and methods rypsin cleavage site) or following Lys or Arg (a trypsin described in U.S. Pat. No. 5,395,490 can be used to produce cleavage site). Chymotrypsin in the intestinal tract will cleave the polypeptides of the present invention. Such polypeptides immediately carboxy terminal to the Trp, Phe or Tyr residue, releasing the dipeptide, AspPhe. This Variant Polypeptides dipeptide has been shown to have analgesic activity is animal 0682. The invention includes variant polypeptides that can models (Abdikkahi et al. 2001 Fundam Clin Pharmacol include one, two, three, four, or five or more (e.g., 6, 7, 8, 9. 15:117-23; Nikfar et al 1997, 29:583-6; Edmundson et al 10, 11, 12, 13, 14, or 15) amino acid substitutions compared US 2012/0283411 A9 Nov. 8, 2012

to any of the polypeptides described above. The substitution encoding the polypeptide may also encode a leader sequence (s) can be conservative or non-conservative. The naturally that permits the secretion of the mature polypeptide from the occurring amino acids can be substituted by D-isomers of any cell. Thus, the sequence encoding the polypeptide can include amino acid, non-natural amino acids, natural and non-natural the pre sequence and the pro sequence of for example, a amino acid analogs, and other groups. A conservative amino naturally-occurring bacterial ST polypeptide. The secreted, acid Substitution results in the alteration of an amino acid for mature polypeptide can be purified from the culture medium. a similar acting amino acid, or amino acid of like charge, 0687. The sequence encoding a polypeptide described polarity, or hydrophobicity. At some positions, even conser herein is can be inserted into a vector capable of delivering vative amino acid substitutions can reduce the activity of the and maintaining the nucleic acid molecule in a bacterial cell. polypeptide. A conservative Substitution can Substitute a The DNA molecule may be inserted into an autonomously naturally-occurring amino acid for a non-naturally-occurring replicating vector (Suitable vectors include, for example, amino acid. Among the naturally occurring amino acid Sub pGEM37 and pcDNA3, and derivatives thereof). The vector stitutions generally considered conservative are: nucleic acid may be a bacterial or bacteriophage DNA such as bacteriophage lambda or M13 and derivatives thereof. Con struction of a vector containing a nucleic acid described hereincan be followed by transformation of a host cell such as For Amino Acid Code Replace with any of a bacterium. Suitable bacterial hosts include but are not lim Alanine Ala Gly, Cys, Ser ited to, E. coli, B subtilis, Pseudomonas, Salmonella. The Arginine Arg Lys, His genetic construct also includes, in addition to the encoding Asparagine ASn Asp, Glu, Gln, Aspartic Acid Asp ASn, Glu, Gln nucleic acid molecule, elements that allow expression, Such Cysteine Cys Met, Thr, Ser as a promoter and regulatory sequences. The expression vec Glutamine Gln ASn, Glu, Asp tors may contain transcriptional control sequences that con Glutamic Acid Glu Asp, ASn, Gln trol transcriptional initiation, Such as promoter, enhancer, Glycine Gly Ala Histidine His Lys, Arg operator, and repressor sequences. A variety of transcrip Isoleucine Ile Val, Leu, Met tional control sequences are well known to those in the art. Leucine Leu Val, Ile, Met The expression vector can also include a translation regula Lysine Lys Arg, His tory sequence (e.g., an untranslated 5' sequence, an untrans Methionine Met Ile, Leu, Val Phenylalanine Phe Tyr, His, Trp lated 3' sequence, or an internal ribosome entry site). The Proline Pro vector can be capable of autonomous replication or it can Serine Ser Thr, Cys, Ala integrate into host DNA to ensure stability during polypeptide Threonine Thr Ser, Met, Val production. Tryptophan Trp Phe, Tyr Tyrosine Tyr Phe, His 0688. The protein coding sequence that includes a Valine Wall Leu, Ile, Met polypeptide described herein can also be fused to a nucleic acid encoding a polypeptide affinity tag, e.g., glutathione S-transferase (GST), maltose E binding protein, protein A, 0683. In some circumstances it can be desirable to treat FLAG tag, hexa-histidine, myc tag or the influenza HA tag, in patients with a variant polypeptide that binds to and activates order to facilitate purification. The affinity tag or reporter intestinal GC-C receptor, but is less active or more active than fusion joins the reading frame of the polypeptide of interest to the non-variant form of the polypeptide. Reduced activity can the reading frame of the gene encoding the affinity tag. Such arise from reduced affinity for the receptor or a reduced that a translational fusion is generated. Expression of the ability to activate the receptor once bound or reduced stability fusion gene results in translation of a single polypeptide that of the polypeptide. Increased activity can arise from includes both the polypeptide of interest and the affinity tag. increased affinity for the receptor or an increased ability to In some instances where affinity tags are utilized, DNA activate the receptor once bound or increased stability of the sequence encoding a protease recognition site will be fused polypeptide. between the reading frames for the affinity tag and the 0684. In some polypeptides one or both members of one or polypeptide of interest. both pairs of Cys residues which normally form a disulfide bond can be replaced by homocysteine, penicillamine, 3-mer 0689 Genetic constructs and methods suitable for produc captoproline (Kolodzie et al. 1996 Int J Pept Protein Res tion of immature and mature forms of the polypeptides and 48:274); BB dimethylcysteine (Hunt et al. 1993 Int Pept variants described herein in protein expression systems other Protein Res 42:249) or diaminopropionic acid (Smith et al. than bacteria, and well known to those skilled in the art, can 1978J Med Chem 21:117) to form alternative internal cross also be used to produce polypeptides in a biological system. links at the positions of the normal disulfide bonds. 0690 Mature polypeptides and variants thereof can be synthesized by the Solid-phase method using an automated Production of Polypeptides polypeptide synthesizer. For example, the polypeptide can be synthesized on Cyc(4-CH Bxl)-OCH-4-(oxymethyl)-phe 0685 Useful polypeptides can be produced either in bac nylacetamidomethyl resin using a double coupling program. teria including, without limitation, E. coli, or in other existing Protecting groups must be used appropriately to create the systems for polypeptide or protein production (e.g., Bacillus correct disulfide bond pattern. For example, the following subtilis, baculovirus expression systems using Drosophila protecting groups can be used: t-butyloxycarbonyl (alpha Sf9 cells, yeast or filamentous fungal expression systems, amino groups); acetamidomethyl (thiol groups of Cys resi mammalian cell expression systems), or they can be chemi dues B and E): 4-methylbenzyl (thiol groups of Cys residues cally synthesized. C and F); benzyl (y-carboxyl of glutamic acid and the 0686. If the polypeptide or variant polypeptide is to be hydroxyl group of threonine, if present); and bromobenzyl produced in bacteria, e.g., E. coli, the nucleic acid molecule (phenolic group of tyrosine, if present). Coupling is effected US 2012/0283411 A9 Nov. 8, 2012 36 with symmetrical anhydride oft-butoxylcarbonylamino acids tide and the test material at various concentrations. The cells or hydroxybenzotriazole ester (for asparagine or glutamine are then washed 4 times with 1 ml of DMEM and solubilized residues), and the polypeptide is deprotected and cleaved with 0.5 ml/well 1N NaOH. The level of radioactivity in the from the solid support in hydrogen fluoride, dimethylsulfide, solubilized material is then determined using standard meth anisole, and p-thiocresol using 8/1/1/0.5 ratio (v/v/v/w) at 0° ods. C. for 60 min. After removal of hydrogen fluoride and dim ethylsulfide by reduced pressure and anisole and p-thiocresol Murine Gastrointestinal Transit (GIT) Assay by extraction with ethyl ether and ethyl acetate sequentially, crude polypeptides are extracted with a mixture of 0.5M 0696. In order to determine whether a test compound or a sodium phosphate buffer, pH 8.0 and N,N-dimethylforma polypeptide, increases the rate of gastrointestinal transit, the mide using 1/1 ratio, V/v. Disulfide bonds between Cys resi test compound can be tested in the murine gastrointestinal dues can be formed using dimethyl sulfoxide (Tam et al. transit (GIT) assay (Moon et al. Infection and Immunity (1991) J. Am. Chem. Soc. 113:6657-62). The resulting 25:127, 1979). In this assay, charcoal, which can be readily polypeptide is the purified by reverse-phase chromatography. visualized in the gastrointestinal tract is administered to mice In some cases it may be necessary to first dissolve the after the administration of a test compound. The distance polypeptide in 50% acetic acid in water before disulfide bond traveled by the charcoal is measured and expressed as a per formation. Saturated iodine Solution in glacial acetic acid is centage of the total length of the colon. added (1 ml iodine solution per 100 ml solution). After incu 0697 Mice are fasted with free access to water for 12 to 16 bation at room temperature for 2 days in closed glass con hours before the treatment with polypeptide or control buffer. tainer, the solution is diluted five-fold with deionized water The polypeptides are orally administered at 1 ug/kg-1 mg/kg and extracted with ethyl ether four times for removal of unre of polypeptide in buffer (20 mM Tris pH 7.5) seven minutes acted iodine. After removal of the residual amount of ethyl before being given an oral dose of 5% Activated Carbon ether by rotary evaporation the solution of crude product is (Aldrich 242276-250G). Control mice are administered lyophilized and purified by Successive reverse-phase chroma buffer only before being given a dose of Activated Carbon. tography. After 15 minutes, the mice are sacrificed and their intestines 0691 Peptides can also be synthesized by many other from the stomach to the cecum are dissected. The total length methods including Solid phase synthesis using traditional of the intestine as well as the distance traveled from the FMOC protection (i.e., coupling with DCC-HOBt and depro stomach to the charcoal front is measured for each animal and tection with piperidine in DMF). Cys thiol groups can be trityl the results are expressed as the percent of the total length of the intestine traveled by the charcoal front. Results are protected. Treatment with TFA can be used for final depro reported as the average of 10 miceitstandard deviation. A tection of the polypeptide and release of the polypeptide from comparison of the distance traveled by the charcoal between the Solid-state resin. In many cases air oxidation is sufficient the mice treated with polypeptide versus the mice treated with to achieve proper disulfide bond formation. vehicle alone is performed using a Student's t test and a Intestinal GC-C Receptor Binding and Activity Assays statistically significant difference is considered for P-0.05. Positive controls for this assay may include commercially 0692. The ability of polypeptides, variant polypeptides available wild-type ST polypeptide (Sigma-Aldrich, St and other compounds to bind to and activate the intestinal Louis, Mo.) and ZelnormR), a drug approved for IBS that is an GC-C receptor can be tested using the T84 human colon agonist for the serotonin receptor 5HT4. carcinoma cell line (American Type Culture Collection (Be 0698 Similar assays can be performed in other rodents, thesda, Md.). for example, rats. In addition, GIT assays can be performed 0693 Briefly, cells are grown to confluency in 24-well and compared in wild-type Versus rodents lacking the guany culture plates with a 1:1 mixture of Ham's F12 medium and late cyclase C receptor (GC-C KO), for example, using the Dulbecco's modified Eagle's medium (DMEM), supple GC-C KO mice described in Mann et al 1997 Biochem and mented with 5% fetal calf serum and are used at between Biophysical Research Communications 239:463. passages 54 and 60. 0694 Monolayers of T84 cells in 24-well plates are Suckling Mouse Model of Intestinal Secretion (SuMi Assay) washed twice with 1 ml/well DMEM, then incubated at 37°C. for 10 min with 0.45 ml DMEM containing 1 mM isobutyl 0699 The polypeptides described herein can be tested for methylxanthine (IBMX), a cyclic nucleotide phosphodi their ability to increase intestinal Secretion using a Suckling esterase inhibitor. Test polypeptides (50 ul) are then added mouse model of intestinal Secretion. In this model a test and incubated for 30 minutes at 37°C. The media is aspirated compound is administered to Suckling mice that are between and the reaction is terminated by the addition of ice cold 0.5 seven and nine days old. After the mice are sacrificed, the ml of 0.1N HC1. The samples are held on ice for 20 minutes gastrointestinal tract from the stomach to the cecum is dis and then evaporated to dryness using a heat gun or vacuum sected (“guts”). The remains (“carcass') as well as the guts centrifugation. The dried samples are resuspended in 0.5 ml are weighed and the ratio of guts to carcass weight is calcu of phosphate buffer provided in the Cayman Chemical Cyclic lated. If the ratio is above 0.09, one can conclude that the test GMP EIA kit (Cayman Chemical, Ann Arbor, Mich.). Cyclic compound increases intestinal Secretion. Controls for this GMP is measured by EIA according to procedures outlined in assay may include wild-type ST polypeptide and ZelnormR). the Cayman Chemical Cyclic GMP EIA kit. 0695 For the binding assay, T84 cell monolayers in Phenylbenzoquinone-Induced Writhing Model 24-well plates are washed twice with 1 ml of binding buffer (0700. The PBQ-induced writhing model can be used to (DMEM containing 0.05% bovine serum albuminand 25 mM assess pain control activity of the polypeptides and GC-C HEPES, pH 7.2), then incubated for 30 min at 37°C. in the receptor agonists described herein. This model is described presence of mature radioactively labeled E. coli ST polypep by Siegmund et al. (1957 Proc. Soc. Exp. Bio. Med. 95:729 US 2012/0283411 A9 Nov. 8, 2012 37

731). Briefly, one hour after oral dosing with a test compound, one way analysis of variance (ANOVA) followed by a post e.g., a polypeptide, morphine or vehicle, 0.02% phenylben hoc (Student or Dunnett tests) and regression analysis for Zoquinone (PBQ) solution (12.5 mL/kg) is injected by intra ED50 if appropriate. peritoneal route into the mouse. The number of stretches and 0706 II. Stress-Induced Hyperalgesia Model writhings are recorded from the 5' to the 10" minute after (0707 Male Wistar Rats (200-250 g) are surgically PBQ injection, and can also be counted between the 35' and implanted with nichrome wire electrodes as in the TNBS 40" minute and between the 60" and 65' minute to provide a model. Ten days post Surgical implantation, partial restraint kinetic assessment. The results are expressed as the number of stress (PRS), is performed as described by Williams et al. for stretches and writhings (meantSEM) and the percentage of two hours (Williams et al. 1988 Gastroenterology 64:611). variation of the nociceptive threshold calculated from the Briefly, under light anaesthesia with ethyl-ether, the fore mean value of the vehicle-treated group. The statistical sig shoulders, upper forelimbs and thoracic trunk are wrapped in nificance of any differences between the treated groups and a confining harness of paper tape to restrict, but not prevent body movements. Control sham-stress animals are anaesthe the control group is determined by a Dunnett's test using the tized but not wrapped. Thirty minutes before the end of the residual variance after a one-way analysis of variance (P<0. PRS session, the animals are administered test-compound or 05) using SigmaStat Software. vehicle. Thirty minutes to one hour after PRS completion, the CRD distension procedure is performed as described above Colonic Hyperalgesia Animal Models for the TNBS model with barostat at pressures of 15, 30, 45 0701 Hypersensitivity to colorectal distension is a com and 60 mm Hg. Statistical analysis on the number of bursts is mon feature in patients with IBS and may be responsible for determined and analyzed as in the TNBS model above. the major symptom of pain. Both inflammatory and non 0708 III. Water Avoidance Stress-Induced Hyperalgesia inflammatory animal models of visceral hyperalgesia to dis Model tension have been developed to investigate the effect of com (0709. The effect of polypeptides/GC-Cagonists described pounds on visceral pain in IBS. herein on basal visceral nociception in a model of water avoidance stress-induced visceral hyperalgesia in adult male (0702 I. Trinitrobenzenesulphonic Acid (TNBS)-Induced Wistar rats can be tested. The stress involves confining rats to Rectal Allodynia Model a platform surrounded by water for a period of 1 hour and then (0703 Male Wistar rats (220-250 g) are premedicated with measuring their visceromotor response to colonic distension 0.5 mg/kg of acepromazine injected intraperitoneally (IP) using electromyography (EMG). and anesthetized by intramuscular administration of 100 0710. At least 7 days prior to stress measurements, ani mg/kg of ketamine. Pairs of nichrome wire electrodes (60 cm mals are deeply anesthetized with pentobarbital sodium (45 in length and 80 Lum in diameter) are implanted in the striated mg/kg) and equipped with electrodes implanted into the muscle of the abdomen, 2 cm laterally from the white line. external oblique musculature, just Superior to the inguinal The free ends of electrodes are exteriorized on the back of the ligament. Electrode leads are then tunneled Subcutaneously neck and protected by a plastic tube attached to the skin. and externalized laterally for future access. Following sur Electromyographic (EMG) recordings are started 5 days after gery, rats are housed in pairs and allowed to recover for at Surgery. Electrical activity of abdominal striated muscle is least 7 days. On the day of the experiment, animals are lightly recorded with an electroencephalograph machine (Mini VIII, anesthetized with halothane, and a lubricated latex balloon (6 Alvar, Paris, France) using a short time constant (0.03 sec.) to cm) is inserted intra-anally into the descending colon. Ani remove low-frequency signals (<3 Hz). mals are allowed to recover for 30 minutes, and colorectal 0704. Ten days post surgical implantation, trinitrobenze distension (CRD) is initiated. The CRD procedure consists of nesulphonic acid (TNBS) is administered to induce rectal graded intensities of phasic CRD (10, 20, 40, 60 mmHg: 20s inflammation. TNBS (80 mg kg in 0.3 ml 50% ethanol) is duration: 4 min inter-stimulus interval). Visceromotor administered intrarectally through a silicone rubber catheter response (VMR) to CRD is quantified by measuring EMG introduced at 3 cm from the anus under light diethyl-ether activity. To determine the effects of polypeptides/GC-C ago anesthesia, as described (Morteau et al. 1994 Dig Dis Sci nists described herein on basal visceral nociception, a base 39:1239). Following TNBS administration, rats are placed in line CRD is recorded. Animals are allowed 1 hour recovery plastic tunnels where they are severely limited in mobility for and then the polypeptide/GC-C agonist described herein or several days before colorectal distension (CRD). Experimen vehicle is orally administered. At 1 hour following adminis tal compound is administered one hour before CRD which is tration of polypeptide/GC-C agonist described herein or performed by insertion into the rectum, at 1 cm of the anus, a vehicle CRD is repeated. 4 cm long balloon made from a latex condom (Gue etal, 1997 0711. To determine the effect of polypeptides/GC-C ago Neurogastroenterol. Motil. 9:271). The balloon is fixed on a nists described herein in a model of water avoidance stress rigid catheter taken from an embolectomy probe (Fogarty). induced visceral hyperalgesia, a baseline CRD is recorded The catheter attached balloon is fixed at the base of the tail. and then the animals were subjected to 1 hour of water avoid The balloon, connected to a barostat is inflated progressively ance stress. For water avoidance stress, the test apparatus by step of 15 mmHg, from 0 to 60 mmHg, each step of consists of a Plexiglas tank with a block affixed to the center inflation lasting 5 min. Evaluation of rectal sensitivity, as of the floor. The tank is filled with fresh room temperature measured by EMG, is performed before (1-2 days) and 3 days water (25° C.) to within 1 cm of the top of the block. The following rectal instillation of TNBS. animals are placed on the block for a period of 1 hour. The 0705 The number of spike bursts that corresponds to sham water avoidance stress consists in placing the rats on the abdominal contractions is determined per 5 min periods. Sta same platform in a waterless container. A second CRD is tistical analysis of the number of abdominal contractions and performed at 24 hours post water avoidance stress. Following evaluation of the dose-effects relationships is performed by a the second CRD, animals are allowed 1 hour recovery and US 2012/0283411 A9 Nov. 8, 2012

then the polypeptide/GC-C agonist described herein or inverted flow path onto the analytical column (Waters Xterra vehicle is orally administered. At 1 hour following adminis MS C85 um IS column, 2.1x20 mm). The sample is eluted tration of polypeptide/GC-C agonist described herein or from the analytical column with a reverse phase gradient vehicle CRD is repeated. Mean+/-SEM is be determined and (Mobile Phase A: 10 mM ammonium hydroxide in dHO, compared in the presence and absence of water avoidance Mobile Phase B: 10 mMammonium hydroxide in 80% aceto stress conditions. nitrile and 20% methanol; 20% B for the first 3 minutes then 0712) Kd Determination and Binding Assays ramping to 95% B over 4 min. and holding for 2 min., all at a 0713 To determine the affinity of polypeptides/GC-C flow rate of 0.4 mL/min.). At 9.1 minutes, the gradient returns agonists described herein for GC-C receptors found in rat to the initial conditions of 20% B for 1 min. polypeptide is intestinal mucosa, a competition binding assay is performed eluted from the analytical column and is detected by triple using rat intestinal epithelial cells. Epithelial cells from the quadrapole mass spectrometry (MRM, 764 (+2 charge State) small intestine of rats are obtained as described by Kessler et >182 (+1 charge state) Da; cone voltage=30V: collision=20 al. (J. Biol. Chem. 245: 5281-5288 (1970)). Briefly, animals eV; parent resolution 2 Da at base peak; daughter resolu are sacrificed and their abdominal cavities exposed. The small tion 2 Da at base peak). Instrument response is converted intestine is rinsed with 300 ml ice cold saline or PBS. 10 cm into concentration units by comparison with a standard curve of the small intestine measured at 10 cm from the pylorus is using known amounts of chemically synthesized polypeptide removed and cut into 1 inch segments. Intestinal mucosa is (s) prepared and injected in mouse plasma using the same extruded from the intestine by gentle pressure between a procedure. piece of parafilm and a P-1000 pipette tip. Intestinal epithelial 0717. Similarly, pharmacokinetic properties are deter cells are placed in 2 ml PBS and pipetted up and down with a mined in rats using LCMS methodology. Rat plasma samples 5 ml pipette to make a Suspension of cells. Protein concen containing the polypeptide are extracted using a Waters Oasis tration in the Suspension is measured using the Bradford MAX 96 well solid phase extraction (SPE) plate. A 200 uL method (Anal. Biochem. 72: 248-254 (1976)). volume of rat plasma is mixed with 200 uL of C, N 0714. A competition binding assay is performed based on labeled polypeptide in the well of a prepared SPE plate. The the method of Giannella et al. (Am. J. Physiol. 245: G492 samples are drawn through the stationary phase with 15 mm G498) between 'I labeled control polypeptide (e.g. wild Hg vacuum. All samples are rinsed with 200LL of 2% ammo type guanylin, uroguanylin or ST polypeptide) and a polypep nium hydroxide in water followed by 200 uL of 20% metha tide/GC-C agonist described herein. The assay mixture nol in water. The samples are eluted with consecutive 100 L contains: 0.5 ml of DME with 20 mM HEPES-KOH pH 7.0, volumes of 5/20/75 formic acid/water/methanol and 100 L 0.9 mg of the cell suspension listed above, 21.4 fmol 'II 5/15/80 formic acid/water/methanol. The samples are dried labeled control polypeptide (42.8 pM), and different concen under nitrogen and resuspended in 100 uL of 20% methanol trations of competitor polypeptide/GC-C agonist described in water. Samples are analyzed by a Waters Quattro Micro herein (0.01 to 1000 nM). The mixture is incubated at room mass spectrometer coupled to a Waters 1525 binary pump temperature for 1 hour, and the reaction stopped by applying with a Waters 2777 autosampler. A 40 uL volume of each the mixture to GF/B glass-fiber filters (Whatman). The filters sample is injected onto a Thermo Hypersil GOLD C18 col are washed with 5 ml ice-cold PBS and radioactivity is mea umn (2.1x50mm, 5um). polypeptide is eluted by a gradient sured. Kd is determined. 96 B/Bo is the percentage of the ratio over 3 minutes with acetonitrile and water containing 0.05% of radioactivity trapped in each sample (B) compared to the trifluoroacetic acid. The Quattro Micro mass spectrometer is radioactivity retained in a control sample with no cold com run in multiple reaction monitoring (MRM) mode using the petitor (Bo). mass transitions of for example 764->182 or 682>136. Using 0715 Similar competition binding assays are performed this methodology, polypeptide is dosed orally and by IV to in intestinal epithelial cells from wild-type and guanylate rats at 10 mg/kg. Pharmacokinetic properties including area cyclase C knockout (GC-C KO: Mann et al. 1997 Biochem under the curve and bioavailability are determined. and Biophysical Research Communications 239:463) mice. Mouse intestinal epithelial cells are prepared identical to that In Vitro Proteolytic Stability above as for rat intestinal epithelial cells except the cells are 0718 The stability of polypeptides/GC-C agonists homogenized with an Omni homogenizer for 20 seconds on described herein in the presence of several mammalian diges the maximum setting to make a suspension of cells. A com tive enzymes is determined. Polypeptide/GC-C agonists petition binding assay is performed identical to that described described herein are exposed to a variety of invitro conditions above between I labeled polypeptide/GC-C agonist including digestive enzymes and low ph environments described herein and unlabeled polypeptide/GC-C agonist designed to simulate gastric fluid. Polypeptide/GC-C ago described herein (competitor). nists described herein are incubated with chymotrypsin, trypsin, pepsin, aminopeptidase, carboxypeptidase A, and Pharmacokinetic Property Determination of Polypeptides/ simulated gastric fluid (sgf) at ph 1.0. Samples are collected at GC-C Agonists Described Herein 0, 3, and 24h for all conditions except pepsin digestion and 0716 Serum samples are extracted from the whole blood the SGF. For the latter two conditions, samples are obtained at of exposed (mice dosed orally or intravenously with polypep 0,1, and 3 h. Negative control samples are prepared for initial tide(s) described herein) and control mice, then injected and final time points. A separate, positive activity control is directly (10 mL) onto an in-line solid phase extraction (SPE) run in parallel for each condition. All samples are analyzed by column (Waters Oasis HLB25um column, 2.0x15 mm direct LC/MS. connect) without further processing. The sample on the SPE column is washed with a 5% methanol, 95% dHO solution Effect on Bowel Habits (2.1 mL/min, 1.0 minute), then loaded onto an analytical 0719. Peptide/GC-C agonists described herein can be column using a valve Switch that places the SPE column in an administered to mammals (e.g. humans) to determine the US 2012/0283411 A9 Nov. 8, 2012 39 effect on bowel habits (including Bristol Stool Form Scale length ratio (W/L) for each loop is calculated to determine the score, stool frequency (number of stools per week), ease of effects of the polypeptide/GC-C agonist described herein on passage and stool weight). polypeptide/GC-C agonist is secretion. administered in a single dose or multiple doses (for example, (0736. To determine the effect of the polypeptide/GC-C once daily over a consecutive 7 day period) and alterations in agonist described herein on coMP activity, fluid from the bowel habit are evaluated (for each collected bowel move loop is collected in ice-cold trichloracetic acid (TCA) and ment), for example, prior to dose, during dosage (for multiple stored at -80° C. for use in an assay to measure ccjMP levels dosing), and postdose. in the fluid. Intestinal fluid samples are TCA extracted, and cyclic GMP is measured by EIA according to procedures The Bristol Stool Form Scale is: outlined in the Cayman Chemical Cyclic GMPEIAkit (Cay man Chemical, Ann Arbor, Mich.) to determine cyclic GMP 0720 1: Separate hard lumps, like nuts levels in the intestinal fluid of the mouse in the presence of 0721 2: Sausage-shaped but lumpy either polypeptide/GC-C agonist described herein or vehicle. 0722 3: Like a sausage or snake but with cracks on its 0737. The effects of polypeptides/GC-C agonists Surface described herein on coMP levels and secretion in ligated 0723 4: Like a sausage or snake, smooth and soft loops in female CD rats can also be determined using proto 0724) 5: Soft blobs with clear-cut edges cols similar to those described above. In the case of the rat, 0725 6: Fluffy pieces with ragged edges, a mushy stool however four loops of intestine are surgically ligated. The first 0726 7: Watery, no solid pieces three loops are distributed equally in the small intestine and the fourth loop is located in colon. Loops are 1 to 3 centime The scale used to determine ease of passage is: ters, and are injected with 200 uL of either polypeptide/ 0727 1. Manual disimpaction agonist described herein (5ug) or vehicle (Krebs Ringer, 10 0728, 2 Enema needed mM glucose, HEPES buffer (KRGH)). 0729) 3. Straining needed (0730. 4. Normal Diuresis Related Experiments (0731 5. Urgent without pain Effect on Diuresis and Natriuresis (0732 6. Urgent with pain 0738. The effect of polypeptides/GC-agonists described 0733 7. Incontinent herein on diuresis and natriuresis can be determined using methodology similar to that described in WO06/001931 (ex Rat Model of Postoperative Ileus. amples 6 (p. 42) and 8 (p. 45)). Briefly, the polypeptide/ 0734. Female CD rats are used to test the effect of agonist described herein (180-pmol) is infused for 60 min polypeptides/GC-C agonists described herein on delayed into a group of 5 anesthetized rats. Given an estimated rat transit induced by abdominal Surgery and manual manipula plasma Volume of 10 mL, the infusion rate is approximately 3 tion of the Small intestine. Groups of at least nine rats undergo pmol/mL/min. Blood pressure, urine production, and sodium abdominal Surgery under isofluraneanesthesia. Surgery con excretion are monitored for approximately 40 minutes prior sists of laparotomy and 5 minutes of gentle manual intestinal to the infusion, during the infusion, and for approximately 50 massage. Following recovery from anesthesia, rats are dosed minutes after the infusion to measure the effect of the orally with either polypeptide/GC-C agonist (for example, 10 polypeptide/GC-C agonist on diuresis and natriuresis. For ug/kg) described herein or vehicle (20 mM Tris) in a volume comparison, a control group offive rats is infused with regular of 300 ul. 1 hour after dosing, intestinal transit rate is mea saline. Urine and sodium excretion can be assessed. Dose sured. Animals are again dosed with 300 ul of the test article response can also be determined. polypeptide/GC-C agonist followed immediately by 500 ul of a charcoal meal (10% described herein is infused intravenously into rats over 60 charcoal, 10% gum arabic in water). To calculate the distance minutes. Urine is collected at 30 minute intervals up to 180 of the small intestine traveled by the charcoal front, after 20 minutes after termination of polypeptide/GC-C agonist infu minutes, the totallength of the intestine as well as the distance Sion, and urine Volume, Sodium excretion, and potassium traveled from the stomach to the charcoal front are measured excretion are determined for each collection interval. Blood for each animal. pressure is monitored continuously. For each dose a dose Effect on coMP Levels and Secretion in Ligated Loops response relationship for urine Volume, Sodium and potas Rodent Models sium excretion can be determined. Plasma concentration of 0735. The effect of polypeptides/GC-Cagonists described the polypeptide/GC-agonist is also determined before and herein on coMP levels and secretion are studied by injecting after iv infusion. polypeptides/GC-C agonists described herein directly into an isolated loop in either wild-type or GC-C KO mice. This is Rat Diuresis Experiment: done by Surgically ligating a loop in the Small intestine of the (0739. Female Sprague-Dawley rats (>170 g, 2-8 per mouse. The methodology for ligated loop formation is similar group) are given 3.0 mL ofiosotonic Saline perorally, and then to that described in London et al. 1997 Am J Physiol p. anesthetized with isoflurane/oxygen. Once an appropriate G93-105. The loop is roughly centered and is a length of 1-3 level of anesthesia has been achieved, a sterile polyurethane cm. The loops are injected with 100 ul of either SEQID NO:3 catheter (~16 cm, 0.6 mm ID, 0.9 mm OD) is inserted 1.5-2.0 (5ug) or vehicle (20 mM Tris, pH 7.5 or Krebs Ringer, 10 mM cm into the urethra and secured using 1-2 drops of Veterinary Glucose, HEPES buffer (KRGH)). Following a recovery time bond adhesive applied to urethra/catheter junction. Rats are of 90 minutes the loops are excised. Weights are recorded for then dosed with either vehicle or test article via the intrave each loop before and after removal of the fluid contained nous or intraperitoneal route. Rats are then placed in appro therein. The length of each loop is also recorded. A weight to priately sized rat restraint tubes, with the catheter protruding US 2012/0283411 A9 Nov. 8, 2012 40 out of the restraint tube into a 10 mL graduated cylinder. Rats 0744 Combination therapy can also include the adminis are allowed to regain consciousness, and the Volume of urine tration of two or more agents via different routes or locations. excreted over a 1-5 hour duration is recorded periodically for For example, (a) one agent is administered orally and another each rat. agents is administered intravenously or (b) one agent is administered orally and another is administered locally. In Administration of Polypeptides and GC-C Receptor Agonists each case, the agents can either simultaneously or sequen 0740 For treatment of gastrointestinal disorders, the tially. Approximated dosages for some of the combination polypeptides and agonists described herein are preferably therapy agents described herein are found in the “BNF Rec administered orally, e.g., as a tablet or cachet containing a ommended Dose” column of tables on pages 11-17 of WO01/ predetermined amount of the active ingredient, pellet, gel. 76632 (the data in the tables being attributed to the March paste, syrup, bolus, electuary, slurry, Sachet, capsule, powder; 2000 British National Formulary) and can also be found in lyophilized powder, granules; as a solution or a suspension in other standard formularies and other drug prescribing direc an aqueous liquid or a non-aqueous liquid; as an oil-in-water tories. For some drugs, the customary prescribed dose for an liquid emulsion or a water-in-oil liquid emulsion, via a lipo indication will vary somewhat from country to country. somal formulation (see, e.g., EP 736299) or in some other 0745. The agents, alone or in combination, can be com form. Orally administered compositions can include binders, bined with any pharmaceutically acceptable carrier or lubricants, inert diluents, lubricating, Surface active or dis medium. Thus, they can be combined with materials that do persing agents, flavoring agents, and humectants. Orally not produce an adverse, allergic or otherwise unwanted reac administered formulations such as tablets may optionally be tion when administered to a patient. The carriers or mediums coated or scored and may be formulated so as to provide used can include Solvents, dispersants, coatings, absorption Sustained, delayed or controlled release of the active ingredi promoting agents, controlled release agents, and one or more ent therein. The polypeptides and agonists can be co-admin inert excipients (which include starches, polyols, granulating istered with other agents used to treat gastrointestinal disor agents, microcrystalline cellulose (e.g. celphere, Celphere ders including but not limited to the agents described herein. Beads(R), diluents, lubricants, binders, disintegrating agents, The polypeptides and agonists can also be administered by and the like), etc. If desired, tablet dosages of the disclosed rectal suppository. For the treatment of disorders outside the compositions may be coated by Standard aqueous or non gastrointestinal tract such as congestive heart failure and aqueous techniques. benign prostatic hypertrophy, polypeptides and agonists are 0746 Compositions of the present invention may also preferably administered parenterally or orally. optionally include other therapeutic ingredients, anti-caking 0741. The polypeptides described herein can be adminis agents, preservatives, Sweetening agents, colorants, flavors, tered alone or in combination with other agents. For example, desiccants, plasticizers, dyes, glidants, anti-adherents, anti the polypeptides can be administered together with an anal static agents, surfactants (wetting agents), anti-oxidants, gesic polypeptide or compound. The analgesic polypeptide or film-coating agents, and the like. Any Such optional ingredi compound can be covalently attached to a polypeptide ent must be compatible with the compound described herein described herein or it can be a separate agent that is admin to insure the stability of the formulation. istered together with or sequentially with a polypeptide 0747 The composition may contain other additives as described herein in a combination therapy. needed, including for example lactose, glucose, fructose, 0742 Combination therapy can be achieved by adminis galactose, trehalose, Sucrose, maltose, raffinose, maltitol, tering two or more agents, e.g., a polypeptide described melezitose, stachyose, lactitol, palatinite, starch, Xylitol, herein and an analgesic polypeptide or compound, each of mannitol, myoinositol, and the like, and hydrates thereof, and which is formulated and administered separately, or by amino acids, for example alanine, glycine and betaine, and administering two or more agents in a single formulation. polypeptides and proteins, for example albumen. Other combinations are also encompassed by combination 0748 Examples of excipients for use as the pharmaceuti therapy. For example, two agents can be formulated together cally acceptable carriers and the pharmaceutically acceptable and administered in conjunction with a separate formulation inert carriers and the aforementioned additional ingredients containing a third agent. While the two or more agents in the include, but are not limited to binders, fillers, disintegrants, combination therapy can be administered simultaneously, lubricants, anti-microbial agents, and coating agents such as: they need not be. For example, administration of a first agent 0749 BINDERS: corn starch, potato starch, other (or combination of agents) can precede administration of a starches, gelatin, natural and synthetic gums such as acacia, second agent (or combination of agents) by minutes, hours, Xanthan, Sodium alginate, alginic acid, otheralginates, pow days, or weeks. Thus, the two or more agents can be admin dered tragacanth, guar gum, cellulose and its derivatives (e.g., istered within minutes of each other or within 1, 2, 3, 6, 9, 12, ethyl cellulose, cellulose acetate, carboxymethyl cellulose 15, 18, or 24 hours of each other or within 1, 2, 3, 4, 5, 6, 7, 8, calcium, Sodium carboxymethyl cellulose), polyvinyl pyr 9, 10, 12, 14 days of each other or within 2, 3, 4, 5, 6, 7, 8, 9. rolidone (e.g., povidone, crospovidone, copovidone, etc), or 10 weeks of each other. In some cases even longerintervals methyl cellulose, Methocel, pre-gelatinized starch (e.g., are possible. While in many cases it is desirable that the two STARCH 1500R and STARCH 1500 LM(R), sold by Color or more agents used in a combination therapy be present in con, Ltd.), hydroxypropyl methyl cellulose, microcrystalline within the patient's body at the same time, this need not be so. cellulose (e.g. AVICELTM, such as, AVICEL-PH-101TM, 0743 Combination therapy can also include two or more -103TM and 105TM, sold by FMC Corporation, Marcus Hook, administrations of one or more of the agents used in the Pa., USA), or mixtures thereof, combination. For example, if agent X and agent Y are used in 0750 FILLERS: talc, calcium carbonate (e.g., granules or a combination, one could administer them sequentially in any powder), dibasic calcium phosphate, tribasic calcium phos combination one or more times, e.g., in the order X-Y-X. phate, calcium sulfate (e.g., granules or powder), microcrys X-X-Y, Y-X-Y, Y-Y-X, X-X-Y-Y, etc. talline cellulose, powdered cellulose, dextrates, kaolin, man US 2012/0283411 A9 Nov. 8, 2012

