USO09533022B2

(12) United States Patent (10) Patent No.: US 9,533,022 B2 Mehta et al. (45) Date of Patent: *Jan. 3, 2017

(54) PEPTIDE ANALOGS FOR TREATING (56) References Cited DISEASES AND DISORDERS U.S. PATENT DOCUMENTS (71) Applicants: Nozer M. Mehta, Randolph, NJ (US); 4,764,589 A 8, 1988 Orlowski William Stern, Tenafly, NJ (US); Amy 9,006,172 B2 * 4/2015 Mehta ...... A61K 38.23 M. Sturmer, Towaco, NJ (US); Morten 514/1.1 Asser Karsdal, Kobenhavn (DK); Kim 2002/0065255 A1* 5/2002 Bay ...... A61K 47, 183 Henriksen, Hillerod (DK) 514,166 2010/0311650 A1* 12/2010 Mehta ...... CO7K 1400 (72) Inventors: Nozer M. Mehta, Randolph, NJ (US); 514,49 William Stern, Tenafly, NJ (US); Amy M. Sturmer, Towaco, NJ (US); Morten FOREIGN PATENT DOCUMENTS Asser Karsdal, Kobenhavn (DK); Kim Henriksen, Hillerod (DK) WO 2010O85700 A2 T 2010 (73) Assignee: KeyBioscience A/S, Stans (CH) OTHER PUBLICATIONS Lee et al., “Metformin Decreases Food Consumption and Induces (*) Notice: Subject to any disclaimer, the term of this Weight Loss in Subjects with Obesity with Type Il Non-Insulin patent is extended or adjusted under 35 Dependent Diabetes,” Obes. Res. 6:47-53 (1998).* U.S.C. 154(b) by 0 days. Feigh et al., “A novel oral form of Salmon calcitonin improves glucose homeostasis and reduces body weight in diet-induced obese This patent is Subject to a terminal dis rats' Diab. Obesity Metab. 13:911-920 (2011).* claimer. Unigene Laboratories Inc. Unigene Laboratories Inc. Corporate Overview 2010, URL: www.getfillings.com/sec-filings/100325/ (21) Appl. No.: 14/634,188 UNIGENE-LABORATORIES-INC 8-K/dex991.htm, whole document. (22) Filed: Feb. 27, 2015 Andreassen, et al. Oral formulation of UGP302, a novel amylin and calcitonin dual receptor agonist, exerts anti-obesity effects in diet Prior Publication Data induced obese (DIO) rats, Diabetologia 2013, vol. 56(Suppl1), p. (65) S344, Abstract 867. US 2015/O196617 A1 Jul. 16, 2015 Feigh, et al. A novel oral dual-action amylin and calcitonin receptor agonist (UGP302) rebalances insulin and glucagon hyperglycaemia in Zucker diabetic fatty rats, Diabetologia 2013, vol. 56(Suppl2), p. Related U.S. Application Data S405-S406, Abstract 1009. (63) Continuation-in-part of application No. 13/667,578, * cited by examiner filed on Nov. 2, 2012, now Pat. No. 9,006,172. Primary Examiner — Julie Ha Assistant Examiner — Kristina M Hellman (60) Provisional application No. 61/578,620, filed on Dec. (74) Attorney, Agent, or Firm — Benjamin Aaron Adler 21, 2011, provisional application No. 61/554,771, filed on Nov. 2, 2011. (57) ABSTRACT Provided herein are methods for the treatment of type I (51) Int. C. diabetes, Type II diabetes, metabolic syndrome, or obesity, A6 IK 38/00 (2006.01) or of appetite Suppression, or for mitigating insulin resis A6 IK 38/23 (2006.01) tance, or for reducing an undesirably high fasting serum A6IP3/04 (2006.01) glucose level, or for reducing an undesirably high peak C07K 5/00 (2006.01) serum glucose level, or for reducing an undesirably high A6 IK 38/16 (2006.01) peak serum insulin level, or for reducing an undesirably A6 IK3I/55 (2006.01) large response to a glucose tolerance test in Synergistic C07K 4/585 (2006.01) combination with metformin. Treatment is effected with a (52) U.S. C. combination therapy of metformin and a peptide with a CPC ...... A61K 38/16 (2013.01); A61K 31/155 sequence selected from SEQ ID NO: 11, SEQ ID NO: 12, (2013.01); A61K 38/23 (2013.01); C07K SEQID NO: 13, SEQ ID NO: 14, SEQID NO: 15, SEQ ID 14/585 (2013.01) NO: 16, SEQID NO: 17, SEQID NO: 19, SEQID NO: 20, (58) Field of Classification Search SEQID NO: 21, SEQID NO: 22, SEQID NO. 23, and SEQ None ID NO: 24 administered to a patient. See application file for complete search history. 22 Claims, 25 Drawing Sheets U.S. Patent Jan. 3, 2017 Sheet 1 of 25 US 9,533,022 B2

2 5

-O- Vehicle -er SC gikg -o- UGP3021 g/kg

5

9 5 O -One Vehicle SCT mg/kg 89 5O OO -o- UGP302 mg/kg

8 O O

FIG. 1B U.S. Patent Jan. 3, 2017 Sheet 2 of 25 US 9,533,022 B2

Vehicle OO sCT mg/kg UGP3021 mg/kg

60

40

FIG. C

Vehicle sCT 1 mgkg UGP302 1 mg/kg

-4 O

FIG. D U.S. Patent Jan. 3, 2017 Sheet 3 of 25 US 9,533,022 B2

2 -O- Vehicle SC1 mg/kg -o- UGP3021 mg/kg

8

4. -30 O 3O 6O 90 20 150 80 21 O 240 Time (min)

FIG. 2A

g 1500 Vehicle sCT 1 mg/kg S. UGP302 1 mg/kg (D - : OOO is 500 O () O 3.

FIG. 2B U.S. Patent Jan. 3, 2017 Sheet 4 of 25 US 9,533,022 B2

s FIG. 3 () . it. e Vehicle sCT 2 mg/kg M UGP3022 mg/kg

o 5. FIG. 4A s g 5 -20 sCT 0.5 mg/kg M. JGP302 0.5 mg/kg

sCT 0.5 mg/kg M UGP302 0.5 mg/kg

5 FIG. 4B

1 O

5 U.S. Patent Jan. 3, 2017 Sheet 5 Of 25 US 9,533,022 B2

W2 O

sCT 1 mg/kg M UGP302 1 mg/kg

F.G. 5A

2 O sCT 1 mg/kg UGP302 1 mg/kg 1 5

O

5

O

FIG. 5B U.S. Patent Jan. 3, 2017 Sheet 6 of 25 US 9,533,022 B2

-3 O

sCT 2 mg/kg UGP3022 mg/kg

F.G. 6A

15 sCT 2 mg/kg M UGP3022 mg/kg O

FIG. 6B U.S. Patent Jan. 3, 2017 Sheet 7 Of 25 US 9,533,022 B2

12

8

-O- Vehicle (ppoied) sCT 1 mg/kg-04 UGP302 1 mg/kg 4 -30 O 3O 6O 90 120 50 18O 21 O 240 Time (min)

FIG. 7A

500 Vehicle (poied) SC ing/kg UGP3021 mg/kg

FIG. 7B U.S. Patent Jan. 3, 2017 Sheet 8 of 25 US 9,533,022 B2

1 2

1 O

8

-O- Vehicle -O- UGP3022 mg/kg SC 2 mg/kg

-30 O 3O 60 90 120 150 18O 21 O 240 Time (min)

FIG. 8A

1500 w SCT2gkg UGP3022 ring/kg

FIG. 8B U.S. Patent Jan. 3, 2017 Sheet 9 Of 25 US 9,533,022 B2

sCT 0.5 mg/kg UGP302 0.5 mg/kg m UGP284 0.5 mg/kg UGP298 0.5 mg/kg

FIG. 9A

sCT 0.5 mg/kg 1 OO " UGP302 0.5 mg/kg

O UGP284 0.5 mg/kg 8 O UGP298 0.5 mg/kg

6 O

4 O

O

FIG. 9B U.S. Patent Jan. 3, 2017 Sheet 10 of 25 US 9,533,022 B2

sCT 1 mg/kg M UGP3021 mg/kg M UGP2841 mg/kg UGP298 1 mg/kg

... 1 O -3-2 OO

FIG. 9C

sCT 1 mg/kg 2 O M UGP302 1 mg/kg 0 UGP2841 mg/kg

f 5 UGP298 1 mg/kg

10

5

O

FIG. 9D U.S. Patent Jan. 3, 2017 Sheet 11 of 25 US 9,533,022 B2

sCT 2 mg/kg M UGP3022 mg/kg O UGP2842 mg/kg UGP298.2 mg/kg

FIG. 9E

5 sCT 2 mg/kg M UGP3022 mg/kg m UGP2842 mg/kg 10 UGP298.2 mg/kg

k es ke

FIG. 9F U.S. Patent Jan. 3, 2017 Sheet 12 of 25 US 9,533,022 B2

-O- Vehicle (pooled) UGP284 0.5 g/kg sCT 0.5 mg/kg -&- UGP298 0.5 mg/kg 1 2 -On UGP3O2 0.5 in gikg

O

8

4. -30 O 30 60 90 120 150 18O 21 O 240 Time (min)

FIG. OA

" Vehicle (pooled) 9. 1500 sCT 0.5 mg/kg

8 M UGP302 0.5 mg/kg 5 UGP284 0.5 mg/kg ge 1000 UGP298 0.5 mg/kg E.O a () - 500 O (...) O s

FIG. OB U.S. Patent Jan. 3, 2017 Sheet 13 Of 25 US 9,533,022 B2

re-Or Vehicle 2 SCT glkg -o- UGP3021 mg/kg -x- UGP284 mg/kg -x- UGP298 1 mg/kg 8

4. -30 O 30 SO 90 120 150 180 20 240 Time (min)

FIG. OC

1500 Vehicle sC 1 mg/kg UGP302 1 mg/kg I UGP2841 mg/kg O O O UGP298 1 mg/kg

e t 5 O O

FIG. OD U.S. Patent Jan. 3, 2017 Sheet 14 of 25 US 9,533,022 B2

8 sCT 2 mg/kg 12, -C- VehicleUGP3022 mg/kg -o--()- UGP298.2UGP2842 mg/kg

8

6

4. -30 O 3O 6O 90 20 SO 80 21 O 240 Time (min)

FIG. OE

1500- Vehicle UGP3022 mg/kg sCT 2 mg/kg UGP2842 mg/kg x UG-298 2. ng fkg

FIG. OF U.S. Patent Jan. 3, 2017 Sheet 15 Of 25 US 9,533,022 B2

250 go

s s s s s s x8% UGP302 (n=7) s sk------'''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''. s s s s a s UGP298 (n=4)

100. x: UGP284 (n-3) 8 a *.x*a- sCT (n-63)

s s s s s s 50.3.s UGP306 (n=4) s s s s s s UGP283 (n=4) UGP303 (n=4) O 250 500 750 1 OOO Peptide (pg/ml) FG.

8

3 . ge 2 : s g a i FIG. 2A & a {} g & - ie:::ce - S -88 -x- 38°382.38 gikg 8 S rix. 328383 gikg - so

gos 9, s {{ ------&isis;i& 8.8 -- 3:2832 gig 9 a. -x- 38°382.38 agikg 5, s { FIG. 12B sg -15 ^^ -2. 28 -3.3 . 8 & s & 8 g U.S. Patent Jan. 3, 2017 Sheet 16 of 25 US 9,533,022 B2

FIG. 3A

-- Vehicle ...«- UGP28310 g/kg)

O

2 FIG. 13B -4

6 --Wehicle. . . UGP28310 g/kg) U.S. Patent Jan. 3, 2017 Sheet 17 Of 25 US 9,533,022 B2

-4 O FIG. 14A s . ------Wehicle -60 ? rx-UGP284(10 uglkg) -8 O

FIG. 4B --Wehicle r x-r UGP284(10 g/kg) U.S. Patent Jan. 3, 2017 Sheet 18 of 25 US 9,533,022 B2

2 O

O

-2 O

-4 O FIG. 5A

-6 O --Wehicle

A& 8 O -x- UGP2984 ag/kg)

FIG. 5B

-H. Vehicle -x-UGP2984 uglkg) U.S. Patent Jan. 3, 2017 Sheet 19 Of 25 US 9,533,022 B2

2 O

O

-2 O

-4 O F.G. 6A

-6 O --Wehicle -8 O -x- UGP302.5 g/kg)

F.G. 6B

x. --Wehicle rx-UGP302.5 g/kg) U.S. Patent Jan. 3, 2017 Sheet 20 of 25 US 9,533,022 B2

as 10

F.G. 7A --Vehicle x: -x-- UGP3035 ug/kg) -50 -x-UGP303(10 uglkg) & rx-UGP30320 ug/kg)

FIG. 7B --Wehicle -x- UGP3035 ug?kg) -x-- UGP303(10 g/kg) sys-X-UGP303(20 ug/kg) U.S. Patent Jan. 3, 2017 Sheet 21 of 25 US 9,533,022 B2

g Sh S sk Ha O

Vehicle sCT 1 mg/kg CM1 1 mg/kg

FIG. 18

5 O) S. s Han O

Vehicle sCT 1 mg/kg CM11 mg/kg

FIG. 19 U.S. Patent Jan. 3, 2017 Sheet 22 of 25 US 9,533,022 B2

- 40 s E. : 30 S. s 20 FIG. 20

O to 0

s - 0 s 8 O 12 14 (12h) Age (weeks)

ox Vehicle x Metformin 400 mg/kg · UGP-3025 g/kg « Metformin 400 mg/kg +UGP-3025 g/kg

FIG. 21

7 8 9 10 11 2 3 4 5 Age (Weeks) U.S. Patent Jan. 3, 2017 Sheet 23 Of 25 US 9,533,022 B2

Vehicle Metformin UGP-302 Metformin 400 mg/kg + 400 mg/kg 5 g/kg UGP-30255 g/kg

FIG. 22 U.S. Patent Jan. 3, 2017 Sheet 24 of 25 US 9,533,022 B2

4.