nitol, silicic acid, Sorbitol, starch, pre-gelatinized starch, glyceryl behenate); softgelatin capsules (like Sorbitol special dextrose, fructose, honey, lactose anhydrate, lactose mono Solution); spheres for coating (like Sugar spheres); spheroni hydrate, lactose and aspartame, lactose and cellulose, lactose Zation agents (like glyceryl behenate and microcrystalline and microcrystalline cellulose, maltodextrin, maltose, man cellulose); Suspending/gelling agents (like carrageenan, gel nitol, microcrystalline cellulose & guar gum, molasses, lan gum, mannitol, microcrystalline cellulose, povidone, Sucrose, or mixtures thereof, Sodium starch glycolate, Xanthan gum); Sweeteners (like (0751. DISINTEGRANTS: agar-agar, alginic acid, cal aspartame, aspartame and lactose, dextrose, fructose, honey, cium carbonate, microcrystalline cellulose, croScarmellose maltodextrin, maltose, mannitol, molasses, Sorbitol crystal Sodium, crospovidone, polacrilin potassium, Sodium starch line, Sorbitol special Solution, Sucrose); wet granulation glycolate, potato or tapioca Starch, other starches, pre-gelati agents (like calcium carbonate, lactose anhydrous, lactose nized starch, clays, other algins, other celluloses, gums (like monohydrate, maltodextrin, mannitol, microcrystalline cel gellan), low-substituted hydroxypropyl cellulose, or mixtures lulose, povidone, starch), caramel, carboxymethylcellulose thereof, Sodium, cherry cream flavor and cherry flavor, citric acid (0752 LUBRICANTS: calcium stearate, magnesium anhydrous, citric acid, confectioner's sugar, D&C Red No. Stearate, mineral oil, light mineral oil, glycerin, Sorbitol, man 33, D&C Yellow #10 Aluminum Lake, disodium edetate, nitol, polyethylene glycol, other glycols, Stearic acid, sodium ethyl alcohol 15%, FD&C Yellow No. 6 aluminum lake, lauryl Sulfate, Sodium Stearyl fumarate, vegetable based fatty FD&C Blue #1 Aluminum Lake, FD&C Blue No. 1, FD&C acids lubricant, talc, hydrogenated vegetable oil (e.g., peanut blue no. 2 aluminum lake, FD&C Green No. 3, FD&C Red oil, cottonseed oil, Sunflower oil, sesame oil, olive oil, corn oil No. 40, FD&C Yellow No. 6 Aluminum Lake, FD&C Yellow and Soybean oil), Zinc Stearate, ethyl oleate, ethyl laurate, No. 6, FD&C Yellow No. 10, glycerol palmitostearate, glyc agar, syloid silica gel (AEROSIL 200, W.R. Grace Co., Bal eryl monostearate, indigo carmine, lecithin, manitol, methyl timore, Md. USA), a coagulated aerosol of synthetic silica and propyl parabens, mono ammonium glycyrrhizinate, natu (Deaussa Co., Plano, Tex. USA), a pyrogenic silicon dioxide ral and artificial orange flavor, pharmaceutical glaze, poloX (CAB-O-SIL, Cabot Co., Boston, Mass. USA), or mixtures amer 188, Polydextrose, polysorbate 20, polysorbate 80, thereof, polyvidone, pregelatinized corn starch, pregelatinized starch, (0753 ANTI-CAKING AGENTS: calcium silicate, mag red iron oxide, Saccharin sodium, Sodium carboxymethyl nesium silicate, silicon dioxide, colloidal silicon dioxide, ether, sodium chloride, sodium citrate, sodium phosphate, talc, or mixtures thereof, strawberry flavor, synthetic black iron oxide, synthetic red 0754). ANTIMICROBIAL AGENTS: benzalkonium chlo iron oxide, titanium dioxide, and white wax. ride, benzethonium chloride, benzoic acid, benzyl alcohol, 0757. Solid oral dosage forms may optionally be treated butyl paraben, cetylpyridinium chloride, cresol, chlorobu with coating systems (e.g. Opadry(R) fix film coating system, tanol, dehydroacetic acid, ethylparaben, methylparaben, phe for example Opadry(R) blue (OY-LS-20921), Opadry(R) white nol, phenylethyl alcohol, phenoxyethanol, phenylmercuric (YS-2-7063), Opadry(R) white (YS-1-7040), and black ink acetate, phenylmercuric nitrate, potassium Sorbate, propylpa (S-1-8106). raben, Sodium benzoate, sodium dehydroacetate, Sodium pro 0758. The agents either in their free form or as a salt can be pionate, Sorbic acid, thimersol, thymo, or mixtures thereof, combined with a polymer Such as polylactic-glycoloic acid and (PLGA), poly-(I)-lactic-glycolic-tartaric acid (P(I)LGT) (0755 COATINGAGENTS: sodium carboxymethylcellu (WO 01/12233), polyglycolic acid (U.S. Pat. No. 3,773.919), lose, cellulose acetate phthalate, ethylcellulose, gelatin, phar polylactic acid (U.S. Pat. No. 4,767,628), poly(e-caprolac maceutical glaze, hydroxypropyl cellulose, hydroxypropyl tone) and poly(alkylene oxide) (U.S. 2003.0068384) to create methylcellulose (hypromellose), hydroxypropyl methyl cel a Sustained release formulation. Such formulations can be lulose phthalate, methylcellulose, polyethylene glycol, poly used to implants that release a polypeptide or another agent vinyl acetate phthalate, shellac, Sucrose, titanium dioxide, over a period of a few days, a few weeks or several months carnauba wax, microcrystalline wax, gellan gum, maltodex depending on the polymer, the particle size of the polymer, trin, methacrylates, microcrystalline cellulose and carrag and the size of the implant (see, e.g., U.S. Pat. No. 6,620,422). eenan or mixtures thereof. Other Sustained release formulations and polymers for use in 0756. The formulation can also include other excipients are described in EP 0 467389 A2, WO 93/24150, U.S. Pat. and categories thereof including but not limited to L-histi No. 5,612,052, WO 97/40085, WO 03/075887, WO dine, Pluronic R, Poloxamers (such as Lutrol(R) and Polox 01/01964A2, U.S. Pat. No. 5,922,356, WO 94/155587, WO amer 188), ascorbic acid, glutathione, permeability enhanc 02/074247A2, WO 98/25642, U.S. Pat. No. 5,968,895, U.S. ers (e.g. lipids, sodium cholate, acylcarnitine, Salicylates, Pat. No. 6,180,608, U.S. 2003.0171296, U.S. 20020176841, mixed bile salts, fatty acid micelles, chelators, fatty acid, U.S. Pat. No. 5,672,659, U.S. Pat. No. 5,893,985, U.S. Pat. Surfactants, medium chain glycerides), protease inhibitors No. 5,134,122, U.S. Pat. No. 5,192,741, U.S. Pat. No. 5,192, (e.g. soybean trypsin inhibitor, organic acids), pH lowering 741, U.S. Pat. No. 4,668,506, U.S. Pat. No. 4,713,244, U.S. agents and absorption enhancers effective to promote bio Pat. No. 5,445,832 U.S. Pat. No. 4,931,279, U.S. Pat. No. availability (including but not limited to those described in 5,980,945, WO 02/058672, WO 9726015, WO 97/04744, U.S. Pat. No. 6,086,918 and U.S. Pat. No. 5,912,014), creams and. US20020019446. In such sustained release formulations and lotions (like maltodextrin and carrageenans); materials microparticles (Delie and Blanco-Prieto 2005 Molecule for chewable tablets (like dextrose, fructose, lactose mono 10:65-80) of polypeptide are combined with microparticles hydrate, lactose and aspartame, lactose and cellulose, malto of polymer. One or more Sustained release implants can be dextrin, maltose, mannitol, microcrystalline cellulose and placed in the large intestine, the small intestine or both. U.S. guar gum, Sorbitol crystalline); parenterals (like mannitol and Pat. No. 6,011,011 and WO 94/06452 describe a sustained povidone); plasticizers (like dibutyl sebacate, plasticizers for release formulation providing either polyethylene glycols coatings, polyvinylacetate phthalate); powder lubricants (like (i.e. PEG 300 and PEG 400) or triacetin. WO 03/053401 US 2012/0283411 A9 Nov. 8, 2012 42 describes a formulation which may both enhance bioavail (administration of powder formulation by Syringe or any ability and provide controlled release of the agent within the other similar device into the lungs) and aerosol inhalation. GI tract. Additional controlled release formulations are Aerosols (e.g., jet or ultrasonic nebulizers, metered-dose described in WO 02/38129, EP326151, U.S. Pat. No. 5,236, inhalers (MDIs), and dry-powder inhalers (DPIs)) can also be 704, WO 02/30398, WO 98/13029; U.S. 20030064105, U.S. used in intranasal applications. Aerosol formulations are 2003.0138488A1, U.S. 20030216307A1, U.S. Pat. No. 6,667, stable dispersions or Suspensions of solid material and liquid 060, WO 01/49249, WO 01/49311, WO 01/49249, WO droplets in a gaseous medium and can be placed into pressur O1/49311, and U.S. Pat. No. 5,877,224. ized acceptable propellants, such as hydrofluoroalkanes 0759. The agents can be administered, e.g., by intravenous (HFAs, i.e. HFA-134a and HFA-227, or a mixture thereof), injection, intramuscular injection, Subcutaneous injection, dichlorodifluoromethane (or other chlorofluocarbon propel intraperitoneal injection, topical, Sublingual, intraarticular (in lants such as a mixture of Propellants 11, 12, and/or 114), the joints), intradermal, buccal, ophthalmic (including propane, nitrogen, and the like. Pulmonary formulations may intraocular), intranasally (including using a cannula), include permeation enhancers such as fatty acids, saccha intraspinally, intrathecally, or by other routes. The agents can rides, chelating agents, enzyme inhibitors (e.g., protease be administered orally, e.g., as a tablet or cachet containing a inhibitors), adjuvants (e.g., glycocholate, Surfactin, span 85. predetermined amount of the active ingredient, gel, pellet, and nafamo.stat), preservatives (e.g., benzalkonium chloride paste, syrup, bolus, electuary, slurry, capsule, powder, lyo or chlorobutanol), and ethanol (normally up to 5% but possi philized powder, granules, Sachet, as a solution or a suspen bly up to 20%, by weight). Ethanol is commonly included in sion in an aqueous liquid or a non-aqueous liquid, as an aerosol compositions as it can improve the function of the oil-in-water liquid emulsion ora water-in-oil liquid emulsion, metering valve and in Some cases also improve the stability of via a micellar formulation (see, e.g. WO 97/11682) via a the dispersion. Pulmonary formulations may also include liposomal formulation (see, e.g., EP 736299, WO 99/59550 surfactants which include but are not limited to bile salts and and WO 97/13500), via formulations described in WO those described in U.S. Pat. No. 6,524,557 and references 03/094886, viabilosome (bile-salt based vesicular system), therein. The surfactants described in U.S. Pat. No. 6,524,557, via a dendrimer, or in some other form. Orally administered e.g., a C8-C16 fatty acid salt, a bile salt, a phospholipid, or compositions can include binders, lubricants, inert diluents, alkyl saccaride are advantageous in that Some of them also lubricating, Surface active or dispersing agents, flavoring reportedly enhance absorption of the polypeptide in the for agents, and humectants. Orally administered formulations mulation. Also suitable in the invention are dry powder for such as tablets may optionally be coated or scored and may be mulations comprising a therapeutically effective amount of formulated so as to provide Sustained, delayed or controlled active compound blended with an appropriate carrier and release of the active ingredient therein. The agents can also be adapted for use in connection with a dry-powder inhaler. administered transdermally (i.e. via reservoir-type or matrix Absorption enhancers which can be added to dry powder type patches, microneedles, thermal poration, hypodermic formulations of the present invention include those described needles, iontophoresis, electroporation, ultrasound or other in U.S. Pat. No. 6,632,456. WO 02/080884 describes new forms of Sonophoresis, jet injection, or a combination of any methods for the surface modification of powders. Aerosol of the preceding methods (Prausnitz et al. 2004, Nature formulations may include U.S. Pat. No. 5,230,884, U.S. Pat. Reviews Drug Discovery 3:115-124)). The agents can be No. 5,292.499, WO 017/8694, WO 01/78696, U.S. administered using high-velocity transdermal particle injec 2003019437, U.S. 20030165436, and WO 96/40089 (which tion techniques using the hydrogel particle formulation includes Vegetable oil). Sustained release formulations Suit described in U.S. 20020061336. Additional particle formula able for inhalation are described in U.S. 20010036481A1, tions are described in WOOO/45792, WOOO/53160, and WO 20030232019A1, and U.S. 20040018243A1 as well as in WO 02/19989. An example of a transdermal formulation contain 01/13891, WO 02/067902, WO 03/072080, and WO ing plaster and the absorption promoter dimethylisosorbide 03/079885. Pulmonary formulations containing micropar can be found in WO 89/04179. WO 96/11705 provides for ticles are described in WO 03/O15750, U.S. 20030008013, mulations Suitable for transdermal administration. The agents and WOOO/OO176. can be administered in the form a Suppository or by other 0761 Pulmonary formulations containing stable glassy vaginal or rectal means. The agents can be administered in a state powder are described in U.S. 2002014 1945 and U.S. Pat. transmembrane formulation as described in WO 90/07923. No. 6,309,671. Other aerosol formulations are described in The agents can be administered non-invasively via the dehy EP 1338272A1 WO 90/09781, U.S. Pat. No. 5,348,730, U.S. drated particles described in U.S. Pat. No. 6,485,706. The Pat. No. 6,436,367, WO 91/04011, and U.S. Pat. No. 6,294, agent can be administered in an enteric-coated drug formu 153 and U.S. Pat. No. 6,290,987 describes a liposomal based lation as described in WO 02/49621. The agents can be formulation that can be administered via aerosol or other administered intranasally using the formulation described in means. Powder formulations for inhalation are described in U.S. Pat. No. 5,179,079. Formulations suitable for parenteral U.S. 2003.0053960 and WO 01/60341. The agents can be injection are described in WO 00/62759. The agents can be administered intranasally as described in U.S. 20010038824. administered using the casein formulation described in U.S. The agents can be incorporated into microemulsions, which 20030206939 and WO 00/06108. The agents can be admin generally are thermodynamically stable, isotropically clear istered using the particulate formulations described in U.S. dispersions of two immiscible liquids, such as oil and water, 2002OO34536. stabilized by an interfacial film of surfactant molecules (En 0760. The agents, alone or in combination with other suit cyclopedia of Pharmaceutical Technology (New York: Mar able components, can be administered by pulmonary route cel Dekker, 1992), volume 9). For the preparation of micro utilizing several techniques including but not limited to emulsions, Surfactant (emulsifier), co-surfactant (co intratracheal instillation (delivery of solution into the lungs emulsifier), an oil phase and a water phase are necessary. by Syringe), intratracheal delivery of liposomes, insufflation Suitable surfactants include any surfactants that are useful in US 2012/0283411 A9 Nov. 8, 2012

the preparation of emulsions, e.g., emulsifiers that are typi U.S. Pat. No. 5,007,790 and U.S. Pat. No. 5,972,389 (sus cally used in the preparation of creams. The co-surfactant (or tained release dosage forms); WOO4112711 (oral extended “co-emulsifer') is generally selected from the group of release compositions); WO05027878, WO02072033, and polyglycerol derivatives, glycerol derivatives and fatty alco WO02072034 (delayed release compositions with natural or hols. Preferred emulsifier/co-emulsifier combinations are synthetic gum); WO05030182 (controlled release formula generally although not necessarily selected from the group tions with an ascending rate of release); WO05048998 (mi consisting of glyceryl monostearate and polyoxyethylene croencapsulation system); U.S. Pat. No. 5,952.314 (biopoly Stearate; polyethylene glycol and ethylene glycol palmito mer); U.S. Pat. No. 5,108,758 (glassy amylose matrix Stearate; and caprilic and capric triglycerides and oleoyl mac delivery); U.S. Pat. No. 5,840,860 (modified starch based rogolglycerides. The water phase includes not only water but delivery). JP10324642 (delivery system comprising chitosan also, typically, buffers, glucose, propylene glycol, polyethyl and gastric resistant material Such as wheat gliadin or Zein); ene glycols, preferably lower molecular weight polyethylene U.S. Pat. No. 5,866.619 and U.S. Pat. No. 6,368,629 (saccha glycols (e.g., PEG 300 and PEG 400), and/or glycerol, and the ride containing polymer); U.S. Pat. No. 6,531,152 (describes like, while the oil phase will generally comprise, for example, a drug delivery system containing a water soluble core (Ca fatty acid esters, modified vegetable oils, silicone oils, mix pectinate or other water-insoluble polymers) and outer coat tures of mono- di- and triglycerides, mono- and di-esters of which bursts (eg hydrophobic polymer-Eudragrit)); U.S. Pat. PEG (e.g., oleoyl macrogol glycerides), etc. No. 6,234,464; U.S. Pat. No. 6,403,130 (coating with poly 0762. The agents described herein can be incorporated mercontaining casein and high methoxy pectin; WOO174175 into pharmaceutically-acceptable nanoparticle, nanosphere, (Maillard reaction product); WO05063206 (solubility and nanocapsule formulations (Delie and Blanco-Prieto 2005 increasing formulation); WOO4019872 (transferring fusion Molecule 10:65-80). Nanocapsules can generally entrap proteins). The agents described herein may be formulated compounds in a stable and reproducible way (Henry-Mich using gastrointestinal retention system technology (GIRES; elland et al., 1987: Quintanar-Guerrero et al., 1998; Douglas Merrion Pharmaceuticals). GIRES comprises a controlled et al., 1987). To avoid side effects due to intracellular poly release dosage form inside an inflatable pouch, which is meric overloading, ultrafine particles (sized around 0.1 um) placed in a drug capsule for oral administration. Upon disso can be designed using polymers able to be degraded in vivo lution of the capsule, a gas-generating system inflates the (e.g. biodegradable polyalkyl-cyanoacrylate nanoparticles). pouch in the stomach where it is retained for 16-24 hours, all Such particles are described in the prior art (Couvreur et al. the time releasing agents described herein. 1980; 1988: Zur Muhlen et al., 1998; Zambaux et al. 1998: 0764. The agents described herein can beformulated in an Pinto-Alphandry et al., 1995 and U.S. Pat. No. 5,145,684). osmotic device including the ones disclosed in U.S. Pat. No. 0763 The agents described herein can be formulated with 4,503,030, U.S. Pat. No. 5,609,590 and U.S. Pat. No. 5,358, pH sensitive materials which may include those described in 502. U.S. Pat. No. 4,503,030 discloses an osmotic device for WOO4041195 (including the seal and enteric coating dispensing a drug to certain pH regions of the gastrointestinal described therein) and pH-sensitive coatings that achieve tract. More particularly, the invention relates to an osmotic delivery in the colon including those described in U.S. Pat. device comprising a wall formed of a semi-permeable pH No. 4,910,021 and WO9001329. U.S. Pat. No. 4,910,021 sensitive composition that Surrounds a compartment contain describes using a pH-sensitive material to coat a capsule. ing a drug, with a passageway through the wall connecting the WO9001329 describes using pH-sensitive coatings on beads exterior of the device with the compartment. The device containing acid, where the acid in the bead core prolongs delivers the drug at a controlled rate in the region of the dissolution of the pH-sensitive coating. U.S. Pat. No. 5,175, gastrointestinal tract having a pH of less than 3.5, and the 003 discloses a dual mechanism polymer mixture composed device self-destructs and releases all its drug in the region of of pH-sensitive enteric materials and film-forming plasticiz the gastrointestinal tract having a pH greater than 3.5, thereby ers capable of conferring permeability to the enteric material, providing total availability for drug absorption. U.S. Pat. Nos. for use in drug-delivery systems; a matrix pellet composed of 5,609.590 and 5.358.502 disclose an osmotic bursting device a dual mechanism polymer mixture permeated with a drug for dispensing a beneficial agent to an aqueous environment. and sometimes covering a pharmaceutically neutral nucleus; The device comprises a beneficial agent and osmagent Sur a membrane-coated pellet comprising a matrix pellet coated rounded at least in part by a semi-permeable membrane. The with a dual mechanism polymer mixture envelope of the same beneficial agent may also function as the osmagent. The semi or different composition; and a pharmaceutical dosage form permeable membrane is permeable to water and substantially containing matrix pellets. The matrix pellet releases acid impermeable to the beneficial agent and osmagent. A trigger soluble drugs by diffusion in acid pH and by disintegration at means is attached to the semi-permeable membrane (e.g., pH levels of nominally about 5.0 or higher. The agents joins two capsule halves). The trigger means is activated by a described herein may be formulated in the pH triggered tar pH of from 3 to 9 and triggers the eventual, but sudden, geted control release systems described in WOO4052339. delivery of the beneficial agent. These devices enable the The agents described herein may be formulated according to pH-triggered release of the beneficial agent core as a bolus by the methodology described in any of WO03105812 (extruded osmotic bursting. hyrdratable polymers); WO0243767 (enzyme cleavable 0765. The agents described herein may be formulated membrane translocators); WO03007913 and WO03086297 based on the invention described in U.S. Pat. No. 5,316,774 (mucoadhesive systems); WO02072075 (bilayer laminated which discloses a composition for the controlled release of an formulation comprising pH lowering agent and absorption active Substance comprising a polymeric particle matrix, enhancer); WOO4064769 (amidated polypeptides); where each particle defines a network of internal pores. The WO05063156 (solid lipid suspension with pseudotropic and/ active Substance is entrapped within the pore network or thixotropic properties upon melting); WO03035029 and together with a blocking agent having physical and chemical WO03035041 (erodible, gastric retentive dosage forms): characteristics selected to modify the release rate of the active US 2012/0283411 A9 Nov. 8, 2012 44 substance from the internal pore network. In one embodi merizable monomers. These pH-sensitive polymers may have ment, drugs may be selectively delivered to the intestines a structure of linear polymer, grafted copolymer, hydrogel or using an enteric material as the blocking agent. The enteric interpenetrating network polymer. material remains intact in the stomach but degrades under the 0769 The agents described herein may be formulated pH conditions of the intestines. In another embodiment, the according U.S. Pat. No. 5,656.292 which discloses a compo Sustained release formulation employs a blocking agent, sition for pH dependent or pH regulated controlled release of which remains stable under the expected conditions of the active ingredients especially drugs. The composition consists environment to which the active substance is to be released. of a compactable mixture of the active ingredient and starch The use of pH-sensitive materials alone to achieve site-spe molecules substituted with acetate and dicarboxylate resi cific delivery is difficult because of leaking of the beneficial dues. The preferred dicarboxylate acid is succinate. The aver agent prior to the release site or desired delivery time and it is age Substitution degree of the acetate residue is at least 1 and difficult to achieve long time lags before release of the active 0.2-1.2 for the dicarboxylate residue. The starch molecules ingredient after exposure to high pH (because of rapid disso can have the acetate and dicarboxylate residues attached to lution or degradation of the pH-sensitive materials). the same starch molecule backbone or attached to separate 0766 The agents may also be formulated in a hybrid sys starch molecule backbones. The present invention also dis tem which combines pH-sensitive materials and osmotic closes methods for preparing said starch acetate dicarboxy delivery systems. These hybrid devices provide delayed ini lates by transesterification or mixing of starch acetates and tiation of Sustained-release of the beneficial agent. In one starch dicarboxylates respectively. device a pH-sensitive matrix or coating dissolves releasing 0770. The agents described herein may be formulated osmotic devices that provide sustained release of the benefi according to the methods described in U.S. Pat. Nos. 5.554, cial agent see U.S. Pat. Nos. 4,578,075, 4,681,583, and 4,851, 147, 5,788,687, and 6,306.422 which disclose a method for 231. A second device consists of a semipermeable coating the controlled release of a biologically active agent wherein made of a polymer blend of an insoluble and a pH-sensitive the agent is released from a hydrophobic, pH-sensitive poly material. As the pH increases, the permeability of the coating mer matrix. The polymer matrix swells when the environment increases, increasing the rate of release of beneficial agent see reaches pH 8.5, releasing the active agent. A polymer of U.S. Pat. Nos. 4,096,238, 4,503,030, 4,522,625, and 4,587, hydrophobic and weakly acidic comonomers is disclosed for 117. use in the controlled release system. Also disclosed is a spe 0767 The agents described herein may be formulated in cific embodiment in which the controlled release system may terpolumers according to U.S. Pat. No. 5,484,610 which dis be used. The pH-sensitive polymer is coated onto a latex closes terpolymers which are sensitive to pH and temperature catheter used in ureteral catheterization. A ureteral catheter which are useful carriers for conducting bioactive agents coated with a pH-sensitive polymer having an antibiotic or through the gastric juices of the stomach in a protected form. urease inhibitor trapped within its matrix will release the The terpolymers swell at the higher physiologic pH of the active agent when exposed to high pH urine. intestinal tract causing release of the bioactive agents into the 0771. The agents can be administered using COLAL(R) intestine. The terpolymers are linear and are made up of 35 to colonic drug delivery technology (U.S. Pat. No. 6,534,549) 99 wt % of a temperature sensitive component, which imparts BTGInternational, Ltd.; Alizyme, plc. Cambridge, UK) in to the terpolymer LCST (lower critical solution temperature) which small pellets containing the agents are coated with properties below body temperatures, 1 to 30 wt % of a pH ethylcellulose and a specific form of amylose. This coating sensitive component having a pKa in the range of from 2 to 8 prevents drug release in the stomach and Small intestine. which functions through ionization or deionization of car When the pellets reach the colon the amylose in the coating is boxylic acid groups to prevent the bioactive agent from being broken down by bacterial enzymes and the agent is released. lost at low pH but allows bioactive agent release at physi The agents described herein may be formulated in/with bio ological pH of about 7.4 and a hydrophobic component which adhesive polymers according to U.S. Pat. No. 6,365,187. stabilizes the LCST below body temperatures and compen Bioadhesive polymers in the form of, or as a coating on, sates for bioactive agent effects on the terpolymers. The ter microcapsules containing drugs or bioactive substances polymers provide for safe bioactive agent loading, a simple which may serve for therapeutic, or diagnostic purposes in procedure for dosage form fabrication and the terpolymer diseases of the gastrointestinal tract, are described in U.S. Pat. functions as a protective carrier in the acidic environment of No. 6,365,187. The polymeric microspheres all have a bio the stomachand also protects the bioactive agents from diges adhesive force of at least 11 mN/cm (110 N/m2) Techniques tive enzymes until the bioactive agent is released in the intes for the fabrication of bioadhesive microspheres, as well as a tinal tract. method for measuring bioadhesive forces between micro 0768. The agents described herein may be formulated in spheres and selected segments of the gastrointestinal tract in pH sensitive polymers according to those described in U.S. vitro are also described. This quantitative method provides a Pat. No. 6,103,865. U.S. Pat. No. 6,103,865 discloses pH means to establish a correlation between the chemical nature, sensitive polymers containing Sulfonamide groups, which the Surface morphology and the dimensions of drug-loaded can be changed in physical properties, such as Swellability microspheres on one hand and bioadhesive forces on the and solubility, depending on pH and which can be applied for other, allowing the screening of the most promising materials a drug-delivery system, bio-material, sensor, and the like, and from a relatively large group of natural and synthetic poly a preparation method therefore. The pH-sensitive polymers mers which, from theoretical consideration, should be used are prepared by introduction of Sulfonamide groups, various for making bioadhesive microspheres. Solutions of medica in pKa, to hydrophilic groups of polymers either through ment in buffered saline and similar vehicles are commonly coupling to the hydrophilic groups of polymers. Such as acry employed to generate an aerosol in a nebulizer. Simple nebu lamide, N,N-dimethylacrylamide, acrylic acid, N-isopropy lizers operate on Bernoulli's principle and employ a stream of lacrylamide and the like or copolymerization with other poly air or oxygen to generate the spray particles. More complex US 2012/0283411 A9 Nov. 8, 2012

nebulizers employ ultrasound to create the spray particles. al., 2001 Innovations in Pharmaceutical Technology 106 Both types are well known in the art and are described in 110). The agent can be administered via a nanocochleate or standard textbooks of pharmacy Such as Sprowls American cochleate delivery vehicle (BioDelivery Sciences Interna Pharmacy and Remington's The Science and Practice of tional). The agents can be delivered transmucosally (i.e. Pharmacy. Other devices for generating aerosols employ across a mucosal Surface Such as the vagina, eye or nose) compressed gases, usually hydrofluorocarbons and chlorof using formulations such as that described in U.S. Pat. No. luorocarbons, which are mixed with the medicament and any 5,204,108. The agents can be formulated in microcapsules as necessary excipients in a pressurized container, these devices described in WO 88/01165. The agent can be administered are likewise described in standard textbooks such as Sprowls intra-orally using the formulations described in U.S. and Remington. 20020055496, WO 00/47203, and U.S. Pat. No. 6,495,120. The agent can be delivered using nanoemulsion formulations 0772. The agents can be a free acid or base, or a pharma described in WOO1/91728A2. cologically acceptable salt thereof. Solids can be dissolved or dispersed immediately prior to administration or earlier. In Controlled Release Formulations Some circumstances the preparations include a preservative to prevent the growth of microorganisms. The pharmaceutical 0773. In general, one can provide for controlled release of forms suitable for injection can include sterile aqueous or the agents described herein through the use of a wide variety organic Solutions or dispersions which include, e.g., water, an of polymeric carriers and controlled release systems includ alcohol, an organic solvent, an oil or other solvent or dispers ing erodible and non-erodible matrices, osmotic control ant (e.g., glycerol, propylene glycol, polyethylene glycol, and devices, various reservoir devices, enteric coatings and mul Vegetable oils). The formulations may contain antioxidants, tiparticulate control devices. buffers, bacteriostats, and solutes that render the formulation 0774. Matrix devices area common device for controlling isotonic with the blood of the intended recipient, and aqueous the release of various agents. In such devices, the agents and non-aqueous sterile Suspensions that can include Sus described herein are generally present as a dispersion within pending agents, solubilizers, thickening agents, stabilizers, the polymer matrix, and are typically formed by the compres and preservatives. Pharmaceutical agents can be sterilized by sion of a polymer/drug mixture or by dissolution or melting. filter sterilization or by other suitable means. The agent can be The dosage release properties of these devices may be depen fused to immunoglobulins or albumin, albumin variants or dent upon the solubility of the agent in the polymer matrix or, fragments thereof, or incorporated into a liposome to improve in the case of porous matrices, the Solubility in the sink half-life. Thus the peptides described herein may be fused solution within the pore network, and the tortuosity of the directly or via a peptide linker, water soluble polymer, or network. In one instance, when utilizing an erodible poly prodrug linker to albumin or an analog, fragment, or deriva meric matrix, the matrix imbibes water and forms an aque tive thereof. Generally, the albumin proteins that are part of ous-swollengel that entraps the agent. The matrix then gradu the fusion proteins of the present invention may be derived ally erodes, Swells, disintegrates or dissolves in the GI tract, from albumin cloned from any species, including human. thereby controlling release of one or more of the agents Human serum albumin (HSA) consists of a single non-gly described herein. In non-erodible devices, the agent is cosylated polypeptide chain of 585 amino acids with a for released by diffusion through an inert matrix. mula molecular weight of 66,500. The amino acid sequence 0775 Agents described herein can be incorporated into an of human HSA is known See Meloun, et al. (1975) FEBS erodible or non-erodible polymeric matrix controlled release Letters 58:136: Behrens, et al. (1975) Fed. Proc. 34:591; device. By an erodible matrix is meant aqueous-erodible or Lawn, et al. (1981) Nucleic Acids Research 9:6102-6114; water-swellable or aqueous-Soluble in the sense of being Minghetti, et al. (1986) J. Biol. Chem. 261:6747, each of either erodible or swellable or dissolvable in pure water or which are incorporated by reference herein. A variety of requiring the presence of an acid or base to ionize the poly polymorphic variants as well as analogs and fragments of meric matrix Sufficiently to cause erosion or dissolution. albumin have been described. See Weitkamp, et al., (1973) When contacted with the aqueous environment of use, the Ann. Hum. Genet. 37:219). For example, in EP 322,094, erodible polymeric matrix imbibes water and forms an aque various shorter forms of HSA. Some of these fragments of ous-swollen gel or matrix that entraps the agent described HSA are disclosed, including HSA (1-373), HSA (1-388), herein. The aqueous-swollen matrix gradually erodes, Swells, HSA (1-389), HSA (1-369), and HSA (1-419) and fragments disintegrates or dissolves in the environment of use, thereby between 1-369 and 1-419. EP 399,666 discloses albumin controlling the release of a compound described herein to the fragments that include HSA (1-177) and HSA (1-200) and environment of use. fragments between HSA (1-177) and HSA (1-200). Methods 0776 The erodible polymeric matrix into which an agent related to albumin fusion proteins can be found in U.S. Pat. described herein can be incorporated may generally be No. 7,056,701, U.S. Pat. No. 6,994,857, U.S. Pat. No. 6,946, described as a set of excipients that are mixed with the agent 134, U.S. Pat. No. 6,926,898, and U.S. Pat. No. 6,905,688 and following its formation that, when contacted with the aqueous the related priority documents and references cited therein. environment of use imbibes water and forms a water-swollen The agent can also be conjugated to polyethylene glycol gel or matrix that entraps the drug form. Drug release may (PEG) chains. Methods for pegylation and additional formu occur by a variety of mechanisms, for example, the matrix lations containing PEG-conjugates (i.e. PEG-based hydro may disintegrate or dissolve from around particles or granules gels, PEG modified liposomes) can be found in Harris and of the agent or the agent may dissolve in the imbibed aqueous Chess, Nature Reviews Drug Discovery 2: 214-221 and the solution and diffuse from the tablet, beads or granules of the references therein. polypeptides can also be modified with device. One ingredient of this water-swollen matrix is the alkyl groups (e.g., C1-C20 straight or branched alkyl groups); water-swellable, erodible, or soluble polymer, which may fatty acid radicals; and combinations of PEG, alkyl groups generally be described as an osmopolymer, hydrogel or and fatty acid radicals (see U.S. Pat. No. 6,309,633; Soltero et water-swellable polymer. Such polymers may be linear, US 2012/0283411 A9 Nov. 8, 2012 46 branched, or crosslinked. The polymers may be homopoly ride, polyethylene), hydrophilic polymers (e.g. ethyl cellu mers or copolymers. In certain embodiments, they may be lose, cellulose acetate, crosslinked polyvinylpyrrolidone synthetic polymers derived from vinyl, acrylate, methacry (also known as crospovidone)), and fatty compounds (e.g. late, urethane, ester and oxide monomers. In other embodi carnauba wax, microcrystalline wax, and triglycerides). Such ments, they can be derivatives of naturally occurring poly devices are described further in Remington: The Science and mers such as polysaccharides (e.g. chitin, chitosan, dextran Practice of Pharmacy, 20th edition (2000). Matrix controlled and pullulan; gum agar, gum arabic, gum karaya, locust bean release devices may be prepared by blending an agent gum, gum tragacanth, carrageenans, gum ghatti, guar gum, described herein and other excipients together, and then form Xanthan gum and Scleroglucan), starches (e.g. dextrin and ing the blend into a tablet, caplet, pill, or other device formed maltodextrin), hydrophilic colloids (e.g. pectin), phosphati by compressive forces. Such compressed devices may be des (e.g. lecithin), alginates (e.g. ammonium alginate, formed using any of a wide variety of presses used in the Sodium, potassium or calcium alginate, propylene glycol fabrication of pharmaceutical devices. Examples include alginate), gelatin, collagen, and cellulosics. Cellulosics are single-punch presses, rotary tablet presses, and multilayer cellulose polymer that has been modified by reaction of at rotary tablet presses, all well known in the art. See for least a portion of the hydroxyl groups on the saccharide repeat example, Remington: The Science and Practice of Pharmacy, units with a compound to form an ester-linked or an ether 20th Edition, 2000. The compressed device may be of any linked substituent. For example, the cellulosic ethyl cellulose shape, including round, oval, oblong, cylindrical, or triangu has an ether linked ethyl substituent attached to the saccharide lar. The upper and lower surfaces of the compressed device repeat unit, while the cellulosic cellulose acetate has an ester may be flat, round, concave, or convex. linked acetate substituent. In certain embodiments, the cellu 0781. In certain embodiments, when formed by compres losics for the erodible matrix comprises aqueous-soluble and sion, the device has a strength of at least 5 Kiloponds (Kp)/ aqueous-erodible cellulosics can include, for example, ethyl cm (for example, at least 7 Kp/cm). Strength is the fracture cellulose (EC), methylethyl cellulose (MEC), carboxymethyl force, also known as the tablet hardness required to fracture a cellulose (CMC), CMEC, hydroxyethyl cellulose (HEC), tablet formed from the materials, divided by the maximum hydroxypropyl cellulose (HPC), cellulose acetate (CA), cel cross-sectional area of the tablet normal to that force. The lulose propionate (CP), cellulose butyrate (CB), cellulose fracture force may be measured using a Schleuniger Tablet acetate butyrate (CAB), CAP CAT, hydroxypropyl methyl Hardness Tester, Model 6D. The compression force required cellulose (HPMC), HPMCP, HPMCAS, hydroxypropyl to achieve this strength will depend on the size of the tablet, methyl cellulose acetate trimellitate (HPMCAT), and ethyl but generally will be greater than about 5 kP/cm. Friability is hydroxyethylcellulose (EHEC). In certain embodiments, the a well-know measure of a device's resistance to Surface abra cellulosics comprises various grades of low viscosity (MW sion that measures weight loss in percentage after Subjecting less than or equal to 50,000 daltons, for example, the Dow the device to a standardized agitation procedure. Friability MethocelTM series E5, E15LV, E50LV and K10OLY) and high values of from 0.8 to 1.0% are regarded as constituting the viscosity (MW greater than 50,000 daltons, for example, upper limit of acceptability. Devices having a strength of E4MCR, E1OMCR, K4M, K15M and K10OM and the greater than 5 kP/cm generally are very robust, having a MethocelTM K series) HPMC. Other commercially available friability of less than 0.5%. Other methods for forming matrix types of HPMC include the Shin Etsu Metolose 90SH series. controlled-release devices are well known in the pharmaceu 0777. The choice of matrix material can have a large effect tical arts. See for example, Remington: The Science and on the maximum drug concentration attained by the device as Practice of Pharmacy, 20th Edition, 2000. well as the maintenance of a high drug concentration. The 0782. As noted above, the agents described herein may matrix material can be a concentration-enhancing polymer, also be incorporated into an osmotic control device. Such for example, as described in WO05/011634. devices generally include a core containing one or more 0778. Other materials useful as the erodible matrix mate agents as described herein and a water permeable, non-dis rial include, but are not limited to, pullulan, polyvinyl pyr Solving and non-eroding coating Surrounding the core which rolidone, polyvinyl alcohol, polyvinyl acetate, glycerol fatty controls the influx of water into the core from an aqueous acid esters, polyacrylamide, polyacrylic acid, copolymers of environment of use so as to cause drug release by extrusion of ethacrylic acid or methacrylic acid (EUDRAGITO, Rohm some or all of the core to the environment of use. In certain America, Inc., Piscataway, N.J.) and other acrylic acid deriva embodiments, the coating is polymeric, aqueous-permeable, tives such as homopolymers and copolymers of butyl and has at least one delivery port. The core of the osmotic methacrylate, methylmethacrylate, ethylmethacrylate, ethy device optionally includes an osmotic agent which acts to lacrylate, (2-dimethylaminoethyl) methacrylate, and imbibe water from the Surrounding environment via Such a (trimethylaminoethyl) methacrylate chloride. semi-permeable membrane. The osmotic agent contained in 0779. The erodible matrix polymer may contain a wide the core of this device may be an aqueous-swellable hydro variety of the same types of additives and excipients known in philic polymer or it may be an osmogen, also known as an the pharmaceutical arts, including osmopolymers, osmagens, oSmagent. Pressure is generated within the device which solubility-enhancing or -retarding agents and excipients that forces the agent(s) out of the device via an orifice (of a size promote stability or processing of the device. designed to minimize solute diffusion while preventing the 0780. Alternatively, the agents of the present invention build-up of a hydrostatic pressure head). may be administered by or incorporated into a non-erodible 0783) Osmotic agents create a driving force for transport matrix device. In such devices, an agent described herein is of water from the environment of use into the core of the distributed in an inert matrix. The agent is released by diffu device. Osmotic agents include but are not limited to water sion through the inert matrix. Examples of materials Suitable Swellable hydrophilic polymers, and osmogens (or osma for the inert matrix include insoluble plastics (e.g. methyl gens). Thus, the core may include water-swellable hydro acrylate-methyl methacrylate copolymers, polyvinyl chlo philic polymers, both ionic and nonionic, often referred to as US 2012/0283411 A9 Nov. 8, 2012 47 osmopolymers and hydrogels. The amount of water thereof), polytetrafluoroethylene), and inorganic materials Swellable hydrophilic polymers present in the core may range (e.g. talc, calcium phosphate), cyclodextrins, Sugars (e.g. lac from about 5 to about 80 wt % (including for example, 10 to tose, Xylitol), sodium starch glycolate). Nonlimiting 50 wt %). Nonlimiting examples of core materials include examples of disintegrants are sodium starch glycolate (e.g., hydrophilic vinyl and acrylic polymers, polysaccharides Such ExplotabTMCLV, (microcrystalline cellulose (e.g., AviceITM), as calcium alginate, polyethylene oxide (PEO), polyethylene microcrystalline silicified cellulose (e.g., ProSolviM), cros glycol (PEG), polypropylene glycol (PPG), poly (2-hydroxy carmellose sodium (e.g., Ac-Di-SolTM). When the agent ethyl methacrylate), poly (acrylic) acid, poly (methacrylic) described herein is a solid amorphous dispersion formed by a acid, polyvinylpyrrolidone (PVP) and crosslinked PVP poly solvent process, such additives may be added directly to the vinyl alcohol (PVA), PVA/PVP copolymers and PVA/PVP spray-drying solution when forming an agent described copolymers with hydrophobic monomers such as methyl herein/concentration-enhancing polymer dispersion Such methacrylate, vinyl acetate, and the like, hydrophilic polyure that the additive is dissolved or suspended in the solution as a thanes containing large PEO blocks, sodium croScarmellose, slurry, Alternatively, such additives may be added following carrageenan, hydroxyethyl cellulose (HEC), hydroxypropyl the spray-drying process to aid informing the final controlled cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), release device. carboxymethyl cellulose (CMC) and carboxyethyl cellulose 0785. A nonlimiting example of an osmotic device con (CEC), sodium alginate, polycarbophil, gelatin, Xanthan sists of one or more drug layers containing an agent described gum, and sodium starch glycolat. Other materials include herein, such as a Solid amorphous drug/polymer dispersion, hydrogels comprising interpenetrating networks of polymers and a Sweller layer that comprises a water-swellable polymer, that may be formed by addition or by condensation polymer with a coating Surrounding the drug layer and Sweller layer. ization, the components of which may comprise hydrophilic Each layer may contain other excipients such as tableting and hydrophobic monomers such as those just mentioned. aids, osmagents, Surfactants, water-soluble polymers and Water-swellable hydrophilic polymers include but are not water-swellable polymers. limited to PEO, PEG, PVP, sodium croscarmellose, HPMC, 0786 Such osmotic delivery devices may be fabricated in Sodium starch glycolate, polyacrylic acid and crosslinked various geometries including bilayer (wherein the core com versions or mixtures thereof. prises a drug layer and a Sweller layer adjacent to each other), 0784 The core may also include an osmogen (or osma trilayer (wherein the core comprises a Sweller layer sand gent). The amount of osmogen present in the core may range wiched between two drug layers) and concentric (wherein the from about 2 to about 70 wt % (including, for example, from core comprises a central Sweller agent surrounded by the drug 10 to 50 wt %). Typical classes of suitable osmogens are layer). The coating of Such a tablet comprises a membrane water-soluble organic acids, salts and Sugars that are capable permeable to water but substantially impermeable to drug and of imbibing water to thereby effect an osmotic pressure gra excipients contained within. The coating contains one or dient across the barrier of the Surrounding coating. Typical more exit passageways or ports in communication with the useful osmogens include but are not limited to magnesium drug-containing layer(s) for delivering the drug agent. The Sulfate, magnesium chloride, calcium chloride, sodium chlo drug-containing layer(s) of the core contains the drug agent ride, lithium chloride, potassium Sulfate, Sodium carbonate, (including optional osmagents and hydrophilic water-soluble Sodium sulfite, lithium sulfate, potassium chloride, sodium polymers), while the sweller layer consists of an expandable Sulfate, mannitol. Xylitol, urea, Sorbitol, inositol, raffinose, hydrogel, with or without additional osmotic agents. Sucrose, glucose, fructose, lactose, citric acid. Succinic acid, 0787 When placed in an aqueous medium, the tablet tartaric acid, and mixtures thereof. In certain embodiments, imbibes water through the membrane, causing the agent to the osmogen is glucose, lactose, Sucrose, mannitol. Xylitol, form a dispensable aqueous agent, and causing the hydrogel Sodium chloride, including combinations thereof. The core layer to expand and push against the drug-containing agent, may include a wide variety of additives and excipients that forcing the agent out of the exit passageway. The agent can enhance the performance of the dosage form or that promote Swell, aiding in forcing the drug out of the passageway. Drug stability, tableting or processing. Such additives and excipi can be delivered from this type of delivery system either ents include tableting aids, Surfactants, water-soluble poly dissolved or dispersed in the agent that is expelled from the mers, pH modifiers, fillers, binders, pigments, disintegrants, exit passageway. antioxidants, lubricants and flavorants. Nonlimiting 0788. The rate of drug delivery is controlled by such fac examples of additives and excipients include but are not lim tors as the permeability and thickness of the coating, the ited to those described elsewhere herein as well as microc osmotic pressure of the drug-containing layer, the degree of rystalline cellulose, metallic salts of acids (e.g. aluminum hydrophilicity of the hydrogel layer, and the surface area of Stearate, calcium Stearate, magnesium Stearate, Sodium Stear the device. Those skilled in the art will appreciate that ate, Zinc Stearate), pH control agents (e.g. buffers, organic increasing the thickness of the coating will reduce the release acids, organic acid salts, organic and inorganic bases), fatty rate, while any of the following will increase the release rate: acids, hydrocarbons and fatty alcohols (e.g. Stearic acid, increasing the permeability of the coating; increasing the palmitic acid, liquid paraffin, Stearyl alcohol, and palmitol), hydrophilicity of the hydrogel layer, increasing the osmotic fatty acid esters (e.g. glyceryl (mono- and di-) Stearates, trig pressure of the drug-containing layer, or increasing the lycerides, glyceryl (palmitics tearic) ester, Sorbitan esters (e.g. device's Surface area. Sorbitan monostearate, saccharose monostearate, saccharose 0789. Other materials useful informing the drug-contain monopalmitate, Sodium Stearyl fumarate), polyoxyethylene ing agent, in addition to the agent described herein itself. Sorbitan esters), Surfactants (e.g. alkyl Sulfates (e.g. sodium include HPMC, PEO and PVP and other pharmaceutically lauryl Sulfate, magnesium lauryl Sulfate), polymers (e.g. acceptable carriers. In addition, osmagents such as Sugars or polyethylene glycols, polyoxyethylene glycols, polyoxyeth salts, including but not limited to Sucrose, lactose, Xylitol, ylene, polyoxypropylene ethers, including copolymers mannitol, or sodium chloride, may be added. Materials which US 2012/0283411 A9 Nov. 8, 2012 48 are useful for forming the hydrogel layer include Sodium Depending upon the particular formulation, some disinte CMC, PEO (e.g. polymers having an average molecular grants work better than others. Several disintegrants tend to weight from about 5,000,000 to about 7,500,000 daltons), form gels as they swell with water, thus hindering drug deliv poly (acrylic acid), sodium (polyacrylate), sodium croscar ery from the device. Non-gelling, non-Swelling disintegrants mellose, sodium starch glycolat, PVP crosslinked PVP, and provide a more rapid dispersion of the drug particles within other high molecular weight hydrophilic materials. the core as water enters the core. In certain embodiments, 0790. In the case of a bilayer geometry, the delivery port(s) non-gelling, non-Swelling disintegrants are resins, for or exit passageway(s) may be located on the side of the tablet example, ion-exchange resins. In one embodiment, the resin containing the drug agent or may be on both sides of the tablet is AmberliteTMIRP88 (available from Rohm and Haas, Phila or even on the edge of the tablet so as to connect both the drug delphia, Pa.). When used, the disintegrant is present in layer and the sweller layer with the exterior of the device. The amounts ranging from about 50-74% of the core agent. exit passageway(s) may be produced by mechanical means or 0793 Water-soluble polymers are added to keep particles by laser drilling, or by creating a difficult-to-coat region on of the agent suspended inside the device before they can be the tablet by use of special tooling during tablet compression delivered through the passageway(s) (e.g., an orifice). High or by other means. Viscosity polymers are useful in preventing settling. How 0791. The osmotic device can also be made with a homo ever, the polymer in combination with the agent is extruded geneous core Surrounded by a semipermeable membrane through the passageway(s) under relatively low pressures. At coating, as in U.S. Pat. No. 3,845,770. The agent described a given extrusion pressure, the extrusion rate typically slows herein can be incorporated into a tablet core and a semiper with increased viscosity. Certain polymers in combination meable membrane coating can be applied via conventional with particles of the agent described hereinform high viscos tablet-coating techniques such as using a pan coater. A drug ity solutions with water but are still capable of being extruded delivery passageway can then be formed in this coating by from the tablets with a relatively low force. In contrast, poly drilling a hole in the coating, either by use of a laser or mers having a low weight-average, molecular weight (

treat constipation, an antidiarrheal agent, an insulin or related 0811 Methods to increase chemical and/or physical sta compound (including those described herein), an anti-hyper bility of the agents the described herein are found in U.S. Pat. tensive agent, an agent useful in the treatment of respiratory No. 6,541,606, U.S. Pat. No. 6,068,850, U.S. Pat. No. 6,124, and other disorders, an anti-obesity agent, an anti-diabetic 261, U.S. Pat. No. 5,904,935, and WO 00/15224, U.S. agents, an agent that activates soluble guanylate cyclase and a 200300.69182 (via the addition of nicotinamide), U.S. pharmaceutically acceptable carrier, vehicle or diluent. The 20030175230A1, U.S. 20030175230A1, U.S. pharmaceutical composition comprising the compound 20030175239A1, U.S. 20020045582, U.S. 20010031726, described herein and the second pharmaceutical composition WO 02/26248, WO 03/014304, WO 98/00152A1, WO contained in the kit may be optionally combined in the same 98/00157A1, WO 90/12029, WO 00/04880, and WO pharmaceutical composition. 91/04743, WO 97/04796 and the references cited therein. 0812 Methods to increase bioavailability of the agents 0808. A kit includes a container or packaging for contain described herein are found in U.S. Pat. No. 6,008,187, U.S. ing the pharmaceutical compositions and may also include Pat. No. 5,424,289, U.S. 20030198619, WO 90/01329, WO divided containers such as a divided bottle or a divided foil 01/49268, WO 00/32172, and WO 02/064166. Glycyrrhiz packet. The container can be, for example a paper or card inate can also be used as an absorption enhancer (see, e.g., boardbox, a glass or plastic bottle orjar, a re-sealable bag (for EP397447). WO 03/004062 discusses Ulex europaeus I example, to hold a “refill of tablets for placement into a (UEAl) and UEAI mimetics which may be used to target the different container), or ablisterpack with individual doses for agents described herein to the GI tract. The bioavailability of pressing out of the pack according to a therapeutic schedule. the agents described herein can also be increased by addition It is feasible that more than one container can be used together of oral bioavailability-enhancing agents such as those in a single package to market a single dosage form. For described in U.S. Pat. No. 6,818,615 including but not limited example, tablets may be contained in a bottle which is in turn to: cyclosporins (including cyclosporins A through Z as contained within a box. defined in Table 1 of U.S. Pat. No. 6,818,615), for example, 0809. An example of a kit is a so-called blister pack. Blis cyclosporin A (cyclosporin), cyclosporin F, cyclosporin D, ter packs are well known in the packaging industry and are dihydro cyclosporin A, dihydro cyclosporin C, acetyl being widely used for the packaging of pharmaceutical unit cyclosporin A, PSC-833. (Me-Ile-4)-cyclosporin (SDZ-NIM dosage forms (tablets, capsules, and the like). Blister packs 811) (both from Sandoz Pharmaceutical Corp.), and related generally consist of a sheet of relatively stiff material covered oligopeptides produced by species in the genus Topycla with a foil of a preferably transparent plastic material. During dium); antifungals including but not limited to ketoconazole; the packaging process, recesses are formed in the plastic foil. cardiovascular drug including but not limited to MS-209 The recesses have the size and shape of individual tablets or (BASF), amiodarone, nifedipine, reserpine, quinidine, nica capsules to be packed or may have the size and shape to rdipine, ethacrynic acid, propafenone, reserpine, amiloride; accommodate multiple tablets and/or capsules to be packed. anti-migraine natural products including but not limited to Next, the tablets or capsules are placed in the recesses accord ergot alkaloids; antibiotics including but not limited to cefop ingly and the sheet of relatively stiff material is sealed against eraZone, tetracycline, chloroquine, fosfomycin; antiparasitics the plastic foil at the face of the foil which is opposite from the including but not limited to ivermectin; multi-drug resistance direction in which the recesses were formed. As a result, the reversers including but not limited to VX-710 and VX-853 tablets or capsules are individually sealed or collectively (Vertex Pharmaceutical Incorporated); tyrosine kinase inhibi sealed, as desired, in the recesses between the plastic foil and tors including but not limited to genistein and related isofla the sheet. Preferably the strength of the sheet is such that the vonoids, quercetin; protein kinase C inhibitors including but tablets or capsules can be removed from the blister pack by not limited to calphostin, apoptosis inducers including but not manually applying pressure on the recesses whereby an open limited to ceramides; and agents active against endorphin ing is formed in the sheet at the place of the recess. The tablet receptors including but not limited to morphine, morphine or capsule can then be removed via said opening. congeners, other opioids and opioid. antagonists including 0810. It maybe desirable to provide a written memory aid (but not limited to) naloxone, naltrexone and nalmefene). containing information and/or instructions for the physician, 0813 The agents described herein can be fused to a modi pharmacist or subject regarding when the medication is to be fied version of the blood serum protein transferrin. U.S. taken. A "daily dose” can be a single tablet or capsule or 20030221201, U.S. 20040023334, U.S. 20030226155, WO several tablets or capsules to be taken on a given day. When 04/020454, and WO 04/019872 discuss the manufacture and the kit contains separate compositions, a daily dose of one or use of transferrin fusion proteins. Transferrin fusion proteins more compositions of the kit can consist of one tablet or may improve circulatory halflife and efficacy, decrease unde capsule while a daily dose of another one or more composi sirable side effects and allow reduced dosage. tions of the kit can consist of several tablets or capsules. A kit 0814. The polypeptides and agonists described herein can can take the form of a dispenser designed to dispense the daily be recombinantly expressed in bacteria. Bacteria expressing doses one at a time in the order of their intended use. The the polypeptide or agonists can be administered orally, rec dispenser can be equipped with a memory-aid, so as to further tally, mucosally or in via Some other mode of administration facilitate compliance with the regimen. An example of such a including but not limited to those described herein. Bacterial memory-aid is a mechanical counter which indicates the hosts suitable for such administration include but are not number of daily doses that have been dispensed. Another limited to certain Lactobacteria (e.g. Lactococcus lactis, Lac example of Such a memory-aid is a battery-powered micro tobacillus plantarum, Lact. rhamnosus and Lact. paracasei chip memory coupled with a liquid crystal readout, or audible ssp. Paracasie and other species found in normal human flora reminder signal which, for example, reads out the date that the (Ahrne et al. Journal of Applied Microbiology 199885:88)), last daily dose has been taken and/or reminds one when the certain Streptococcus sp. (e.g. S. gordonii), and certain B. next dose is to be taken. subtilis strains (including pSM539 described in Porzio et al. US 2012/0283411 A9 Nov. 8, 2012 52

BMC Biotechnology 2004 4:27). The polypeptides and ago 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 nists described herein can be administered using the Helio ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, bacter based preparation methods described in WO06/ 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 O15445. ug, 4750 ug. 5000 ug of a polypeptide or GC-C agonist described herein. In certain embodiments the dosage unit and Dosage daily dose are equivalent. In various embodiments, the dos 0815. The dose range for adult humans is generally from age unit is administered with food at anytime of the day, 0.005 mg to 10 g/day orally. Tablets or otherforms of presen without food at anytime of the day, with food after an over tation provided in discrete units may conveniently contain an night fast (e.g. with breakfast), at bedtime after a low fat amount of compound described herein which is effective at Snack. In various embodiments, the dosage unit is adminis Such dosage or as a multiple of the same, for instance, units tered once a day, twice a day, three times a day, four times a containing 5 mg to 500 mg, usually around 10 mg to 200 mg. day, five times a day, six times a day. The dosage unit can The precise amount of compound administered to a patient optionally comprise other agents. will be the responsibility of the attendant physician. However, 0817. A dosage unit (e.g. an oral dosage unit) can include, the dose employed will depend on a number of factors, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 including the age and sex of the patient, the precise disorder ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 being treated, and its severity. ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 0816. A dosage unit (e.g. an oral dosage unit) can include 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 from, for example, 1 to 30 ug. 1 to 40 ug, 1 to 50 ug, 1 to 100 to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 g, 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, ug, 4750 ug, 5000 ug of a polypeptide or agonist described US 2012/0283411 A9 Nov. 8, 2012

herein and from 50 mg to 650 mg (e.g. 50 mg, 100 mg, 150 ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, mg, 200 mg, 250 mg. 300 mg. 350 mg. 400 mg. 450 mg, 500 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, mg, 550 mg, 600 mg) of Modulon R (trimebutine maleate). 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 0818. A dosage unit (e.g. an oral dosage unit) can include, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 500 to 2250 ug, 500 to 2500 g, 500 to 2750 ug,500 to 3000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, herein and from 10 mg to 600 mg (e.g. 10 mg, 20 mg, 30 mg. 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg. ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 200 mg, 250 mg. 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 300 mg, 350 mg. 400 mg. 450 mg, 500 mg, 550 mg. 600 mg) ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, of Bentylr/Bentylol R (diciclomine). 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 0820. A dosage unit (e.g. an oral dosage unit) can include, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to herein and from 1 mg to 80 mg (e.g. 1 mg, 5 mg, 10 mg, 15 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 mg, 60 mg. 65 mg, 70 mg, 75 mg, 80 mg) of Propulsider) ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, (cisapride). 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 0819. A dosage unit (e.g. an oral dosage unit) can include, 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 US 2012/0283411 A9 Nov. 8, 2012 54 to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug. 700 ug, 750 ug. 800 ug. 850 ug, 900 g, 950 ug, 1000 ug, hereinand from 100 mg to 3000mg (e.g. 100 mg, 200mg,300 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 mg, 400 mg, 500 mg, 600 mg. 625 mg, 700 mg. 800 mg,900 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, mg, 1000 mg, 1250 mg, 1300 mg, 1400 mg, 1500 mg, 1600 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 mg, 1700 mg, 1800 mg, 1875 mg, 1900 mg, 2000 mg, 2100 ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, mg, 2200 mg, 2300 mg, 2400 mg, 2500 mg.) of Equalactin R/ 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 Fibercon R (Calcium Polycarbophil). ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, 0822. A dosage unit (e.g. an oral dosage unit) can include, 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 herein and from 1 mg to 25 mg (e.g. 1 mg, 2 mg, 3 mg, 4 mg. ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 5 mg, 6 mg, 7 mg. 8 mg.9 mg, 10 mg, 11 mg, 12 mg, 13 mg. 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 mg, 23 mg, 24 mg, 25 mg) of QuestranR (cholestyramine). ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 0821. A dosage unit (e.g. an oral dosage unit) can include, 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 US 2012/0283411 A9 Nov. 8, 2012

to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, herein and from 1 mg to 250 mg (e.g. 1 mg, 2 mg, 3 mg, 4 mg. 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 20 mg. 30 mg, 40 mg. ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 50 mg. 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg. 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg. ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg) of 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 Ondansetron HCl (ZofranR). ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, 0824. A dosage unit (e.g. an oral dosage unit) can include, 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 herein and from 1 mg to 20 mg (e.g. 1 mg, 2 mg, 2.5 mg.3 mg. ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 4 mg, 5 mg, 6 mg, 7 mg, 7.5 mg. 8 mg, 9 mg, 10 mg, 11 mg. 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 12 mg, 12.5 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17.5 mg, 18 mg, to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 19 mg, 20 mg) of darifenacin (EnableX(R). ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 0823. A dosage unit (e.g. an oral dosage unit) can include, 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 ug, 1 to 700 ug, 1 to 800 ug, 1 to 900 ug, 1 to 1000 ug, 10 to ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 US 2012/0283411 A9 Nov. 8, 2012 56 ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, mg, 100 mg, 150 mg, 200 mg, 250 mg. 300 mg, 350 mg. 400 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 mg, 450 mg, 500 mg, 550 mg. 600 mg, 650 mg, 700 mg, 750 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, mg, 800 mg. 850 mg, 900 mg, 950 mg, 1000 mg) of Dolas 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 etron (Anzemet(R). ug, 4750 ug, 5000 ug of a polypeptide or agonist described 0826. A dosage unit (e.g. an oral dosage unit) can include, herein and from 1 mg to 3000 mg (e.g. 1 mg, 2 mg, 3 mg. 4 for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 20 mg. 30 mg, 40 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 mg, 50 mg. 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 200 mg, 250 ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to mg, 300 mg, 350 mg, 400 mg. 450 mg, 500 mg, 750 mg, 1000 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 mg, 1250 mg, 1500 mg, 1750 mg, 2000 mg, 2250 mg, 2500 to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 mg, 2750 mg, 3000 mg) of Cimetropium (Alginor R). ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 0825. A dosage unit (e.g. an oral dosage unit) can include, 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, herein and from 1 mg to 180 mg (e.g. 1 mg, 2 mg, 3 mg, 4 mg. 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 20 mg. 30 mg, 40 mg. ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, 50 mg. 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg. 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg) of ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, Zelnorm R (tegaserod). 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 0827. A dosage unit (e.g. an oral dosage unit) can include, ug, 4750 ug, 5000 ug of a polypeptide or agonist described for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 herein and from 1 mg to 1000 mg (e.g. 1 mg, 5 mg, 10 mg, 15 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 75 ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to US 2012/0283411 A9 Nov. 8, 2012 57

30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 300 g, 350 ug, 400 ug, 450 Lig, 500 g, 550 ug. 600 ug, 650 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, herein and from 50 mg to 500 mg (e.g. 50 mg, 60 mg, 70 mg. 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 80 mg. 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, mg, 250 mg, 275 mg, 300 mg. 325 mg, 350 mg, 375 mg. 400 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 mg, 425 mg. 450 mg, 500 mg) of Dicetel(R) (pinaverium ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, bromide). 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 0829. A dosage unit (e.g. an oral dosage unit) can include, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to herein and from 1 Jug to 500 Lug (e.g. lug, 5ug. 101g, 50 ug, 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 75ug. 100 ug, 125ug. 150 ug, 175ug. 200g, 225ug. 250 ug, to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 275ug. 300 lug. 325 ug, 350 lug. 375ug, 400 ug, 425 ug, 450 ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, ug, 475 ug, 500 ug) of Levsin(R) (hyoscyamine Sulfate). 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 0828. A dosage unit (e.g. an oral dosage unit) can include, 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to US 2012/0283411 A9 Nov. 8, 2012

1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, ug, 4750 ug, 5000 ug of a polypeptide or agonist described 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 herein and from 1 mg to 120 mg (e.g. 1 mg, 2.5 mg, 5 mg, 7.5 ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg. 30 mg, 40 mg, 50 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg) ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, of Propanthiline bromide (Pro-Banthine(R). 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 0831. A dosage unit (e.g. an oral dosage unit) can include, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 3000 ug,3250 ug,3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 ug, 1 to 600 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to herein and from 50 mg to 500 mg (e.g. 50 mg, 75 mg, 100 mg. 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 125 mg, 135 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg. to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 275 mg, 300 mg. 325 mg, 350 mg. 375 mg, 400 mg. 425 mg. ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 450 mg, 475 mg, 500 mg) of mebeverine (DUSPATAL(R), 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, DUSPATALINR, COLOFAC MRR, COLOTAL(R). 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 0830. A dosage unit (e.g. an oral dosage unit) can include, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 US 2012/0283411 A9 Nov. 8, 2012 59 to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, ug, 4750 ug, 5000 ug of a polypeptide or agonist described 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 herein and from 50g to 3000 Lug (e.g. 50 Lig, 100 ug. 200 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 300 ug, 400 ug, 500 g. 600 ug. 700 ug, 800 ug,900 ug. 1000 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug. 1250 lug. 1500 lug. 1750 lug. 2000 ug. 2250 ug. 2500 ug, ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, 2750 lug. 3000 ug) of Lotronex R (alosetron hydrochloride). 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 0833. A dosage unit (e.g. an oral dosage unit) can include, ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to herein and from 100 ug to 5000 ug (e.g. 100 lug. 200 ug. 300 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 ug, 10 to 200 ug. 10 ug, 400 ug, 500 ug. 600 lug. 700 lug. 800 ug,900 g, 1000 ug, to 300 ug. 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 1250 lug. 1500 ug. 1750 ug, 2000 ug. 2250 ug. 2500 ug. 2750 ug. 10 to 800 lug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, ug. 3000 ug. 3500 lug, 4000 ug, 4500 g, 5000 ug) of Gran 100 to 300 ug, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, isetron (KytrilR). 100 to 700 ug, 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 0832 A dosage unit (e.g. an oral dosage unit) can include, 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug. 100 to 2000 for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 ug. 100 to 2250 ug, 100 to 2500 g, 100 to 2750 ug, 100 to ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 600 g, 200 to 700 ug, 200 to 800 ug, 200 to 900 ug, 200 to 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 to 2000 ug. 200 to 2250 ug. 200 to 2500 ug. 200 to 2750 ug, ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 200 to 3000 ug, 300 to 400 ug,300 to 500 ug,300 to 600 ug, 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug. 300 to 2000 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 ug. 300 to 2250 ug, 300 to 2500 g, 300 to 2750 ug, 300 to ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug, 500 to 3000 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug. 600 ug, 650 ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 1050 lug, 1100 g, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 1700 ug, 1750 ug, 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 ug. 2050 ug. 2100 g, 2150 ug. 2200 ug. 2250 ug. 2300 ug, to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 2350 ug. 2400 ug. 2450 lug. 2500 ug. 2550 ug. 2600 ug. 2650 ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, 3000 ug,3250 ug, 3500 ug, 3750 ug, 4000 ug, 4250 ug, 4500 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 ug, 4750 ug, 5000 ug of a polypeptide or agonist described ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, herein and from 10 mg to 600 mg (e.g. 10 mg, 20 mg, 30 mg. 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 125 mg. ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, 150 mg, 175 mg, 200 mg, 250 mg. 300 mg, 350 mg, 400 mg. 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 450 mg, 500 mg, 550 mg. 600 mg) of Xifaxan R (rifaximin). US 2012/0283411 A9 Nov. 8, 2012 60