30 & 5 * Vehicle 8

3 20 UGP-3025 g/kg CSs Metformin 400 mg/kg 3 10 . Metformin 400 mg/kg+ UGP-3025 g/kg

5. O 50 O 50 20 ite after itstsii itection is FIG. 23A 6000

4000 E o s es O O 5, 2000

O Wehicle Metformin UGP-302 Metformin 400 mg/kg + 400 mg/kg 5 g/kg UGP-30255 g/kg

FIG. 23B U.S. Patent Jan. 3, 2017 Sheet 25 Of 25 US 9,533,022 B2

-50 O 50 OO 150 Time after insulin injection (min) x- Vehicle « Metformin 400 mg/kg 8. UGP-3025 g/kg .x. Metformin 400 mg/kg + UGP-3025 g/kg FIG. 24A

Vehicle Metformin UGP-302 Metformin 400 mg/kg 400 mg/kg 5 g/kg UGP-3025 g/kg

FIG. 24B US 9,533,022 B2 1. 2 PEPTIDE ANALOGS FOR TREATING Inflammatory or degenerative diseases, including diseases DISEASES AND DISORDERS of the joints, e.g. osteoarthritis (OA), rheumatoid arthritis (RA) or juvenile rheumatoid arthritis (JRA), and including CROSS-REFERENCE TO RELATED inflammation that results from autoimmune response, e.g. APPLICATIONS 5 lupus, ankylosing spondylitis (AS) or multiple Sclerosis (MS), can lead to substantial loss of mobility due to pain and This application is a continuation-in-part under 35 U.S.C. joint destruction. Cartilage that covers and cushions bone S120 of non-provisional application U.S. Ser. No. 13/667, within joints may become degraded over time thus undesir 578, filed Nov. 2, 2012, which claims benefit of priority ably permitting direct contact of two bones that can limit under 35 U.S.C. S 119(e) to provisional application U.S. Ser. 10 motion of one bone relative to the other and/or cause damage No. 61/578,620, filed Dec. 21, 2011, and to provisional to one by the other during motion of the joint. Subchondral bone just beneath the cartilage may also degrade. Adminis application U.S. Ser. No. 61/554,771, filed Nov. 2, 2011, the tering an effective amount of an anti-resorptive agent. Such entirety of all of which are hereby incorporated herein by as calcitonin, may prevent resorption of bone. reference. 15 SUMMARY OF THE INVENTION BACKGROUND OF THE INVENTION Calcitonin mimetics are disclosed herein. Field of the Invention According to aspects illustrated herein, there is disclosed The embodiments disclosed herein relate to mimetics of a peptide having a sequence selected from SEQID NO: 11, calcitonin, and more particularly to their use in the treatment SEQID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID of various diseases and disorders, including, but not limited NO: 15, SEQ ID NO: 17 and SEQ ID NO: 18. to diabetes (Type I and Type II), excess bodyweight, exces According to aspects illustrated herein, there is disclosed sive food consumption and metabolic syndrome, the regu a method that includes administering to a patient an effective lation of blood glucose levels, the regulation of response to 25 amount of a peptide selected from the group consisting of glucose tolerance tests, the regulation of food intake, the SEQID NO: 11, SEQID NO: 12, SEQID NO: 13, SEQ ID treatment of osteoporosis and the treatment of osteoarthritis. NO: 14, SEQ ID NO: 15, and SEQ ID NO: 17 to affect a Description of the Related Art weight reduction in the patient. Worldwide, there are about 250 million diabetics and the According to aspects illustrated herein, there is disclosed number is projected to double in the next two decades. Over 30 a method that includes administering to a patient an effective 90% of this population suffers from type 2 diabetes mellitus amount of a peptide selected from the group consisting of (T2DM). It is estimated that only 50-60% of persons SEQID NO: 11, SEQID NO: 12, SEQID NO: 13, SEQID affected with T2DM or in stages preceding overt T2DM are NO: 14, SEQ ID NO: 15, and SEQ ID NO: 17 to affect currently diagnosed. T2DM is a heterogeneous disease char postprandial glycemic control in the patient. acterized by abnormalities in carbohydrate and fat metabo 35 According to aspects illustrated herein, there is disclosed lism. The causes of T2DM are multi-factorial and include a method that includes administering to a patient an effective both genetic and environmental elements that affect B-cell amount of a peptide selected from the group consisting of function and insulin sensitivity in tissues such as muscle, SEQID NO: 11, SEQID NO: 12, SEQID NO: 13, SEQ ID liver, pancreas and adipose tissue. As a consequence NO: 14, SEQID NO: 15, and SEQID NO: 17 to affect an impaired insulin secretion is observed and paralleled by a 40 improvement in glycemic control in the patient. progressive decline in B-cell function and chronic insulin According to aspects illustrated herein, there is disclosed resistance. The inability of the endocrine pancreas to com a method that includes administering to a patient an effective pensate for peripheral insulin resistance leads to hypergly amount of a peptide of SEQID NO: 18 having the sequence caemia and onset of clinical diabetes. Tissue resistance to CSNLSTCVLGKLSQELHKLQTYPRTDVGANXaaXaa, insulin-mediated glucose uptake is now recognized as a 45 So as to reduce at least one of bone resorption and cartilage major pathophysiologic determinant of T2DM. Type I dia degradation in the patient. betes is characterised by a loss of the ability to produce According to aspects illustrated herein, there is disclosed insulin in response to food intake and hence an inability to a method for the treatment of type I diabetes, Type II regulate blood glucose to a normal physiological level. diabetes, metabolic syndrome, or obesity, or of appetite A successful criterion for an optimal T2DM intervention 50 Suppression, or for mitigating insulin resistance, or for is the lowering of blood glucose levels, which can be both reducing an undesirably high fasting serum glucose level, or chronic lowering of blood glucose levels and increased for reducing an undesirably high peak serum glucose level. ability to tolerate high glucose levels after food intake, or for reducing an undesirably high peak serum insulin level. described by lower peak glucose levels and faster clearance. or for reducing an undesirably large response to a glucose Both of these situations exert less strain on B-cell insulin 55 tolerance test comprising administering to a patient as a output and function. combination therapy an effective amount metformin and of The physical structure of bone may be compromised by a a peptide of SEQID NOS: 11-24. It has been found that the variety of factors, including disease and injury. One of the Subject peptides show a synergistic activity when adminis most common bone diseases is osteoporosis, which is char tered in combination with metformin. acterized by low bone mass and structural deterioration of 60 bone tissue, leading to bone fragility and an increased BRIEF DESCRIPTION OF THE DRAWINGS Susceptibility to fractures, particularly of the hip, spine and wrist. Osteoporosis develops when there is an imbalance The presently disclosed embodiments will be further such that the rate of bone resorption exceeds the rate of bone explained with reference to the attached drawings, which formation. Administering an effective amount of an anti 65 illustrate the principles thereof. resorptive agent, such as calcitonin, has shown to prevent FIG. 1A, FIG. 1B, FIG. 1C, and FIG. 1D show the effect resorption of bone. of chronic oral salmon calcitonin (“sCT) versus oral UGP US 9,533,022 B2 3 4 302 administration food intake (FIG. 1A), on body weight as plots of blood glucose against time (FIG. 23A) and as (FIG. 1B), cumulative food intake (FIG. 1C), and change in total Area Under Curve tAUC measurements (FIG. 23B). body weight (FIG. 1D) in DIO rats as measured in Example FIG.24A and FIG.24B show results obtained in Example 1; 7 showing the results of insulin tolerance tests (IPITT) as FIG. 2A and FIG. 2B Show the effect of oral SCT versus plots of blood glucose against time (FIG. 24A) and as total oral UGP302 on glucose tolerance during OGTT in DIO rats Area Under Curve tAUC measurements (FIG. 24B). as measured in Example 1. While the above-identified drawings set forth presently FIG. 3 shows the effect of oral SCT versus oral UGP 302 disclosed embodiments, other embodiments are also con on fasting glycemia in DIO rats as measured in Example 1. templated, as noted in the discussion. This disclosure pres FIG. 4A and FIG. 4B show the effect of oral SCT versus 10 oral UGP 302 on body weight (FIG. 4A) and food intake ents illustrative embodiments by way of representation and (FIG. 4B) in DIO rats observed in Example 2 at a first not limitation. Numerous other modifications and embodi dosage. ments can be devised by those skilled in the art which fall FIG. 5A and FIG. SB Show the effect of oral SCT versus within the scope and spirit of the principles of the presently disclosed embodiments. oral UGP 302 on body weight (FIG. 5A) and food intake 15 (FIG. 5B) in DIO rats observed in Example 2 at a second DETAILED DESCRIPTION OF THE dosage. INVENTION FIG. 6A and FIG. 6B show the effect of oral SCT versus oral UGP 302 on body weight (FIG. 6A) and food intake (FIG. 6B) in DIO rats observed in Example 2 at a third Calcitonins are highly conserved over a wide range of dosage. species. Full-length native calcitonin is 32 amino acids in FIG. 7A and FIG. 7B show the effect of oral SCT versus length. The sequences of examples of calcitonins are set out oral UGP302 at a first dosage on glucose tolerance during below: OGTT in DIO rats as measured in Example 2. FIG. 8A and FIG. 8B show the effect of oral SCT versus 25 Salmon oral UGP 302 at a second dosage on glucose tolerance SEQ ID NO: 1 during OGTT in DIO rats as measured in Example 2. CSNLSTCVLGKLSOELHKLOTYPRTNTGSGTP FIG.9A, FIG.9B, FIG. 9C, FIG. 9D, FIG.9E, and FIG. Mouse 9F show the effect of oral SCT versus three oral UGPs on SEQ ID NO: 2 body weight (FIGS.9A, 9C,9E) and food intake (FIGS.9B, 30 CGNLSTCMLGTYTODLNKFHTFPOTSIGWEAP 9C) in DIO rats as measured in Example 3. Chicken FIG. 10A, FIG. 10B, FIG. 10C, FIG. 10D, FIG. 10E, and SEQ ID NO: 3 FIG. 10F show the effect of oral SCT versus three oral UGPS CASLSTCVLGKLSOELHKLOTYPRTDVGAGTP on glucose levels in a glucose tolerance test in DIO rats as measured in Example 3. 35 Eel FIG. 11 shows binding results for six UGP compounds to SEQ ID NO: 4 T47D cell calcitonin receptors as measured in Example 4. CSNLSTCVLGKLSOELHKLOTYPRTDVGAGTP FIG. 12A and FIG. 12B show food consumption (FIG. Rat 12A) and weight change measurements (FIG. 12B) for UGP SEO ID NO : 5 282 as measured in Example 5. 40 CGNLSTCMLGTYTODLNKFHTFPOTSIGWGAP FIG. 13A and FIG. 13B show food consumption (FIG. Horse 13A) and weight change measurements (FIG. 13B) for UGP SEQ ID NO: 6 283 as measured in Example 5. CSNLSTCVLGTYTODLNKFHTFPOTAIGVGAP FIG. 14A and FIG. 14B show food consumption (FIG. Canine-1 14A) and weight change measurements (FIG. 14B) for UGP 45 SEO ID NO : 7 284 as measured in Example 5. CSNLSTCWLGTYSKDLNNFHTFSGIGFGAETP FIG. 15A and FIG. 15B show food consumption (FIG. Canine-2 15A) and weight change measurements (FIG. 15B) for UGP SEQ ID NO: 8 298 as measured in Example 5. CSNLSTCVLGTYTODLNKFHTFPOTAIGVGAP FIG. 16A and FIG. 16B show food consumption (FIG. 50 16A) and weight change measurements (FIG. 16B) for UGP Porcine 302 as measured in Example 5. SEO ID NO : 9 FIG. 17A and FIG. 17B show food consumption (FIG. CSNLSTCWLSAYWRNLNNFHRFSGMGFGPETP 17A) and weight change measurements (FIG. 17B) for UGP Human 303 as measured in Example 5: 55 SEO ID NO: 10 FIG. 18 shows the reduction of bone resorption produced CGNLSTCMLGTYTODFNKFHTFPOTAIGVGAP by treatment with UGP302 in rats. Embodiments of the present disclosure relate to calcitonin FIG. 19 shows cartilage resorption produced by treatment mimetics. The amino acid sequence of the calcitonin mimet with UGP302 in rats. ics of the present disclosure are found in Table 1A below. FIG. 20 shows results obtained in Example 7 showing 60 fasting blood glucose (FBG) of tested rats. TABLE 1A FIG. 21 shows results obtained in Example 7 showing non-fasted blood glucose (PPG) of subject rats. Calcitonin SEQ FIG. 22 shows results obtained in Example 7 showing Mimetic (CM) Amino Acid Sequence ID NO : HbA1c levels of subject rats. 65 UGP281 AcOSNLSTCWLGKLSOELHKLOTY 11 FIG.23A and FIG. 23B show results obtained in Example PRTDVGANTY-NH 7 showing the results of oral glucose tolerance tests (OGTT) US 9,533,022 B2 6 TABLE 1.A - continued As those of skill in the art will appreciate, peptides having a plurality of cysteine residues frequently form a disulfide Calcitonin SEQ bridge between two such cysteine residues. All Such pep Mimetic (CM) Amino Acid Sequence ID NO : tides set forth herein are defined as optionally including one or more such disulfide bridges. While calcitonin mimetics of