0834. A dosage unit (e.g. an oral dosage unit) can include, 10 to 1000 ug, 100 to 200 ug, 100 to 300 ug, 100 to 400 ug, for example, from 1 to 30 ug, 1 to 40 ug, 1 to 50 g, 1 to 100 100 to 500 ug, 100 to 600 ug, 100 to 700 ug, 100 to 800 ug, ug, 1 to 200 ug, 1 to 300 ug, 1 to 400 ug, 1 to 500 lug. 1 to 600 100 to 900 ug, 100 to 1000 ug. 100 to 1250 ug, 100 to 1500 ug, ug, 1 to 700 ug, 1 to 800 lug. 1 to 900 ug, 1 to 1000 ug. 10 to 100 to 1750 ug, 100 to 2000 ug, 100 to 2250 ug. 100 to 2500 30 ug. 10 to 40 ug. 10 to 50 ug. 10 to 100 lug. 10 to 200 ug. 10 ug. 100 to 2750 ug, 100 to 3000 ug, 200 to 300 ug,200 to 400 to 300 ug, 10 to 400 ug, 10 to 500 ug, 10 to 600 ug, 10 to 700 ug. 200 to 500g, 200 to 600g, 200 to 700 ug. 200 to 800 ug, ug. 10 to 800 ug. 10 to 900 ug. 10 to 1000 ug. 100 to 200 ug, 200 to 900ug,200 to 1000 ug. 200 to 1250 ug,200 to 1500 g, 100 to 300 g, 100 to 400 ug, 100 to 500 ug, 100 to 600 ug, 200 to 1750 ug, 200 to 2000 ug, 200 to 2250 ug. 200 to 2500 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, ug. 200 to 2750 ug, 200 to 3000 ug,300 to 400 ug,300 to 500 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 ug,300 to 600 ug,300 to 700 ug,300 to 800 ug,300 to 900 ug, ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to 300 to 1000 ug,300 to 1250 ug, 300 to 1500 ug. 300 to 1750 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ug. 300 to 2000 ug, 300 to 2250 ug, 300 to 2500 ug, 300 to 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to 2750 ug,300 to 3000 ug, 400 to 500 g, 400 to 600g, 400 to 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 700 g, 400 to 800 ug, 400 to 900 ug, 400 to 1000 ug, 400 to to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, 1250 ug, 400 to 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, to 2250 ug, 400 to 2500 ug, 400 to 2750 ug, 400 to 3000 ug, 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, 500 to 600 ug, 500 to 700 ug, 500 to 800 ug, 500 to 900 ug, 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 500 to 1000 ug, 500 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to ug, 500 to 2000 ug, 500 to 2250 ug, 500 to 2500 ug, 500 to 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to 2750 ug, 500 to 3000 ug, 600 to 700 ug, 600 to 800g, 600 to 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to 900 ug, 600 to 1000 ug, 600 to 1250 ug, 600 to 1500 g, 600 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 to 1750 ug, 600 to 2000 ug. 600 to 2250 ug, 600 to 2500 ug, to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 600 to 2750 ug, 600 to 3000 ug,700 to 800 ug,700 to 900 ug, to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 700 to 1000 ug, 700 to 1250 pg. 700 to 1500 ug, 700 to 1750 to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, ug. 700 to 2000 ug, 700 to 2250 ug, 700 to 2500 ug, 700 to 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 2750 ug, 700 to 3000 ug. 800 to 900 ug. 800 to 1000 ug, 800 ug, 600 to 700 ug. 600 to 800 ug. 600 to 900 ug. 600 to 1000 to 1250 ug, 800 to 1500 ug. 800 to 1750 ug, 800 to 2000 ug, ug, 600 to 1250 ug, 600 to 1500 ug, 600 to 1750 ug, 600 to 800 to 2250 ug,800 to 2500 g, 800 to 2750 ug, 800 to 3000 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 ug,900 to 1000 ug,900 to 1250 ug,900 to 1500 ug,900 to to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 1750 ug,900 to 2000 ug,900 to 2250 ug,900 to 2500 ug,900 to 1250 ug, 700 to 1500 ug, 700 to 1750 ug, 700 to 2000 ug, to 2750 ug,900 to 3000 ug. 1000 to 1250 ug. 1000 to 1500 ug, 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 1000 to 1750 ug, 1000 to 2000 ug, 1000 to 2250 ug, 1000 to ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 2500 ug. 1000 to 2750 ug, 1000 to 3000 ug, 2 to 500 ug, 50 to ug, 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to 500g, 3 to 100 ug. 5 to 20 ug. 5 to 100 ug. 50 ug, 100 ug. 150 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 ug. 200 ug, 250 ug. 300 g, 350 ug, 400 ug, 450 ug, 500 ug, to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, 550 g. 600 ug, 650 ug. 700 ug, 750 lug. 800 ug. 850 ug,900 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 ug, 950 ug, 1000 ug. 1050 ug, 1100 lug. 1150 lug. 1200 ug, ug, 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 1250 lug. 1300 g, 1350 lug. 1400 ug. 1450 ug. 1500 ug. 1550 to 2000 ug, 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 ug. 1600 ug. 1650 lug. 1700 lug. 1750 ug, 1800 ug, 1850 ug, ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 1900 g, 1950 ug. 2000 ug. 2050 ug. 2100 ug. 2150 lug. 2200 to 20 Jug, 5 to 100 ug, 50 Lig, 100 ug. 150 ug. 200 ug. 250 ug, ug. 2250 ug. 2300 ug. 2350 Lig, 2400 ug. 2450 ug. 2500 ug, 300 ug, 350 ug, 400 ug, 450 ug, 500 ug. 550 ug. 600 ug, 650 2550 ug. 2600 ug. 2650 lug. 2700 lug. 2750 ug. 2800 ug. 2850 ug. 700 ug, 750 ug. 800 lug. 850 ug,900 ug,950 ug, 1000 ug, ug. 2900 ug. 2950 ug. 3000 ug. 3250 ug, 3500 ug, 3750 ug, 1050 lug, 1100 ug, 1150 lug. 1200 ug. 1250 ug. 1300 ug. 1350 4000 ug, 4250 ug, 4500 ug, 4750 ug. 5000 ug of a polypeptide ug. 1400 ug. 1450 ug. 1500 Lig, 1550 ug. 1600 ug. 1650 jug, oragonist described herein and from 0.2 mg to 10 mg (e.g. 0.2 1700 ug, 1750 ug. 1800 ug, 1850 ug, 1900 ug, 1950 ug, 2000 mg, 0.4 mg. 0.5 mg. 0.75 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg. 3 ug. 2050 ug. 2100 ug. 2150 ug. 2200 ug. 2250 ug. 2300 ug, mg, 3.5 mg, 4 mg. 4.5 mg, 5 mg, 5.5 mg, 6 mg. 6.5 mg, 7 mg. 2350 ug. 2400 ug. 2450 ug. 2500 ug. 2550 ug. 2600 ug. 2650 7.5 mg, 8 mg, 8.5 mg, 9 mg, 9.5 mg, 10 mg) of bumetanide ug. 2700 lug. 2750 lug. 2800 ug. 2850 ug. 2900 ug. 2950 ug, (Bumex (R). 3000 ug,3250 ug, 3500 ug,3750 ug, 4000 ug, 4250 ug, 4500 0836. The precise amount of each of the two or more active ug, 4750 ug, 5000 ug of a polypeptide or agonist described ingredients in a dosage unit will depend on the desired dosage herein and from 10 mg to 600 mg (e.g. 10 mg, 20 mg, 30 mg. of each component. Thus, it can be useful to create a dosage 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 120 mg. unit that will, when administered according to a particular 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg. dosage schedule (e.g., a dosage schedule specifying a certain 280 mg. 300 mg. 320 mg, 340 mg. 360 mg, 380 mg, 400 mg. number of units and a particular timing for administration), 420 mg. 440 mg., 460 mg, 480 mg, 500 mg, 520 mg, 540 mg. deliver the same dosage of each component as would be 560 mg, 580 mg. 600 mg) of furosemide (Lasix). administered if the patient was being treated with only a 0835. A dosage unit (e.g. an oral, intravenous or intramus single component. In other circumstances, it might be desir cular dosage unit) can include, for example, from 1 to 30 Jug, able to create a dosage unit that will deliver a dosage of one or 1 to 40 ug, 1 to 50 ug, 1 to 100 g, 1 to 200 ug, 1 to 300 lug. 1 more components that is less than that which would be admin to 400 ug, 1 to 500 ug, 1 to 600 ug, 1 to 700 ug, 1 to 800 ug, istered if the patient was being treated only with a single 1 to 900 ug. 1 to 1000 lug. 10 to 30 ug. 10 to 40 ug, 10 to 50 ug, component. Finally, it might be desirable to create a dosage 10 to 100 ug. 10 to 200 ug. 10 to 300 lug. 10 to 400 lug. 10 to unit that will deliver a dosage of one or more components that 500 ug. 10 to 600 ug. 10 to 700 ug. 10 to 800 ug. 10 to 900 ug, is greater than that which would be administered if the patient US 2012/0283411 A9 Nov. 8, 2012

was being treated only with a single component. The phar 0841. The therapeutic combinations described herein can maceutical composition can include additional ingredients beformulated as a capsule comprising one or more microtab including but not limited to the excipients described herein. In lets and powder, or one or more microtablets and granules or certain embodiments, one or more therapeutic agents of the beads. In order to avoid interactions between drugs, some dosage unit may exist in an extended or control release for active ingredients of a said combination can be formulated as mulation and additional therapeutic agents may not exist in microtablets and the others filled into capsules as a powder, extended release formulation. For example, a polypeptide or granules, or beads. The microtablets may be film coated or agonist described herein may exist in a controlled release enteric coated. At least one active ingredient can be presented formulation or extended release formulation in the same dos in controlled release form. age unit with another agent that may or may not be in either a 0842. The therapeutic combinations described herein can controlled release or extended release formulation. Thus, in be formulated wherein the active ingredients are distributed certain embodiments, it may be desirable to provide for the in the inner and outer phase of tablets. In an attempt to divide immediate release of one or more of the agents described chemically incompatible components of proposed combina herein, and the controlled release of one or more other agents. tion, few interacting components are converted in granules or 0837. In certain embodiments the dosage unit and daily beads using well known pharmaceutical procedures in prior dose are equivalent. In certain embodiments the dosage unit art. The prepared granules or beads (inner phase) are then and the daily dose are not equivalent. In various embodi mixed with outer phase comprising the remaining active ments, the dosage unit is administered twenty minutes prior to ingredients and at least one pharmaceutically acceptable food consumption, twenty minutes after food consumption, excipient. The mixture thus comprising inner and outer phase with food at anytime of the day, without food at anytime of the is compressed into tablets or molded into tablets. The gran day, with food after an overnight fast (e.g. with breakfast), at ules or beads can be controlled release or immediate release bedtime after a low fat snack. In various embodiments, the beads or granules, and can further be coated using an enteric dosage unit is administered once a day, twice a day, three polymer in an aqueous or non-aqueous system, using meth times a day, four times a day, five times a day, six times a day. ods and materials that are known in the art. 0838. When two or more active ingredients are combined 0843. The therapeutic combinations described herein can in single dosage form, chemical interactions between the be formulated as single dosage unit comprising Suitable buff active ingredients may occur. For example, acidic and basic ering agent. All powdered ingredients of said combination are active ingredients can react with each other and acidic active mixed and a suitable quantity of one or more buffering agents ingredients can facilitate the degradation of acid labile sub is added to the blend to minimize possible interactions. stances. Thus, in certain dosage forms, acidic and basic Sub 0844. The agents described herein, alone or in combina stances can be physically separated as two distinct or isolated tion, can be combined with any pharmaceutically acceptable layers in a compressed tablet, or in the core and shell of a carrier or medium. Thus, they can be combined with materials press-coated tablet. Additional agents that are compatible that do not produce an adverse, allergic or otherwise with acidic as well as basic substances, have the flexibility of unwanted reaction when administered to a patient. The car being placed in either layer. In certain multiple layer compo riers or mediums used can include solvents, dispersants, coat sitions at least one active ingredient can be enteric-coated. In ings, absorption promoting agents, controlled release agents, certain embodiments thereofat least one active ingredient can and one or more inert excipients (which include starches, be presented in a controlled release form. In certain embodi polyols, granulating agents, microcrystalline cellulose, dilu ments where a combination of three or more active substances ents, lubricants, binders, disintegrating agents, and the like), are used, they can be presented as physically isolated seg etc. If desired, tablet dosages of the disclosed compositions ments of a compressed mutlilayer tablet, which can be option may be coated by standard aqueous or nonaqueous tech ally film coated. niques. 0839. The therapeutic combinations described herein can be formulated as a tablet or capsule comprising a plurality of Analgesic Agents in Combitherapy beads, granules, or pellets. All active ingredients including 0845 The polypeptides and agonists described herein can the vitamins of the combination are formulated into granules be used in combination therapy with an analgesic agent, e.g., or beads or pellets that are further coated with a protective an analgesic compound or an analgesic polypeptide. These coat, an enteric coat, or a film coat to avoid the possible polypeptides and compounds can be administered with the chemical interactions. Granulation and coating of granules or polypeptides described herein (simultaneously or sequen beads is done using techniques well known to a person skilled tially). They can also be optionally covalently linked or in the art. At least one active ingredient can present in a attached to an agent described herein to create therapeutic controlled release form. Finally these coated granules or conjugates. Among the useful analgesic agents are: Cachan beads are filled into hard gelatin capsules or compressed to nel blockers, 5HT receptor antagonists (for example 5HT3, form tablets. 5HT4 and 5HT1 receptor antagonists), opioid receptor ago 0840. The therapeutic combinations described herein can nists (loperamide, fedotozine, and fentanyl), NK1 receptor be formulated as a capsule comprising microtablets or minit antagonists, CCK receptor agonists (e.g., loxiglumide), NK1 ablets of all active ingredients. Microtablets of the individual receptor antagonists, NK3 receptor antagonists, norepineph agents can be prepared using well known pharmaceutical rine-serotonin reuptake inhibitors (NSRI), Vanilloid and can procedures of tablet making like direct compression, dry nabanoid receptoragonists, and sialorphin. Analgesics agents granulation or wet granulation. Individual microtablets can in the various classes are described in the literature. be filled into hard gelatin capsules. A final dosage form may 0846 Among the useful analgesic polypeptides are sialor comprise one or more microtablets of each individual com phin-related polypeptides, including those comprising the ponent. The microtablets may be film coated or enteric amino acid sequence QHNPR (SEQ ID NO:), including: coated. VQHNPR (SEQ ID NO:); VRQHNPR (SEQ ID NO:); US 2012/0283411 A9 Nov. 8, 2012 62

VRGQHNPR (SEQID NO:); VRGPQHNPR (SEQID NO:); 0854. Other useful analgesic agents include 5-HT4 ago VRGPRQHNPR (SEQID NO:); VRGPRRQHNPR (SEQID nists such as tegaserod (Zelnorm R), mosapride, metoclopra NO:); and RQHNPR (SEQ ID NO:). Sialorphin-related mide, Zacopride, cisapride, renzapride, benzimidazolone polypeptides bind to neprilysin and inhibit neprily sin-medi derivatives such as BIMU 1 and BIMU 8, and lirexapride. ated breakdown of substance P and Met-enkephalin. Thus, Such agonists are described in: EP1321142 A1, WO compounds or polypeptides that are inhibitors of neprilysin 03/053432A1, EP505322 A1, EP505322 B1, U.S. Pat. No. are useful analgesic agents which can be administered with 5,510,353, EP 507672A1 EP 507672 B1, and U.S. Pat. No. the polypeptides described herein in a co-therapy or linked to the polypeptides described herein, e.g., by a covalent bond. 5,273,983. Sialophin and related polypeptides are described in U.S. Pat. 0855 Calcium channel blockers such as Ziconotide and No. 6,589,750; U.S. 20030078200 A1; and WO 02/051435 related compounds described in, for example, EP625162B1, A2. U.S. Pat. No. 5,364,842, U.S. Pat. No. 5,587,454, U.S. Pat. 0847 Opioid receptor antagonists and agonists can be No. 5,824,645, U.S. Pat. No. 5,859,186, U.S. Pat. No. 5,994, administered with the polypeptides described herein in co 305, U.S. Pat. No. 6,087,091, U.S. Pat. No. 6,136,786, WO therapy or linked to the agent described herein, e.g., by a 93/13128A1, EP1336409 A1 EP 835126A1, EP 835126 B1, covalent bond. For example, opioid receptor antagonists Such U.S. Pat. No. 5,795,864, U.S. Pat. No. 5,891,849, U.S. Pat. as naloxone, naltrexone, methyl naloZone, nalmefene, cypri No. 6,054,429, WO 97/01351 A1, can be used with or linked dime, beta funaltrexamine, naloxonazine, naltrindole, and to the polypeptides described herein. nor-binal torphimine are thought to be useful in the treatment 0856 Various antagonists of the NK-1, NK-2, and NK-3 of IBS. It can be useful to formulate opioidantagonists of this receptors (for a review see Giardina et al. 2003 Drugs 6:758) type is a delayed and Sustained release formulation Such that can be can be used with or linked to the polypeptides initial release of the antagonist is in the mid to distal Small described herein. intestine and/or ascending colon. Such antagonists are described in WO 01/32180 A2. Enkephalin pentapeptide 0857 NK1 receptor antagonists such as: aprepitant (HOE825: Tyr-D-Lys-Gly-Phe-L-homoserine) is an agonist (Merck & Co Inc), Vofopiitant, ezlopitant (Pfizer, Inc.), R-673 of the mu and delta opioid receptors and is thought to be (Hoffmann-La Roche Ltd), SR-48968 (Sanofi Synthelabo), useful for increasing intestinal motility (Eur: J. Pharm. 219: CP-122,721 (Pfizer, Inc.), GW679769 (GlaxoSmith Kline), 445, 1992), and this polypeptide can be used in conjunction TAK-637 (Takeda/Abbot), SR-14033, and related com with the polypeptides described herein. Also useful is trime pounds described in, for example, EP 873753 A1, US butine which is thought to bind to mu/delta/kappa opioid 20010006972 A1, US 2003.0109417 A1, WO 01/52844 A1, receptors and activate release of motilin and modulate the can be used with or linked to the polypeptides described release of gastrin, vasoactive intestinal polypeptide, gastrin herein. and glucagons. Kappa opioid receptoragonists such as fedot 0858 NK-2 receptor antagonists such as nepadutant (Me ozine, asimadoline, and ketocyclazocine, and compounds narini Ricerche SpA), saredutant (Sanofi-Synthelabo), described in WOO3/097051 and WOO5/007626 can be used GW5975.99 (Glaxo Smith Kline), SR-144190 (Sanofi-Syn with or linked to the polypeptides described herein. In addi tion, mu opioid receptoragonists Such as morphine, dipheny thelabo) and UK-290795 (Pfizer Inc) can be used with or loxylate, frakefamide (H-Tyr-D-Ala-Phe(F)-Phe-NH. WO linked to the polypeptides described herein. 01/019849 A1) and loperamide can be used. 0859 NK3 receptor antagonists such as osanetant (SR 0848 Tyr-Arg (kyotorphin) is a dipeptide that acts by 142801; Sanofi-Synthelabo), SSR-241586, talnetant and stimulating the release of met-enkephalins to elicit an anal related compounds described in, for example, WO 02/094187 gesic effect (J. Biol. Chem. 262:8165, 1987). Kyotorphin can A2, EP 876347 A1, WO 97/21680 A1, U.S. Pat. No. 6,277, be used with or linked to the polypeptides described herein. 862, WO 98/11090, WO95/28418, WO 97/19927, and Boden 0849 Chromogranin-derived polypeptide (CgA 47-66: et al. (J Med Chem. 39:1664-75, 1996) can be used with or see, e.g., Ghia et al. 2004 Regulatory polypeptides 119:199) linked to the polypeptides described herein. can be used with or linked to the polypeptides described 0860 Norepinephrine-serotonin reuptake inhibitors herein. (NSRI) such as milnacipran and related compounds 0850 CCK receptor agonists such as caerulein from described in WO 03/077897 A1 can be used with or linked to amphibians and other species are useful analgesic agents that the polypeptides described herein. can be used with or linked to the polypeptides described 0861 Vanilloid receptor antagonists such as arvanil and herein. related compounds described in WO 01/64212 A1 can be 0851 Conotoxin polypeptides represent a large class of used with or linked to the polypeptides described herein. analgesic polypeptides that act at Voltage gated Ca channels, 0862 The analgesic polypeptides and compounds can be NMDA receptors or nicotinic receptors. These polypeptides administered with the polypeptides and agonists described can be used with or linked to the polypeptides described herein (simultaneously or sequentially). The analgesic agents herein. can also be covalently linked to the polypeptides and agonists 0852. Peptide analogs of thymulin (FR Application described herein to create therapeutic conjugates. Where the 2830451) can have analgesic activity and can be used with or analgesic is a polypeptide and is covalently linked to an agent linked to the polypeptides described herein. described herein the resulting polypeptide may also include at 0853 CCK (CCKa or CCKb) receptor antagonists, least one trypsin cleavage site. When present within the including loxiglumide and dexloxiglumide (the R-isomer of polypeptide, the analgesic polypeptide may be preceded by loxiglumide) (WO 88/05774) can have analgesic activity and (if it is at the carboxy terminus) or followed by (if it is at the can be used with or linked to the polypeptides described amino terminus) a trypsin cleavage site that allows release of herein. the analgesic polypeptide. US 2012/0283411 A9 Nov. 8, 2012

0863. In addition to sialorphin-related polypeptides, anal fiber (e.g. FIBERCONR) (Calcium Polycarbophil), an gesic polypeptides include: Aspphe, endomorphin-1, endo osmotic laxative, a stimulant laxative (such as diphenyl morphin-2, nocistatin, dalargin, lupron, Ziconotide, and Sub methanes (e.g. bisacodyl), anthraquinones (e.g. cascara, stance P. Senna), and Surfactant laxatives (e.g. castor oil, docusates), an emollient/lubricating agent (Such as mineral oil, glycerine, Diabetes, Obesity and Other Disorders and docusates), Mirallax (Braintree Laboratories, Braintree 0864 Pharmaceutical compositions comprising at least Mass.), dexloxiglumide (Forest Laboratories, also known as two of: 1) an agent that stimulates the production of cAMP CR 2017 Rottapharm (Rotta Research Laboratorium SpA)), (e.g., glucagon-like polypeptide 1 (GLP-1)); 2) an agent that saline laxatives, enemas, suppositories, and CR 3700 (Rot inhibits the degradation of a cyclic nucleotide (e.g., a phos tapharm (Rotta Research Laboratorium SpA); phodiesterase inhibitor); and 3) a polypeptide or agonist acid reducing agents such as proton pump inhibitors (e.g., described herein useful for treating diabetes and obesity. Such omeprazole (PrilosecR), esomeprazole (Nexium(R), lanso compositions may also be useful for treating secondary prazole (PrevacidR), pantoprazole (Protonix(R) and rabepra hyperglycemias in connection with pancreatic diseases Zole (AcipheX(R)) and Histamine H2-receptor antagonist (chronic pancreatitis, pancreasectomy, hemochromatosis) or (also known as H2 receptor blockers including cimetidine, endocrine diseases (acromegaly, Cushing's syndrome, pheo ranitidine, famotidine and nizatidine); chromocytoma or hyperthyreosis), drug-induced hyperglyce prokinetic agents including itopride, octreotide, bethanechol, mias (benzothiadiazine Saluretics, diaZoxide or glucocorti metoclopramide (ReglanR), domperidone (Motilium(R), coids), pathologic glucose tolerance, hyperglycemias, erythromycin (and derivatives thereof) or cisapride (Propul dyslipoproteinemias, adiposity, hyperlipoproteinemias and/ sider): or hypotensions. The phosphodiesterase inhibitor can be spe Prokineticin polypeptides homologs, variants and chimeras cific for a particular phosphodiesterase (e.g., Group III or thereof including those described in U.S. Pat. No. 7,052,674 Group IV) or a non-specific phosphodiesterase inhibitor, such which can be used with or linked to the polypeptides as papaverine, theophylline, enprofyllines and/or IBMX. described herein; Specific phosphodiesterase inhibitors which inhibit group III pro-motility agents such as the vasostatin-derived polypep phosphodiesterases (cGMP-inhibited phosphodiesterases), tide, chromogranin A (4-16) (see, e.g., Ghia et al. 2004 Regu including indolidane (LY195115), cilostamide (OPC 3689), latory polypeptides 121:31) or motilin agonists (e.g., lixazinone (RS 82856), Y-590, imazodane (CI914), SKF GM-611 or mitemcinal fumarate) or nociceptin/Orphanin FQ 94120, quazinone, ICI 153,110, cilostazole, bemorandane receptor modulators (US20050169917); (RWJ 22867), siguaZodane (SK&F 94-836), adibendane other peptides which can bind to and/or activate GC-C includ (BM 14.478), milrinone (WIN 47203), enoximone (MDL ing those described in US20050287067; 17043), pimobendane (UD-CG 115), MCI-154, saterinone complete or partial 5HT (e.g. 5HT1, 5HT2, 5HT3, 5HT4) (BDF 8634), sulmazole (ARL 115), UD-CG 212, motapi receptor agonists or antagonists (including 5HT1A antago Zone, piroximone, and ICI 118233 can be useful. In addition, nists (e.g. AGI-001 (AGI therapeutics), 5HT2B antagonists phosphodiesterase inhibitors which inhibit group IV phos (e.g. PGN1091 and PGN1164 (Pharmagene Laboratories phodiesterases (cAMP-specific phosphodiesterases). Such as Limited), and 5HT4 receptor agonists (such as tegaserod rolipram ZK 62711; pyrrolidone), imidazolidinone (RO (ZELNORMR), prucalopride, mosapride, metoclopramide, 20-1724), etazolate (SQ 65442), denbufylline (BRL 30892), Zacopride, cisapride, renzapride, benzimidazolone deriva ICI63197, and RP73401 can be used. tives such as BIMU 1 and BIMU 8, and lirexapride). Such agonists/modulatos are described in: EP1321142 A1, WO Other Agents for Use in Combitherapy 03/053432A1, EP505322 A1, EP505322 B1, U.S. Pat. No. 5,510,353, EP 507672 A1 EP 507672 B1, U.S. Pat. No. 0865 Also within the invention are pharmaceutical com 5,273,983, and U.S. Pat. No. 6,951.867): 5HT3 receptorago positions comprising a polypeptide or agonists described nists such as MKC-733; and 5HT3 receptor antagonists such hereinanda second therapeutic agent. The second therapeutic as DDP-225 (MCI-225; Dynogen Pharmaceuticals, Inc.), agent can be administered to treat any condition for which it cilansetron (CalmactinR), alosetron (Lotronex(R), is useful, including conditions that are not considered to be Ondansetron HCl (Zofran R), Dolasetron (ANZEMETR), the primary indication for treatment with the second thera palonosetron (Aloxi(R), Granisetron (KytrilR), YMO60 (ra peutic agent. The second therapeutic agent can be adminis mosetron; Astellas Pharma Inc.; ramosetron may be given as tered simultaneously or sequentially. The second therapeutic a daily dose of 0.002 to 0.02 mgas described in EP01588707) agent can be covalently linked to the polypeptides and ago and ATI-7000 (Aryx Therapeutics, Santa Clara Calif.); nists described hereinto create atherapeutic conjugate. When muscarinic receptor agonists; the second therapeutic agent is another polypeptide, a linker anti-inflammatory agents; including those described herein may be used between the antispasmodics including but not limited to anticholinergic polypeptide described herein and the second therapeutic drugs (like dicyclomine (e.g. Colimex R, FormuleX(R), Lom polypeptide. ine(R), Protylol R, Viscerol R, Spasmoban R., Bentyl(R), Benty 0866. Examples of additional therapeutic agents to treat lolR), hyoscyamine (e.g. IB-Stat(R), Nulev(R), Levsin(R), Lev gastrointestinal and other disorders include bid R, Levsinex Timecaps(R), Levsin/SL(R), AnaspaZR), A-Spas agents to treat constipation (e.g., a chloride channel activator S/L(R), CystospaZR), Cystospaz-M(R), Donnamar R, Colidrops such as the bicylic fatty acid, Lubiprostone (formerly known Liquid Pediatric(R), GastrosedR), Hyco Elixir R, Hyosol(R), as SPI-0211; Sucampo Pharmaceuticals, Inc.; Bethesda, HyospaZR), Hyosyne R, Losamine(R), MedispaZ(R), Neosol(R), Md.), a laxative (eg. a bulk-forming laxative (e.g. nonstarch Spacol R, Spasdel R. Symax(R), Symax SL(R), Donnatal (e.g. polysaccharides, Colonel Tablet (polycarbophil calcium), Donnatal Extentabs(R), clidinium (e.g. Quarzan, in combina Plantago Ovata(R), EqualactinR (Calcium Polycarbophil)), tion with Librium=Librax), methantheline (e.g. Banthine), US 2012/0283411 A9 Nov. 8, 2012 64

MepenZolate (e.g. Cantil), homatropine (e.g. hycodan, (E)-4 (1.3bis(cyclohexylmethyl)-1,2,34-tetrahydro-2,6-di Homapin), Propantheline bromide (e.g. Pro-Banthine), Gly ono-9H-purin-8-yl)cinnamic acid nonaethylene glycol copyrrolate (e.g. Robinul R. Robinul ForteR), scopolamine methyl ether ester and related compounds described in WO (e.g. Transderm-ScopR, Transderm-V(R), hyosine-N-butyl 02/067942: bromide (e.g. Buscopan R), Pirenzepine (e.g. Gastrozepin(R) the probiotic PROBACTRIX(R) (The BioBalance Corpora Propantheline Bromide (e.g. Propanthel(R), dicycloverine tion; New York, N.Y.) which contains microorganisms useful (e.g. Merbenty1R), glycopyrronium bromide (e.g. Glycopy in the treatment of gastrointestinal disorders; rrolate(R), hyoscine hydrobromide, hyoscine methobromide, antidiarrheal drugs including but not limited to loperamide methanthelinium, and octatropine); peppermint oil; and (Imodium, Pepto Diarrhea), diphenoxylate with atropine (Lo direct Smooth muscle relaxants like cimetropium bromide, motil, Lomocot), cholestyramine (Questran, Cholybar), atro mebeverine (DUSPATAL(R), DUSPATALINR, COLOFAC pine (Co-Phenotrope, Diarsed, Diphenoxylate, Lofene, MRR), COLOTAL(R), otilonium bromide (octilonium), Logen, Lonox, Vi-Atro, atropine Sulfate injection) and pinaverium (e.g. Dicetel(R) (pinaverium bromide: Solvay Xifaxan R (rifaximin; Salix Pharmaceuticals Ltd), TZP-201 S.A.)), SpasfonR (hydrated phloroglucinol and trimeth (Tranzyme Pharma Inc.), the neuronal acetylcholine receptor (nAChR) blocker AGI-004 (AGI therapeutics), and bismuth ylphloroglucinol) and trimebutine (including trimebutine subsalicylate (Pepto-bismol); maleate (Modulon R.); anxiolytic drugs including but not limited to Ativan antidepressants, including but not limited to those listed (lorazepam), alprazolam (Xanax(R), chlordiazepoxide/clid herein, as well as tricyclic antidepressants like amitriptyline inium (Librium(R), Librax(R), clonazepam (Klonopin(R), clo (ElavilR), desipramine (Norpramin(R), imipramine (Tofra razepate (Tranxene.R.), diazepam (Valium(R), estazolam nil.R.), amoxapine (Asendinr), nortriptyline; the selective (ProSomr), flurazepam (Dalmane(R), oxazepam (Serax(R), serotonin reuptake inhibitors (SSRIs) like paroxetine prazepam (CentraX(R), temazepam (Restoril.R.), triazolam (Paxil.R.), fluoxetine (ProzacR), sertraline (Zoloft(R), and (Halcion(R): citralopram (CelexaR); and others like doxepin (Sinequan R.) Bedelix(R) (Montmorillonite beidellitic; Ipsen Ltd), Solvay and trazodone (Desyrel(R): SLV332 (ArCule Inc), YKP (SK Pharma), Asimadoline centrally-acting analgesic agents such as opioid receptorago (Tioga Pharmaceuticals/Merck), AGI-003 (AGI Therapeu nists, opioid receptor antagonists (e.g., naltrexone); tics); agents for the treatment of Inflammatory bowel disease; neurokinin antagonists including those described in agents for the treatment of Crohn's disease and/or ulcerative US20060040950; colitis (e.g., alequel (Enzo Biochem, Inc.; Farmingsale, potassium channel modulators including those described in N.Y.), the anti-inflammatory polypeptide RDP58 (Genzyme, US7,002,015; Inc.; Cambridge, Mass.), and TRAFICET-ENTM the serotonin modulator AZD7371 (AstraZeneca Plc); (ChemoCentryx, Inc.; San Carlos, Calif.); M3 muscarinic receptor antagonists such as darifenacin (En agents that treat gastrointestinal or visceral pain; ablex; Novartis AG and Zamifenacin (Pfizer); agents that increase coMP levels (as described in herbal and natural therapies including but not limited to aci US20040121994) like adrenergic receptor antagonists, dophilus, chamomile tea, evening primrose oil, fennel seeds, dopamine receptor agonists and PDE (phosphodiesterase) wormwood, comfrey, and compounds of Bao-Ji-Wan (mag nolol, honokiol, imperatorin, and isoimperatorin) as in U.S. inhibitors including but not limited to those disclosed herein; Pat. No. 6,923,992; and purgatives that draw fluids to the intestine (e.g., VISICOLR), compositions comprising lysine and an anti-stress agent for a combination of sodium phosphate monobasic monohydrate the treatment of irritable bowel syndrome as described in and sodium phosphate dibasic anhydrate); EPO1550443. Corticotropin Releasing Factor (CRF) receptor antagonists 0867. The polypeptides and agonists described herein can (including NBI-34041 (Neurocrine Biosciences, San Diego, be used in combination therapy with insulin and related com Calif.), CRH9-41, astressin, R121919 (Janssen Pharmaceu pounds including primate, rodent, or rabbit insulin including tica), CP154,526, NBI-27914, Antalarmin, DMP696 (Bris biologically active variants thereof including allelic variants, tol-Myers Squibb) CP-316,311 (Pfizer, Inc.), SB723620 more preferably human insulin available in recombinant (GSK), GW876008 (Neurocrine/GlaxoSmithKline), ONO form. Sources of human insulin include pharmaceutically 2333Ms (Ono Pharmaceuticals), TS-041 (Janssen), AAG561 acceptable and sterile formulations such as those available (Novartis) and those disclosed in U.S. Pat. No. 5,063,245, from Eli Lilly (Indianapolis, Ind. 46285) as HumulinTM (hu U.S. Pat. No. 5,861,398, US20040224964, US20040198726, man insulin rDNA origin). See the THE PHYSICIANS US20040176400, US20040171607, US20040110815, DESK REFERENCE, 55.sup.th Ed. (2001) Medical Eco US20040006066, and US20050209253); nomics, Thomson Healthcare (disclosing other Suitable glucagon-like polypeptides (glp-1) and analogues thereof (in human insulins). The polypeptides and agonists described cluding exendin-4 and GTP-010 (Gastrotech Pharma A)) and herein can also be used in combination therapy with agents inhibitors of DPP-IV (DPP-IV mediates the inactivation of that can boost insulin effects or levels of a subject upon glp-1); administration, e.g. glipizide and/or rosiglitaZone. The tofisopam, enantiomerically-pure R-tofisopam, and pharma polypeptides and agonists described herein can be used in ceutically-acceptable salts thereof (US 20040229867): combitherapy with SYMLINR) (pramlintide acetate) and tricyclic anti-depressants of the dibenzothiazepine type Exenatide(R) (synthetic exendin-4; a 39 aa polypeptide). including but not limited to DextofisopamR (Vela Pharma 0868. The polypeptides and agonists described herein can ceuticals), tianeptine (Stablon R) and other agents described also be used in combination therapy with agents (e.g., in U.S. Pat. No. 6,683,072: EnteregTM (alvimopan; formerly called adolor/ADL 8-2698), US 2012/0283411 A9 Nov. 8, 2012

conivaptain and related agents describe in U.S. Pat. No. 6,645, benzthiazide (CAS RN 91-33-8, which may be prepared as 959) used for the treatment of postoperative ileus and other disclosed in U.S. Pat. No. 3,108,097), buthiazide (which may disorders. be prepared as disclosed in British Patent Nos. 861.367), and 0869. The polypeptides and agonists described herein can hydrochlorothiazide), loop diuretics (e.g. bumetanide, be used in combination therapy with an anti-hypertensive ethacrynic acid, furosemide, and torasemide), potassium agent including but not limited to: sparing agents (e.g. amiloride, and triamterene (CAS Number (1) diuretics, such as thiazides, including chlorthalidone, 396-01-0)), and aldosterone antagonists (e.g. spironolactone chlorthiazide, dichlorophenamide, hydroflumethiazide, inda (CAS Number 52-01-7), epirenone, and the like); B-adrener pamide, polythiazide, and hydrochlorothiazide; loop diuret gic blockers such as Amiodarone (Cordarone, Pacerone), ics, such as bumetanide, ethacrynic acid, furosemide, and bunolol hydrochloride (CAS RN31969-05-8, Parke-Davis), torsemide; potassium sparing agents, such as amiloride, and acebutolol (N-3-Acetyl-4-2-hydroxy-3-(1 methylethyl) triamterene; carbonic anhydrase inhibitors, osmotics (such as glycerin) and aldosterone antagonists, such as spironolac aminopropoxyphenyl-butanamide, or (t)-3'-Acetyl-4-2- tone, epirenone, and the like; hydroxy-3-(isopropylamino) propoxybutyranilide), acebu (2) beta-adrenergic blockers such as acebutolol, atenolol. tolol hydrochloride (e.g. Sectral(R), Wyeth-Ayerst), alprenolol betaxolol, bevantolol, bisoprolol, bopindolol, carteolol. hydrochloride (CAS RN 13707-88-5 see Netherlands Patent carvedilol, celiprolol, esmolol, indenolol, metaprolol, nad Application No. 6,605,692), atenolol (e.g. Tenormin R, Astra olol, nebivolol, penbutolol, pindolol, propanolol, Sotalol, ter Zeneca), carteolol hydrochloride (e.g. Cartrol(R) Filmtab(R), tatolol, tilisolol, and timolol, and the like: Abbott), Celiprolol hydrochloride (CAS RN 57470-78-7, (3) calcium channel blockers such as amlodipine, aranid also see in U.S. Pat. No. 4,034,009), cetamolol hydrochloride ipine, azelnidipine, barnidipine, benidipine, bepridil, cinal (CAS RN 77590-95-5, see also U.S. Pat. No. 4,059,622), dipine, clevidipine, diltiazem, efonidipine, felodipine, gallo labetalol hydrochloride (e.g. Normodyne(R), Schering), pamil. isradipine, lacidipine, lemildipine, lercanidipine, esmolol hydrochloride (e.g. BreviblocR, Baxter), levobetax nicardipine, nifedipine, nilvadipine, nimodepine, nisol olol hydrochloride (e.g. BetaxonTM Ophthalmic Suspension, dipine, nitrendipine, manidipine, pranidipine, and Verapamil, Alcon), levobunolol hydrochloride (e.g. Betagan R. Liqui and the like; filmR) with C CAP(R) Compliance Cap, Allergan), nadolol (4) angiotensin converting enzyme (ACE) inhibitors such as (e.g. Nadolol, Mylan), practolol (CAS RN 6673-35-4, see benazepril; captopril; ceranapril; cilaZapril; delapril; enala also U.S. Pat. No. 3,408,387), propranolol hydrochloride pril; enalopril; fosinopril; imidapril: lisinopril; losinopril; (CAS RN 318-98-9), Sotalol hydrochloride (e.g. Betapace moexipril; quinapril; quinaprilat, ramipril; perindopril; per AFTM, Berlex), timolol (2-Propanol, 1-(1,1-dimethylethyl) indropril; quanipril; spirapril; tenocapril; trandolapril, and amino-3-4-4(4-morpholinyl)-1,2,5-thiadiazol-3-yloxy-, hemihydrate, (S)- CAS RN 91524-16-2), timolol maleate Zofenopril, and the like: (S)-1-(1,1-dimethylethyl)amino-3-4-(4-morpholinyl)-1, (5) neutral endopeptidase inhibitors such as omapatrilat, 2.5-thiadiazol-3-yl)oxy-2-propanol (Z)-2-butenedioate cadoxatril and ecadotril, fosidotril, sampatrilat, AVE7688, (1:1) salt, CAS RN 26921-17-5), bisoprolol (2-Propanol, ER4030, and the like: 1-4-2-(1-methylethoxy)ethoxy-methylphenoxyl-3-(1- (6) endothelin antagonists such as tezosentan, A3081.65, and meth-ylethyl)amino-, (+), CAS RN 66722-44-9), bisoprolol YM62899, and the like: fumarate (such as (+)-1-4-2-(1-Methylethoxy)ethoxyme (7) vasodilators such as hydralazine, clonidine, minoxidil, thylphenoxy-3-(1-methylethyl)amino-2-propanol (E)-2- and nicotinyl alcohol, and the like; butenedioate (2:1) (salt), e.g., ZebetaTM, Lederle Consumer), (8) angiotensin II receptor antagonists such as aprosartan, nebivalol (2H-1-Benzopyran-2-methanol, C.C.'-(iminobis candesartan, eprosartan, irbesartan, losartan, olmesartan, pra (methylene)bis 6-fluoro-3,4-dihydro-, CAS RN99200-09-6 tosartan, tasosartan, telmisartan, Valsartan, and EXP-3137. see also U.S. Pat. No. 4,654.362), cicloprolol hydrochloride, FI6828K, and RNH6270, and the like: Such 2-Propanol. 1-4-2-(cyclopropylmethoxy)ethoxyphe (9) C/B adrenergic blockers such as nipradillol, arotinolol and noxy-3-1-methylethyl)amino-, hydrochloride, A.A.S. RN amoSulalol, and the like; 63686-79-3), dexpropranolol hydrochloride (2-Propano-1,1- (10) alpha 1 blockers, such as teraZosin, urapidil, praZosin, 1-methylethy)-amino-3-(1-naphthalenyloxy)-hydrochlo tamsulosin, bunaZosin, trimaZosin, doxazosin, naftopidil, ride (CASRN 13071-11-9), diacetololhydrochloride (Aceta indoramin, WHP 164, and XENO10, and the like: mide, N-3-acetyl-4-2-hydroxy-3-(1-methyl-ethyl)amino (11) alpha 2 agonists such as lofexidine, tiamenidine, moX propoxyphenyl-, monohydrochloride CAS RN 69796-04 onidine, rillmenidine and guanobenz, and the like; 9), dilevalol hydrochloride (Benzamide, 2-hydroxy-5-1- (12) aldosterone inhibitors, and the like; and hydroxy-2-1-methyl-3-phenylpropyl)aminoethyl-, (13) angiopoietin-2-binding agents such as those disclosed in monohydrochloride, CAS RN 75659-08-4), exaprolol hydro WOO3/O3O833. chloride (2-Propanol, 1-(2-cyclohexylphenoxy)-3-(1-meth 0870 Specific anti-hypertensive agents that can be used in ylethyl)amino-, hydrochloride CAS RN 59333-90-3), fles combination with polypeptides and agonists described herein tolol sulfate (Benzoic acid, 2-fluoro-3-2-aminocarbonyl) include, but are not limited to: amino-dimethylethylamino-2-hydroxypropyl ester, (+)- diuretics, such as thiazides (e.g., chlorthalidone, cyclothiaz sulfate (1:1) (salt), CAS RN 88844-73-9; metalol ide (CAS RN 2259-96-3), chlorothiazide (CAS RN 72956 hydrochloride (Methanesulfonamide, N-(4-1-hydroxy-2- 09-3, which may be prepared as disclosed in US2809194), (methylamino)propylphenyl-, monohydrochloride CAS dichlorophenamide, hydroflumethiazide, indapamide, poly RN 7701-65-7), metoprolol 2-Propanol. 1-4-(2-methoxy thiazide, bendroflumethazide, methyclothazide, polythiaz ethyl)phenoxy-3-1-methylethyl)amino-: CAS RN 37350 ide, trichlormethazide, chlorthalidone, indapamide, metola 58-6), metoprolol tartrate (such as 2-Propanol. 1-4-(2-meth Zone, quinethazone, althiazide (CAS RN 5588-16-9, which oxyethyl)phenoxy-3-(1-methylethyl)amino-, C.9. may be prepared as disclosed in British Patent No. 902,658), Lopressor(R), Novartis), pamatolol sulfate (Carbamic acid, US 2012/0283411 A9 Nov. 8, 2012 66

2-4-2-hydroxy-3-(1-methylethyl)aminopropoxylphe 1-(2S)-3-mercapto-2-methylpropionyl-L-proline, C.9. nyl-ethyl-, methyl ester, (+) sulfate (salt) (2:1), CAS RN Captopril, Mylan, CAS RN 62571-86-2 and others disclosed 59954-01-7), penbutolol sulfate (2-Propanol, 1-(2-cyclopen in U.S. Pat. No. 4,046,889), ceranapril (and others disclosed tylphenoxy)-3-1,1-dimethylethyl)amino1, (S) , sulfate in U.S. Pat. No. 4,452.790), cetapril (alacepril, Dainippon (2:1) (salt), CAS RN 38363-32-5), practolol (Acetamide, disclosed in Eur. Therap. Res. 39:671 (1986): 40:543 (1986)), N-4-2-hydroxy-3-(1-methylethyl)amino-propoxyphe cilazapril (Hoffman-LaRoche) disclosed in J. Cardiovasc. nyl-, CAS RN 6673-35-4) tiprenolol hydrochloride (Pro Pharmacol. 9:39 (1987), indalapril (delapril hydrochloride panol, 1-(1-methylethyl)amino-3-2-(methylthio)-phe (2H-1,2,4-Benzothiadiazine-7-sulfonamide, 3-bicyclo[2.2. noxy-, hydrochloride, (+), CAS RN39832-43-4), tolamolol 1 hept-5-en-2-yl-6-chloro-3,4-dihydro-, 1,1-dioxide CAS (BenZamide, 4-2-2-hydroxy-3-(2-methylphenoxy)-pro RN2259-96-3); disclosed in U.S. Pat. No. 4,385,051), enala pylaminoethoxyl-, CAS RN 38.103-61-6), bopindolol. pril (and others disclosed in U.S. Pat. No. 4.374,829), enalo indenolol, pindolol, propanolol, tertatolol, and tilisolol, and pril, enaloprilat, fosinopril, ((such as L-proline, 4-cyclo the like; calcium channel blockers such as besylate salt of hexyl-1-(2-methyl-1-(1-oxopropoxy)propoxy (4- amlodipine (such as 3-ethyl-5-methyl-2-(2-aminoethoxym phenylbutyl)phosphinyl)acetyl-, Sodium salt, trans—, e.g., ethyl)-4-(2-chlorophenyl)-1,4-dihydro-6-methyl-3,5-py Monopril, Bristol-Myers Squibb and others disclosed in U.S. ridinedicarboxylate benzenesulphonate, e.g., Norvasc(R), Pat. No. 4,168.267), fosinopril sodium (L-Proline, 4-cyclo Pfizer), clentiazem maleate (1.5-Benzothiazepin-4(5H)-one, hexyl-1-(R)-(1S)-2-methyl-1-(1-ox-opropoxy)propox), 3-(acetyloxy)-8-chloro-5-2-(dimethylamino)ethyl-2,3-di imidapril, indolapril (Schering, disclosed in J. Cardiovasc. hydro-2-(4-methoxyphenyl)-(2S-cis)-, (Z)-2-butenedioate Pharmacol. 5:643,655 (1983)), lisinopril (Merck), losinopril, (1:1), see also U.S. Pat. No. 4,567,195), isradipine (3.5-Py moexipril, moexipril hydrochloride (3-Isoquinolinecarboxy ridinedicarboxylic acid, 4-(4-benzofurazanyl)-1,4-dihydro lic acid, 2-(2S)-2-(1S)-1-(ethoxycarbonyl)-3-phenylpro 2,6-dimethyl-, methyl 1-methylethyl ester, (+)-4-(4-benzo pylamino-1-oxopropyl-1-2,3,4-tetrahydro-6,7- furazanyl)-1,4-dihydro-2,6-dimethyl-3,5- dimethoxy-, monohydrochloride, (3S)-CAS RN 82586-52 pyridinedicarboxylate, see also U.S. Pat. No. 4,466.972); 5), quinapril, quinaprilat, ramipril (Hoechsst) disclosed in EP nimodipine (such as is isopropyl (2-methoxyethyl) 1,4-dihy 79022 and Curr. Ther. Res. 40:74 (1986), perindopril erbu dro-2,6-dimethyl-4-(3-nitrophenyl)-3,5-pyridine-dicarboxy mine (such as 2S,3aS,7aS-1-(S)- N -(S)-1-Carboxybutyl late, e.g. Nimotop(R), Bayer), felodipine (such as ethyl methyl alanylhexahydro-2-indolinecarboxylic acid, 1-ethyl ester, 4-(2,3-dichlorophenyl)-1,4-dihydro-2,6-dimethyl-3,5-py compound with tert-butylamine (1:1), e.g., Aceon(R), Solvay), ridinedicarboxylate-, e.g. Plendil R. Extended-Release, Astra perindopril (Servier, disclosed in Eur. J. din. Pharmacol. Zeneca LP), nilvadipine (3.5-Pyridinedicarboxylic acid, 31:519 (1987)), quanipril (disclosed in U.S. Pat. No. 4.344, 2-cyano-1,4-dihydro-6-methyl-4-(3-nitrophenyl)-3-methyl 949), spirapril (Schering, disclosed in Acta. Pharmacol. Toxi 5-(1-methylethyl) ester, also see U.S. Pat. No. 3,799,934), col. 59 (Supp. 5):173 (1986)), tenocapril, trandolapril, nifedipine (such as 3.5-pyridinedicarboxylic acid, 1,4-dihy Zofenopril (and others disclosed in U.S. Pat. No. 4.316,906), dro-2,6-dimethyl-4-(2-nitrophenyl)-, dimethyl ester, e.g., rentiapril (fentiapril, disclosed in Clin. Exp. Pharmacol. Procardia XLR) Extended Release Tablets, Pfizer), diltiazem Physiol. 10:131 (1983)), pivopril, YS980, teprotide (Brady hydrochloride (such as 1.5-Benzothiazepin-4(5H)-one.3- kinin potentiator BPP9a CAS RN35115-60-7), BRL 36,378 (acetyloxy)-52-(dimethylamino)ethyl-2-3-dihydro-2(4- (SmithKline Beecham, see EP80822 and EP60668), MC-838 methoxyphenyl)-, monohydrochloride, (+)-cis., e.g., (Chugai, see C.A. 102:72588v and Jap. J. Pharmacol. 40:373 TiazacR, Forest), Verapamil hydrochloride (such as benzene (1986), CGS 14824 (Ciba-Geigy, 3-(1-ethoxycarbonyl-3- acetronitrile, (alpha)-3-2-(3,4-dimethoxyphenyl)ethyl phenyl-(15)-propylamino)-2,3,4,5-tetrahydro-2-ox-o-1- methylaminopropyl-3,4-dimethoxy-(alpha)-(1-methyl (3S)-benzazepine-1 acetic acid HCl, see U.K. Patent No. ethyl) hydrochloride, e.g., Isoptin R. SR, Knoll Labs), 2103614), CGS 16,617 (Ciba-Geigy, 3(S)-(1S)-5-amino-1- teludipine hydrochloride (3.5-Pyridinedicarboxylic acid, carboxypentyl)amino-2,3,4,-5-tetrahydro-2-oxo-1H-1-ben 2-(dimethylamino)methyl4-2-(1E)-3-(1,1-dimethyl zazepine-1-ethanoic acid, see U.S. Pat. No. 4.473,575), Ru ethoxy)-3-oxo-1-propenylphenyl-1,4-dihydro-6-methyl-, 44570 (Hoechst, see Arzneimittelforschung 34:1254 (1985)), diethyl ester, monohydrochloride) CAS RN 108700-03-4), R 31-2201 (Hoffman-LaRoche see FEBS Lett. 165:201 belfosdil (Phosphonic acid, 2-(2-phenoxyethyl)-1,3-pro (1984)), C1925 (Pharmacologist 26:243, 266 (1984)), pane-diyl)bis-, tetrabutyl ester CAS RN 103486-79–9), foste WY-44221 (Wyeth, see J. Med. Chem. 26:394 (1983)), and dil (Phosphonic acid, 4-(2-benzothiazolyl)phenylme those disclosed in US2003006922 (paragraph 28), U.S. Pat. thyl-, diethyl ester CAS RN 75889-62-2), aranidipine, No. 4,337,201, U.S. Pat. No. 4,432,971 (phosphonamidates): aZelnidipine, barnidipine, benidipine, bepridil, cinaldipine, neutral endopeptidase inhibitors such as omapatrilat (Van clevidipine, efonidipine, gallopamil, lacidipine, lemildipine, lev(R), CGS 30440, cadoxatril and ecadotril, fasidotril (also lercanidipine, monatepil maleate (1-piperazinebutanamide, known as aladotril oralatriopril), Sampatrilat, mixanpril, and N-(6,11-dihydrodibenzo(b.e)thiepin-11-yl)-(4-fluorophe gemopatrilat, AVE7688, ER4030, and those disclosed in U.S. nyl)-, (+)-, (Z)-2-butene dioate (1:1) (+)-N-(6,11-Dihy Pat. No. 5,362,727, U.S. Pat. No. 5,366,973, U.S. Pat. No. drodibenzo(b.e)thiep-in-11-yl)-4-(p-fluorophenyl)-1-pip 5,225,401, U.S. Pat. No. 4,722,810, U.S. Pat. No. 5,223,516, erazinebutyramide maleate (1:1) CAS RN 132046-06-1), U.S. Pat. No. 4,749,688, U.S. Pat. No. 5,552,397, U.S. Pat. nicardipine, nisoldipine, nitrendipine, manidipine, pranid No. 5,504,080, U.S. Pat. No. 5,612,359, U.S. Pat. No. 5,525, ipine, and the like: T-channel calcium antagonists such as 723, EP0599444, EP0481522, EP0599444, EP0595610, mibefradil; angiotensin converting enzyme (ACE) inhibitors EP0534363, EP534396, EP534492, EP0629627; such as benazepril, benazepril hydrochloride (such as 3-1- endothelin antagonists such as tezosentan, A308165, and (ethoxycarbonyl)-3-phenyl-(1S)-propylamino-2,3,4,5-tet YM62899, and the like; vasodilators such as hydralazine rahydro-2-oxo-1H-1-(3S)-benzazepine-1-acetic acid mono (apresoline), clonidine (clonidine hydrochloride (1H-Imida hydrochloride, e.g., Lotrel (R), Novartis), captopril (such as Zol-2-amine, N-(2,6-dichlorophenyl)4,5-dihydro-, monohy

US 2012/0283411 A9 Nov. 8, 2012

5-ethyl-3-2'-(1H-tetrazole-5-yl)biphenyl-4-yl)methyl-1,3, No. 5,187,159, U.S. Pat. No. 5,198,438, U.S. Pat. No. 5,182, 4-thiadiazoline, 2-5-ethyl-3-2-(1H-tetrazole-5-yl) 288, U.S. Pat. No. 5,036,048, U.S. Pat. No. 5,140,036, U.S. biphenyl-4-yl)methyl-1,3,4-thiazoline-2-ylidene Pat. No. 5,087,634, U.S. Pat. No. 5,196,537, U.S. Pat. No. aminocarbonyl-1-cyclopentencarboxylic acid dipotassium 5,153,347, U.S. Pat. No. 5,191,086, U.S. Pat. No. 5,190,942, salt, and 2-butyl-4-N-methyl-N-(3-methylcrotonoyl) U.S. Pat. No. 5,177,097, U.S. Pat. No. 5,212,177, U.S. Pat. amino-1-2'-(1H-tetrazol-5-yl)biphenyl-4-yl)methyl)-1H No. 5,208,234, U.S. Pat. No. 5,208,235, U.S. Pat. No. 5,212, imidzole-5-carboxylic acid 1-ethoxycarbonyloxyethyl ester, 195, U.S. Pat. No. 5,130,439, U.S. Pat. No. 5,045,540, U.S. those disclosed in patent publications EP475206, EP497150, Pat. No. 5,041,152, and U.S. Pat. No. 5,210,204, and phar EP539086, EP539713, EP535463, EP535465, EP542059, maceutically acceptable salts and esters thereof; C/B adren EP497.121, EP535420, EP407342, EP415886, EP424317, ergic blockers such as nipradillol, arotinolol, amoSulalol, EP435827, EP433983, EP475898, EP490820, EP528762, bretylium tosylate (CAS RN: 61-75-6), dihydroergtamine EP324377, EP323841, EP420237, EP500297, EP426021, mesylate (such as ergotaman-3',6', 18-trione.9,-10-dihydro EP480204, EP429257, EP430709, EP434249, EP446062, 12'-hydroxy-2-methyl-5'-(phenylmethyl)-(5'(O))-. EP505954, EP524217, EP514197, EP514198, EP514193, monomethanesulfonate, e.g., DHE 45(R) Injection, Novartis), EP514192, EP450566, EP468372, EP485929, EP503162, carvedilol (such as (+)-1-(Carbazol-4-yloxy)-3-2-(o-meth EP533058, EP467207 EP399731, EP399732, EP412848, oxyphenoxy)ethylamino-2-propanol, e.g., Coreg R, Smith EP453210, EP456442, EP470794, EP470795, EP495626, Kline Beecham), labetalol (such as 5-1-hydroxy-2-(1-me EP495627, EP499414, EP499416, EP49.9415, EP511791, thyl-3-phenylpropyl)aminoethylsalicylamide EP516392, EP520723, EP520724, EP539066, EP438869, monohydrochloride, e.g., Normodyne R, Schering), brety EP505893, EP530702, EP400835, EP400974, EP401030, lium tosylate (Benzenemethanaminium, 2-bromo-N-ethyl EP407102, EP411766, EP409332, EP4 12594, EP419048, N,N-dimethyl-, salt with 4-methylbenzenesulfonic acid (1:1) EP480659, EP481614, EP490587, EP467715, EP479479, CAS RN 61-75-6), phentolamine mesylate (Phenol, 3-(4.5- EP502725, EP503838, EP505098, EP505111 EP513,979 dihydro-1H-imidazol-2-yl)methyl(4-methylphenyl) EP507594, EP510812, EP511767, EP512675, EP512676, amino-, monomethanesulfonate (salt) CAS RN 65-28-1), EP512870, EP517357, EP537937, EP534706, EP527534, solypertine tartrate (5H-1,3-Dioxolo4.5-findole, 7-2-4- EP540356, EP461040, EP540039, EP465368, EP498723, (2-methoxyphenyl)-1-piperazinylethyl-, (2R,3R)-2,3-di EP498722, EP498721, EP515265, EP503785, EP501892, hydroxybutanedioate (1:1) CAS RN 5591-43-5), Zolertine EP519831, EP532410, EP498361, EP432737, EP504888, hydrochloride (piperazine, 1-phenyl-4-2-(1H-tetrazol-5-yl) EP508393, EP508445, EP403159, EP403158, EP425211, ethyl-, monohydrochloride (8C1,9CI) CAS RN 7241-94-3) EP427463, EP437103, EP481448, EP488532, EP501269, and the like; EP5004.09, EP540400, EP005528, EP028834, EP028833, C. adrenergic receptor blockers, such as alfuzosin (CAS RN: EP411507, EP425921, EP430300, EP434038, EP442473, 81403-68-1), terazosin, urapidil, prazosin (Minipress.(R), EP443568, EP445811, EP459136, EP483.683, EP518033, tamsulosin, bunaZosin, trimaZosin, doxazosin, naftopidil, EP520423, EP531876, EP531874, EP392317, EP468470, indoramin, WHP 164, XENO10, fenspiride hydrochloride EP470543, EP502314, EP529253, EP543263, EP540209, (which may be prepared as disclosed in U.S. Pat. No. 3,399, EP449699, EP465323, EP521768, EP415594, WO92/14468, 192), proroxan (CAS RN33743-96-3), and labetalol hydro WO93/08171, WO93/08169, WO91/00277, WO91/00281, chloride and combinations thereof. C.2 agonists such as meth WO91/14367, WO92/00067, WO92/00977, WO92/20342, yldopa, methyldopa HCL, lofexidine, tiamenidine, WO93/04045, WO93/04046, WO91/15206, WO92/14714, moXonidine, rillmenidine, guanobenz, and the like; WO92/09600, WO92/16552, WO93/05025, WO93/03018, aldosterone inhibitors, and the like; renin inhibitors including WO91/07404, WO92/02508, WO92/13853, WO91/19697, Aliskiren (SPP 100; Novartis/Speedel); angiopoietin-2-bind WO91/11909, WO91/12001, WO91/11999, WO91/15209, ing agents such as those disclosed in WO03/030833; anti WO91/15479, WO92/20687, WO92/20662, WO92/20661, angina agents such as ranolazine (hydrochloridel-pipera WO93/01177, WO91/14679, WO91/13063, WO92/13564, zineacetamide, N-(2,6-dimethylphenyl)-4-2-hydroxy-3-(2- WO91/17148, WO91/18888, WO91/19715, WO92/02257, methoxyphenoxy)propyl-, dihydrochloride CAS RN95635 WO92/04335, WO92/05161, WO92/07852, WO92/15577, 56-6), betaxolol hydrochloride (2-Propanol. 1-4-2 WO93/03033, WO91/16313, WO92/00068, WO92/02510, (cyclopropylmethoxy)ethylphenoxy-3-(1-methylethyl) WO92/09278, WO9210179, WO92/10180, WO92/10186, amino-, hydrochloride CAS RN 63659-19-8), butoprozine WO92/10181, WO92/10097, WO92/10183, WO92/10182, hydrochloride (Methanone, 4-3(dibutylamino)propoxy WO92/10187, WO92/10184, WO92/10188, WO92/10180, phenyl (2-ethyl-3-indolizinyl)-, monohydrochloride CAS WO92/10185, WO92/20651, WO93/03722, WO93/06828, RN 62134-34-3), cinepazet maleatel-piperazineacetic acid, WO93/03040, WO92/19211, WO92/22533, WO92/06081, 4-1-oxo-3-(3,4,5-trimethoxyphenyl)-2-propenyl-, ethyl WO92/05784, WO93/00341, WO92/04343, WO92/04059, ester, (2Z)-2-butenedioate (1:1) CAS RN 50679-07-7), U.S. Pat. No. 5,104,877, U.S. Pat. No. 5,187,168, U.S. Pat. tosifen (Benzenesulfonamide, 4-methyl-N-(1S)-1-me No. 5,149,699, U.S. Pat. No. 5,185,340, U.S. Pat. No. 4,880, thyl-2-phenylethylaminocarbonyl-CAS RN 32295-184), 804, U.S. Pat. No. 5,138,069, U.S. Pat. No. 4,916,129, U.S. Verapamilhydrochloride (Benzeneacetonitrile, C-3-2-(3,4- Pat. No. 5,153,197, U.S. Pat. No. 5,173,494, U.S. Pat. No. dimethoxyphenyl)ethylmethylaminopropyl-3,4- 5,137,906, U.S. Pat. No. 5,155,126, U.S. Pat. No. 5,140,037, dimethoxy-O-(1-methylethyl)-, monohydrochloride CAS U.S. Pat. No. 5,137,902, U.S. Pat. No. 5,157,026, U.S. Pat. RN 152-114), molsidomine (1,2,3-Oxadiazolium, No. 5,053,329, U.S. Pat. No. 5,132,216, U.S. Pat. No. 5,057, 5-(ethoxycarbonyl)amino-3-(4-morpholinyl)-, inner salt 522, U.S. Pat. No. 5,066,586, U.S. Pat. No. 5,089,626, U.S. CAS RN25717-80-0), and ranolazine hydrochloride (1-pip Pat. No. 5,049,565, U.S. Pat. No. 5,087,702, U.S. Pat. No. erazineacetamide, N-(2,6-dimethylphenyl)-2-hydroxy-3- 5,124,335, U.S. Pat. No. 5,102,880, U.S. Pat. No. 5,128,327, (2-meth-oxyphenoxy)propyl-, dihydrochloride CAS RN U.S. Pat. No. 5,151,435, U.S. Pat. No. 5,202,322, U.S. Pat. 95635-56-6); tosifen (Benzenesulfonamide, 4-methyl-N- US 2012/0283411 A9 Nov. 8, 2012 69

(1S)-1-methyl-2-phenylethylaminocarbonyl-CAS RN Zole, acrivastine, antazoline, azatadine, azelastine, 32295-184); adrenergic stimulants such as guanfacine hydro astamizole, bromopheniramine, bromopheniramine maleate, chloride (such as N-amidino-2-(2,6-dichlorophenyl)aceta carbinoxamine, carebastine, cetirizine, chlorpheniramine, mide hydrochloride, e.g., Tenex R. Tablets available from chloropheniramine maleate, cimetidine, clemastine, cycliz Robins); methyldopa-hydrochlorothiazide (such as levo-3- ine, cyproheptadine, descarboethoxyloratadine, dexchlor (3,4-dihydroxyphenyl)-2-methylalanine) combined with pheniramine, dimethindene, diphenhydramine, diphe Hydrochlorothiazide (such as 6-chloro-3,4-dihydro-2H-1.2. nylpyraline, doxylamine Succinate, doxylamine, ebastine, 4-benzothiadiazine-7-sulfonamide 1,1-dioxide, e.g., the efletirizine, epinastine, farnotidine, feXofenadine, hydrox combination as, e.g., Aldoril R. Tablets available from Merck), yZine, hydroxy Zine, ketotifen, levocabastine, levocetirizine, methyldopa-chlorothiazide (such as 6-chloro-2H-1,2,4-ben levocetirizine, loratadine, meclizine, mepyramine, meduita Zothiadiazine-7-sulfonamide 1,1-dioxide and methyldopa as Zine, methdilazine, mianserin, mizolastine, noberastine, described above, e.g., Aldoclor(R), Merck), clonidine hydro norasternizole, noraZtemizole, phenindamine, pheniramine, chloride (such as 2-(2,6-dichlorophenylamino)-2-imidazo picumast, promethazine, pynlamine, pyrilamine, ranitidine, line hydrochloride and chlorthalidone (such as 2-chloro-5-(1- temelastine, terfenadine, trimeprazine, tripelenamine, and hydroxy-3-oxo-1-isoindolinyl) benzenesulfonamide), e.g., triprolidine: Combipres(R), Boehringer Ingelheim), clonidine hydrochlo (7) an anticholinergic including but not limited to: atropine, ride (such as 2-(2,6-dichlorophenylamino)-2-imidazoline benztropine, biperiden, flutropium, hyoscyamine (e.g. hydrochloride, e.g., Catapres(R), Boehringer Ingelheim), Levsin(R); LevbidR; Levsin/SL(R), AnaspazR), Levsinex Time clonidine (1H-Imidazol-2-amine, N-(2,6-dichlorophenyl)4, caps(R), NuLev(R), ilutropium, ipratropium, ipratropium bro 5-dihydro-CAS RN 4205-90-7), Hyzaar (Merck; a combina mide, methScopolamine, oxybutinin, rispenzepine, Scopola tion of losartan and hydrochlorothiazide), Co-Diovan (No mine, and tiotropium; vartis; a combination of Valsartan and hydrochlorothiazide, (8) an anti-tussive including but not limited to: dextrometho Lotrel (Novartis; a combination of benazepril and amlo rphan, codeine, and hydromorphone; dipine) and Caduet (Pfizer; a combination of amlodipine and (9) a decongestant including but not limited to: pseudoephe atorvastatin), and those agents disclosed in US20030069221. drine and phenylpropanolamine; 0871. The polypeptides and agonists described herein can (10) an expectorant including but not limited to: guafenesin, be used in combination therapy with one or more of the guaicolsulfate, terpin, ammonium chloride, glycerol guaico following agents useful in the treatment of respiratory and late, and iodinated glycerol; other disorders including but not limited to: (11) a bronchodilator including but not limited to: theophyl (1) B-agonists including but not limited to: albuterol line and aminophylline; (PROVENTILR, SALBUTAMO1(R, VENTOLINR), bam (12) an anti-inflammatory including but not limited to: buterol, bitoterol, clenbuterol, fenoterol, formoterol, isoet fluribiprofen, diclophenac, indomethacin, ketoprofen, S-ket harine (BRONKOSOLR, BRONKOMETER(R), metaproter roprophen, tenoxicam, enol (ALUPENTR), METAPRELR), pirbuterol (13) a PDE (phosphodiesterase) inhibitor including but not (MAXAIRR), reproterol, rimiterol, salmeterol, terbutaline limited to those disclosed herein; (BRETHAIRE(R), BRETHINER, BRICANYL(R), adrenalin, (14) a recombinant humanized monoclonal antibody e.g. isoproterenol (ISUPREL(R), epinephrine bitartrate (PRI Xolair (also called omalizumab), rhuMab, and talizumab: MATENER), ephedrine, orciprenline, fenoterol and isoet (15) a humanized lung Surfactant including recombinant harine; forms of surfactant proteins SP-B, SP-C or SP-D e.g. SUR (2) Steroids, including but not limited to beclomethasone, FAXINR, formerly known as disc-104 (Discovery Laborato beclomethasone dipropionate, betamethasone, budesonide, ries). bunedoside, butiXocort, dexamethasone, flunisolide, fluocor (16) agents that inhibit epithelial sodium channels (ENaC) tin, fluticaSone, hydrocortisone, methylprednisone, mometa Such as amiloride and related compounds; Sone, predonisolone, predonisone, tipredane, tiXocortal, tri (17) antimicrobial agents used to treat pulmonary infections amcinolone, and triamcinolone acetonide; Such as acyclovir, amikacin, amoxicillin, doxycycline, trime (3) B2-agonist-corticosteroid combinations e.g., salmeterol thoprin Sulfamethoxazole, amphotericin B, azithromycin, fluticasone (ADVAIRR), formoterol-budesonid (SYM clarithromycin, roXithromycin, clarithromycin, cephalospor BICORTR)); ins (ceffoxitin, cefimetazole etc), ciprofloxacin, ethambutol, (4) leukotriene D4 receptor antagonists/leukotriene antago gentimycin, ganciclovir, imipenem, iSoniazid, itraconazole, nists/LTD4 antagonists (i.e., any compound that is capable of penicillin, ribavirin, rifampin, rifabutin, amantadine, riman blocking, inhibiting, reducing or otherwise interrupting the tidine, Streptomycin, tobramycin, and Vancomycin; interaction between leukotrienes and the Cys LTI receptor) (18) agents that activate chloride secretion through Ca++ including but not limited to: Zafirlukast, montelukast, mon dependent chloride channels (such as purinergic receptor telukast sodium (SINGULAIRR), pranlukast, iralukast, (P2Y(2) agonists); pobilukast, SKB-106,203 and compounds described as hav (19) agents that decrease sputum viscosity, Such as human ing LTD4 antagonizing activity described in U.S. Pat. No. recombinant DNase 1. (PulmozymeR); 5,565,473; (20) nonsteroidal anti-inflammatory agents (acemetacin, (5) 5-lipoxygenase inhibitors and/or leukotriene biosynthesis acetaminophen, acetyl salicylic acid, alclofenac, alminopro inhibitors e.g., zileuton and BAY1005 (CA registry 128253 fen, apaZone, aspirin, benoxaprofen, bezpiperylon, bucloxic 31-6): acid, carprofen, clidanac, diclofenac, diclofenac, diflunisal, (6) histamine H1 receptor antagonists/antihistamines (i.e., diflusinal, etodolac, fenbufen, fenbufen, fenclofenac, fen any compound that is capable of blocking, inhibiting, reduc clozic acid, fenoprofen, fentiazac, feprazone, flufenamic ing or otherwise interrupting the interaction between hista acid, flufenisal, flufenisal, fluprofen, flurbiprofen, flurbipro mine and its receptor) including but not limited to: astemi fen, furofenac, ibufenac, ibuprofen, indomethacin, US 2012/0283411 A9 Nov. 8, 2012 70 indomethacin, indoprofen, isoxepac, isoxicam, ketoprofen, CNTF (Ciliary neurotrophic factors), such as GI-181771 ketoprofen, ketorolac, meclofenamic acid, meclofenamic (Glaxo-SmithKline), SR 146131 (Sanofi Synthelabo), butab acid, mefenamic acid, mefenamic acid, miroprofen, mofeb indide, PD 170,292, and PD 149164 (Pfizer); utaZone, nabumetone oxaprozin, naproxen, naproxen, niflu CNTF derivatives, such as AxokineR (Regeneron), and those mic acid, Oxaprozin, Oxpinac, oxyphenbutaZone, phenacetin, disclosed in WO94/09134, WO98/22 128, and WO99/43813; phenylbutaZone, phenylbutaZone, piroxicam, piroXicam, pir dipeptidyl peptidase IV (DP-IV) inhibitors, such as isoleu profen, pranoprofen, Sudoxicam, tenoxican, Sulfasalazine, cine thiazolidide, valine pyrrolidide, NVP-DPP728, Sulindac, Sulindac, Suprofen, tiaprofenic acid, tiopinac, tioX LAF237, P93/01, P3298, TSL 225 (tryptophyl-1,2,3,4-tet aprofen, tolfenamic acid, tolmetin, tolmetin, Zidometacin, rahydroisoquinoline-3-carboxylic acid; disclosed byYamada Zomepirac, and Zomepirac); and et al, Bioorg. & Med. Chem. Lett. 8 (1998) 1537-1540), (21) aerosolized antioxidant therapeutics such as S-Nitroso TMC-2A/2B/2C, CD26 inhibtors, FE 999011, P9310/K364, glutathione. VIP 0177. SDZ 274-444, 2-cyanopyrrolidides and 4-cyan 0872. The polypeptides and agonists described herein can opyrrolidides as disclosed by Ashworth etal, Bioorg. & Med. be used in combination therapy with an anti-obesity agent. Chem. Lett. Vol. 6, No. 22, pp 1163-1166 and 2745-2748 Suitable such agents include, but are not limited to: (1996) and the compounds disclosed patent publications. 11 BHSD-1 (11-beta hydroxy steroid dehydrogenase type 1) WO99/38501, WO99/46272, WO99/67279 (Probiodrug), inhibitors, such as BVT 3498, BVT 2733, 3-(1-adamantyl)- WO99/67278 (Probiodrug), WO99/61431 (Probiodrug), 4-ethyl-5-(ethylthio)-4H-1,2,4-triazole, 3-(1-adamantyl)-5- WO02/083128, WO02/062764, WO03/000180, WO03/ (3,4,5-trimethoxyphenyl)-4-methyl-4H-1,2,4-triazole, 000181, WO03/000250, WO03/002530, WO03/002531, 3-adamantanyl-4,5,6,7,8,9,10,11,12.3a-decahydro-1,2,4- WO03/002553, WO03/002593, WO03/004498, WO03/ triazolo 4.3-all annulene, and those compounds disclosed 004496, WO03/017936, WO03/024942, WO03/024965, in WO01/90091, WO01/90090, WO01/90092 and WO02/ WO03/033524, WO03/037327 and EP1258476; 072084; growth hormone secretagogue receptoragonists/antagonists, 5HT antagonists such as those in WO03/037871, WO03/ such as NN703, hexarelin, MK-0677 (Merck), SM-130686, 037887, and the like: CP-424391 (Pfizer), LY 444,711 (Eli Lilly), L-692,429 and 5HT1a modulators such as carbidopa, benserazide and those L-163,255, and such as those disclosed in U.S. Ser. No. disclosed in U.S. Pat. No. 6,207,699, WO03/031439, and the 09/662,448, U.S. provisional application 60/203,335, U.S. like: Pat. No. 6,358,951, US20020491.96, US2002/022637, 5HT2c (serotonin receptor 2c) agonists, such as BVT933, WO01/56592 and WO02/32888; DPCA37215, 1K264, PNU 22394, WAY161503, R-1065, SB H3 (histamine H3) antagonist/inverse agonists, such as thio 243213 (GlaxoSmithKline) andYM348 and those disclosed peramide, 3-(1H-imidazol-4-yl)propyl N-(4-pentenyl)car in U.S. Pat. No. 3,914,250, WO00/77010, WO02/36596, bamate), clobenpropit, iodophenpropit, imoproxifan, WO02/48124, WO02/10169, WO01/66548, WO02/44152, GT2394 (Gliatech), and A331440, O-3-(1H-imidazol-4-yl) WO02/.51844, WO02/40456, and WO02/40457; propanol carbamates (Kiec-Kononowicz, K. et al., Pharma 5HT6 receptor modulators, such as those in WO03/030901, Zie, 55:349-55 (2000)), piperidine-containing histamine WO03/035061, WO03/039547, and the like: H3-receptor antagonists (Lazewska, D. et al., Pharmazie, acyl-estrogens. Such as oleoyl-estrone, disclosed in del Mar 56:927-32 (2001), benzophenone derivatives and related Grasa, M. et al., Obesity Research, 9:202-9 (2001) and Japa compounds (Sasse, A. et al., Arch. Pharm. (Weinheim) 334: nese Patent Application No.JP 2000256190; 45-52 (2001)), substituted N-phenylcarbamates (Reidemeis anorectic bicyclic compounds such as 1426 (Aventis) and ter, S. et al., Pharmazie, 55: 83-6 (2000)), and proxifan deriva 1954 (Aventis), and the compounds disclosed in WO00/ tives (Sasse, A. et al., J. Med. Chem.43:3335-43 (2000)) and 18749, WOO1/32638, WO01/62746, WO01/62747, and histamine H3 receptor modulators such as those disclosed in WO03/015769; CB 1 (cannabinoid-1 receptor) antagonist/inverse agonists WO02/15905, WO03/024928 and WO03/024929; such as rimonabant (Acomplia; Sanofi), SR-147778 (Sanofi), leptin derivatives, such as those disclosed in U.S. Pat. No. SR-141716 (Sanofi), BAY 65-2520 (Bayer), and SLV 319 5,552,524, U.S. Pat. No. 5,552,523, U.S. Pat. No. 5,552,522, (Solvay), and those disclosed in patent publications U.S. Pat. U.S. Pat. No. 5,521,283, WO96/23513, WO96/23514, No. 4,973,587, U.S. Pat. No. 5,013,837, U.S. Pat. No. 5,081, WO96/23515, WO96/23516, WO96/23517, WO96/23518, 122, U.S. Pat. No. 5,112,820, U.S. Pat. No. 5,292,736, U.S. WO96/23519, and WO96/23520; Pat. No. 5,532,237, U.S. Pat. No. 5,624,941, U.S. Pat. No. leptin, including recombinant human leptin (PEG-OB. Hoff 6,028,084, U.S. Pat. No. 6,509,367, U.S. Pat. No. 6,509,367, man La Roche) and recombinant methionyl human leptin WO96/33159, WO97/29079, WO98/31227, WO98/33765, (Amgen); WO98/37061, WO98/41519, WO98/43635, WO98/43636, 0873 lipase inhibitors, such as tetrahydrolipstatin (orl WO99/02499, WO00/10967, WO00/10968, WO01/09120, istat/Xenical(R), Triton WR1339, RHC80267, lipstatin, WO01/58869, WO01/64.632, WO01/64633, WO01/64634, teasaponin, diethylumbelliferyl phosphate, FL-386, WAY WO01/70700, WO01/96330, WO02/076949, WO03/ 121898, Bay-N-3176, valilactone, esteracin, ebelactone A, 006007, WO03/007887, WO03/020217, WO03/026647, ebelactone B, and RHC 80267, and those disclosed in patent WO03/026648, WO03/027069, WO03/027076, WO03/ publications WO01/77094, U.S. Pat. No. 4,598,089, U.S. Pat. 027114, WO03/037332, WO03/040 107, WO03/086940, No. 4452,813, U.S. Pat. No. 5,512,565, U.S. Pat. No. 5,391, WO03/084943 and EP658546; 571, U.S. Pat. No. 5,602,151, U.S. Pat. No. 4,405,644, U.S. CCK-A (cholecystokinin-A) agonists, such as AR-R 15849, Pat. No. 4,189,438, and U.S. Pat. No. 4,242,453; GI 181771 (GSK), JMV-180, A-71378, A-71623 and lipid metabolism modulators such as maslinic acid, erythro SR146131 (Sanofi), and those described in U.S. Pat. No. diol, ursolic acid uvaol, betulinic acid, betulin, and the like 5,739,106; and compounds disclosed in WO03/01 1267; US 2012/0283411 A9 Nov. 8, 2012