UGP283 AcOSNLSTCWLGRLSOELHRLOTF 2 the present disclosure may exist in free acid form, it is PRTDWGANTAcy preferred that the C-terminal amino acid be amidated. Appli UGP284 ProSNLSTCVLGKLSOELHKLOTY 3 cants expect that such amidation may contribute to the PRTNTGSGTP-NH, effectiveness and/or bioavailability of the peptide. A pre 10 ferred technique for manufacturing amidated versions of the UGP298 SuccCSNLSTCWLGKLSOELHKLQ 4. calcitonin mimetics of the present disclosure is to react TYPRTNTGSGTP-NH, precursors (having glycine in place of the C-terminal amino UGP3 O2 AcOSNLSTCWLGKLSOELHKLOTY 5 group of the desired amidated product) in the presence of PRTDVGANAP-NH, peptidylglycine alpha-amidating monooxygenase in accor 15 dance with known techniques wherein the precursors are UGP3O3 KCSNLSTCVLGKLSOELHKLOTYP 6 converted to amidated products in reactions described, for RTDVGANTY-NH, example, in U.S. Pat. No. 4,708,934 and European Patent UGP3 O6 SuccCSNLSTCWLGKLSOELHKLQ 7 Publication Nos. 0308 067 and 0 382 403. Recombinant TYPRTDVGANAY-NH, production is preferred for both the precursor and the UGP1 OOO CSNLSTCVLGKLSQELHKLQTYP 8 enzyme that catalyzes the conversion of the precursor to RTDVGANXaaXaa, salmon calcitonin. Such recombinant production is dis cussed in Biotechnology, Vol. 11 (1993) pp. 64–70, which In some embodiments, the cysteine at position 1 of the further describes a conversion of a precursor to an amidated calcitonin mimetics discussed supra is modified (“C.”) to product. The recombinant product reported there is identical reduce the positive charge of the first amino acid. For 25 to natural salmon calcitonin, and to salmon calcitonin pro example, an acetyl group (SEQ ID NOs: 11, 12 and 15), duced using solution and Solid phase chemical peptide propionyl group (SEQID NO: 13), or succinyl group (SEQ synthesis. Production of amidated products may also be ID NOs: 14 and 17) may be substituted on cysteine-1. In accomplished using the process and amidating enzyme set Some embodiments, the amino acid at the last position forth by Consalvo, et al. in U.S. Pat. No. 7,445,911; Miller (“Xaa) (position 32 in SEQID Nos: 11, 13-15 and 17-18 30 et al., U.S. Patent Publication No. 2006/0292672; Ray et al. or position 33 in SEQID NO: 16) may include an amidated 2002, Protein Expression and Purification, 26:249-259; and group “NH. Alternative ways of reducing positive charge Mehta, 2004, Biopharm. International, July, pp. 44-46. include, but are not limited to, polyethylene glycol-based The production of the preferred amidated peptides may PEGylation, or the addition of another amino acid such as proceed, for example, by producing glycine-extended pre glutamic acid or aspartic acid at the N-terminus. Alterna 35 cursor in E. coli as a soluble fusion protein with glutathione tively, other amino acids may be added to the N-terminus of S-transferase, or by direct expression of the precursor in peptides discussed Supra including, but not limited to, accordance with the technique described in U.S. Pat. No. lysine, glycine, formylglycine, leucine, alanine, acetyl ala 6,103,495. Such a glycine extended precursor has a molecu nine, and dialanyl. An example of an amino acid added to the lar structure that is identical to the desired amidated product N-terminus of peptides includes SEQ ID NO:16, where a 40 except at the C-terminus (where the product terminates lysine has been added. —X—NH2, while the precursor terminates —X-gly, X “Xaa' in SEQID NO: 18 in Table 1 can be any naturally being the C-terminal amino acid residue of the product). An occurring amino acid. In an embodiment Xaa at position 31 alpha-amidating enzyme described in the publications above is selected from one of threonine or alanine. In an embodi catalyzes conversion of precursors to product. That enzyme ment Xaa at position 32 is selected from one of tyrosine or 45 is preferably recombinantly produced, for example, in Chi proline. Thus, SEQID NOs: 11, 15, 16 and 17, are encom nese Hamster Ovary (CHO) cells), as described in the passed by SEQ ID NO: 18. Biotechnology and Biopharm. articles cited above. Other amino acid sequences of the calcitonin mimetics of Free acid forms of peptide active agents of the present the present disclosure are found in Table 1B below. In these disclosure may be produced in like manner, except without sequences the R Substituent may be an acylation moiety or 50 including a C-terminal glycine on the “precursor, which may be absent. precursor is instead the final peptide product and does not require the amidation step. TABLE 1B Except where otherwise stated, the preferred dosage of the calcitonin mimetics of the present disclosure is identical for SEQ 55 both therapeutic and prophylactic purposes. Desired dosages Amino Acid Sequence ID NO : are discussed in more detail, infra, and differ depending on R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANAP-NH, 19 mode of administration. Except where otherwise noted or where apparent from R-CSNLSTCVLGKLSOELHKLQTYPRTNTGSGTP-NH, 2O context, dosages herein refer to weight of active compounds 60 unaffected by pharmaceutical excipients, diluents, carriers R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANTY-NH, 21 or other ingredients, although such additional ingredients are R-CSNLSTCVLGRLSOELHRLOTFPRTDVGANTAcY 22 desirably included, as shown in the examples herein. Any dosage form (capsule, tablet, injection or the like) com R-KCSNLSTCVLGKLSQELHKLQTYPRTDVGANTY-NH, 23 monly used in the pharmaceutical industry for delivery of R-CSNLSTCVLGKLSOELHKLQTYPRTDVGANAY-NH, 24 65 peptide active agents is appropriate for use herein, and the terms “excipient”, “diluent’, or “carrier includes such non-active ingredients as are typically included, together US 9,533,022 B2 7 8 with active ingredients in Such dosage form in the industry. Said method may include a preliminary step of determin A preferred oral dosage form is discussed in more detail, ing whether the patient Suffers from a said condition, and/or infra, but is not to be considered the exclusive mode of a Subsequent step of determining to what extent said treat administering the active agents of the present disclosure. ment is effective in mitigating the condition in said patient, The calcitonin mimetics of the present disclosure can be e.g. in each case, carrying out an oral glucose tolerance test administered to a patient to treat a number of diseases or or a resting blood Sugar level. disorders. As used herein, the term "patient’ means any For improved control over the weight of the patient, to organism belonging to the kingdom Animalia. In an embodi produce a loss of weight or an avoidance of weight gain, the ment, the term “patient” refers to vertebrates, more prefer active compound is preferably administered at least twice ably, mammals including humans. 10 per day, e.g. from 2-4 times per day. Formulations of the Accordingly, the present disclosure provides a method of active compound may contain a unit dosage appropriate for treatment of type I diabetes, Type II diabetes or metabolic Such an administration schedule. The active compounds may syndrome, obesity, or of appetite Suppression, or for miti be administered with a view to controlling the weight of a gating insulin resistance, or for reducing an undesirably high patient undergoing treatment for diabetes or metabolic Syn fasting serum glucose level, or for reducing an undesirably 15 drome. high peak serum glucose level, or for reducing an undesir Oral enteral formulations are for ingestion by Swallowing ably high peak serum insulin level, or for reducing an for subsequent release in the intestine below the stomach, undesirably large response to a glucose tolerance test, or for and hence delivery via the portal vein to the liver, as opposed treating osteoporosis, or for treating osteoarthritis. to formulations to be held in the mouth to allow transfer to As used herein, the term “glycemic control” refers to the the bloodstream via the sublingual or buccal routes. typical levels of blood Sugar (glucose) in a person with Suitable dosage forms for use in the present disclosure diabetes mellitus. The percentage of hemoglobin which is include tablets, mini-tablets, capsules, granules, pellets, glycosolated (measured as hemoglobin Alc) is used as a powders, effervescent solids and chewable solid formula proxy measure of long-term glycemic control. tions. Such formulations may include gelatin which is pref As used herein, the term “improved glycemic control 25 erably hydrolysed gelatin or low molecular weight gelatin. refers to the ability of a calcitonin mimetic of the present Such formulations may be obtainable by freeze drying a disclosure to reduce the percentage of hemoglobin which is homogeneous aqueous solution comprising calcitonin or a glycosolated. fragment or conjugate thereof and hydrolysed gelatin or low There are a number of art-recognized measures of normal molecular weight gelatin and further processing the resulting range for body weight in view of a number of factors such 30 Solid material into said oral pharmaceutical formulation, and as gender, age and height. A patient in need of treatment or wherein the gelatin may have a mean molecular weight from prevention regimens set forth herein include patients whose 1000 to 15000 Daltons. Such formulations may include a body weight exceeds recognized norms or who, due to protective carrier compound such as 5-CNAC or others as heredity, environmental factors or other recognized risk disclosed herein. factor, are at higher risk than the general population of 35 Whilst oral formulations such as tablets and capsules are becoming overweight or obese. In accordance with the preferred, compositions for use in the present disclosure present disclosure, it is contemplated that the calcitonin may take the form of syrups, elixirs or the like and Supposi mimetics may be used to treat diabetes where weight control tories or the like. Oral delivery is generally the delivery is an aspect of the treatment. route of choice since it is convenient, relatively easy and In an embodiment, the method includes enteral adminis 40 generally painless, resulting in greater patient compliance tration to a patient in need thereof for treatment of a said relative to other modes of delivery. However, biological, condition of a pharmaceutically effective amount of any one chemical and physical barriers such as varying pH in the of the peptides described herein. gastrointestinal tract, powerful digestive enzymes, and In an embodiment, the method includes parenteral admin active agent impermeable gastrointestinal membranes, istration to a patient in need thereof for treatment of a said 45 makes oral delivery of calcitonin like peptides to mammals condition of a pharmaceutically effective amount of any one problematic, e.g. the oral delivery of calcitonins, which are of the peptides described herein. For parenteral administra long-chain polypeptide hormones secreted by the parafolli tion (including intraperitoneal, Subcutaneous, intravenous, cular cells of the thyroid gland in mammals and by the intradermal or intramuscular injection), Solutions of a pep ultimobranchial gland of birds and fish, originally proved tide of the present disclosure in either sesame or peanut oil 50 difficult due, at least in part, to the insufficient stability of or in aqueous propylene glycol may be employed, for calcitonin in the gastrointestinal tract as well as the inability example. The aqueous solutions should be suitably buffered of calcitonin to be readily transported through the intestinal (preferably pH greater than 8) if necessary and the liquid walls into the blood stream. diluent first rendered isotonic. These aqueous solutions are Suitable oral formulations are however described below. Suitable for intravenous injection purposes. The oily solu 55 Treatment of Patients tions are Suitable for intraarticular, intramuscular and Sub In an embodiment, a calcitonin mimetic of the present cutaneous injection purposes. The preparation of all these disclosure is administered at adequate dosage to maintain Solutions under Sterile conditions is readily accomplished by serum levels of the mimetic in patients between 5 and 500 standard pharmaceutical techniques well known to those picograms per milliliter, preferably between 10 and 250 skilled in the art. For parenteral application, examples of 60 picograms per milliliter. The serum levels may be measured Suitable preparations include solutions, preferably oily or by radioimmunoassay techniques known in the art. The aqueous solutions as well as Suspensions, emulsions, or attending physician may