Mc4r (melanocortin 4 receptor) agonists, such as WO02/20488, WO02/22592, WO02/48152, WO02/49648, CHIR86036 (Chiron), ME-10142, ME-10145, and HS-131 WO02/051806, WO02/094789, WO03/009845, WO03/ (Melacure), and those disclosed in PCT publication Nos. O14083, WO03/022849, WO03/028726 and Norman et al., J. WO99/64002, WO00/74679, WO01/991752, WO01/25192, Med. Chem. 43:4288-4312 (2000); WO01/52880, WO01/74844, WO01/70708, WO01/70337, opioid antagonists, such as nalmefene (REVEXR), 3-meth WO01/91752, WO02/059095, WO02/059107, WO02/ oxynaltrexone, methylmaltrexone, naloxone, and naltrexone 059108, WO02/059117, WO02/06276, WO02/12166, (e.g. PT901; Pain Therapeutics, Inc.) and those disclosed in WO02/11715, WO02/12178, WO02/15909, WO02/38544, US20050004155 and WO00/21509; WO02/068387, WO02/068388, WO02/067869, WO02/ orexin antagonists, such as SB-334867-A and those disclosed 081430, WO03/06604, WO03/007949, WO03/009847, in patent publications WO01/96302, WO01/68609, WO02/ WO03/009850, WO03/013509, and WO03/031410; 44172, WO02/51232, WO02/51838, WO02/089800, WO02/ Mc5r (melanocortin 5 receptor) modulators, such as those 090355, WO03/023561, WO03/032991, and WO03/037847: disclosed in WO97/19952, WO00/15826, WO00/15790, PDE inhibitors (e.g. compounds which slow the degradation US20030092041: of cyclic AMP (cAMP) and/or cyclic GMP (cGMP) by inhi melanin-concentrating hormone 1 receptor (MCHR) antago bition of the phosphodiesterases, which can lead to a relative nists, such as T-226296 (Takeda), SB 568849, SNP-7941 increase in the intracellular concentration of cAMP and (Synaptic), and those disclosed in patent publications WO01/ cGMP; possible PDE inhibitors are primarily those sub 21169, WO01/82925, WO01/87834, WO02/051809, WO02/ stances which are to be numbered among the class consisting 06245, WO02/076929, WO02/076947, WO02/04433, of the PDE3 inhibitors, the class consisting of the PDE4 WO02/.51809, WO02/083134, WO02/094799, WO03/ inhibitors and/or the class consisting of the PDE5 inhibitors, 004027, WO03/13574, WO03/15769, WO03/028641, in particular those Substances which can be designated as WO03/035624, WO03/033476, WO03/033480, mixed types of PDE3/4 inhibitors or as mixed types of PDE3/ JP13226269, and JP1437059; 4/5 inhibitors) such as those disclosed in patent publications mGluR5 modulators such as those disclosed in WOO3/ DE1470341, DE2108438, DE2123328, DE2305339, 029210, WO03/047581, WO03/048137, WO03/051315, DE2305575, DE2315801, DE2402908, DE2413935, WO03/051833, WO03/053922, WO03/059904, and the like; DE2451417, DE2459090, DE2646469, DE2727481, serotoninergic agents, such as fenfluramine (such as Pondi DE2825048, DE2837161, DE2845220, DE2847621, min(R) (Benzeneethanamine, N-ethyl-alpha-methyl-3-(trif DE2934747, DE3021792, DE3038166, DE3044568, luoromethyl)-, hydrochloride), Robbins), dexfenfluramine EP000718, EP0008.408, EP0010759, EP0059948, (such as Redux(R) (Benzeneethanamine, N-ethyl-alpha-me EP0075436, EP00965.17, EPO112987, EP0116948, thyl-3-(trifluoromethyl)-, hydrochloride), Interneuron) and EPO150937, EP0158380, EPO161632, EPO161918, sibutramine ((MeridiaR), Knoll/ReductiltM) including race EP0167121, EP019.9127, EP0220044, EP0247725, mic mixtures, as optically pure isomers (+) and (-), and EP0258191 EP0272910, EP0272914, EP0294647, pharmaceutically acceptable salts, solvents, hydrates, clath EP0300726, EP0335386, EP0357788, EP0389282, rates and prodrugs thereof including sibutramine hydrochlo EP0406958, EP0426180, EP0428302, EP0435811, ride monohydrate salts thereof, and those compounds dis EP0470805, EP0482208, EP0490823, EP0506194, closed in U.S. Pat. No. 4,746,680, U.S. Pat. No. 4,806,570, EP0511865, EP0527117, EP0626939, EP0664289, and U.S. Pat. No. 5,436,272, US20020006964, WO01/ EP0671389, EP0685474, EP0685475, EP0685479, 27068, and WOO1/623.41: JP92234389, JP94329652, JP950.10875, U.S. Pat. No. 4,963, NE (norepinephrine) transport inhibitors, such as GW 561, U.S. Pat. No. 5,141,931, WO9117991, WO9200968, 320659, despiramine, talsupram, and nomifensine; WO9212961, WO9307146, WO9315044, WO9315045, NPY 1 antagonists, such as BIBP3226, J-115814, BIBO WO9318024, WO9319068, WO9319720, WO931.9747, 3304, LY-357897, CP-671906, GI-264879A, and those dis WO9319749, WO9319751, WO9325517, WO9402465, closed in U.S. Pat. No. 6,001,836, WO96/14307, WO01/ WO9406423, WO9412461, WO9420455, WO9422852, 23387, WO99/51600, WO01/85690, WO01/85098, WO01/ WO9425437, WO9427947, WO9500,516, WO950.1980, 85173, and WO01/89528; WO9503794, WO9504045, WO9504046, WO9505386, NPY5 (neuropeptide Y Y5) antagonists, such as 152,804, WO9508534, WO9509623, WO9509624, WO9509627, GW-569180A, GW-594884A, GW-587081X, WO9509836, WO9514667, WO9514680, WO9514681, GW-548118X, FR235208, FR226928, FR240662, WO9517392, WO9517399, WO9519362, WO9522520, FR252384, 1229U91, GI-264879A, CGP71683A, WO9524381, WO9527692, WO9528926, WO9535281, LY-377897, LY-366377, PD-160170, SR-120562A, WO9535282, WO9600218, WO9601825, WO9602541, SR-120819A, JCF-104, and H409/22 and those compounds WO961 1917, DE3142982, DE 1116676, DE2162096, disclosed in patent publications U.S. Pat. No. 6,140,354, U.S. EP0293063, EP0463756, EP0482208, EP0579496, Pat. No. 6, 191,160, U.S. Pat. No. 6,218,408, U.S. Pat. No. EP0667345 U.S. Pat. No. 6,331,543, US20050004222 (in 6,258,837, U.S. Pat. No. 6,313,298, U.S. Pat. No. 6,326,375, cluding those disclosed in formulas I-XIII and paragraphs U.S. Pat. No. 6,329,395, U.S. Pat. No. 6,335,345, U.S. Pat. 37-39, 85-0545 and 557-577), WO9307124, EPO163965, No. 6,337,332, U.S. Pat. No. 6,329,395, U.S. Pat. No. 6,340, EP0393500, EP0510562, EP0553174, WO950.1338 and 683, EP01010691, EP-01044970, WO97/19682, WO97/ WO9603399, as well as PDE5 inhibitors (such as RX-RA-69, 20820, WO97/20821, WO97/20822, WO97/20823, WO98/ SCH-51866, KT-734, vesnarinone, Zaprinast, SKF-96231, 27063, WO00/107409, WO00/185714, WO00/185730, ER-21355, BF/GP-385, NM-702 and sildenafil (ViagraTM)), WO00/64880, WO00/68197, WO00/69849, WO/0113917, PDE4 inhibitors (such as etazolate, ICI63197, RP73401, ima WO01/09120, WO01/14376, WO01/85714, WO01/85730, Zolidinone (RO-20-1724), MEM 1414 (R1533/R1500; Phar WO01/07409, WO01/02379, WO01/23388, WO01/23389, macia Roche), denbufylline, rolipram, oxagrelate, nitradua WO01/44201, WO01/62737, WO01/62738, WO01/09120, Zone, Y-590, DH-6471, SKF-94120, motapizone, lixazinone, US 2012/0283411 A9 Nov. 8, 2012 72 indolidan, olprinone, atizoram, KS-506-G, dipamfylline, termine (or Phenol, 3-4,5-duhydro-1H-imidazol-2-yl) BMY-43351, atizoram, arofylline, filaminast, PDB-093, ethyl(4-methylpheny-1)amino, monohydrochloride) Such UCB-29646, CDP-840, SKF-107806, piclamilast, as Adipex-PR), Lemmon, FASTINR, Smith-Kline Beecham RS-17597, RS-25344-000, SB-207499, TIBENELAST, and Ionamin(R), Medeva), phendimetrazine (or (2S,3S)-3,4- SB-210667, SB-211572, SB-211600, SB-212066, Dimethyl-2-phenylmorpholine L-(+)-tartrate (1:1)) such as SB-212179, GW-3600, CDP-840, mopidamol, anagrelide, Metra R. (Forest), PlegineR (Wyeth-Ayerst), Prelu-2(R) (Boe ibudilast, aminone, pimobendan, cilostaZol, quaZinone and hringer Ingelheim), and StatobeXR (Lemmon), phendamine N-(3,5-dichloropyrid-4-yl)-3-cyclopropylmethoxy-4-difluo tartrate (such as ThephorinR) (2.3.4.9-Tetrahydro-2-methyl romethoxybenzamide, PDE3 inhibitors (such as ICI153, 100, 9-phenyl-1H-indenol2,1-cpyridine L-(+)-tartrate (1:1)), bemorandane (RWJ 22867), MCI-154, UD-CG 212, sulma Hoffmann-LaRoche), methamphetamine (such as Des Zole, ampiZone, cilostamide, carbaZeran, piroXimone, ima oxynR), Abbot (S)- N, (alpha)-dimethylbenzeneetha Zodan, CI-930, siguaZodan, adibendan, saterinone, SKF namine hydrochloride)), and phendimetrazine tartrate (such 95654, SDZ-MKS-492, 349-U-85, emoradan, EMD-53998, as Bontril.R Slow-Release Capsules, Amarin (-3,4-Dim EMD-57033, NSP-306, NSP-307, revizinone, NM-702, ethyl-2-phenylmorpholine Tartrate); WIN-62582 and WIN-63291, enoximone and milrinone, fatty acid oxidation upregulator/inducers such as Famoxin R PDE3/4 inhibitors (such as benafentrine, trequinsin, ORG (Genset); 3.0029, Zardaverine, L-686398, SDZ-ISQ-844, ORG-20241, monamine oxidase inhibitors including but not limited to EMD-54622, and tolafentrine) and other PDE inhibitors befloxatone, moclobemide, brofaromine, phenoxathine, esu (such as Vinpocetin, papaverine, enprofylline, cilomilast, prone, befol, toloxatone, pirlindol, amiflamine, Sercloremine, fenoximone, pentoxifylline, roflumilast, tadalafil (Clalis(R), baZinaprine, lazabemide, millacemide, caroXaZone and other theophylline, and Vardenafil (Levitra(R): certain compounds as disclosed by WO01/12176; and Neuropeptide Y2 (NPY2) agonists include but are not limited other anti-obesity agents such as 5HT-2 agonists, ACC to: polypeptide YY and fragments and variants thereof (e.g. (acetyl-CoA carboxylase) inhibitors such as those described YY3-36 (PYY3-36) (N. Engl. J. Med. 349:941, 2003; in WO03/072197, alpha-lipoic acid (alpha-LA), AOD9604, IKPEAPGE DASPEELNRY YASLRHYLNL VTRQRY appetite suppressants such as those in WO03/401.07, ATL (SEQ ID NO:XXX)) and PYY agonists such as those dis 962 (Alizyme PLC), benzocaine, benzphetamine hydrochlo closed in WO02/47712, WO03/026591, WO03/057235, and ride (Didrex), bladderwrack (focus vesiculosus), BRS3 WO03/027637; (bombesin receptor Subtype 3) agonists, bupropion, caffeine, serotonin reuptake inhibitors, such as, paroxetine, fluoxetine CCKagonists, chitosan, chromium, conjugated linoleic acid, (ProzacTM), fluvoxamine, sertraline, citalopram, and imi corticotropin-releasing hormone agonists, dehydroepi pramine, and those disclosed in U.S. Pat. No. 6,162,805, U.S. androsterone, DGAT1 (diacylglycerol acyltransferase 1) Pat. No. 6,365,633, WO03/00663, WO01/27060, and WO01/ inhibitors, DGAT2 (diacylglycerol acyltransferase 2) inhibi 162341: tors, dicarboxylate transporter inhibitors, ephedra, exendin-4 thyroid hormone B agonists, such as KB-2611 (Karo (an inhibitor of glp-1) FAS (fatty acid synthase) inhibitors BioBMS), and those disclosed in WO02/15845, WO97/ (such as Cerulenin and C75), fat resorption inhibitors (such as 21993, WO99/00353, GB98/284425, U.S. Provisional Appli those in WO03/053451, and the like), fatty acid transporter cation No. 60/183.223, and Japanese Patent Application No. inhibitors, natural water soluble fibers (such as psyllium, JP 2000256190; plantago, guar, oat, pectin), galanin antagonists, galega UCP-1 (uncoupling protein-1), 2, or 3 activators, such as (Goat's Rue, French Lilac), garcinia cambogia, germander phytanic acid, 4-(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetram (teucrium chamaedrys), ghrelin antibodies and ghrelin ethyl-2-napthalenyl)-1-propenylbenzoic acid (TTNPB), ret antagonists (such as those disclosed in WOO1/87335, and inoic acid, and those disclosed in WO99/00123; WO02/08250), polypeptide hormones and variants thereof B3 (beta adrenergic receptor 3) agonists, such as AJ9677/ which affect the islet cell secretion, such as the hormones of TAK677 (Dainippon/Takeda), L750355 (Merck), CP331648 the secretin/gastric inhibitory polypeptide (GIP)/vasoactive (Pfizer), CL-316,243, SB 418790, BRL-37344, L-796568, intestinal polypeptide (VIP)/pituitary adenylate cyclase acti BMS-196085, BRL-35135A, CGP12177A, BTA-243, GW Vating polypeptide (PACAP)/glucagon-like polypeptide II 427353, Trecadrine, Zeneca D7114, N-5984 (Nisshin (GLP-II)/glicentin/glucagon gene family and/or those of the Kyorin), LY-377604 (Lilly), SR 59119A, and those disclosed adrenomedullinfamylin/calcitonin gene related polypeptide in U.S. Pat. No. 5,541,204, U.S. Pat. No. 5,770,615, U.S. Pat. (CGRP) gene family including GLP-1 (glucagon-like No. 5,491,134, U.S. Pat. No. 5,776,983, US488.064, U.S. Pat. polypeptide 1) agonists (e.g. (1) exendin-4, (2) those GLP-1 No. 5,705,515, U.S. Pat. No. 5,451,677, WO94/18161, molecules described in US20050130891 including GLP-1 (7- WO95/2915.9, WO97/.46556, WO98/04526 and WO98/ 34), GLP-1 (7-35), GLP-1 (7-36) or GLP-1 (7-37) in its 32753, WO01/74782, WO02/32897, WO03/014113, WO03/ C-terminally carboxylated or amidated form or as modified 016276, WO03/016307, WO03/024948, WO03/024953 and GLP-1 polypeptides and modifications thereof including WO03/037881: those described in paragraphs 17-44 of US20050130891, and noradrenergic agents including, but not limited to, diethyl derivatives derived from GLP-1-(7-34)COOH and the corre propion (such as Tenuate R (1-propanone, 2-(diethylamino)- sponding acid amide are employed which have the following 1-phenyl-, hydrochloride), Merrell), dextroamphetamine general formula: (also known as dextroamphetamine Sulfate, dexamphet amine, dexedrine, Dexampex, Ferndex. Oxydess II, Robese, Spancap #1), mazindol (or 5-(p-chlorophenyl)-2,5-dihydro R-NH-HAEGTFTSDVSYLEGOAAKEFIAWLVK- CONH, 3H-imidazo[2,1-aisoindol-5-ol) such as Sanorex R, Novartis wherein R=H or an organic compound having from 1 to 10 or Mazanor R, Wyeth Ayerst), phenylpropanolamine (or Ben carbon atoms. Preferably, R is the residue of a carboxylic Zenemethanol, alpha-(1-aminoethyl)-, hydrochloride), phen acid. Particularly preferred are the following carboxylic acid US 2012/0283411 A9 Nov. 8, 2012 residues: formyl, acetyl, propionyl, isopropionyl, methyl, naphthalenesulphonate, 1-glucosephosphate, nitrate, Sul ethyl, propyl, isopropyl. n-butyl, sec-butyl, tert-butyl.) and phite, dithionate and phosphate), and phenformin; glp-1 (glucagon-like polypeptide-1), glucocorticoid antago protein tyrosine phosphatase-1B (PTP-1B) inhibitors, such as nists, glucose transporter inhibitors, growth hormone secre A-401,674, KR 61639, OC-060062, OC-83839, OC-297962, tagogues (such as those disclosed and specifically described MC52445, MC52453, ISIS113715, and those disclosed in in U.S. Pat. No. 5,536,716), interleukin-6 (IL-6) and modu WO99/585521, WO99/58518, WO99/58522, WO99/61435, lators thereof (as in WO03/057237, and the like), L-carnitine, WO03/032916, WO03/032982, WO03/041729, WO03/ Mc3r (melanocortin 3 receptor) agonists, MCH2R (melanin 055883, WO02/26707, WO02/26743, JP2002114768, and concentrating hormone 2R) agonist/antagonists, melanin pharmaceutically acceptable salts and esters thereof; concentrating hormone antagonists, melanocortin agonists Sulfonylureas such as acetohexamide (e.g. Dymelor, Eli (such as Melanotan II or those described in WO99/64002 and Lilly), carbutamide, chlorpropamide (e.g. DiabineseR, WO 00/74.679), nomame herba, phosphate transporter inhibi Pfizer), gliamilide (Pfizer), gliclazide (e.g. Diamcron, Servier tors, phytopharm compound 57 (CP 644,673), pyruvate, Canada Inc), glimepiride (e.g. disclosed in U.S. Pat. No. SCD-1 (stearoyl-CoA desaturase-1) inhibitors, T71 (Tularik, 4,379,785, such as AmarylTM, Aventis), glipentide, glipizide Inc., Boulder Colo.), Topiramate (Topimax(R), indicated as an (e.g. Glucotrol or Glucotrol XL Extended Release, Pfizer), anti-convulsant which has been shown to increase weight gliquidone, glisolamide, glyburide/glibenclamide (e.g. Mic loss), transcription factor modulators (such as those disclosed ronase or Glynase Prestab, Pharmacia & Upjohn and Diabeta, in WO03/026576), B-hydroxy steroid dehydrogenase-1 Aventis), tolaZamide (e.g. Tolinase), and tolbutamide (e.g. inhibitors (B-HSD-1), B-hydroxy-3-methylbutyrate, p57 Orinase), and pharmaceutically acceptable salts and esters (Pfizer), Zonisamide (ZonegranTM, indicated as an anti-epi thereof; leptic which has been shown to lead to weight loss), and the meglitinides such as repaglinide (e.g. Pranidin R, Novo Nor agents disclosed in US200301 19428 paragraphs 20-26. disk), KAD1229 (PF/Kissei), and nateglinide (e.g. Starlix(R), 0874. The polypeptides and agonists described herein can Novartis), and pharmaceutically acceptable salts and esters be used in therapeutic combination with one or more anti thereof; a glucoside hydrolase inhibitors (or glucoside inhibi diabetic agents, including but not limited to: tors) such as acarbose (e.g. PrecoseTM, Bayer disclosed in PPARY agonists such as glitazones (e.g., WAY-120,744. AD U.S. Pat. No. 4,904.769), miglitol (such as GLYSETTM, Phar 5075, balaglitazone, ciglitazone, dargilitazone (CP-86325, macia & Upjohn disclosed in U.S. Pat. No. 4,639.436), cam Pfizer), englitazone (CP-68722, Pfizer), isaglitazone (MIT/ iglibose (Methyl 6-deoxy-6-(2R.3R.4R,5S)-3,4,5-trihy J&J), MCC-555 (Mitsibishi disclosed in U.S. Pat. No. 5,594, droxy-2-(hydroxymethyl)piperidino-alpha-D- 016), pioglitaZone (such as Such as ActosTM pioglitaZone; glucopyranoside, Marion Merrell Dow), Voglibose (Takeda), Takeda), rosiglitazone (AvandiaTM. Smith Kline Beecham), adiposine, emiglitate, pradimicin-Q, salbostatin, CKD-711, rosiglitaZone maleate, troglitaZone (ReZulin(R), disclosed in MDL-25,637, MDL-73,945, and MOR 14, and the com U.S. Pat. No. 4,572,912), rivoglitazone (CS-011, Sankyo), pounds disclosed in U.S. Pat. No. 4,062,950, U.S. Pat. No. GL-262570 (Glaxo Welcome), BRL49653 (disclosed in 4,174,439, U.S. Pat. No. 4,254,256, U.S. Pat. No. 4,701,559, WO98/05331), CLX-0921,5-BTZD, GW-0207, LG-100641, U.S. Pat. No. 4,639,436, U.S. Pat. No. 5,192,772, U.S. Pat. JJT-501 (JPNT/P&U), L-895.645 (Merck), R-1 19702 (San No. 4,634,765, U.S. Pat. No. 5,157,116, U.S. Pat. No. 5,504, kyo/Pfizer), NN-2344 (Dr. Reddy/NN), YM-440 (Yamanou 078, U.S. Pat. No. 5,091,418, U.S. Pat. No. 5,217,877, chi), LY-300512, LY-519818, R483 (Roche), T131 (Tularik), US51091 and WO01/47528 (polyamines): and the like and compounds disclosed in U.S. Pat. No. 4,687, C.-amylase inhibitors such as tendamistat, trestatin, and 777, U.S. Pat. No. 5,002,953, U.S. Pat. No. 5,741,803, U.S. A1-3688, and the compounds disclosed in U.S. Pat. No. Pat. No. 5,965,584, U.S. Pat. No. 6,150,383, U.S. Pat. No. 4,451,455, U.S. Pat. No. 4,623,714, and U.S. Pat. No. 4,273, 6,150,384, U.S. Pat. No. 6,166,042, U.S. Pat. No. 6,166,043, 765; U.S. Pat. No. 6,172,090, U.S. Pat. No. 6,211,205, U.S. Pat. SGLT2 inhibtors including those disclosed in U.S. Pat. No. No. 6,271,243, U.S. Pat. No. 6,288,095, U.S. Pat. No. 6,303, 6,414,126 and U.S. Pat. No. 6,515,117: 640, U.S. Pat. No. 6,329,404, U.S. Pat. No. 5,994,554, an aP2 inhibitor such as disclosed in U.S. Pat. No. 6,548,529; WO97/10813, WO97/27857, WO97/28115, WO97/28137, insulin secreatagogues such as linogliride, A-4166, forskilin, WO97/27847, WO00/76488, WO03/000685, WO03/ dibutyrl cAMP isobutylmethylxanthine (IBMX), and phar 027112, WO03/035602, WO03/048130, WO03/055867, and maceutically acceptable salts and esters thereof. pharmaceutically acceptable salts thereof; fatty acid oxidation inhibitors, such as clomoxir, and eto biguanides such as metformin hydrochloride (N,N-dimeth moxir, and pharmaceutically acceptable salts and esters ylimidodicarbonimidic diamide hydrochloride, such as Glu thereof; cophageTM, Bristol-Myers Squibb); metformin hydrochlo A2 antagonists, such as midaglizole, isaglidole, deriglidole, ride with glyburide, such as GlucovanceTM, Bristol-Myers idazoxan, earoxan, and fluparoxan, and pharmaceutically Squibb); buformin (Imidodicarbonimidic diamide, N-bu acceptable salts and esters thereof; tyl-): etoformine (1-Butyl-2-ethylbiguanide, Schering A.G.); insulin and related compounds (e.g. insulin mimetics) such as other metformin salt forms (including where the salt is chosen biota, LP-100, novarapid, insulin detemir, insulin lispro, insu from the group of acetate, benzoate, citrate, ftimarate, lin glargine, insulin Zinc suspension (lente and ultralente), embonate, chlorophenoxyacetate, glycolate, palmoate, Lys-Pro insulin, GLP-1 (1-36) amide, GLP-1 (73-7) (insulin aspartate, methanesulphonate, maleate, parachlorophenoxy tropin, disclosed in U.S. Pat. No. 5,614,492), LY-315902 isobutyrate, formate, lactate. Succinate, Sulphate, tartrate, (Lilly), GLP-1 (7-36)-NH2), AL-401 (AutoImmune), certain cyclohexanecarboxylate, hexanoate, octanoate, decanoate, compositions as disclosed in U.S. Pat. No. 4.579.730, U.S. hexadecanoate, octodecanoate, benzenesulphonate, tri Pat. No. 4,849,405, U.S. Pat. No. 4,963,526, U.S. Pat. No. methoxybenzoate, paratoluenesulphonate, adamantanecar 5,642,868, U.S. Pat. No. 5,763,396, U.S. Pat. No. 5,824,638, boxylate, glycoxylate, glutamate, pyrrolidonecarboxylate, U.S. Pat. No. 5,843.866, U.S. Pat. No. 6,153,632, U.S. Pat. US 2012/0283411 A9 Nov. 8, 2012 74

No. 6,191,105, and WO 85/05029, and primate, rodent, or 11603, 1999), P32/98, NVP-LAF-237, P3298, TSL225 (tryp rabbit insulin including biologically active variants thereof tophyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid, including allelic variants, more preferably human insulin disclosed by Yamada et al. Bioorg. & Med. Chem. Lett. 8 available in recombinant form (sources of human insulin (1998) 1537-1540), valine pyrrolidide, TMC-2A/2B/2C, include pharmaceutically acceptable and sterile formulations CD-26 inhibitors, FE999011, P9310/K364, VIP 0177, DPP4, such as those available from Eli Lilly (Indianapolis, Ind. SDZ 274-444, 2-cyanopyrrolidides and 4-cyanopyrrolidides 46285) as HumulinTM (human insulin rDNA origin), also see as disclosed by Ashworth et al. Bioorg. & Med. Chem. Lett. the THE PHYSICIAN'S DESK REFERENCE,55.sup...th Ed. Vol. 6, No. 22, pp 1163-1166 and 2745-2748 (1996), and the (2001) Medical Economics, Thomson Healthcare (disclosing compounds disclosed in U.S. Pat. No. 6,395,767, U.S. Pat. other Suitable human insulins): No. 6,573.287, U.S. Pat. No. 6,395,767 (compounds dis non-thiazolidinediones such as JT-501 and fargilitazar (GW closed include BMS-4771 18, BMS-471211 and BMS 538, 2570/GI-262579), and pharmaceutically acceptable salts and 305), WO99/38501, WO99/46272, WO99/67279, WO99/ esters thereof; 67278, WO99/61431 WO03/004498, WO03/004496, PPARO/Y dual agonists such as AR-HO39242 (AztraZeneca), EP1258476, WO02/083128, WO02/062764, WO03/000250, GW-409544 (Glaxo-Wellcome), BVT-142, CLX-0940, WO03/002530, WO03/002531, WO03/002553, WO03/ GW-1536, GW-1929, GW-2433, KRP-297 (Kyorin Merck; 002593, WO03/000180, and WO03/000181; 5-(2,4-Dioxo thiazolidinyl)methylmethoxy-N-(4-(trifluo GLP-1 agonists such as exendin-3 and exendin-4 (including romethyl)phenylmethylbenzamide), L-796449, LR-90, the 39 aa polypeptide synthetic exendin-4 called MK-0767 (Merck/Kyorin/Banyu), SB 219994, muraglitazar Exenatide(R), and compounds disclosed in US2003087821 (BMS), tesaglitzar (AstraZeneca), reglitazar (JTT-501) and and NZ 504256, and pharmaceutically acceptable salts and those disclosed in WO99/16758, WO99/19313, WO99/ esters thereof 20614, WO99/38850, WO00/23415, WO00/23417, WO00/ peptides including amlintide and Symlin R (pramlintide 23445, WO00/50414, WO01/00579, WO01/79150, WO02/ acetate); and 062799, WO03/004.458, WO03/016265, WO03/018010, glycokinase activators such as those disclosed in WO03/033481, WO03/033450, WO03/033453, WO03/ US2002103199 (fused heteroaromatic compounds) and 043985, WO 031053976, U.S. application Ser. No. 09/664, WO02/48106 (isoindolin-1-one-substituted propionamide 598, filed Sep. 18, 2000, Murakami et al. Diabetes 47, 1841 compounds). 1847 (1998), and pharmaceutically acceptable salts and The polypeptides and agonists described herein useful in the esters thereof; treatment of obesity can be administered as a cotherapy with other insulin sensitizing drugs; electrostimulation (US2004.0015201). VPAC2 receptor agonists: The polypeptides and agonists described herein can be used GLK modulators, such as those disclosed in WO03/015774; in combination therapy with agents that activate soluble gua retinoid modulators such as those disclosed in WOO3/ nylate cyclase, for example those described in 000249; US2004O19268O. GSK3 B/GSK3 inhibitors such as 4-2-(2-bromophenyl)-4- 0875. The polypeptides and agonists described herein can (4-fluorophenyl-1H-imidazol-5-ylpyridine and those com be used in combination therapy with a phosphodiesterase pounds disclosed in WO03/024447, WO03/037869, WO03/ inhibitor. PDE inhibitors are those compounds which slow 037877, WO03/037891, WO03/068773, EP1295884, the degradation of cyclic AMP (cAMP) and/or cyclic GMP EP1295885, and the like: (cGMP) by inhibition of the phosphodiesterases, which can glycogen phosphorylase (HGLPa) inhibitors such as CP-368, lead to a relative increase in the intracellular concentration of 296, CP-316,819, BAYR3401, and compounds disclosed in cAMP and/or cGMP Possible PDE inhibitors are primarily WO01/94300, WO02/20530, WO03/037864, and pharma those Substances which are to be numbered among the class ceutically acceptable salts or esters thereof; consisting of the PDE3 inhibitors, the class consisting of the ATP consumption promotors such as those disclosed in PDE4 inhibitors and/or the class consisting of the PDE5 WO03/007990; inhibitors, in particular those Substances which can be desig TRB3 inhibitors; nated as mixed types of PDE3/4 inhibitors or as mixed types vanilloid receptor ligands such as those disclosed in WO03/ of PDE3/4/5 inhibitors. By way of example, those PDE 049702: inhibitors may be mentioned such as are described and/or hypoglycemic agents such as those disclosed in WO03/ claimed in the following patent applications and patents: O15781 and WO03/040114; DE1470341, DE2108438, DE2123328, DE2305339, glycogen synthase kinase 3 inhibitors such as those disclosed DE2305575, DE2315801, DE2402908, DE2413935, in WOO3FO35663 DE2451417, DE2459090, DE2646469, DE2727481, agents such as those disclosed in WO99/51225, DE2825048, DE2837161, DE2845220, DE2847621, US2003.0134890, WO01/24786, and WO03/059870; DE2934747, DE3021792, DE3038166, DE3044568, insulin-responsive DNA binding protein-1 (IRDBP-1) as dis EP000718, EP0008.408, EP0010759, EP0059948, closed in WO03/057827, and the like: EP0075436, EP00965.17, EPO112987, EP0116948, adenosine A2 antagonists such as those disclosed in WO03/ EPO150937, EP0158380, EPO161632, EPO161918, 035639, WO03/035640, and the like: EP0167121, EP019.9127, EP0220044, EP0247725, PPARö agonists such as GW 501516, GW 590735, and com EP0258191 EP0272910, EP0272914, EP0294647, pounds disclosed in JP10237049 and WO02/14291; EP0300726, EP0335386, EP0357788, EP0389282, dipeptidyl peptidase IV (DP-IV) inhibitors, such as isoleu EP0406958, EP0426180, EP0428302, EP0435811, cine thiazolidide, NVP-DPP728A (1-2-(5-cyanopyridin EP0470805, EP0482208, EP0490823, EP0506194, 2-yl)aminoethylaminoacetyl-2-cyano-(S)-pyrrolidine, EP0511865, EP0527117, EP0626939, EP0664289, disclosed by Hughes et al. Biochemistry, 38(36), 11597 EP0671389, EP0685474, EP0685475, EP0685479, US 2012/0283411 A9 Nov. 8, 2012

JP92234389, JP94329652, JP950.10875, U.S. Pat. Nos. agonists described herein can be used in combination therapy 4,963,561, 5,141,931, WO9117991, WO9200968, (for example as in a chemotherapeutic composition) with an WO9212961, WO9307146, WO9315044, WO9315045, antiviral and monoclonal antibody therapies. WO9318024, WO9319068, WO9319720, WO931.9747, 0878 The polypeptides and agonists described herein can WO9319749, WO9319751, WO9325517, WO9402465, be used in combination therapy (for example, in prevention/ WO9406423, WO9412461, WO9420455, WO9422852, treatment of congestive heart failure or another method WO9425437, WO9427947, WO9500,516, WO950.1980, described herein) with the partial agonist of the nociceptin WO9503794, WO9504045, WO9504046, WO9505386, receptor ORL1 described by Dooley et al. (The Journal of WO9508534, WO9509623, WO9509624, WO9509627, Pharmacology and Experimental Therapeutics, 283 (2): 735 WO9509836, WO9514667, WO9514680, WO9514681, 741, 1997). The agonist is a hexapeptide having the amino WO9517392, WO9517399, WO9519362, WO9522520, acid sequence Ac-RYY (RK) (WI) (RK)-NH2 (“the Dooley WO9524381, WO9527692, WO9528926, WO9535281, polypeptide'), where the brackets show allowable variation WO9535282, WO9600218, WO9601825, WO9602541, of amino acid residue. Thus Dooley polypeptide can include WO961 1917, DE3142982, DE 1116676, DE2162096, but are not limited to KYYRWR, RYYRWR, KWRYYR, EP0293063, EP0463756, EP0482208, EP0579496, RYYRWK, RYYRWK (all-D amin acids), RYYRIK, EP0667345 U.S. Pat. No. 6,331,543, US20050004222 (in RYYRIR, RYYKIK, RYYKIR, RYYKWR, RYYKWK, cluding those disclosed in formulas I-XIII and paragraphs RYYRWR, RYYRWK, RYYRIK, RYYKWR, RYYKWK, 37-39, 85-0545 and 557-577) and WO9307124, EP0163965, RYYRWK and KYYRWK, wherein the amino acid residues EP0393500, EP0510562, EP0553174, WO950.1338 and are in the L-form unless otherwise specified. The polypep WO9603399. PDE5 inhibitors which may be mentioned by tides and agonists described herein can also be used in com way of example are RX-RA-69, SCH-51866, KT-734, bination therapy with polypeptide conjugate modifications of vesnarinone, Zaprinast, SKF-96231, ER-21355, BF/GP-385, the Dooley polypeptide described in WO0198324. NM-702 and sildenafil (Viagra R). PDE4 inhibitors which may be mentioned by way of example are RO-20-1724, MEM Methods of Treatment 1414 (R1533/R1500; Pharmacia Roche), DENBUFYL 0879. A number of disorders might be treated with GC-C LINE, ROLIPRAM, OXAGRELATE, NITRAQUAZONE, receptor agonists and agents that increase coMP levels Y-590, DH-6471, SKF-94120, MOTAPIZONE, LIXAZI including the polypeptides and agonists described herein. NONE, INDOLIDAN, OLPRINONE, ATIZORAM, 0880. The polypeptides and agonists described herein can KS-506-G, DIPAMFYLLINE, BMY-43351, ATIZORAM, be used alone or in combination therapy for the treatment or AROFYLLINE, FILAMINAST, PDB-093, UCB-29646, prevention of congestive heart failure. Such agents can be CDP-840, SKF-107806, PICLAMILAST, RS-17597, used in combination with natriuretic polypeptides (e.g., atrial RS-25344-000, SB-207499, TIBENELAST, SB-210667, natriuretic polypeptide, brain natriuretic polypeptide or SB-211572, SB-211600, SB-212066, SB-212179, C-type natriuretic polypeptide), a diuretic, or an inhibitor of GW-3600, CDP-840, MOPIDAMOL, ANAGRELIDE, angiotensin converting enzyme. IBUDILAST, AMRINONE, PIMOBENDAN, CILOSTA 0881. The polypeptides and agonists described herein can ZOL, QUAZINONE and N-(3,5-dichloropyrid-4-yl)-3-cy be used alone or in combination therapy for the treatment or clopropylmethoxy-4-difluoromethoxybenzamide. PDE3 prevention of benign prostatic hyperplasia (BPH). Such inhibitors which may be mentioned by way of example are agents can be used in combination with one or more agents for SULMAZOLE, AMPIZONE, CILOSTAMIDE, CARBAZ treatment of BPH, for example, a 5-alpha reductase inhibitor ERAN, PIROXIMONE, IMAZODAN, CI-930, SIGUA (e.g., finasteride) or an alpha adrenergic inhibitor (e.g., dox ZODAN, ADIBENDAN, SATERINONE, SKF-95654, SDZ aZosine). MKS-492, 349-U-85, EMORADAN, EMD-53998, EMD 0882. The polypeptides and agonists described herein can 57033, NSP-306, NSP-307, REVIZINONE, NM-702, WIN be used alone or in combination therapy for the treatment, 62582 and WIN-63291, ENOXIMONE and MILRINONE. prevention or reduction of visceral pain associated with a PDE3/4 inhibitors which may be mentioned by way of gastrointestinal disorder or pain associated with another dis example are BENAFENTRINE, TREQUINSIN, ORG order. 3.0029, ZARDAVERINE, L-686398, SDZ-ISQ-844, ORG 0883. The polypeptides and agonists described herein can 20241, EMD-54622, and TOLAFENTRINE. Other PDE be used alone or in combination therapy for the treatment or inhibitors include: cilomilast, pentoxifylline, roflumilast, tad prevention of obesity-related disorders (e.g. disorders that are alafil (Clalis(R), theophylline, and Vardenafil (Levitra(R), Zap associated with, caused by, or result from obesity). Examples rinast (PDE5 specific). of obesity-related disorders include overeating and bulimia, 0876 The polypeptides and agonists described herein can hypertension, diabetes, elevated plasma insulin concentra be used in combination therapy (for example, in order to tions and insulin resistance, dyslipidemias, hyperlipidemia, decrease or inhibituterine contractions) with a tocolytic agent endometrial, breast, prostate and colon cancer, osteoarthritis, including but not limited to beta-adrenergic agents, magne obstructive sleep apnea, cholelithiasis, gallstones, heart dis sium Sulfate, prostaglandin inhibitors, and calcium channel ease, abnormal heart rhythms and arrhythmias, myocardial blockers. infarction, congestive heart failure, coronary heart disease, 0877. The polypeptides and agonists described herein can Sudden death, stroke, polycystic ovarian disease, craniophar be used in combination therapy with an anti-neoplastic agents yngioma, the Prader-Willi Syndrome. Frohlich's syndrome, including but not limited to alkylating agents, epipodophyl GH-deficient subjects, normal variant short stature, Turner's lotoxins, nitrosoureas, antimetabolites, Vinca alkaloids, syndrome, and other pathological conditions showing anthracycline antibiotics, nitrogen mustard agents, and the reduced metabolic activity or a decrease in resting energy like. Particular anti-neoplastic agents may include tamoxifen, expenditure as a percentage of total fat-free mass, e.g., chil taxol, etoposide and 5-fluorouracil. The polypeptides and dren with acute lymphoblastic leukemia. The agents US 2012/0283411 A9 Nov. 8, 2012 76 described herein may be used to reduce or control body been described as the passage of stool less than a certain weight (or fat) or to prevent and/or treat obesity or other number (e.g. 3) of times per week. A number of conditions appetite related disorders related to the excess consumption can be associated with constipation. Constipation can be of food, ethanol and other appetizing Substances. The agents associated with numerous disorders and conditions. For may be used to modulate lipid metabolism, reduce body fat example, constipation can be (1) associated with the use of a (e.g. via increasing fat utilization) or reduce (or Suppress) therapeutic agent (e.g. antihypertensives, anticonvulsants, appetite (e.g. via inducing satiety). Further examples of obe antispasmodics, analgesics, anticholinergics, antidepres sity-related disorders are metabolic syndrome, also known as sants, antipsychotics, cation-containing agents, anticonvul syndrome X, insulin resistance syndrome, sexual and repro sants, ganglion blockers, Vinca alkaloids); (2) associated with ductive dysfunction, such as infertility, hypogonadism in a muscular, neuropathic, metabolic or endocrine disorder (in males and hirsutism in females, gastrointestinal motility dis cluding but not limited to myotonic dystrophy, dermamyosi orders, such as obesity-related gastroesophageal reflux, res tis, systemic Sclerosis, Sclerodoma, amyloidosis (neurologic piratory disorders, such as obesity-hypoVentilation syndrome (Pickwickian syndrome), cardiovascular disorders, inflam or muscular), ischemia, tumor of the central nervous system, mation, Such as systemic inflammation of the vasculature, autonomic neuropathy, Chagas disease, cystic fibrosis, dia arteriosclerosis, hypercholesterolemia, hyperuricaemia, betes mellitus, Hirschsprung disease, hyperthyroidism, lowerback pain, gallbladder disease, gout, and kidney cancer. hypocalcaemia, hypothyroidism, Multiple Sclerosis, neurofi The agents of the present invention are also useful for reduc bromatosis, Parkinson's disease, and spinal cord lesions (for ing the risk of secondary outcomes of obesity, Such as reduc example, related to sacral nerve damage related to trauma or ing the risk of left ventricular hypertrophy. a tumor or the enteric nervous system)); (3) post-Surgical 0884 The polypeptides and agonists described herein can constipation (postoperative ileus); (4) associated with a struc be used alone or in combination therapy for the treatment or tural colon alteration (for example that associated with Neo prevention of gastrointestinal related disorders including: plasm, stricture, Volvulus, anorectal, inflammation, prolapse, chronic intestinal pseudo-obstruction (Ogilvie’s syndrome), rectocele, or fissure); (5) associated with the agastrointestinal colonic pseudoobstruction, Crohn's disease, dyspepsia (in disorder; (6) associated with a systemic illness or disorder cluding functional dyspepsia or nonulcer dyspepsia), duode (for example, electrolyte abnormalities, thyroid disease, dia nogastric reflux, functional bowel disorder, functional gas betes mellitus, panhypopituitarism, Addison's disease, pheo trointestinal disorders, functional heartburn, chromocytoma, uremia, porphyria); (7) chronic constipation; gastroesophageal reflux disease (GERD), gastrointestinal (8) associated with the use of analgesic drugs (e.g. opioid motility disorders, gastroparesis (e.g. idopathic gastropare induced constipation); (9) associated with megacolon; and sis), hypertrophic pyloric Stenosis, Inflammatory bowel dis (10) idiopathic constipation (functional constipation). Func ease, irritable bowel syndrome (IBS), post-operative ileus, tional constipation can be associated with normal transit, and ulcerative colitis. The polypeptides and agonists slow transit (e.g. one or fewer bowel movements per week) described herein can be used alone or in combination therapy and pelvic floor dyssynergia. Pelvic floor dyssynergia is con to patient suffering from or susceptible to GI disorders relat sidered a disorder of the rectum and anus although these ing to damage to the GI tract stemming from impact or Sur patients also have abnormal contractions throughout the gical intervention. The polypeptides and agonists described colon. Patients with pelvic floor dyssynergia have abnormal herein can be used alone or in combination therapy to patients at risk for or having particular diseases associated with hypo colonic pressure waves prior to defecation and present with motility (e.g. colonic inertia) or stasis in the GI tract. For symptoms that may include a sensation of incomplete evacu example, diabetic neuropathy, anorexia nervosa, and achlo ation, excessive straining, a need for digital disimpaction, rhydria are frequently accompanied by gastric hypomotility. perianal heaviness, and tenesmus. Constipation can be asso Damage to the GI tract following Surgical intervention, for ciated with bloating and abdominal pain. The polypeptides instance, can result in Substantial gastric Stasis. The polypep and agonists described herein can be used to prevent and/or tides and agonists described herein can be administered alone treat low stool frequency or poor stool consistency. or in combination therapy to patients Susceptible to or having 0886. The polypeptides and agonists described herein can a GI disorder associated with diabetes (e.g. diabetic gastropa be used to treat decreased intestinal motility, slow digestion or thy). The polypeptides and agonists described herein can be slow stomach emptying. The polypeptides and agonists can used alone or in combination therapy to prevent and/or treat be used to relieve one or more symptoms of IBS (bloating, GI disorders characterized by at least one of nausea, vomit pain, constipation), GERD (acid reflux into the esophagus), ing, heartburn, postprandial discomfort, diarrhea, constipa duodenogastric reflux, functional dyspepsia, or gastroparesis tion, indigestion or related symptoms. The polypeptides and (nausea, vomiting, bloating, delayed gastric emptying) and agonists described herein can be used alone or in combination other disorders described herein. The polypeptides and ago therapy to prevent and/or treat GI disorders associated with at nists described herein can be used to treat flatulence. least one of diabetes, anorexia nervosa, bulimia, achlorhy 0887. The polypeptides and agonists described herein can dria, achalasia, anal fissure, haemorrhoids, irritable bowel be used to increase intestinal motility, slow colonic transit, syndrome, intestinal pseudoobstruction, Scleroderma and and to prevent and/or treat gastrointestinal immotility and gastrointestinal damage. other conditions calling for laxative or stool softener therapy. 0885. The polypeptides and agonists described herein can Gastrointestinal immotility can include constipation, and also be used to prevent and/or treat constipation. Constipation can includes delayed oral cecal transit time, irregular Taxation, be used to describe bowel patterns which include one or more and other related gastrointestinal motility disfunction includ of hard, Small, infrequent stools; the sensation of difficulty in ing impaction. Impaction is a condition where a large mass of passing stool, specifically excessive or ineffectual straining; dry, hard stool develops in the rectum, often due to chronic the sensation of incomplete evacuation. Constipation has also constipation. This mass may be so hard that it cannot be US 2012/0283411 A9 Nov. 8, 2012 77 excreted. The Subjects affected by constipation or gas of which are herein incorporated by reference. Pyrazol-1-yl) trointestinal immotility can be refractory to laxative therapy benzenesulfonamides have also been described as inhibitors and/or stool softener therapy. of cyclooxygenase-2. 0888. The polypeptides and agonists described herein can 0891. The polypeptides and agonists described herein can be used for the treatment or prevention of cancer, pre-cancer be used in the treatment or prevention of inflammation. Thus, ous growths, or metastatic growths. For example, they can be they can be used alone or in combination with an inhibitor of used for the prevention or treatment of: colorectal/local cGMP-dependent phosphodiesterase or a selective cyclooxy metastasized colorectal cancer, intestinal polyps, gastrointes genase-2 inhibitor for treatment of organ inflammation, IBD tinal tract cancer, lung cancer, cancer or pre-cancerous (e.g., Crohn's disease, ulcerative colitis), asthma, nephritis, growths or metastatic growths of epithelial cells, polyps, hepatitis, pancreatitis, bronchitis, cystic fibrosis, ischemic breast, colorectal, lung, ovarian, pancreatic, prostatic, renal, bowel diseases, intestinal inflammations/allergies, coeliac stomach, bladder, liver, esophageal and testicular carcinoma, disease, proctitis, eosinophilic gastroenteritis, mastocytosis, carcinoma (e.g., basal cell, basosquamous, Brown-Pearce, and other inflammatory disorders. The polypeptides and ago ductal carcinoma, Ehrlichtumor, Krebs, Merkel cell, small or nists described herein can be used alone or in combination non-Small cell lung, oat cell, papillary, bronchiolar, Squamous therapy in the treatment or prevention of gastrointestinal tract cell, transitional cell, (Walker), leukemia (e.g., B-cell, T-cell, inflammation (e.g. inflammation associated with a gas HTLV. acute or chronic lymphocytic, mast cell, myeloid), trointestinal disorder, gastrointestinal tract infection, or histiocytonia, histiocytosis, Hodgkin's disease, non another disorder). They can be used alone or in combination Hodgkin's lymphoma, plasmacytoma, reticuloendotheliosis, therapy with phenoxyalkycarboxylic acid derivatives for the adenoma, adeno-carcinoma, adenofibroma, adenolym treatment of interstitial cystitis, irritable bowel syndrome, phoma, ameloblastoma, angiokeratoma, angiolymphoid ulcerative colitis, and other inflammatory conditions, as men hyperplasia with eosinophilia, Sclerosing angioma, angioma tioned in US200502399.02A1. tosis, apudoma, branchionia, malignant carcinoid syndrome, 0892. The polypeptides and agonists described herein can carcinoid heart disease, carcinosarcoma, cementoma, cholan also be used to treat or prevent insulin-related disorders, for gioma, cholesteatoma, chondrosarcoma, chondroblastoma, example: II diabetes mellitus, hyperglycemia, obesity, disor chondrosarcoma, chordoma, choristoma, craniopharyn ders associated with disturbances in glucose or electrolyte gioma, chrondrorna, cylindroma, cystadenocarcinoma, cys transport and insulin secretion in cells, or endocrine disor tadenoma, cystosarconia phyllodes, dysgenninoma, ependy ders. They can be also used in insulin resistance treatment and moma, Ewing sarcoma, fibroma, fibrosarcoma, giant cell post-Surgical and non-post Surgery decrease in insulin tumor, ganglioneuroma, glioblastoma, glomangioma, granu responsiveness. losa cell tumor, gynandroblastoma, hamartoma, heman 0893. The polypeptides and agonists described herein can gioendothelioma, hemangioma, hemangio-pericytoma, be used to prevent and/or treat pulmonary and respiratory hemangiosarcoma, hepatoma, islet cell tumor, Kaposi sar related disorders, including, inhalation, ventilation and coma, leiomyoma, leiomyosarcoma, leukosarcoma, Leydig mucus secretion disorders, pulmonary hypertension, chronic cell tumor, lipoma, liposarcoma, lymphaugioma, lymphangi obstruction of vessels and airways, acute respiratory failure, omyoma, lymphangiosarcoma, medulloblastoma, menin and irreversible obstructions of vessels and bronchi. One may gioma, mesenchymoma, mesonephroma, mesothelioma, administer an agent described herein for treating broncho myoblastoma, myoma, myosarcoma, myxoma, myxosar spasm, for inducing bronchodilation, for treating chronic coma, neurilemmoma, neuroma, neuroblastoma, neuroepi obstructive pulmonary disease (including chronic bronchitis thelioma, neurofibroma, neurofibromatosis, odontoma, with normal airflow), for treating asthma (including bronchial osteoma, osteosarcoma, papilloma, paraganglioma, paragan asthma, intrinsic asthma, extrinsic asthma, acute asthma, glionia. nonchromaffin, pinealoma, rhabdomyoma, rhab chronic or inveterate asthma (e.g. late asthma and airways domyosarcoma, Sertoli cell tumor, teratoma, theca cell hyper-responsiveness), dust-induced asthma, allergen-in tumor, and other diseases in which cells have become dys duced asthma, viral-induced asthma, cold-induced asthma, plastic, immortalized, or transformed. pollution-induced asthma and exercise-induced asthma) and 0889. The polypeptides and agonists described herein can for treating rhinitis (including acute-, allergic, hatrophic be used for the treatment or prevention of Familial Adenoma rhinitis or chronic rhinitis (such as rhinitis caseosa, hyper tous Polyposis (FAP) (autosomal dominant syndrome) that trophic rhinitis, rhinitis purulenta, rhinitis sicca), rhinitis precedes colon cancer, hereditary nonpolyposis colorectal medicamentosa, membranous rhinitis (including croupous, cancer (HNPCC), and inherited autosomal dominant syn fibrinous and pseudomembranous rhinitis), scrofulous rhini drome. tis, perennial allergic rhinitis, seasonal rhinitis (including 0890 For treatment or prevention of cancer, pre-cancer rhinitis nervosa (hay fever) and vasomotor rhinitis). The ous growths and metastatic growths, the polypeptides and polypeptides described herein may also be useful in the treat agonists described herein can be used in combination therapy ment of dry eye disease and chronic sinusitis. The polypep with radiation or chemotherapeutic agents, an inhibitor of a tides described herein may also be used to prevent and/or treat cGMP-dependent phosphodiesterase or a selective cyclooxy disorders characterized by acute pulmonary vasoconstriction genase-2 inhibitor. A number of selective cyclooxygenase-2 Such as may result from pneumonia, traumatic injury, aspira inhibitors are described in US2001.0024664, U.S. Pat. No. tion or inhalation injury, fat embolism in the lung, acidosis 5,380,738, U.S. Pat. No. 5,344,991, U.S. Pat. No. 5,393,790, inflammation of the lung, adult respiratory distress syndrome, U.S. Pat. No. 5,434,178, U.S. Pat. No. 5,474,995, U.S. Pat. acute pulmonary edema, acute mountain sickness, post-car No. 5,510,368, WO02/062369, WO 96/06840, WO diac Surgery, acute pulmonary hypertension, persistent pull 96/03388, WO 96/03387, WO 96/19469, WO 96/25405, WO monary hypertension of the newborn, perinatal aspiration 95/15316, WO 94/15932, WO 94/27980, WO95/00501, WO syndrome, hyaline membrane disease, acute pulmonary 94/13635, WO 94/20480, and WO 94/26731, the disclosures thromboembolism, herapin-protamine reactions, sepsis, sta US 2012/0283411 A9 Nov. 8, 2012 tus asthmaticus or hypoxia (including iatrogenic hypoxia) sis, polyp syndrome, intestinal adenocarcinoma, Liddle Syn and other forms of reversible pulmonary vasoconstriction. drome, Brody myopathy, infantile convulsions, and chore Such pulmonary disorders also are also characterized by oathetosis inflammation of the lung including those associated with the 0897. The polypeptides and agonists described herein can migration into the lung of nonresident cell types including the be conjugated to another molecule (e.g., a diagnostic orthera various leucocyte subclasses. Also included in the respiratory peutic molecule) to target cells bearing the GC-C receptor, disorders contemplated are: bullous disease, cough, chronic e.g., cystic fibrosis lesions and specific cells lining the intes cough associated with inflammation or iatrogenic induced, tinal tract. Thus, they can be used to target radioactive moi airway constriction, pigeon fancier's disease, eosinophilic eties or therapeutic moieties (active moieties like a radionu bronchitis, asthmatic bronchitis, chronic bronchitis with air clide, an enzyme, a fluorescent label, a metal chelating group, way obstruction (chronic obstructive bronchitis), eosino a chemiluminescent label, a bioluminescent label, a chemo philic lung disease, emphysema, farmer's lung, allergic eye therapeutic, a toxin, an inactive prodrug, a radiosensitizing diseases (including allergic conjunctivitis, Vernal conjunc agent, a photodynamic agent) to the intestine to aid in imag tivitis, Vernal keratoconjunctivitis, and giant papillary con ing and diagnosing or treating colorectal/metastasized or junctivitis), idiopathic pulmonary fibrosis, cystic fibrosis, dif local colorectal cancer. In addition, they can be used to deliver antisense molecules or nucleic acid molecules (like normal fuse pan bronchiolitis and other diseases which are copies of the p53 tumor suppressor gene) to the intestinal characterized by inflammation of the lung and/or excess tract. The polypeptides and agonists described herein can also mucosal secretion. Other physiological events which are con be used to increase the number of GC-C molecules on the templated to be prevented, treated or controlled include plate Surface of a cell. In some embodiments the cell is a metasta let activation in the lung, chronic inflammatory diseases of the sized colorectal cancer cell. In one embodiment the polypep lung which result in interstitial fibrosis, such as interstitial tide or agonist described herein is therapeutically conjugated lung diseases (ILD) (e.g., idiopathic pulmonary fibrosis, or to a second agent. In certain embodiments, the second agent ILD associated with rheumatoid arthritis, or other autoim can be radioactive or radiostable. In certain embodiments the mune conditions), chronic obstructive pulmonary disease second agent can be selected from the group consisting of a (COPD) (such as irreversible COPD), chronic sinusitis, compound that causes cell death, a compound that inhibits fibroid lung, hypersensitivity lung diseases, hypersensitivity cell division, a compound that induces cell differentiation, a pneumonitis, idiopathic interstitial pneumonia, nasal conges chemotherapeutic, a toxin and a radiosensitizing agent. In tion, nasal polyposis, and otitis media. certain embodiments the second agent can be selected from 0894. The polypeptides and agonists described herein can the group consisting of methotrexate, doxorubicin, daunoru be used alone or in combitherapy to prevent or treat: retin bicin, cytosinarabinoside, etoposide, 5-4 fluorouracil, mel opathy, nephropathy, diabetic angiopathy, and edema forma phalan, chlorambucil, cis-platin, vindesine, mitomycin, bleo tion mycin, purothionin, macromomycin, 1,4-benzoquinone derivatives, trenimon, ricin, ricin A chain, Pseudomonas exo 0895. The polypeptides and agonists described herein can toxin, diphtheria toxin, Clostridium perfiringens phospholi be used alone or in combitherapy to prevent or treat neuro pase C. bovine pancreatic ribonuclease, pokeweed antiviral logical disorders, for example, headache, tension-type head protein, abrin, abrin A chain, cobra Venom factor, gelonin, ache, migraines, anxiety, stress, cognitive disorders, cerebral saporin, modeccin, Viscumin, Volkensin, nitroimidazole, ischemia, brain trauma, movement disorders, aggression, metronidazole and misonidazole. In certain embodiments the psychosis, seizures, panic attacks, hysteria, sleep disorders, second agent can be a cytoxic agent selected from the group depression, Schizoaffective disorders, sleep apnea, attention consisting of cemadotin, a derivative of cemadotin, a deriva deficit syndromes, memory loss, dementia, memory and tive of hemiasterlin, esperamicin C, neocarzinostatin, may learning disorders as discussed in Moncada and Higgs 1995 tansinoid DM1, 7-chloromethyl-10, 11 methylenedioxy FASEB.J. 9:1319-1330; Severina 1998 Biochemistry 63:794; camptothecin, rhizoxin, and the halichondrin B analog, Lee et al. 2000 PNAS 97: 10763-10768; Hobbs 1997 TIPS ER-086526. 18:484-491; Murad 1994 Adv. Pharmacol. 26:1-335; and 0898. The polypeptides and agonists described herein can Denningeret al. 1999 Biochim. Biophys. Acta 1411:334-350 be used alone or in combination therapy to prevent and/or and narcolepsy. They may also be used as a sedative. treat inner ear disorders, e.g., to prevent and/or treat 0896. The polypeptides and detectably polypeptides and Meniere's disease (including symptoms thereof Such as Ver agonists described herein can be used as markers to identify, tigo, hearing loss, tinnitus, sensation of fullness in the ear), detect, stage, or diagnosis diseases and conditions of Small Mal de debarquement syndrome, otitis externa, otitis media, intestine, including, without limitation: Crohn's disease, coli otorrhea, acute mastoiditis, otosclerosis, otic pain, otic bleed tis, inflammatory bowel disease, tumors, benign tumors. Such ing, otic inflammation, Lermoyez's syndrome, Vestibular as benign stromal tumors, adenoma, angioma, adenomatous neuronitis, benign paroxysmal positional vertigo (BPPV), (pedunculated and sessile) polyps, malignant, carcinoid herpes Zosteroticus, Ramsay Hunt's syndrome, herpes, laby tumors, endocrine cell tumors, lymphoma, adenocarcinoma, rinthitis, purulent labyrinthitis, perilymph fistulas, presbycu foregut, midgut, and hindgut carcinoma, gastroinstestinal sis, ototoxicity (including drug-induced ototoxicity), neuro stromal tumor (GIST), such as leiomyoma, cellular lei mias (including acoustic neuromas), aerotitis media, omyoma, leiomyoblastoma, and leiomyosarcoma, gas infectious myringitis, bullous myringitis, squamous cell car trointestinal autonomic nerve tumor, malabsorption syn cinoma, basal cell carcinoma, pre-cancerous otic conditions, dromes, celiac diseases, diverticulosis, Meckel's nonchromaffin paragangliomas, chemodectomas, glomus diverticulum, colonic diverticula, megacolon, Hirschs jugulare tumors, glomus tympanicum tumors, perichondritis, prung's disease, irritable bowel syndrome, mesenteric aural eczematoid dermatitis, malignant external otitis, Sub ischemia, ischemic colitis, colorectal cancer, colonic polypo perichondrial hematoma, ceruminomas, impacted cerumen, US 2012/0283411 A9 Nov. 8, 2012 79 sebaceous cysts, osteomas, keloids, otalgia, tinnitus, tym and recessive autosomal recessive polycystic kidney disease panic membrane infection, tympanitis, otic furuncles, (ADPKD), in all stages of development, regardless of the petrositis, conductive and sensorineural hearing loss, epidu underlying cause. ral abscess, lateral sinus thrombosis, Subdural empyema, 0902. The polypeptides and agonists described herein can otitic hydrocephalus, Dandy's syndrome, bullous myringitis, be used alone or in combination therapy to prevent and/or diffuse external otitis, foreign bodies, keratosis obturans, otic treat disorders associated with bicarbonate secretion, e.g., neoplasm, otomycosis, trauma, acute barotitis media, acute Cystic Fibrosis. eustachian tube obstruction, postSurgical otalgia, choleste 0903. The polypeptides and agonists described herein can atoma, infections related to an otic Surgical procedure, and be used alone or in combination therapy to prevent and/or complications associated with any of said disorders. The treat disorders associated with bile secretion. In addition, they polypeptides and agonists described herein can be used alone can be used to facilitate or control chloride and bile fluid or in combination therapy to maintain fluid homeostasis in the secretion in the gallbladder. inner ear, neuronitis (including viral neuronitis), ganglionitis, 0904. The polypeptides and agonists described herein can geniculate. be used alone or in combination therapy to prevent and/or treat disorders associated with liver cell regeneration. This 0899. The polypeptides and agonists described herein can may include administration of the polypeptides and agonists be used alone or in combination therapy to prevent and/or to liver transplant recipients and to patients with drug or treat disorders associated with fluid and sodium retention, alcohol induced-liver damage. Furthermore, the polypeptides e.g., diseases of the electrolyte-water/electrolyte transport and agonists may be useful to treat liver damage as in the case system within the kidney, gut and urogenital system, conges of viral mediated hepatitis. The polypeptides and agonists tive heart failure, hypertension, hypotension, salt dependent described herein may be used alone or in combination to forms of high blood pressure, hepatic edema, and liver cir prevent and/or treat liver abscess, liver cancer (either primary rhosis. In addition they can be used to facilitate diuresis or or metastatic), cirrhosis (such as cirrhosis caused by the alco control intestinal fluid. The polypeptides and agonists hol consumption or primary biliary cirrhosis), amebic liver described herein can also be used to treat disorders where abscess, autoimmune hepatitis, biliary atresia, coccidioido there is abnormal proliferation of epithelial cells within the mycosis disseminated, 6 agent (hepatitis Ö), hemochromato kidney (e.g. as in the case of renal cancer). In some cases, the sis, hepatitis a, hepatitis b, hepatitis c, or any other acute, subacute, fulminant or chronic hepatitis of viral, metabolic or methods entail treatment of an individual that is saltsensitive, toxic etiology, hepatocellular carcinoma, pyogenic liver e.g., an individual that exhibits greater than average changes abscess, Reye's syndrome, Sclerosing cholangitis, Wilson's in blood pressure in response to changes in Salt intake. Some disease, drug induced hepatotoxicity, or fulminant or acute salt sensitive individuals may have a greater than 5-mm Hg liver failure. The polypeptides and agonists may be used in decrease in mean blood pressure when daily salt intake is stimulating hepatic regeneration after Surgical hepatectomy. reduced from 260 mmol to 20 mmol. There are various tests 0905. The polypeptides and agonists described herein can for assessing salt sensitivity (see, for example, de la Sierra et be used alone or in combination therapy to prevent and/or al. 2002 Journal of Human Hypertension 16:256). treat myocardial infraction, coronary artery disease, nitrate 0900. The polypeptides and agonists described herein can induced tolerance, nitrate tolerance, diastolic dysfunction, be used alone or in combination therapy to prevent and/or angina pectoris, stable, unstable and variant (Prinzmetal) treat kidney disease. "Kidney disease' includes renal failure angina, atherosclerosis, thrombosis, endothelial dysfunction, (including acute renal failure), renal insufficiency, nephrotic cardiac edema, stroke, conditions of reduced blood vessel edema, glomerulonephritis, pyelonephritis, kidney failure, patency, e.g., postpercutaneous transluminal coronary angio chronic renal failure, nephritis, nephrosis, azotemia, uremia, plasty (post-PTCA), and peripheral vascular disease. immune renal disease, acute nephritic syndrome, rapidly pro 0906. The polypeptides and agonists described herein can gressive nephritic syndrome, nephrotic syndrome, Berger's be used alone or in combination therapy to prevent and/or Disease, chronic nephritic/proteinuric syndrome, tubuloint treat glaucoma. erstital disease, nephrotoxic disorders, renal infarction, athe 0907. The polypeptides and agonists described herein can roembolic renal disease, renal cortical necrosis, malignant be used alone or in combination therapy to prevent and/or nephroangiosclerosis, renal vein thrombosis, renal tubular treat immunodeficiency. acidosis, renal glucosuria, nephrogenic diabetes insipidus, 0908. The polypeptides and agonists described herein can Bartter's Syndrome, Liddle's Syndrome, polycystic kidney be used alone or in combination therapy to prevent and/or disease, medullary cystic disease, medullary sponge kidney, treat bladder outlet obstruction and incontinence. hereditary nephritis, and nail-patella syndrome, along with 0909. The polypeptides and agonists described herein can any disease or disorder that relates to the renal system and be used alone or in combination therapy to prevent and/or related disorders, as well as symptoms indicative of, or treat male (e.g. erectile dysfunction) or female sexual dys related to, renal or kidney disease and related disorders. function, dysmenorrhea, endometriosis, polycystic ovary 0901. The polypeptides and agonists described herein can syndrome, vaginal dryness, uterine pain, or pelvic pain. These be used alone or in combination therapy to prevent or treat polypeptides and agonists described herein can be utilized as polycystic kidney disease. Polycystic kidney disease” “PKD' tocolytic agents that decrease or arrest uterine contractions. (also called “polycystic renal disease') refers to a group of The polypeptides and agonists described herein can be used disorders characterized by a large number of cysts distributed to prevent/treat premature/preterm labor. Premature or pre throughout dramatically enlarged kidneys. The resultant cyst term labor can be associated with, for example, an illness/ development leads to impairment of kidney function and can disorder/condition of the mother (such as pre-eclampsia, high eventually cause kidney failure. “PKD’specifically includes blood pressure or diabetes, abnormal shape or size of the autosomal dominant polycystic kidney disease (ADPKD) uterus, weak or short cervix, hormone imbalance, vaginal US 2012/0283411 A9 Nov. 8, 2012