monitor patient response, and may implants, including Suppositories. Peptides may be formu then alter the dosage somewhat to account for individual lated in sterile form in multiple or single dose formats Such patient metabolism and response. Near simultaneous release as being dispersed in a fluid carrier Such as sterile physi 65 is best achieved by administering all components of the ological saline or 5% saline dextrose solutions commonly present disclosure as a single pill or capsule. However, the used with injectables. disclosure also includes, for example, dividing the required US 9,533,022 B2 10 amount of the calcitonin mimetic among two or more tablets wherein R', R. R. and R are independently hydrogen, or capsules which may be administered together such that —OH, -NR'R'', halogen, C-C alkyl, or C-C alkoxy; R they together provide the necessary amount of all ingredi is a Substituted or unsubstituted C-C alkylene, Substituted ents. "Pharmaceutical composition, as used herein includes but is not limited to a complete dosage appropriate to a 5 or unsubstituted C-C alkenylene, Substituted or unsubsti particular administration to a patient regardless of whether tuted C-C alkyl(arylene), or substituted or unsubstituted one or more tablets or capsules (or other dosage forms) are aryl(C-C alkylene); and R and R are independently recommended at a given administration. hydrogen, oxygen, or C-C alkyl; and hydrates and Solvates A calcitonin mimetic of the present disclosure may be thereof as particularly efficacious for the oral delivery of formulated for oral administration using the methods active agents. Such as calcitonins, e.g. salmon calcitonin, employed in the Unigene Enteripep(R) products. These may 10 and these may be used in the present disclosure. include the methods as described in U.S. Pat. No. 5,912,014, Preferred enteric formulations using optionally U.S. Pat. No. 6,086,918, U.S. Pat. No. 6,673,574, U.S. Pat. micronised 5-CNAC may be generally as described in No. 7,316,819, U.S. Pat. No. 8,093,207, and US Publication WO2005/O14031. No. 2009/0317462. In particular, it may include the use of The compound may be formulated for oral administration conjugation of the compound to a membrane translocator 15 using the methods employed in the Capsitonin product of such as the protein transduction domain of the HIV TAT Bone Medical Limited. These may include the methods protein, co-formulation with one or more protease inhibitors, incorporated in AXcess formulations. More particularly, the and/or a pH lowering agent which may be coated and/or an active ingredient may be encapsulated in an enteric capsule acid resistant protective vehicle and/or an absorption capable of withstanding transit through the stomach. This enhancer which may be a surfactant. may contain the active compound together with a hydro In an embodiment, a calcitonin mimetic of the present philic aromatic alcohol absorption enhancer, for instance as disclosure is preferably formulated for oral delivery in a described in WO02/028436. In a known manner the enteric manner known in U.S. Patent Publication No. 2009/ 0317462. One preferred oral dosage form in accordance with coating may become permeable in a pH sensitive manner, 25 e.g. at a pH of from 3 to 7. WO2004/091584 also describes the present disclosure is set forth in Table 2 below: Suitable formulation methods using aromatic alcohol absorption enhancers. TABLE 2 The compound may be formulated using the methods seen ACTIVE AGENT OR EXCIPIENT FUNCTION in the Oramed products, which may include formulation 30 with omega-3 fatty acid as seen in WO2007/029238 or as A Calcitonin Mimetic selected from Active agent one of SEQ ID NO: 11 described in U.S. Pat. No. 5,102,666. SEQ ID NO: 18 Generally, the pharmaceutically acceptable salts (espe Coated Citric Acid Particles Protease Inhibitor cially mono or di Sodium salts), Solvates (e.g. alcohol Lauroylcarnitine Absorption Enhancer Solvates) and hydrates of these carriers or delivery agents Nonionic Polymer Subcoat 35 may be used. Eudragit L3OD-55 Enteric Coat Oral administration of the pharmaceutical compositions according to the disclosure can be accomplished regularly, In an embodiment, a calcitonin mimetic of the present e.g. once or more on a daily or weekly basis; intermittently, disclosure may be formulated for enteral, especially oral, e.g. irregularly during a day or week; or cyclically, e.g. administration by admixture with a suitable carrier com 40 regularly for a period of days or weeks followed by a period pound. Suitable carrier compounds include those described without administration. The dosage form of the pharmaceu in U.S. Pat. No. 5,773,647 and U.S. Pat. No. 5,866,536 and tical compositions of the presently disclosed embodiments amongst these, 5-CNAC (N-(5-chlorosalicyloyl)-8-amin can be any known form, e.g. liquid or solid dosage forms. ocaprylic acid, commonly as its disodium salt) is particu The liquid dosage forms include Solution emulsions, Sus larly effective. Other preferred carriers or delivery agents are 45 pensions, syrups and elixirs. In addition to the active com SNAD (sodium salt of 10-(2-Hydroxybenzamido)decanoic pound and carrier such as 5-CNAC, the liquid formulations acid) and SNAC (sodium salt of N-(8-2-hydroxybenzoyl may also include inert excipients commonly used in the art amino)caprylic acid). In an embodiment, a pharmaceutical Such as, solubilizing agents e.g. ethanol; oils such as cot composition of the present disclosure comprises a delivery tonseed, castor and sesame oils; wetting agents; emulsifying effective amount of carrier such as 5-CNAC, i.e. an amount 50 agents; Suspending agents; Sweeteners; flavourings; and sufficient to deliver the compound for the desired effect. Solvents such as water. The solid dosage forms include Generally, the carrier such as 5-CNAC is present in an capsules, soft-gel capsules, tablets, caplets, powders, gran amount of 2.5% to 99.4% by weight, more preferably 25% ules or other solid oral dosage forms, all of which can be to 50% by weight of the total composition. prepared by methods well known in the art. The pharma In addition, WO 00/0598.63 discloses the disodium salts 55 ceutical compositions may additionally comprise additives of formula I in amounts customarily employed including, but not limited to, a pH adjuster, a preservative, a flavorant, a taste-masking agent, a fragrance, a humectant, a tonicifier, a colorant, a Surfactant, a plasticizer, a lubricant Such as magnesium 60 Stearate, a flow aid, a compression aid, a solubilizer, an excipient, a diluent such as microcrystalline cellulose, e.g. Avicel PH 102 supplied by FMC corporation, or any com bination thereof. Other additives may include phosphate OH buffer salts, citric acid, glycols, and other dispersing agents. 65 The composition may also include one or more enzyme inhibitors, such as actinonin or epiactinonin and derivatives thereof: aprotinin, Trasylol and Bowman-Birk inhibitor. US 9,533,022 B2 11 12 Further, a transport inhibitor, i.e. a rhol-glycoprotein use of thiolated polymers as carrier matrix in oral peptide Such as Ketoprofin, may be present in the compositions of delivery—Proof of concept. J. Control. Release, 106, the present disclosure. The Solid pharmaceutical composi 26-33. tions of the instant disclosure can be prepared by conven The active compound may be formulated in seamless tional methods e.g. by blending a mixture of the active micro-spheres as described in WO2004/084870 where the compound, the carrier such as 5-CNAC, and any other active pharmaceutical ingredient is solubilised as an emul ingredients, kneading, and filling into capsules or, instead of Sion, microemulsion or Suspension, formulated into mini filling into capsules, molding followed by further tableting spheres; and variably coated either by conventional or novel or compression-molding to give tablets. In addition, a solid coating technologies. The result is an encapsulated drug in dispersion may be formed by known methods followed by 10 further processing to form a tablet or capsule. Preferably, the “pre-solubilised form which when administered orally pro ingredients in the pharmaceutical compositions of the instant vides for predetermined instant or sustained release of the disclosure are homogeneously or uniformly mixed through active drug to specific locations and at specific rates along out the Solid dosage form. the gastrointestinal tract. In essence, pre-solubilization of the Alternatively, the active compound may be formulated as 15 drug enhances the predictability of its kinetic profile while a conjugate with said carrier, which may be an oligomer as simultaneously enhancing permeability and drug stability. described in US2003/0069170, e.g. One may employ chitosan coated nanocapsules as described in US2009/0074824. The active molecule admin istered with this technology is protected inside the nanocap O Sules since they are stable against the action of the gastric fluid. In addition, the mucoadhesive properties of the system compound--C- (CH2)7(OCH)OCH3)2. enhances the time of adhesion to the intestine walls (it has been verified that there is a delay in the gastrointestinal Such conjugates may be administered in combination with a transit of these systems) facilitating a more effective absorp fatty acid and a bile salt as described there. 25 tion of the active molecule. Conujugates with polyethylene glycol (PEG) may be Methods developed by TSRI Inc. may be used. These used, as described for instance in Mansoor et al. include Hydrophilic Solubilization Technology (HST) in Alternatively, active compounds may be admixed with which gelatin, a naturally derived collagen extract carrying nitroso-N-acetyl-D.L-penicillamine (SNAP) and Carbopol both positive and negative charges, coats the particles of the solution or with taurocholate and Carbapol solution to form 30 active ingredient contained in lecithin micelles and prevents a mucoadhesive emulsion. their aggregation or clumping. This results in an improved The active compound may be formulated by loading into wettability of hydrophobic drug particles through polar chitosan nanocapsules as disclosed in Prego et al. (option interactions. In addition, the amphiphilic lecithin reduces ally PEG modified as in Prego Prego C, Torres D. Fernan surface tension between the dissolution fluid and the particle dez-Megia E, Novoa-Carballal R. Quiñoá E. Alonso M.J.) or 35 Surface. chitosan or PEG coated lipid nanoparticles as disclosed in The active ingredient may be formulated with cucurbi Garcia-Fuentes et al. Chitosan nanoparticles for this purpose turils as excipients. may be iminothiolane modified as described in Guggi et al. Alternatively, one may employ the GIPET technology of They may be formulated in waterfoil/water emulsions as Merrion Pharmaceuticals to produce enteric coated tablets described in Dogru et al. The bioavailability of active 40 containing the active ingredient with an absorption enhancer compounds may be increased by the use of taurodeoxy which may be a medium chain fatty acid or a medium chain cholate or lauroyl carnitine as described in Sinko et al. or in fatty acid derivative as described in US2007/0238707 or a Song et al. Generally, Suitable nanoparticles as carriers are membrane translocating peptide as described in U.S. Pat. discussed in de la Fuente etal and may be used in the present No. 7,268,214. disclosure. 45 One may employ GIRESTM technology which consists of Other suitable strategies for oral formulation include the a controlled-release dosage form inside an inflatable pouch, use of a transient permeability enhancer (TPE) system as which is placed in a drug capsule for oral administration. described in WO2005/094785 of Chiasma Ltd. TPE makes Upon dissolution of the capsule, a gas-generating system use of an oily Suspension of Solid hydrophilic particles in a inflates the pouch in the stomach. In clinical trials the pouch hydrophobic medium to protect the drug molecule from 50 has been shown to be retained in the stomach for 16-24 inactivation by the hostile gastrointestinal (GI) environment hours. and at the same time acts on the GI wall to induce perme Alternatively, the active may be conjugated to a protective ation of its cargo drug molecules. modifier that allows it to withstand enzymatic degradation in Further included is the use of glutathione or compounds the stomach and facilitate its absorption. The active may be containing numerous thiol groups as described in US2008/ 55 conjugated covalently with a monodisperse, short-chain 0200563 to inhibit the action of efflux pumps on the mucous methoxy polyethylene glycol glycolipids derivative that is membrane. Practical examples of Such techniques are crystallized and lyophilized into the dry active pharmaceu described also in Caliceti, P. Salmaso, S., Walker, G. and tical ingredient after purification. Such methods are Bernkop-Schnurch, A. (2004) Development and in vivo described in U.S. Pat. No. 5,438,040. evaluation of an oral insulin-PEG delivery system. Eur. J. 60 One may also employ a hepatic-directed vesicle (HDV) Pharm. Sci., 22, 315-323, in Guggi, D., Krauland, A. H., and for active delivery. An HDV may consist of liposomes (s.150 Bernkop-Schnurch, A. (2003) Systemic peptide delivery nm diameter) encapsulating the active, which also contain a via the stomach: in vivo evaluation of an oral dosage form hepatocyte-targeting molecule in their lipid bilayer. The for salmon calcitonin. J. Control. Rel. 92, 125-135, and in targeting molecule directs the delivery of the encapsulated Bernkop-Schnurch, A., Pinter. Y., Guggi, D., Kahlbacher, 65 active to the liver cells and therefore relatively minute H., Schöffmann, G., Schuh, M., Schmerold, I., Del Curto, M. amounts of active are required for effect. Such technology is D., D'Antonio, M., Esposito, P. and Huck, Ch. (2005) The described in US2009/0087479. US 9,533,022 B2 13 14 The active may be incorporated into a composition con tration, e.g. for treating type I diabetes, type II diabetes, or taining additionally a Substantially non-aqueous hydrophilic metabolic syndrome, or for mitigating insulin resistance, or medium comprising an alcohol and a cosolvent, in associa for reducing an undesirably high fasting serum glucose tion with a medium chain partial glyceride, optionally in level, or for reducing an undesirably high peak serum admixture with a long-chain PEG species as described in glucose level, or for reducing an undesirably high peak US2002/0115592 in relation to insulin. serum insulin level, or for reducing an undesirably high Alternatively, use may be made of intestinal patches as response to a glucose tolerance test, or for treating osteo described in Shen Z. Mitragotri S, Pharm Res. 2002 April; porosis, or for treating osteoarthritis. The formulation may 19(4):391-5 Intestinal patches for oral drug delivery. comprise also a carrier serving to enable effective enteral The active may be incorporated into an erodible matrix 10 formed from a hydrogel blended with a hydrophobic poly administration of said active compound. mer as described in U.S. Pat. No. 7,189,414. Preferably, said formulation is formulated for oral admin Suitable oral dosage levels for adult humans to be treated istration to the digestive tract. may be in the range of 0.05 to 5 mg, preferably about 0.1 to Preferably, said carrier comprises 5-CNAC, SNAD, or SNAC 2.5 mg. 15 The frequency of dosage treatment of patients may be Additionally, the present disclosure includes said peptides from 1 to six times daily, for instance from two to four times as new compounds. daily. Treatment will desirably be maintained over a pro The presently disclosed embodiments is described in the longed period of at least 6 weeks, preferably at least 6 following Examples, which are set forth to aid in the months, preferably at least a year, and optionally for life. understanding of the disclosure, and should not be construed Combination treatments for relevant conditions may be to limit in any way the scope of the disclosure as defined in carried out using a composition according to the present the claims which follow thereafter. The following examples disclosure and separate administration of one or more other are put forth so as to provide those of ordinary skill in the therapeutics. Alternatively, the composition according to the art with a complete disclosure and description of how to present disclosure may incorporate one or more other thera 25 make and use the described embodiments, and are not peutics for combined administration. intended to limit the scope of the present disclosure nor are Combination therapies according to the present disclosure they intended to represent that the experiments below are all include combinations of an active compound as described or the only experiments performed. Efforts have been made with insulin, GLP-2, GLP-1, GIP, or amylin, or generally to ensure accuracy with respect to numbers used (e.g. with other anti-diabetics. Thus combination therapies 30 amounts, temperature, etc.) but some experimental errors including co-formulations may be made with insulin sensi and deviations should be accounted for. Unless indicated tizers including biguanides such as Metformin, Buformin otherwise, parts are parts by weight, molecular weight is and Phenformin, TZD's (PPAR) such as Balaglitazone, weight average molecular weight, temperature is in degrees PioglitaZone, RivoglitaZone, RosiglitaZone and Troglita Centigrade, and pressure is at or near atmospheric. Zone, dual PPAR agonists such as Aleglitazar, Muraglitazar 35 and Tesaglitazar, or secretagogues including Sulphonylureas EXAMPLE 1. such as Carbutamide, Chloropropamide, Gliclazide, Tolbu tamide, Tolazamide, Glipizide, Glibenclamide, Glyburide, Chronic Effect of Calcitonin Mimetic 1 (CM1) Compared to Gliquidone, Glyclopyramide and Glimeprinide, Megli SCT tinides/glinides (K--) such as Nateglinide, Repaglinide and 40 Animals Mitiglinide, GLP-1 analogs such as Exenatide, Liraglutide The study was performed in male Levin-DIO rats (diet and Albiglutide, DPP-4 inhibitors such as Alogliptin, Lina sensitive) and Levin-DR (diet-resistant) (TacLevin: CD gliptin, Saxagliptin, Sitagliptin and Vildagliptin, insulin ana (SD) DDIO) (Taconic, Hudson, N.Y., U.S.A.) obtained at logs or special formulations such as (fast acting) Insulin age 6-7 weeks. On arrival, DIO rats were given high fat diet lispro, Insulin aspart, Insulin glulisine, (long acting) Insulin 45 (60 kcal %) (#D12495, Research Diets Inc., New Bruns glargine, Insulin detemir), inhalable insulin-Exubra and wick, N.J., USA) and kept on the same diet for 16 weeks NPH insulin, and others including alpha-glucosidase inhibi prior to and during the experiment. DR rats were given tors such as Acarbose, Miglitol and Voglibose, amylin low-fat diet and served as control group. Animals were analogues such as Pramlintide, SGLT2 inhibitors such as pair-wise housed throughout the study. Rats were handled Dapagliflozin, Remogliflozin and Sergliflozin as well as 50 and pre-dosed once daily with MilliO HO for 2-3 weeks miscellaneous ones including Benfluorex and Tolrestat. prior to experimental start to reduce stress-induced hyper Further combinations include co-administration or co glycaemia. Baseline parameters were recorded in an fasting formulation with leptins. Leptin resistance is a well-estab (6 h) condition. Rats were randomized into treatment groups lished component of type 2 diabetes; however, injections of based on fasting body weight (BW) and fasting plasma leptin have so far failed to improve upon this condition. In 55 glucose (FPG). Body weight, food and water intake were contrast, there is evidence Supporting that amylin, and recorded once weekly during the study period. thereby molecules with amylin-like abilities, as the salmon Compound calcitonin mimetics, are able to improve leptin sensitivity. Oral sGT or calcitonin mimetic 1 solution was prepared Amylin/leptin combination has shown a synergistic effect on on the day of dosing by mixing a carrier with the given body weight and food intake, and also insulin resistance 60 compound based on the following calculations: Kusakabe T et al. Accordingly, the present disclosure 5-CNAC (Vehicle): provides a compound of the formula Ac-CSNLSTCVLG Animals treated with oral 5-CNAC received a dose of 150 KLSQELHKLQ TYPRTDVGAN AP-NH. (SEQ ID NO: mg/kg dissolved in milliO H20. 15), which will be referred to herein as calcitonin mimetic Dosage-level for 5-CNAC: 150 mg/kg 1 or UGP3O2. 65 Dosing Volume: 5 ml/kg Accordingly, the present disclosure includes a pharma Compound concentration: 30 mg/ml ceutical formulation of Such a peptide for enteral adminis sCT/Calcitonin Mimetic 1: US 9,533,022 B2 15 16 Animals treated with oral SCT or oral calcitonin mimetic parison. Student's t-test was performed to compare two 1 received doses of 1.0 mg/kg combined with 150 mg/kg paired group. All analysis was performed using GRAPH 5-CNAC all dissolved in milliO H20. PAD PRISM software (GraphPad Prism, San Diego, Calif. Dosage-level for SCT/calcitonin mimetic 1: 1.0 mg/kg U.S.A). Incremental area under curve (iAUC) during OGTT Dosing Volume: 5 ml/kg was calculated by the trapezoidal method. A value of P-0.05 Compound concentration: 0.2 mg/ml was considered to be significant. All data are presented as Drug administration by per oral (p.o.) gavage b.i.d. (7-8 meanistandard error of the mean (SEM). am and 3-4 pm) during the study period and as single dose Results in the morning prior to start of OGTT. Baseline Characteristics: Oral gavage of glucose during OGTT was prepared by the 10 Results are summarized in FIGS. 1A-1D (Food intake and following calculation: body weight), FIGS. 2A-2B (OGTT) and FIG.3 (FPG). FIG. D-Glucose: Animals were given 2 g/kg single dose dissolved in 1A, FIG. 1B, FIG. 1C, and FIG. 1D show the effect of milliO H20. chronic oral salmon calcitonin (“sCT) versus oral UGP302 Dosage-level for D-Glucose: 2 g/kg 15 administration on body weight and food intake in DIO rats Dosing Volume: 4 ml/kg as measured in Example 1. FIG. 2A and FIG. 2B show the Compound concentration: 500 mg/ml effect of oral sGT versus oral UGP302 on glucose tolerance Experimental Setup: during OGTT in DIO rats as measured in Example 1. FIG. 3 shows the effect of oral SCT versus oral UGP 302 on fasting glycemia in DIO rats as measured in Example 1: One dose of oral SCT/calcitonin mimetic 1 containing 1 Baseline Week 1 Week 2 Week 3 Week 4 mg/kg compound was applied by gavage twice daily to four FPG BW BW BW BW BW Food Food Food Food groups of rats for 4 weeks. An oral vehicle group served as B FPG FPG dosing regimen control, respectively. * P-0.05, ** P<0.01, B B 25 *** P<0.001 vs Vehicle. Results are presented as OGTT means-SEM. FPG = Fasting Plasma Glucose; The 16-weeks ad libitum high-fat diet induced a pro BW = Body Weight; nounced obese phenotype in the diet-sensitive (D10) rats B = Blood; when comparing body weight to their diet-resistant (DR) OGTT = Oral Glucose Tolerance Test 30 littermates (P<0.001) (Table 3). 6-hrs Fasting glycemia was not different between DIO and DR. In contrast, area under OGTT Following Overnight Fasting (16 h): curve (AUC) calculations during OGTT was significantly higher in DIO rats compared to DR rats, demonstrating the -30 O 15 30 60 120 240 min high-fat diet-induced glucose intolerance (Table 3). 35 D G B B B B B TABLE 3 B B BG BG BG BG BG BG BG Metabolic parameters in DIO and DR rats D = Drug; Diet-resistant (DR) Diet-sensitive (DIO) BG = Blood glucose; 40 B = Blood; Body Weight (g) 609.5 24.5 841.8 22.9*** G = Glucose Fasting plasma glucose (mM) 6.5 - 0.1 6.8 O2 AUC in OGTT 625.1 20.5 914.3 44.6*** Blood sampling and glycemia were measured by heated Blood glucose (mM*min) tail venous puncture. AUC, area under curve; OGTT, oral glucose tolerance test, Whole blood glucose levels were determined with an 45 ACCU-CHEKR) Avia blood glucose meter (Roche Diagnos Data are means + SEM (n = 12/DR) n = 24/DIO). tics, Rotkreuz, Switzerland). Blood (approx 300 ul) is col Body Weight and Food Intake lected in 1 ml MiniCollect K3EDTA plasma-tube (Greiner During the first week of treatment administration of oral Bio-One GmbH, Frickenhausen, Germany), inverted, and SCT significantly reduced food intake compared to oral stored on ice. Tubes are centrifuged 3000xg (5000 rpm in 50 vehicle treated rats. Furthermore, oral sGT protected against table centrifuge) for 10 min at 4° C. and plasma obtained. further gain in body weight as observed for oral vehicle Plasma samples are stored at -20°C. until analysis. A total group (FIGS. 1A-1D). Thus, these observations confirm the of -2.5 ml blood is obtained during OGTT (-0.3% of body acute strong anorectic action induced by application of oral weight). SCT in DIO rats. Interestingly, from week 2 of treatment and Experimental Groups 55 throughout the study period, food intake normalized in oral SCT treated rats and resembled ingestion by oral vehicle resulting in a lack of difference in regards to cumulative food Intervention Compound Conc. Number intake at study end. This confirms previously reports Sug Oral vehicle S-CNAC 150 mg/kg n = 10 gesting a transient effect of oral SCT upon energy intake. Oral SCT 5-CNAC - SCT 150 mg/kg + n = 10 60 