infection that spreads to the uterus, abnormalities of the pla 0913. The polypeptide can be administered to maintain centa, Such as placenta previa, and excessive amniotic fluid), regular bowel movements in a patient who is a chronic opioid premature rupture of the amniotic membranes (“water user (e.g., a terminally-ill patient). The administration can be breaks”), large fetus, and more than one fetus. The polypep via any convenient route (e.g., Sublingual, parenteral, intra tides or agonists described herein can be used to prevent venous, Subcutaneous). uterine rupture. The polypeptides or agonists described 0914. Thus, the polypeptides described herein can be herein can be used treat rapid uterine contractions (for administered to a patient that is taking one or more of the example, associated with placental abruption wherein the following opioids: Acetorphine, Acetyldihydrocodeine, placental abruption is associated with hypertension, diabetes, Acetylmorphone, Alfentanil, Allylprodine, Anileridine, a multiply pregnancy, an unusually large amount of amniotic Bemidone, Benzylmorphine, Bezitramide, Buprenorphine, fluid, numerous previous deliveries, or advanced maternal Butorphanol, Carfentanil/Carfentanyl, ClonitaZene, age (e.g. D40 years old). In certain embodiments they can be Codeine, Codeine-N-Oxide, Codeinone, Cyclazocine, used in combination with a phosphodiesterase inhibitor. The Cyclorphan, Desomorphine, Dextromoramide, Dextropro polypeptides and agonists described herein can be used alone poxyphene, Dezocine, Diacetyldihydromorphine, Diamor or in combination therapy to prevent and/or treat infertility, phine/Diacetylmorphine (Heroin), Diethylthiambutene, for example, male infertility due to poor sperm quality, Difenoxin, Dihydrocodeine, Dihydrocodeinone Enol decreased sperm motility or low sperm count. Acetate, Dihydroetorphine, Dihydroisocodeine, Dihydro 0910. The polypeptides and agonists described herein can morphine, Dimethylthiambutene, Diphenoxylate, Dipro be used alone or in combination therapy to prevent and/or panoylmorphine, Drobetabol, Ethylketocyclazocine, Ethyl treat osteopenia disorders (bone loss disorders). “Bone loss morphine, EtonitaZene, Etorphine, Fentanyl. Hydrocodone, disorders' include conditions and diseases wherein the inhi Hydromorphone, Isomethadone, Ketobemidone, Laudanum, bition of bone loss and/or the promotion of bone formation is Lefetamine, Levallorphan, Levo-Alphacetylmethadol desirable. Among such conditions and diseases are (LAAM), Levomethorphan, Levorphanol, Loperamide, osteoporosis, osteomyelitis, Paget's disease (osteitis defor Meptazinol, Metazocine, Methadone, Monoacetylmorphine, mans), periodontitis, hypercalcemia, osteonecrosis, osteosa Morphine, Morphine-6-Glucuronide, Morphine-N-Oxide, rcoma, osteolyic metastases, familial expansile osteolysis, Morphinone, MPPP (1-Methyl 4-Phenyl 4-Propionoxypip prosthetic loosening, periprostetic osteolysis, bone loss atten eridine), Myorphine, Nalbuphine/Nalbufine, Nicocodeine, dant rheumatoid arthritis, and cleiodocranial dysplasia Nicodicodeine, Nicomorphine, Norcodeine, Ohmefentanyl, (CCD). Osteoporosis includes primary osteoporosis, endo Oxycodone, Oxymorphone, Pentazocine, PEPAP (1-Phen crine osteoporosis (hyperthyroidism, hyperparathyroidism, ethyl-4-Phenyl-4-Piperidinol Acetate (Ester)), Pethidine Cushing's syndrome, and acromegaly), hereditary and con (Meperidine), Phenadoxone, Phenazocine, Phenoperidine, genital forms of osteoporosis (osteogenesis imperfecta, Pholcodeine, Piminodine, Piritramide, Prodine, Propiram, homocystinuria, Menkes syndrome, and Rile-Day Syn Propoxyphene, Racemethorphan, Remifentanil, Sufentanil, drome) and osteoporosis due to immobilization of extremi Thebaine. Thiofentanil/Thiofentanyl. Tilidine, and Trama tiesosteomyelitis, or an infectious lesion in bone leading to dol. The peptide can be co-administered with or co-formu bone loss. The polypeptides and agonists can be used alone or lated with any of the preceeding peptides. in combination therapy to stimulating bone regeneration. The (0915. Where the polypeptide described herein is co-for bone regeneration may be following reconstruction of bone mulated with an opioid the composition may further include defects in cranio-maxillofacial Surgery, or following an one or more other active ingredients that may be convention implant into bone, for example a dental implant, bone Sup ally employed in analgesic and/or cough-cold-antitussive porting implant, or prosthesis. The bone regeneration may combination products. Such conventional ingredients also be following a bone fracture. include, for example, aspirin, acetaminophen, phenylpro 0911. The polypeptides and agonists described herein panolamine, phenylephrine, chlorpheniramine, caffeine, and/ may be used alone or in combination therapy (for example, or guaifenesin. Typical or conventional ingredients that may with other agents that increase coiMP) to prevent or treat be included in the opioid component are described, for disorders related to an alteration in coMP including, but not example, in the Physicians’ Desk Reference, 1999, the dis limited to Alzheimer's disease, psoriasis, skin necrosis, scar closures of which are hereby incorporated herein by refer ring, fibrosis, baldness, Kawasaki's Disease, nutcracker ence, in their entirety. esophagus (US20050245544), septic shock, NSAID-induced 0916. In addition, the composition may further include gastric disease or disorder, ischemic renal disease or disorder, one or more compounds that may be designed to enhance the peptic ulcer, sickle cell anemia, epilepsy, and a neuroinflam analgesic potency of the opioid and/or to reduce analgesic matory disease or disorder (for example as described in tolerance development. Such compounds include, for WO05105765). example, dextromethorphan or other NMDA antagonists (Mao, M.J. et al., Pain 1996, 67,361), L-364,718 and other Treatment of the Side-Effects of Opioid Administration CCKantagonists (Dourish, C.T. et al., EurJ Pharmacol 1988, 0912. The polypeptides described herein (e.g. the GCC 147, 469), NOS inhibitors (Bhargava, H.N. et al., Neuropep agonist polypeptides described herein) may be useful in the tides 1996, 30, 219), PKC inhibitors (Bilsky, E. J. et al., J treatment of one or more side effects of opioid administration, Pharmacol Exp Ther 1996, 277, 484), and dynorphin antago e.g., opioid induced constipation, nausea and/or vomiting. In nists or antisera (Nichols, M. L. et al., Pain 1997, 69,317). the case of constipation, the polypeptide can be administered The disclosures of each of the foregoing documents are at a dosage to induce laxation within a desired time (e.g., hereby incorporated herein by reference, in their entireties. within 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 0917. The combination products, such as pharmaceutical hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 12 compositions comprising opioids in combination with a hours, 18 hours or 24 hours). polypeptide described herein may be in any dosage form, US 2012/0283411 A9 Nov. 8, 2012

Such as those described herein, and can also be administered ug. 600 to 700 ug. 600 to 800 ug, 600 to 900 ug, 600 to 1000 in various ways, as described herein. In a preferred embodi ug. 600 to 1250 ug, 600 to 1500 g, 600 to 1750 ug, 600 to ment, the combination products of the disclosure are formu 2000 ug, 600 to 2250 ug, 600 to 2500 g, 600 to 2750 ug, 600 lated together, in a single dosage form (that is, combined to 3000 ug,700 to 800 ug,700 to 900 ug,700 to 1000 ug. 700 together in one capsule, tablet, powder, or liquid, etc.). When to 1250 ug. 700 to 1500 ug. 700 to 1750 ug. 700 to 2000 ug, the combination products are not formulated together in a 700 to 2250 ug, 700 to 2500 pig, 700 to 2750 ug, 700 to 3000 single dosage form, the opioid compounds and the polypep ug,800 to 900 ug,800 to 1000 ug,800 to 1250 ug,800 to 1500 tides described herein may be administered at the same time ug. 800 to 1750 ug, 800 to 2000 ug, 800 to 2250 ug, 800 to (that is, together), or in any order. When not administered at 2500 ug,800 to 2750 ug,800 to 3000 ug,900 to 1000 ug,900 the same time, preferably the administration of an opioid and to 1250 ug,900 to 1500 ug,900 to 1750 ug,900 to 2000 ug, a polypeptide described herein occurs less than about one 900 to 2250 ug,900 to 2500 ug,900 to 2750 ug,900 to 3000 hour apart, less than about 30 minutes apart, less than about ug. 1000 to 1250 ug, 1000 to 1500 ug, 1000 to 1750 ug, 1000 15 minutes apart, and less than about 5 minutes apart. Admin to 2000 ug. 1000 to 2250 ug, 1000 to 2500 ug, 1000 to 2750 istration of the combination of an opioid and a polypeptide ug. 1000 to 3000 g, 2 to 500 ug, 50 to 500 lug. 3 to 100 ug. 5 described herein can be, for example, oral, although other to 20 Jug, 5 to 100 ug, 10g, 201g, 30 Jug, 40 ug, 50g, 60 ug, routes of administration, as described above, are contem 70 ug, 75ug, 80 ug,90 ug. 100 ug. 150 lug. 200g, 250 ug,300 plated to be within the scope of the present disclosure. ug, 350 ug, 400 ug, 450 ug, 500 ug, 550 ug, 600 g. 650 jug, Although it is the opioids and polypeptides described herein 700 ug, 750 ug. 800 ug. 850 ug,900 ug,950 ug. 1000 ug. 1050 may both be administered in the same fashion (that is, for ug, 1100 ug. 1150 ug, 1200 Lig, 1250 ug. 1300 ug. 1350 ug, example, both orally), if desired, they may each be adminis 1400 lug. 1450g, 1500 lug. 1550 ug. 1600 ug. 1650 ug. 1700 tered in different fashions (that is, for example, one compo ug. 1750 lug. 1800 ug, 1850 ug. 1900 ug. 1950 ug. 2000 ug, nent of the combination product may be administered orally, 2050 ug. 2100 ug. 2150 lug. 2200 lug. 2250 ug. 2300 ug. 2350 and another component may be administered intravenously). ug, 2400 ug. 2450 ug, 2500 g, 2550 ug. 2600 ug. 2650 ug, The dosage of the combination products of the disclosure 2700 g, 2750 ug, 2800 ug. 2850 ug, 2900 ug, 2950 ug, 3000 may vary depending upon various factors such as the phar ug. 3250 lug. 3500 g, 3750 ug, 4000 ug, 4250 ug, 4500 ug, macodynamic characteristics of the particular agent and its 4750 ug. 5000 ug of a polypeptide described herein. mode and route of administration, the age, health and weight 0919. When provided as a single dosage form, the poten of the recipient, the nature and extent of the symptoms, the tial exists for a chemical interaction between the combined kind of concurrent treatment, the frequency of treatment, and active ingredients (for example, an opioid and a polypeptide the effect desired. described herein). For this reason, the preferred dosage forms 0918. Although the proper dosage of the combination of the combination products of this disclosure are formulated products of this disclosure will be readily ascertainable by Such that although the active ingredients are combined in a one skilled in the art, by way of general guidance, where an single dosage form, the physical contact between the active opioid compounds is combined with a polypeptide described ingredients is minimized (that is, reduced). herein, for example, typically a daily dosage may range from 0920. In order to minimize contact, one embodiment of about 0.01 to about 100 milligrams, 0.1 to about 10 milli this disclosure where the product is orally administered pro grams of the opioid, 15 to about 200 milligrams, 1, 2, 3, 4, 5, vides for a combination product wherein one active ingredi 6,7,8,9, 10 milligrams of opioid per kilogram of patient body ent is enteric coated. By enteric coating one or more of the weight. The opioid-GCC agonist combination product can active ingredients, it is possible not only to minimize the include, for example, from 1 to 30 Lig, 1 to 40 ug, 1 to 50 ug, contact between the combined active ingredients, but also, it 1 to 100 ug, 1 to 200 ug. 1 to 300 ug, 1 to 400 lug. 1 to 500 ug, is possible to control the release of one of these components 1 to 600 ug. 1 to 700 ug, 1 to 800 ug, 1 to 900 ug, 1 to 1000 ug, in the gastrointestinal tract Such that one of these components 10 to 30 Jug, 10 to 40 ug. 10 to 50 lug. 10 to 100 ug. 10 to 200 is not released in the stomach but rather is released in the ug. 10 to 300 g, 10 to 400 ug. 10 to 500 ug. 10 to 600 ug. 10 intestines. Another embodiment of this disclosure where oral to 700 ug. 10 to 800 ug, 10 to 900 ug, 10 to 1000 ug, 100 to 200 administration is desired provides for a combination product ug. 100 to 300 ug. 100 to 400 ug. 100 to 500 ug. 100 to 600 ug, wherein one of the active ingredients is coated with a Sus 100 to 700 ug. 100 to 800 ug, 100 to 900 ug, 100 to 1000 ug, tained-release material which effects a Sustained-release 100 to 1250 ug, 100 to 1500 ug, 100 to 1750 ug, 100 to 2000 throughout the gastrointestinal tract and also serves to mini ug, 100 to 2250 ug, 100 to 2500 ug, 100 to 2750 ug, 100 to mize physical contact between the combined active ingredi 3000 ug, 200 to 300 ug, 200 to 400 ug, 200 to 500 ug, 200 to ents. Furthermore, the Sustained-released component can be 600 ug, 200 to 700 ug. 200 to 800 ug, 200 to 900 ug, 200 to additionally enteric coated such that the release of this com 1000 ug. 200 to 1250 ug, 200 to 1500 ug, 200 to 1750 ug, 200 ponent occurs only in the intestine. Still another approach to 2000 ug, 200 to 2250 ug, 200 to 2500 ug, 200 to 2750 ug, would involve the formulation of a combination product in 200 to 3000 ug, 300 to 400 ug, 300 to 500 ug, 300 to 600 ug, which the one component is coated with a Sustained and/or 300 to 700 ug,300 to 800 ug,300 to 900 ug,300 to 1000 ug, enteric release polymer, and the other component is also 300 to 1250 ug,300 to 1500 g, 300 to 1750 ug,300 to 2000 coated with a polymer Such as a low-viscosity grade of ug, 300 to 2250 ug, 300 to 2500 ug. 300 to 2750 ug, 300 to hydroxypropyl methylcellulose (HPMC) or other appropriate 3000 ug, 400 to 500 ug, 400 to 600 ug, 400 to 700 ug, 400 to materials as known in the art, in order to further separate the 800 ug, 400 to 900 ug, 400 to 1000 ug,400 to 1250 ug, 400 to active components. The polymer coating serves to form an 1500 ug, 400 to 1750 ug, 400 to 2000 ug, 400 to 2250 ug, 400 additional barrier to interaction with the other component. to 2500 ug, 400 to 2750 ug,400 to 3000 ug,500 to 600 ug,500 0921 Dosage forms of the combination products include to 700 ug, 500 to 800 ug, 500 to 900 ug, 500 to 1000 ug, 500 those wherein one active ingredient is enteric coated can be in to 1250 ug, 500 to 1500 ug, 500 to 1750 ug, 500 to 2000 ug, the form of tablets such that the enteric coated component and 500 to 2250 pig, 500 to 2500 g, 500 to 2750 ug,500 to 3000 the other active ingredient are blended together and then US 2012/0283411 A9 Nov. 8, 2012

compressed into a tablet or Such that the enteric coated com d) a polypeptide comprising amino acids 1-72 of SEQID ponent is compressed into one tablet layer and the other active NO:X; ingredient is compressed into an additional layer. Optionally, e) a polypeptide comprising amino acids 17-47 of SEQID in order to further separate the two layers, one or more pla NO:X; cebo layers may be present Such that the placebo layer is f) a polypeptide comprising amino acids 17-61 of SEQID between the layers of active ingredients. In addition, dosage NO:X; forms of the present disclosure can be in the form of capsules g) a polypeptide comprising amino acids 17-72 of SEQID wherein one active ingredient is compressed into a tablet or in NO:X; the form of a plurality of microtablets, particles, granules or h) a polypeptide comprising amino acids 17-94 of SEQID non-perils, which are then enteric coated. These enteric NO:X; coated microtablets, particles, granules or non-perils are then i) a polypeptide comprising amino acids 48-61 of SEQID placed into a capsule or compressed into a capsule along with NO:X; a granulation of the other active ingredient. j) a polypeptide comprising amino acids 48-72 of SEQID 0922. These as well as other ways of minimizing contact NO:X; between the components of combination products of the k) a polypeptide comprising amino acids 48-94 of SEQID present disclosure, whether administered in a single dosage NO:X; form or administered in separate forms but at the same time 1) a polypeptide comprising amino acids 62-72 of SEQID by the same manner, will be readily apparent to those skilled NO:X; in the art in light of the present disclosure. m) a polypeptide comprising amino acids 62-94 of SEQID NO:X; Peptides as Immunogens n) a polypeptide comprising amino acids 73-94 of SEQID NO:X; 0923. The polypeptides described herein can be used as o) a polypeptide comprising amino acids 1-23 of SEQID immunogens to create antibodies for immunoassays. The NO:X; polypeptides described herein can be used as immunogens to p) a polypeptide comprising amino acids 48-66 of SEQID treat and/or prevent one or more disease symptoms associated NO:X; with traveler's diarrhea and for vaccination against patho q) a polypeptide comprising amino acids 48-71 of SEQID gens, including but not limited to enterotoxigenic E. coli NO:X; (ETEC). They may also be used in vaccines which also com r) a polypeptide comprising amino acids 48-76 of SEQID prise interleukin 18 and either saponin adjuvant or CpGadu NO:X; vant for example as described in WO05039634 and s) a polypeptide comprising amino acids 43-61 of SEQID WO05039630. The methods described in US20040146534, NO:X; U.S. Pat. No. 4,220,584, U.S. Pat. No. 4,285,391, U.S. Pat. t) a polypeptide comprising amino acids 38-61 of SEQID No. 5,182,109, U.S. Pat. No. 4,603,049, U.S. Pat. No. 4,545, 931, U.S. Pat. No. 4,886,663, U.S. Pat. No. 4,758,655, NO:X; WO08402700, FR2525592, and FR2532850 can be similarly u) a polypeptide comprising amino acids 33-61 of SEQID used to create immunogens comprising the polypeptides NO:X; described herein. U.S. Pat. No. 6,043,057, U.S. Pat. No. v) a polypeptide comprising amino acids 43-66 of SEQID 5,834,246, U.S. Pat. No. 5,268.276, and EP368819, specifi NO:X; cally describe an expression system containing CTB (cholera w) a polypeptide comprising amino acids 38-66 of SEQID toxin Beta subunit) fused to an ST-like polypeptide under a NO:X; foreign promoter for use as a vaccine. The nucleic acids that x) a polypeptide comprising amino acids 33-66 of SEQID encode the polypeptides described herein may be use as NO:X; genetic vaccines as described in US20050260605 and y) a polypeptide comprising amino acids 43-71 of SEQID WO0148018. The nucleic acid molecules may also be used NO:X; for the manufacture of a functional ribonucleic acid, wherein Z) a polypeptide comprising amino acids 38-71 of SEQID the functional ribonucleic acid is selected from the group NO:X; comprising ribozymes, antisense nucleic acids and siRNA (as aa) a polypeptide comprising amino acids 33-71 of SEQID described in WO05103073). NO:X; ab) a polypeptide comprising amino acids 43-76 of SEQID 1. A purified polypeptide comprising at least 10 contiguous NO:X; amino acids of SEQID NO:X ac) a polypeptide comprising amino acids 38-76 of SEQID TVQDGNFSFSLES NO:X; and VKKLKDLOEPQEPRVGKLRN ad) a polypeptide comprising amino acids 33-76 of SEQID FAPIPGEPVVPILCSNPNFPEE LKPLCKEP NOX. NAQEILQRLEEIAEDPGTCEICAYAACTGC (SEQ 3. The purified polypeptide of claim 1 wherein the ID NO:X; human proguanylin). polypeptide is selected from: 2. The purified polypeptide of claim 1 wherein the a) a polypeptide consisting of amino acids 1-16 of SEQID polypeptide is selected from: NO:X; a) a polypeptide comprising amino acids 1-16 of SEQID b) a polypeptide consisting of amino acids 1-47 of SEQID NO:X; NO:X; b) a polypeptide comprising amino acids 1-47 of SEQID c) a polypeptide consisting of amino acids 1-61 of SEQID NO:X; NO:X; c) a polypeptide comprising amino acids 1-61 of SEQID d) a polypeptide consisting of amino acids 1-72 of SEQID NO:X; NO:X; US 2012/0283411 A9 Nov. 8, 2012

e) a polypeptide consisting of amino acids 17-47 of SEQID f) a polypeptide consisting essentially of amino acids NO:X; 17-61 of SEQID NO:X; f) a polypeptide consisting of amino acids 17-61 of SEQID g) a polypeptide consisting essentially of amino acids NO:X; 17-72 of SEQID NO:X; g) a polypeptide consisting of amino acids 17-72 of SEQ h) a polypeptide consisting essentially of amino acids ID NO:X; 17-94 of SEQID NO:X; h) a polypeptide consisting of amino acids 17-94 of SEQ i) a polypeptide consisting essentially of amino acids 48-61 ID NO:X; of SEQID NO:X; i) a polypeptide consisting of amino acids 48-61 of SEQID j) a polypeptide consisting essentially of amino acids 48-72 NO:X; of SEQID NO:X; j) a polypeptide consisting of amino acids 48-72 of SEQID k) a polypeptide consisting essentially of amino acids NO:X; 48-94 of SEQID NO:X; k) a polypeptide consisting of amino acids 48-94 of SEQ 1) a polypeptide consisting essentially of amino acids 62-72 ID NO:X; of SEQID NO:X; 1) a polypeptide consisting of amino acids 62-72 of SEQID m) a polypeptide consisting essentially of amino acids NO:X; 62-94 of SEQID NO:X; m) a polypeptide consisting of amino acids 62-94 of SEQ n) a polypeptide consisting essentially of amino acids ID NO:X; 73-94 of SEQID NO:X; n) a polypeptide consisting of amino acids 73-94 of SEQ o) a polypeptide consisting essentially of amino acids 1-23 ID NO:X; of SEQID NO:X; o) a polypeptide consisting of amino acids 1-23 of SEQID p) a polypeptide consisting essentially of amino acids NO:X; 48-66 of SEQID NO:X; p) a polypeptide consisting of amino acids 48-66 of SEQ q) a polypeptide consisting essentially of amino acids ID NO:X; 48-71 of SEQID NO:X; q) a polypeptide consisting of amino acids 48-71 of SEQ r) a polypeptide consisting essentially of amino acids ID NO:X; 48-76 of SEQID NO:X; r) a polypeptide consisting of amino acids 48-76 of SEQID S) a polypeptide consisting essentially of amino acids NO:X; 43-61 of SEQID NO:X; s) a polypeptide consisting of amino acids 43-61 of SEQID t) a polypeptide consisting essentially of amino acids 38-61 NO:X; of SEQID NO:X; t) a polypeptide consisting of amino acids 38-61 of SEQID u) a polypeptide consisting essentially of amino acids NO:X; 33-61 of SEQID NO:X; u) a polypeptide consisting of amino acids 33-61 of SEQ V) a polypeptide consisting essentially of amino acids ID NO:X; 43-66 of SEQID NO:X; V) a polypeptide consisting of amino acids 43-66 of SEQ w) a polypeptide consisting essentially of amino acids ID NO:X; 38-66 of SEQID NO:X; w) a polypeptide consisting of amino acids 38-66 of SEQ X) a polypeptide consisting essentially of amino acids ID NO:X; 33-66 of SEQID NO:X; x) a polypeptide consisting of amino acids 33-66 of SEQ y) a polypeptide consisting essentially of amino acids ID NO:X; 43-71 of SEQID NO:X; y) a polypeptide consisting of amino acids 43-71 of SEQ Z) a polypeptide consisting essentially of amino acids ID NO:X; 38-71 of SEQID NO:X; Z) a polypeptide consisting of amino acids 38-71 of SEQID aa) a polypeptide consisting essentially of amino acids NO:X; 33-71 of SEQID NO:X; aa) a polypeptide consisting of amino acids 33-71 of SEQ ab) a polypeptide consisting essentially of amino acids ID NO:X; 43-76 of SEQID NO:X; ab) a polypeptide consisting of amino acids 43-76 of SEQ ac) a polypeptide consisting essentially of amino acids ID NO:X; 38-76 of SEQID NO:X; and ac) a polypeptide consisting of amino acids 38-76 of SEQ ad) a polypeptide consisting essentially of amino acids ID NO:X; and 33-76 of SEQID NO:X. ad) a polypeptide consisting of amino acids 33-76 of SEQ 5. A purified polypeptide comprising at least 0.10 contigu ID NOX. ous amino acids of SEQID NO:X1 4. The purified polypeptide of claim 1 wherein the XXX XXs XX, Xs Xo Xio XL, EXV KX7 L. polypeptide is selected from: X19X20 LX22X23X24X2s X26X27X28X29Xso X3X32 a) a polypeptide consisting essentially of amino acids 1-16 Xss X34 Xss Xso X37 Xss X39 X40 X4, X42 X4s X44 X4s of SEQID NO:X; X46 X47 X4s X49 Xso C Xs2 Xss XS4 Xss Xso X57 P Xso b) a polypeptide consisting essentially of amino acids 1-47 Xso X61 X62 PX64 CX66 X67X6's Xso X7o X7 X72 X7s of SEQID NO:X; XX-7s RLX-7s X, Xso Xs, Xs, Xs PG TC EICAY c) a polypeptide consisting essentially of amino acids 1-61 AACT G CX9 (SEQID NO:X1-guanylin consensus of SEQID NO:X; 1) wherein: d) a polypeptide consisting essentially of amino acids 1-72 X is V or S; of SEQID NO:X; X is T, L, I, Y or E: e) a polypeptide consisting essentially of amino acids X is V or F: 17-47 of SEQID NO:X; X is Q or K.

US 2012/0283411 A9 Nov. 8, 2012 86

Xs is H or L.; d) a polypeptide comprising amino acids 1-72 of SEQID X is R. K or is missing: NO:X; X-7 is N. K or is missing: e) a polypeptide comprising amino acids 17-47 of SEQID Xss is For is missing: NO:X; Xso is A or is missing: f) a polypeptide comprising amino acids 17-61 of SEQID Xao is P or is missing: NO:X; X is I, R or is missing: g) a polypeptide comprising amino acids 17-72 of SEQID X is L or P; NO:X; X is G or L. h) a polypeptide comprising amino acids 17-94 of SEQID X is G, E or K, NO:X; Xs is P or S; i) a polypeptide comprising amino acids 48-61 of SEQID X is V or A: NO:X; Xa, is A or V: j) a polypeptide comprising amino acids 48-72 of SEQID Xs is P or is missing: NO:X; Xao is I or Q. k) a polypeptide comprising amino acids 48-94 of SEQID Xso is L.; NO:X; Xs2 is S. 1) a polypeptide comprising amino acids 62-72 of SEQID Xs is missing: NO:X; Xs is H, N, or D: m) a polypeptide comprising amino acids 62-94 of SEQID Xss is P or S; NO:X; Xs is A, K or N: n) a polypeptide comprising amino acids 73-94 of SEQID Xs, is For L.; NO:X; Xso is E. o) a polypeptide comprising amino acids 1-23 of SEQID Xeo is E or A: NO:X; X is L.; p) a polypeptide comprising amino acids 48-66 of SEQID X is K or R; NO:X; X is L. I or V. q) a polypeptide comprising amino acids 48-71 of SEQID X is K, E, Q, Tor R: NO:X; X, is E or K. r) a polypeptide comprising amino acids 48-76 of SEQID Xs is P. NO:X; X is N: s) a polypeptide comprising amino acids 43-61 of SEQID Xzo is A: NO:X; X-7 is E or Q; t) a polypeptide comprising amino acids 38-61 of SEQID X72 is E. NO:X; X7 is I; u) a polypeptide comprising amino acids 33-61 of SEQID X74 is L.; NO:X; X-7s is Q or E: v) a polypeptide comprising amino acids 43-66 of SEQID X-7s is E or A: NO:X; X7 is E or A: w) a polypeptide comprising amino acids 38-66 of SEQID Xso is I; NO:X; Xs is A: x) a polypeptide comprising amino acids 33-66 of SEQID Xs is E or Q; NO:X; Xss is D; y) a polypeptide comprising amino acids 43-71 of SEQID Xss is G, or S. NO:X; Xs is T. Z) a polypeptide comprising amino acids 38-71 of SEQID X is Y: NO:X; Xo is T. and aa) a polypeptide comprising amino acids 33-71 of SEQID Xoo is missing. NO:X; 9-11. (canceled) ab) a polypeptide comprising amino acids 43-76 of SEQID 12. A purified polypeptide consisting of a polypeptide frag NO:X; ment of SEQ ID NO:X comprising at least 10 contiguous ac) a polypeptide comprising amino acids 38-76 of SEQID amino acids of SEQ ID NO:X VTVQDGNFSFSLES NO:X; and VKKLKDLOEPQEPRVGKLRN ad) a polypeptide comprising amino acids 33-76 of SEQID FAPIPGEPVVPILCSNPNFPEELKPLC KEPNAQEILQR NOX. LEEIAEDPGTCEICAYAACTGC (SEQ ID NO:X; human 14. The purified polypeptide of claim 12 wherein the proguanylin). polypeptide fragment of SEQID NO:X is selected from: 13. The purified polypeptide of claim 12 wherein the a) a polypeptide consisting of amino acids 1-16 of SEQID polypeptide fragment of SEQID NO:X is selected from: NO:X; a) a polypeptide comprising amino acids 1-16 of SEQID b) a polypeptide consisting of amino acids 1-47 of SEQID NO:X; NO:X; b) a polypeptide comprising amino acids 1-47 of SEQID c) a polypeptide consisting of amino acids 1-61 of SEQID NO:X; NO:X; c) a polypeptide comprising amino acids 1-61 of SEQID d) a polypeptide consisting of amino acids 1-72 of SEQID NO:X; NO:X; US 2012/0283411 A9 Nov. 8, 2012

e) a polypeptide consisting of amino acids 17-47 of SEQID c) a polypeptide consisting essentially of amino acids 1-61 NO:X; of SEQID NO:X; f) a polypeptide consisting of amino acids 17-61 of SEQID d) a polypeptide consisting essentially of amino acids 1-72 NO:X; of SEQID NO:X; g) a polypeptide consisting of amino acids 17-72 of SEQ e) a polypeptide consisting essentially of amino acids ID NO:X; 17-47 of SEQID NO:X; h) a polypeptide consisting of amino acids 17-94 of SEQ f) a polypeptide consisting essentially of amino acids ID NO:X; 17-61 of SEQID NO:X; i) a polypeptide consisting of amino acids 48-61 of SEQID g) a polypeptide consisting essentially of amino acids NO:X; 17-72 of SEQID NO:X; j) a polypeptide consisting of amino acids 48-72 of SEQID h) a polypeptide consisting essentially of amino acids NO:X; 17-94 of SEQID NO:X; k) a polypeptide consisting of amino acids 48-94 of SEQ i) a polypeptide consisting essentially of amino acids 48-61 ID NO:X; of SEQID NO:X; 1) a polypeptide consisting of amino acids 62-72 of SEQID j) a polypeptide consisting essentially of amino acids 48-72 NO:X; of SEQID NO:X; m) a polypeptide consisting of amino acids 62-94 of SEQ k) a polypeptide consisting essentially of amino acids ID NO:X; 48-94 of SEQID NO:X; n) a polypeptide consisting of amino acids 73-94 of SEQ 1) a polypeptide consisting essentially of amino acids 62-72 ID NO:X; of SEQID NO:X; o) a polypeptide consisting of amino acids 1-23 of SEQID m) a polypeptide consisting essentially of amino acids NO:X; 62-94 of SEQID NO:X; p) a polypeptide consisting of amino acids 48-66 of SEQ n) a polypeptide consisting essentially of amino acids ID NO:X; 73-94 of SEQID NO:X; q) a polypeptide consisting of amino acids 48-71 of SEQ o) a polypeptide consisting essentially of amino acids 1-23 ID NO:X; of SEQID NO:X; r) a polypeptide consisting of amino acids 48-76 of SEQID p) a polypeptide consisting essentially of amino acids NO:X; 48-66 of SEQID NO:X; s) a polypeptide consisting of amino acids 43-61 of SEQID q) a polypeptide consisting essentially of amino acids NO:X; 48-71 of SEQID NO:X; t) a polypeptide consisting of amino acids 38-61 of SEQID r) a polypeptide consisting essentially of amino acids NO:X; 48-76 of SEQID NO:X; u) a polypeptide consisting of amino acids 33-61 of SEQ S) a polypeptide consisting essentially of amino acids ID NO:X; 43-61 of SEQID NO:X; V) a polypeptide consisting of amino acids 43-66 of SEQ t) a polypeptide consisting essentially of amino acids 38-61 ID NO:X; of SEQID NO:X; w) a polypeptide consisting of amino acids 38-66 of SEQ u) a polypeptide consisting essentially of amino acids ID NO:X; 33-61 of SEQID NO:X; x) a polypeptide consisting of amino acids 33-66 of SEQ V) a polypeptide consisting essentially of amino acids 43-66 of SEQID NO:X; ID NO:X; w) a polypeptide consisting essentially of amino acids y) a polypeptide consisting of amino acids 43-71 of SEQ 38-66 of SEQID NO:X; ID NO:X; X) a polypeptide consisting essentially of amino acids Z) a polypeptide consisting of amino acids 38-71 of SEQID 33-66 of SEQID NO:X; NO:X; y) a polypeptide consisting essentially of amino acids aa) a polypeptide consisting of amino acids 33-71 of SEQ 43-71 of SEQID NO:X; ID NO:X; Z) a polypeptide consisting essentially of amino acids ab) a polypeptide consisting of amino acids 43-76 of SEQ 38-71 of SEQID NO:X; ID NO:X; aa) a polypeptide consisting essentially of amino acids ac) a polypeptide consisting of amino acids 38-76 of SEQ 33-71 of SEQID NO:X; ID NO:X; and ab) a polypeptide consisting essentially of amino acids ad) a polypeptide consisting of amino acids 33-76 of SEQ 43-76 of SEQID NO:X; ID NOX. ac) a polypeptide consisting essentially of amino acids 15. The purified polypeptide of claim 12 wherein the 38-76 of SEQID NO:X; and polypeptide fragment of SEQID NO:X is selected from: ad) a polypeptide consisting essentially of amino acids a) a polypeptide consisting essentially of amino acids 1-16 33-76 of SEQID NO:X. of SEQID NO:X; 16-148. (canceled) b) a polypeptide consisting essentially of amino acids 1-47 of SEQID NO:X;