However, throughout the study period, oral SCT sustained 1 mg/kg the protecting effect on body weight gain and significantly Oral calcitonin mimetic 1 5-CNAC + 150 mg/kg n = 10 reduced body weight from baseline when compared to oral calcitonin mimetic 1 1 mg/kg vehicle at study end (FIGS. 1A-1B). This is in line with a possibly endogenous effect of oral SCT upon energy expen Statistics 65 diture to chronically regulate energy balance. Statistical analysis was performed by one-way ANOVA Generally, oral application of calcitonin mimetic 1 followed by the Dunnett's post hoc test for multiple com resembled the strong anorectic action of oral SCT during the US 9,533,022 B2 17 18 initial week of treatment and significantly reduced food Dosage-level for 5-CNAC: 150 mg/kg intake and protected against gain in body weight compared Dosing Volume: 5 ml/kg to oral vehicle group (FIG. 1). As observed for oral SCT, Compound concentration: 30 mg/ml calcitonin mimetic 1 exerted a transient effect on food sCT/Calcitonin Mimetic 1: intake, although food intake trended reduced when com 5 Animals treated with oral SCT or oral calcitonin mimetic pared to oral SCT during the study period. Thus, following 1 received doses of 0.5 mg/kg, 1.0 mg/kg or 2.0 mg/kg four weeks of treatment cumulative food intake was signifi combined with 150 mg/kg. 5-CNAC all dissolved in milliO cantly reduced in calcitonin mimetic 1 when compared to H2O. oral vehicle. Furthermore, when compared to oral SCT, a Dosage-level for SCT/calcitonin mimetic 1: 0.5 mg/kg more pronounced significant reduction in body weight was 10 Dosing Volume: 5 ml/kg observed suggesting Superiority in regards to effect on Compound concentration: 0.1 mg/ml energy balance. Dosage-level for SCT/calcitonin mimetic 1: 1.0 mg/kg Glucose Tolerance: Dosing Volume: 5 ml/kg Results are shown in FIGS. 2A-2B. One dose of oral Compound concentration: 0.2 mg/ml 15 Dosage-level for SCT/calcitonin mimetic 1: 2.0 mg/kg SCT/calcitonin mimetic 1 containing 1 mg/kg compound Dosing Volume: 5 ml/kg were applied by gavage twice daily to four groups of rats for Compound concentration: 0.4 mg/ml 4 weeks. An oral vehicle group served as dosing regimen Drug administration were given by per oral (p.o.) gavage control. OGTT performed following 2 weeks of treatment b.i.d. during the study period and as single dose in the after overnight-fasting.*** P<0.001 vs Vehicle. Results are morning prior to start of OGTT. presented as means-SEM. Oral gavage of glucose during OGTT was prepared by the Oral SCT significantly reduced glucose iAUC during following calculation: OGTT after 2 weeks of treatment compared to oral vehicle, D-Glucose: thus confirming the postprandial glycemic control exerted Animals were given 2 g/kg single dose dissolved in by oral application of SCT as previously demonstrated. In 25 milliO H20. general, calcitonin mimetic 1 demonstrated a similar sig Dosage-level for D-Glucose: 2 g/kg nificant reduction in iAUC as observed for oral SCT, Dosing Volume: 4 ml/kg although with no clear superiority to oral SCT in this respect. Compound concentration: 500 mg/ml Fasting Glycaemia: Experimental Setup Following 2 and 4 weeks of treatment, oral SCT applica 30 Acute Testing Treatment Period for 0.5 mg/kg, 1 mg/kg tion was not significantly different from oral vehicle treated and 2 mg/kg: rats, which is in contrast with previously observations in male DIO rats, in where a 1-1.5 mM reduction in fasting blood glucose typically is observed following chronic treat Day O Day 1-2 Day 3 Day 4 Day 5 Day 6 Day 7 35 ment. For calcitonin mimetic 1, a trend towards Superiority 1st Rest Pre-dose Treat- Treat- Treat- 2' in fasting glycaemia was observed throughout the study OGTT ment ment ment OGTT period when compared to oral vehicle or oral SCT. All No All (b.i.d) (b.i.d) (b.i.d) Single vehicle handling vehicle dose EXAMPLE 2 40 Following the initial (1) OGTT, animals are randomized Acute and Short Term Effects of Oral SCT Versus Oral into treatment groups based on FBG and BW. Animals will Calcitonin Mimetic 1 be pre-treated 3 days (b.i.d.) prior to 2" OGTT. Dosing will Animals be performed in the morning (7-8 am) and afternoon (3-4 The study was performed in male Levin-DIO rats (diet pm). sensitive) and Levin-DR (diet-resistant) (TacLevin: CD 45 The study was performed in an X-over design with each (SD) D10) (Taconic, Hudson, N.Y., U.S.A.) obtained at age animal being its own control. 6-7 weeks. On arrival, DIO rats were given high fat diet (60 OGTT Following Overnight Fasting (16 h): kcal %) (#D12495, Research Diets Inc., New Brunswick, N.J., USA) and kept on the same diet for 12 weeks prior to and during the experiment. DR rats were given low-fat diet 50 -30 O 15 3O 60 120 240 min and served as control group. Animals were pair-wise housed throughout the study. Rats were handled and pre-dosed once D G B B B B B daily with MilliOHO for 2-3 weeks prior to experimental B B BG BG BG BG BG start to reduce stress-induced hyperglycaemia. On the day BG BG prior to study start animals were given a single dose of 55 D = Drug; vehicle. Baseline parameters were recorded in an overnight BG = Blood glucose; fasting (16 h) condition. Rats were randomized into treat B = Blood; ment groups based on fasting body weight (BW) and fasting G = Glucose plasma glucose (FPG). Body weight, food and water intake Blood sampling and glycemia were measured by heated were recorded prior to and at study end. 60 tail venous puncture. Compounds Whole blood glucose levels were determined with an Oral SCT/calcitonin mimetic 1 solution was prepared on ACCU-CHEKR) Avia blood glucose meter (Roche Diagnos the day of dosing by mixing the carrier with the given tics, Rotkreuz, Switzerland). Blood (approx 300 ul) is col compound based on the following calculations: lected in 1 ml MiniCollect K3EDTA plasma-tube (Greiner 5-CNAC (Vehicle): 65 Bio-One GmbH, Frickenhausen, Germany), inverted, and Animals treated with oral 5-CNAC received a dose of 150 stored on ice. Tubes are centrifuged 3000xg (5000 rpm in mg/kg dissolved in milliO H20. table centrifuge) for 10 min at 4° C. and plasma obtained. US 9,533,022 B2 19 20 Plasma samples are stored at -20°C. until analysis. A total the effect of oral sGT versus oral UGP302 on body weight of -2.5 ml blood is obtained during OGTT (-0.3% of body and food intake in DIO rats observed in Example 2 at a first weight). dosage. FIG. 5A and FIG. 5B show the effect of oral sGT Experimental Groups versus oral UGP302 on body weight and food intake in DIO rats observed in Example 2 at a second dosage. FIG. 6A and FIG. 6B show the effect of oral SCT versus oral UGP302 on Intervention Compound Conc. Number body weight and food intake in DIO rats observed in Oral vehicle S-CNAC 150 mg/kg (4 groups of n = 8) Example 2 at a third dosage; X-over design Oral SCT dose-dependently decreased body weight and to 0.5 mg/kg 10 Oral SCT S-CNAC - SCT 150 mg/kg + n = 8 food intake following the short-term treatment period and 0.5 mg/kg thus confirmed the anorectic action induced by targeting the Oral calcitonin S-CNAC - 150 mg/kg n = 8 amylin receptor as previously observed. In general, the mimetic 1 calcitonin 0.5 mg/kg mimetic demonstrated dose-dependent Superiority to oral mimetic 1 Oral vehicle S-CNAC 150 mg/kg (4 groups of n = 8) SCT in regards to reduction in body weight as illustrated in X-over design 15 FIGS. 4A-4B, FIGS. 5A-5B and FIGS. 6A-6B. Application to 1 mg/kg of calcitonin mimetic 1 at 0.5 mg/kg demonstrated signifi Oral SCT S-CNAC - SCT 150 mg/kg + n = 8 cantly difference to oral SCT 0.5 mg/kg. The food intake for 1 mg/kg Oral calcitonin S-CNAC - 150 mg/kg n = 8 the mimetic trended dose-dependently reduced compared to mimetic 1 calcitonin 1 mg/kg oral SCT. mimetic 1 Glucose Tolerance Oral vehicle S-CNAC 150 mg/kg (4 groups of n = 8) Three different doses of oral SCT/calcitonin mimetic 1 X-over design to 2 mg/kg containing 0.5, 1 and 2 mg/kg compound were applied by Oral SCT S-CNAC - SCT 150 mg/kg + n = 8 gavage twice daily to 4 groups of rats for 3 days prior to 2 mg/kg OGTT. The experimental set-up was a cross-over design. * Oral calcitonin S-CNAC - 150 mg/kg n = 8 25 P<0.05, ** P-0.01, *** P<0.001 vs oral vehicle. Results are mimetic 1 calcitonin 2 mg/kg presented in FIGS. 7A-7B and FIGS. 8A-8B as mimetic 1 means-SEM. FIG. 7A and FIG. 7B show the effect of oral SCT versus oral UGP 302 at a first dosage on glucose Statistics tolerance during OGTT in DIO rats as measured in Example Statistical analysis was performed by one-way ANOVA 30 2. FIG. 8A and FIG. 8B show the effect of oral SCT versus followed by the Dunnett's post hoc test for multiple com oral UGP 302 at a second dosage on glucose tolerance parison. Student's t-test was performed to compare two during OGTT in DIO rats as measured in Example 2. paired group. All analysis was performed using GRAPH Oral sGT significantly reduced glucose iAUC during PAD PRISM software (GraphPad Prism, San Diego, Calif. OGTT for 0.5, 1 and 2 mg/kg doses compared to oral U.S.A). Incremental area under curve (iAUC) during OGTT 35 vehicle, thus confirming the postprandial glycemic control was calculated by the trapezoidal method. A value of P-0.05 exerted by oral application of SCT as previously demon was considered to be significant. All data are presented as strated. Calcitonin mimetic 1 demonstrated a similar signifi meanistandard error of the mean (SEM). cantly reduction in iAUC as observed for oral SCT, although Results with no clear superiority to oral sGT within the various Baseline Characteristics 40 UGPS. The 12-weeks ad libitum high-fat diet induced a pro nounced obese phenotype in the diet-sensitive (D10) rats EXAMPLE 3 when comparing body weight to their diet-resistant (DR) littermates (P<0.001) (Table 1). Fasting glycemia was not Acute and Short Term Effects of Oral SCTVersus UGP284, different between DIO and DR. In contrast, area under curve 45 UGP298 and UGP3O2 (AUC) calculations during OGTT was significantly higher Animals in DIO rats compared to DR rats, demonstrating the high-fat The study was performed in male Levin-DIO rats (diet diet-induced glucose intolerance (Table 4). sensitive) and Levin-DR (diet-resistant) (TacLevin: CD (SD) D10) (Taconic, Hudson, N.Y., U.S.A.) obtained at age TABLE 4 50 6-7 weeks. On arrival, DIO rats were given high fat diet (60 kcal %) (#D12495, Research Diets Inc., New Brunswick, Metabolic parameters in DIO and DR rats N.J., USA) and kept on the same diet for 12 weeks prior to Diet-resistant (DR) Diet-sensitive (DIO) and during the experiment. DR rats were given low-fat diet Body Weight (g) 609.5 24.5 813.69.8*** and served as control group. Animals were pair-wise housed Fasting plasma glucose (mM) 5.8 0.1 5.8 O2 55 throughout the study. Rats were handled and pre-dosed once AUC in OGTT 648.8 27.3 888.4 64.3*** daily with MilliO H2O for 2-3 weeks prior to experimental Blood glucose (mM*min) start to reduce stress-induced hyperglycemia. On the day prior to study start animals were given a single dose of AUC, area under curve; OGTT, oral glucose tolerance test, Data are means + SEM (n = 12/DR, n = 24.DIO). vehicle. Baseline parameters were recorded in an overnight 60 fasting (16 h) condition. Rats were randomized into treat Body Weight and Food Intake ment groups based on fasting body weight (BW) and fasting Three different doses of oral SCT/calcitonin mimetic 1 plasma glucose (FPG). Body weight, food and water intake containing 0.5, 1 and 2 mg/kg compound were applied by were recorded prior to and at study end. gavage twice daily to 4 groups of rats for 3 days. * PK0.05, Compound ** P-3O.O1 VS Oral SCT. 65 Oral SCT/UGP solution was prepared on the day of dosing Results are presented in FIGS. 4A-4B, FIGS. 5A-5B, and by mixing the carrier with the given compound based on the FIGS. 6A-6B as means-SEM. FIG. 4A and FIG. 4B show following calculations: US 9,533,022 B2 21 22 5-CNAC (Vehicle): plasma-tube (Greiner-Bio-One GmbH, Frickenhausen, Ger Animals treated with oral 5-CNAC received a dose of 150 many), inverted, and stored on ice. Tubes are centrifuged mg/kg dissolved in milliO H20. 3000xg (5000 rpm in table centrifuge) for 10 min at 4° C. Dosage-level for 5-CNAC: 150 mg/kg and plasma obtained. Plasma samples are stored at -20°C. Dosing Volume: 5 ml/kg until analysis. A total of -2.5 ml blood is obtained during Compound concentration: 30 mg/ml OGTT (-0.3% of body weight). (sCT/UGP284/UGP298/UGP302) Experimental Groups: Animals treated with oral SCT or oral UGP284/UGP298/ UGP302 received doses of 0.5 mg/kg, 1.0 mg/kg or 2.0 10 mg/kg combined with 150 mg/kg. 5-CNAC all dissolved in Intervention Compound Conc. Number milliO H.O. Dosage-level for SCTAUGP284/UGP298/UGP3O2: O.5 Oral vehicle S-CNAC 50 mg/kg (4 groups of n = 8) X-Over design to 0.5 mg/kg mg/kg Oral SCT S-CNAC -- 150 mg/kg + n = 8 Dosing Volume: 5 ml/kg SCT 0.5 mg/kg Compound concentration: 0.1 mg/ml 15 Oral UGP284 S-CNAC -- 50 mg/kg n = 8 1.O UGP284 0.5 mg/kg Dosage-level for SCT/UGP284/UGP298/UGP302: Oral UGP298 S-CNAC - 50 mg/kg n = 8 mg/kg UGP298 0.5 mg/kg Dosing Volume: 5 ml/kg Oral UGP3O2 S-CNAC - 50 mg/kg n = 8 Compound concentration: 0.2 mg/ml UGP3O2 0.5 mg/kg Oral vehicle S-CNAC 50 mg/kg (4 groups of n = 8) Dosage-level for SCT/UGP284/UGP298/UGP302: 2.0 X-Over design to 1 mg/kg mg/kg Oral SCT S-CNAC -- 150 mg/kg + n = 8 Dosing Volume: 5 ml/kg SCT 1 mg/kg Compound concentration: 0.4 mg/ml Oral UGP284 S-CNAC -- 50 mg/kg n = 8 Drug administration were given by per oral (p.o.) gavage UGP284 1 mg/kg 25 Oral UGP298 S-CNAC - 50 mg/kg n = 8 b.i.d. during the study period and as single dose in the UGP298 1 mg/kg morning prior to start of OGTT. Oral UGP3O2 S-CNAC - 50 mg/kg n = 8 Oral gavage of glucose during OGTT was prepared by the UGP3O2 1 mg/kg following calculation: Oral vehicle S-CNAC 50 mg/kg (4 groups of n = 8) X-Over design to 2 mg/kg D-Glucose: Oral SCT S-CNAC -- 150 mg/kg + n = 8 Animals were given 2 g/kg single dose dissolved in milliO 30 SCT 2 mg/kg H2O. Oral UGP284 S-CNAC -- 50 mg/kg n = 8 Dosage-level for D-Glucose: 2 g/kg UGP284 2 mg/kg Oral UGP298 S-CNAC - 50 mg/kg n = 8 Dosing Volume: 4 ml/kg UGP298 2 mg/kg Compound concentration: 500 mg/ml Oral UGP3O2 S-CNAC - 50 mg/kg n = 8 Experimental Setup 35 UGP3O2 2 mg/kg Acute Testing Treatment Period for 0.5 mg/kg, 1 mg/kg and 2 mg/kg: Statistics Statistical analysis was performed by one-way ANOVA Day O Day 1-2 Day 3 Day 4 Day 5 Day 6 Day 7 40 followed by the Dunnett's post hoc test for multiple com 1st Rest Pre-dose Treat- Treat- Treat- 2" parison. Student's t-test was performed to compare two OGTT ment ment ment OGTT paired group. All analysis was performed using GRAPH All No All (b.i.d) (b.i.d.) (b.i.d) Single PAD PRISM software (GraphPad Prism, San Diego, Calif. vehicle handling vehicle dose U.S.A). Incremental area under curve (iAUC) during OGTT 45 was calculated by the trapezoidal method. A value of P-0.05 Following the initial (1) OGTT, animals were random was considered to be significant. All data are presented as ized into treatment groups based on FBG and BW. Animals meanistandard error of the mean (SEM). were pre-treated 3 days (b.i.d.) prior to 2" OGTT. The study was performed in an X-over design with each Results animal being its own control. 50 Baseline Characteristics OGTT Following Overnight Fasting (16 h): The 12-weeks ad libitum high-fat diet induced a pro nounced obese phenotype in the diet-sensitive (D10) rats when comparing body weight to their diet-resistant (DR) -30 O 15 30 60 120 240 min littermates (P<0.001) (Table 5). Fasting glycemia was not 55 D G B B B B B different between DIO and DR. In contrast, area under curve B B BG BG BG BG BG (AUC) calculations during OGTT were significantly higher BG BG in DIO rats compared to DR rats, demonstrating the high-fat D = Drug; diet-induced glucose intolerance (Table 5). BG = Blood glucose; 60 B = Blood; TABLE 5 G = Glucose Metabolic parameters in DIO and DR rats Blood sampling and glycemia were measured by heated tail venous puncture. Whole blood glucose levels were Diet-resistant (DR) Diet-sensitive (DIO) determined with an ACCU-CHEKR) Avia blood glucose 65 Body Weight (g) 609.5 24.5 813.69.8*** meter (Roche Diagnostics, Rotkreuz, Switzerland). Blood Fasting plasma glucose (mM) 5.8 O.1 5.8 O2 (approx 300 ul) is collected in 1 ml MiniCollect K3EDTA US 9,533,022 B2 23 24 TABLE 5-continued allowed to eat ad libitum. Food consumption and rat weights were monitored daily during each study. Rats were injected Metabolic parameters in DIO and DR rats intramuscularly with a saline placebo or the indicated pep Diet-resistant (DR) Diet-sensitive (DIO) tide at the specified dose in saline. The data in the following 5 tables is Summarized as the mean change in food consump AUC in OGTT 648.8 27.3 888.4 64.3*** tion relative to the day before treatment began (day -1) and Blood glucose (mM*min) as the mean change in weight relative to the day before AUC, area under curve; OGTT, oral glucose tolerance test, treatment began. Data are means + SEM (n = 12/DR, n = 24.DIO). The results are shown in FIGS. 12A-12B, 13 A-13B, 10 Body Weight and Food Intake 14A-14B, 15A-15B, 16A-16B, and 17A-17B. FIG. 12A and Oral SCT dose-dependently decreased body weight and FIG. 12B show food consumption and weight change mea food intake following the short-term treatment period and surements, respectively, for UGP 282 as measured in thus confirmed the anorectic action induced by targeting the Example 5. FIG. 13A and FIG. 13B show food consumption amylin receptor as previously observed. In general, all UGP 15 and weight change measurements, respectively, for UGP mimetics demonstrated dose-dependently Superiority to oral 283 as measured in Example 5. FIG. 14A and FIG. 14B SCT in regards to reduction in body weight as illustrated in show food consumption and weight change measurements, FIGS. 9A-9F. Application of UGP302 at 0.5 mg/kg demon respectively, for UGP 284 as measured in Example 5. FIG. strated significantly difference to oral sGT 0.5 mg/kg. For 15A and FIG. 15B show food consumption and weight UGP284, significantly difference at 2 mg/kg dose was change measurements, respectively, for UGP 298 as mea observed when compared to oral sGT 2 mg/kg. Finally, sured in Example 5. FIG. 16A and FIG. 16B show food UGP298 at both 1 mg/kg and 2 mg/kg doses were signifi consumption and weight change measurements, respec cantly different when compared with oral sGT at similar tively, for UGP302 as measured in Example 5. FIG. 17A doses. FIG. 9A, FIG.9B, FIG. 9C, FIG. 9D, FIG. 9E, and and FIG. 17B show food consumption and weight change FIG. 9F show the effect of three different doses of oral 25 measurements, respectively, for UGP 303 as measured in sCT/UGP284/UGP298/UGP302 containing 0.5, 1 and 2 Example 5. mg/kg compound were applied by gavage twice daily to 4 It can be seen that all of the tested compounds induce groups of rats for 3 days. * P-0.05, ** P<0.01 vs oral sGT. weight loss and reduce feed intake. Results are presented as means-SEM. EXAMPLE 6 Glucose Tolerance 30 FIG. 10A, FIG. 10B, FIG. 10C, FIG. 10D, FIG. 10E, and Markers of Osteoporosis and Osteoarthritis FIG. 10F show the effect of oral SCT versus oral UGPs on The effect of SCT/calcitonin mimetic on bone and carti glucose tolerance during OGTT in DIO rats. Three different lage loss was studied in DIO rats. The animals were dosed doses of oral sGT/UGP284/UGP298/UGP302 containing as described in Table 6 below, and 2 hours after treatment 0.5, 1 and 2 mg/kg compound were applied by gavage twice 35 blood sampling was done by heated tail venous puncture. daily to 4 groups of rats for 3 days prior to OGTT. The Serum CTX-I levels, as an indication of bone resorption, experimental set-up was a cross-over design. PK0.05, ** were measured using the RatapsTM ELISA, and serum P<0.01, *** P<0.001 vs oral vehicle. Results are presented CTX-II levels, as an indication of cartilage degradation, as means-SEM. All UGPs demonstrated a similar signifi cant reduction in iAUC as observed for oral SCT. 40 were measured using the Serum PC CartilapsTM ELISA. In conclusion, application of UGP284, UGP298 and UGP302 at 0.5, 1 and 2 mg/kg doses demonstrated superi TABLE 6 ority to equivalent doses of oral SCT in regards to energy Experimental groups balance in male DIO rats. Furthermore, UGP284, UGP298 and UGP302 at doses of 0.5, 1 and 2 mg/kg produced an 45 Intervention Compound Conc. Number improvement in glucose tolerance during OGTT. Oral vehicle S-CNAC 150 mg/kg n = 8 Oral SCT 5-CNAC - 150 mg/kg + n = 8 EXAMPLE 4 SCT 1 mg/kg Oral calcitonin mimetic 5-CNAC - 150 mg/kg n = 8 of SEQ ID NO: 18 SEQ ID NO: 15 1 mg/kg Binding of SCT Analogs to T47D Cell Calcitonin Receptors 50 SCT analogs at various concentrations were tested in a T47D (human breast epithelial cell line) bioassay. This cell The results are seen in FIG. 18 and FIG. 19, where a line is known to have the following receptors: calcitonin, calcitonin mimetic of SEQ ID NO: 18 shows a stronger androgen, progesterone, glucocrticoid, prolactin and estro effect in reduction of both bone resorption and cartilage gen. The results are presented in FIG. 11 as % cAMP binding 55 degradation than does SCT. relative to SCT which was set at 100% cAMP binding at a In some embodiments, a peptide of the present disclosure concentration of 1000 pg/mL. It can be seen that UGP302 has a sequence selected from SEQID NO: 11, SEQID NO: provides the highest level of binding of all the tested 12, SEQID NO: 13, SEQID NO: 14, SEQID NO: 15, SEQ compounds and that it provides a higher level of binding ID NO: 17 and SEQ ID NO: 18. than SCT. 60 In some embodiments, a method includes administering to a patient an effective amount of a peptide selected from EXAMPLE 5 the group consisting of: SEQ ID NO: 11, SEQ ID NO: 12, SEQID NO: 13, SEQID NO: 14, SEQID NO: 15, and SEQ Food Consumption and Weight Change in Rats Fed sGT ID NO: 17 to affect a weight reduction in the patient. Analogs 65 In some embodiments, a method includes administering Male Sprague-Dawley rats were housed individually in to a patient an effective amount of a peptide selected from cages in which the light/dark cycle was reversed. Rats were the group consisting of: SEQ ID NO: 11, SEQ ID NO: 12, US 9,533,022 B2 25 26 SEQID NO: 13, SEQID NO: 14, SEQID NO: 15, and SEQ As can be seen in FIGS. 20, 21 and 22, both metformin ID NO: 17 to affect postprandial glycemic control in the and UGP302 alone reduced both FPG and PPG, with patient. UGP302 providing a pronounced reduction throughout the study, while metformin lost the ability to control blood In some embodiments, a method includes administering glucose with time. In the combination group, a markedly to the patient an effective amount of a peptide selected from superior response was observed in both FPG and PPG, the group consisting of: SEQID NO: 11, SEQ ID NO: 12, clearly showing that these molecules work as adjunct thera SEQID NO: 13, SEQID NO: 14, SEQID NO: 15, and SEQ pies. This synergistic effect is also seen in the HbA1c levels, ID NO: 17 to affect an improvement in glycemic control in where the combination leads to a Superior improvement the patient. when compared to both vehicle and either molecule alone. In some embodiments, a method includes administering 10 Particularly notably, the Superior effect of the combination to a patient an effective amount of a peptide of SEQID NO: as compared to UGP302 alone persisted throughout the test, 18 having the sequence CSNLSTCVLGKLSQELH KLQ even though the individual effect of metformin did not. TYPRTDVGANXaaXaa, so as to reduce at least one of To further assess the effect of both single doses of each bone resorption and cartilage degradation in the patient. 15 molecule and the combination of these at those same doses, the rats underwent an oral glucose tolerance test (OGTT) EXAMPLE 7 and an insulin tolerance test (ITT). In these tests the activity of both molecules is apparent, as they individually improve Combination Treatment with Metformin both glucose and insulin tolerance. However, the combina To assess the potential of UGP302 (SEQ ID NO: 15) in tion of UGP302 and metformin was superior to either combination with metformin, Zucker Diabetic Fatty rats molecule alone, both in terms of glucose tolerance and were used. At the age of 6 weeks 40 rats were randomized insulin tolerance (FIGS. 23A-23B), clearly indicating the into four groups according to Hb A1c, FPG and BW. The benefit of the combination. groups were: Control (Vehicle), Metformin 400 mg/kg/day, It will be appreciated that several of the above-disclosed UGP302 5 g/kg/day and the combination of the two 25 and other features and functions, or alternatives thereof, may molecules in the Stated respective amounts. The treatments be desirably combined into many other different systems or were given once daily, and metformin was dosed orally, applications. Various presently unforeseen or unanticipated while UGP302 was given subcutaneously, and the animals alternatives, modifications, variations, or improvements were treated for 8 weeks. Fasting blood glucose (FPG) and therein may be subsequently made by those skilled in the art non-fasted blood glucose (PPG) were monitored biweekly, which are also intended to be encompassed by the following while HbA1c was measured at termination. claims.

SEQUENCE LISTING

<16 Os NUMBER OF SEO ID NOS: 24

<21 Oc SEO ID NO 1 <211 LENGTH: 32 <212> TYPE PRT ORGANISM: salmon

<4 OOs SEQUENCE: 1

Cys Ser Asn Lieu Ser Thr Cys Wall Lieu Gly Lys Luell Ser Glin Glu Lell 1. 5 15

His Lys Lieu Gln Thr Tyr Pro Arg Thir Asn. Thir Gly Ser Gly Thir Pro 25 3 O

<21 Os SEQ ID NO 2 &211s LENGTH: 32 212s. TYPE: PRT &213s ORGANISM: Mouse

<4 OOs SEQUENCE: 2

Cys Gly Asn Lieu Ser Thr Cys Met Leu. Gly Thir Tyr Thir Glin Asp Lieu. 1. 5 15

Asn Llys Phe His Thr Phe Pro Gn. Thir Ser Ile Gly Wall Glu Ala Pro 25 3 O

<21 Os SEQ ID NO 3 &211s LENGTH: 32 212s. TYPE: PRT <213> ORGANISM: Chicken

<4 OOs SEQUENCE: 3

Cys Ala Ser Leu Ser Thr Cys Wall Lieu. Gly Lys Luell Ser Glin Glu Lell 1. 5 15 US 9,533,022 B2 27 28 - Continued

His Llys Lieu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro 2O 25 3O

<210s, SEQ ID NO 4 &211s LENGTH: 32 212. TYPE: PRT <213s ORGANISM: Eel

<4 OOs, SEQUENCE: 4 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Llys Lieu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro 2O 25 3O

<210s, SEQ ID NO 5 &211s LENGTH: 32 212. TYPE: PRT &213s ORGANISM: Rat

<4 OOs, SEQUENCE: 5 Cys Gly Asn Lieu Ser Thr Cys Met Leu Gly Thr Tyr Thr Glin Asp Leu 1. 5 1O 15 Asn Llys Phe His Thr Phe Pro Gln Thr Ser Ile Gly Val Gly Ala Pro 2O 25 3O

<210s, SEQ ID NO 6 &211s LENGTH: 32 212. TYPE: PRT & 213 ORGANISM; Horse

<4 OOs, SEQUENCE: 6 Cys Ser Asn Lieu Ser Thr Cys Val Lieu. Gly Thr Tyr Thr Glin Asp Leu 1. 5 1O 15 Asn Llys Phe His Thr Phe Pro Gln Thr Ala Ile Gly Val Gly Ala Pro 2O 25 3O

<210s, SEQ ID NO 7 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Canine

<4 OO > SEQUENCE: 7 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Thr Tyr Ser Lys Asp Lieu. 1. 5 1O 15 Asn Asn Phe His Thr Phe Ser Gly Ile Gly Phe Gly Ala Glu. Thr Pro 2O 25 3O

<210s, SEQ ID NO 8 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Canine

<4 OOs, SEQUENCE: 8 Cys Ser Asn Lieu Ser Thr Cys Val Lieu. Gly Thr Tyr Thr Glin Asp Leu 1. 5 1O 15

Asn Llys Phe His Thr Phe Pro Gln Thr Ala Ile Gly Val Gly Ala Pro 2O 25 3O

<210s, SEQ ID NO 9 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Porcine US 9,533,022 B2 29 30 - Continued

<4 OOs, SEQUENCE: 9 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Ser Ala Tyr Trp Arg Asn Lieu. 1. 5 1O 15 Asn Asn Phe His Arg Phe Ser Gly Met Gly Phe Gly Pro Glu. Thr Pro 2O 25 3O

<210s, SEQ ID NO 10 &211s LENGTH: 32 212. TYPE: PRT &213s ORGANISM: Human

<4 OOs, SEQUENCE: 10 Cys Gly Asn Lieu Ser Thr Cys Met Leu Gly Thr Tyr Thr Glin Asp Phe 1. 5 1O 15 Asn Llys Phe His Thr Phe Pro Gln Thr Ala Ile Gly Val Gly Ala Pro 2O 25 3O

<210s, SEQ ID NO 11 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: <223> OTHER INFORMATION: Synthetic Construct 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1. ... 1 223 OTHER INFORMATION: ACETYLATION 22 Os. FEATURE: <221s NAME/KEY: MOD RES <222. LOCATION: 32 . .32 223 OTHER INFORMATION: AMIDATION

<4 OOs, SEQUENCE: 11 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Llys Lieu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Asn Thr Tyr 2O 25 3O

<210s, SEQ ID NO 12 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: <223> OTHER INFORMATION: Synthetic Construct 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1. ... 1 223 OTHER INFORMATION: ACETYLATION 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 . .32 223 OTHER INFORMATION: ACETYLATION

<4 OOs, SEQUENCE: 12 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Arg Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Arg Lieu Gln Thr Phe Pro Arg Thr Asp Val Gly Ala Asn Thr Tyr 2O 25 3O

<210s, SEQ ID NO 13 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: <223> OTHER INFORMATION: Synthetic Construct 22 Os. FEATURE: US 9,533,022 B2 31 32 - Continued NAME/KEY: MOD RES LOCATION: 1. ... 1 OTHER INFORMATION: propionyl FEATURE: NAME/KEY: MOD RES LOCATION: 32 . .32 OTHER INFORMATION: AMIDATION

SEQUENCE: 13 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Llys Lieu Gln Thr Tyr Pro Arg Thr Asn Thr Gly Ser Gly Thr Pro

SEQ ID NO 14 LENGTH: 32 TYPE PRT ORGANISM: Artificial Sequence FEATURE: OTHER INFORMATION: Synthetic construct FEATURE: NAME/KEY: MOD RES LOCATION: 1. ... 1 OTHER INFORMATION: Succinyl FEATURE: NAME/KEY: MOD RES LOCATION: 32 . .32 OTHER INFORMATION: AMIDATION

SEQUENCE: 14 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Llys Lieu Gln Thr yr Pro Arg Thr Asn Thr Gly Ser Gly Thr Pro 2O 25 3O

SEO ID NO 15 LENGTH: 32 TYPE PRT ORGANISM: Artificial Sequence FEATURE: OTHER INFORMATION: Synthetic construct FEATURE: NAME/KEY: MOD RES LOCATION: 1. ... 1 OTHER INFORMATION: ACETYLATION FEATURE: NAME/KEY: MOD RES LOCATION: 32 . .32 OTHER INFORMATION: AMIDATION

SEQUENCE: 15 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Llys Lieu Gln Thr yr Pro Arg Thir Asp Val Gly Ala Asn Ala Pro 2O 25 3O

SEQ ID NO 16 LENGTH: 33 TYPE PRT ORGANISM: Artificial Sequence FEATURE: OTHER INFORMATION: Synthetic Construct FEATURE: NAME/KEY: MOD RES LOCATION: 33 33 OTHER INFORMATION: AMIDATION

SEQUENCE: 16 Lys Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Llys Lieu. Ser Glin Glu US 9,533,022 B2 33 34 - Continued

1. 5 1O 15 Lieu. His Llys Lieu. Glin Thr Tyr Pro Arg Thr Asp Val Gly Ala Asn Thr 2O 25 Tyr

<210s, SEQ ID NO 17 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: 223 OTHER INFORMAT ON: Synthetic Construct 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1. ... 1 223 OTHER INFORMAT ON: Succinyl 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 . .32 223 OTHER INFORMAT ON: AMIDATION

<4 OOs, SEQUENCE: 17 Cys Ser Asn Lieu. Ser hir Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu 1. 5 1O 15

His Llys Lieu Gln Thr yr Pro Arg Thr Asp Val Gly Ala Asn Ala Tyr 2O 25 3O

<210s, SEQ ID NO 18 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence & 22 O FEATURE; 223 OTHER INFORMATION: Synthetic Construct 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1. ... 1 <223> OTHER INFORMATION: Cysteine can be modified with an acetyl group, propionyl group, or succinyl group. 22 Os. FEATURE: <221s NAME/KEY: SITE &222s. LOCATION: 31 31 223 OTHER INFORMAT ON: Xaa can be any naturally occurring amino acid 22 Os. FEATURE: <221s NAME/KEY: SITE &222s. LOCATION: 32 . .32 223 OTHER INFORMAT ON: Xaa can be any naturally occurring amino acid 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 . .32 223 OTHER INFORMAT ON: Optionally amidated

<4 OOs, SEQUENCE: 18 Cys Ser Asn Lieu. Ser hir Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu 1. 5 1O 15

His Llys Lieu Gln Thr yr Pro Arg Thr Asp Val Gly Ala Asn. Xaa Xala 2O 25 3O

<210s, SEQ ID NO 19 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: 223s OTHER INFORMATION: Calcitonin mimetic 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1 <223> OTHER INFORMATION: Optional acylation 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 <223> OTHER INFORMATION: Amidation US 9,533,022 B2 35 36 - Continued

<4 OOs, SEQUENCE: 19 Cys Ser Asn Lieu Ser Thr Cys Val Lieu Gly Lys Lieu. Ser Glin Glu Lieu 1. 5 1O 15 His Llys Lieu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Asn Ala Pro 25 3O

<210s, SEQ ID NO 2 O &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: 223 OTHER INFORMAT ON: Calcitonin mimetic 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1 223 OTHER INFORMAT ON: Optional acylation 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 223 OTHER INFORMAT ON: Amidation

<4 OOs, SEQUENCE: 2O Cys Ser Asn Lieu. Ser hir Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu 1. 5 1O 15 His Llys Lieu Gln Thr yr Pro Arg Thr Asn Thr Gly Ser Gly Thr Pro 2O 25 3O

<210s, SEQ ID NO 21 &211s LENGTH: 32 212. TYPE PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: 223 OTHER INFORMAT ON: Calcitonin mimetic 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1 223 OTHER INFORMAT ON: Optional acylation 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 223 OTHER INFORMAT ON: Amidation

<4 OOs, SEQUENCE: 21 Cys Ser Asn Lieu. Ser hir Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu 1. 5 1O 15 His Llys Lieu Gln Thr yr Pro Arg Thr Asp Val Gly Ala Asn. Thir Tyr 2O 25 3O

<210s, SEQ ID NO 22 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: 223s OTHER INFORMATION: Calcitonin mimetic 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 1 223 OTHER INFORMATION: Optional amidation 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 223 OTHER INFORMATION: Acetylation

<4 OOs, SEQUENCE: 22 Cys Ser Asn Lieu Ser Thr Cys Val Lieu Gly Arg Lieu. Ser Glin Glu Lieu 1. 5 1O 15

His Arg Lieu Gln Thr Phe Pro Arg Thr Asp Val Gly Ala Asn. Thir Tyr US 9,533,022 B2 37 38 - Continued

2O 25

SEQ ID NO 23 LENGTH: 33 TYPE PRT ORGANISM: Artificial Sequence FEATURE: OTHER INFORMATION: Calcitonin mimetic FEATURE: NAME/KEY: MOD RES &222s. LOCATION: 1 <223> OTHER INFORMATION: Optional acylation 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 33 <223> OTHER INFORMATION: Amidation

<4 OOs, SEQUENCE: 23 Lys Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Llys Lieu. Ser Glin Glu 1. 5 1O 15 Lieu. His Llys Lieu. Glin Thr Tyr Pro Arg Thr Asp Val Gly Ala Asn Thr 2O 25 Tyr

<210s, SEQ ID NO 24 &211s LENGTH: 32 212. TYPE: PRT <213> ORGANISM: Artificial Sequence 22 Os. FEATURE: <223> OTHER INFORMATION: Calcitonin mimetic 22 Os. FEATURE: <221. NAME/KEY: MOD RES &222s. LOCATION: 1 <223> OTHER INFORMATION: Optional acylation 22 Os. FEATURE: <221s NAME/KEY: MOD RES &222s. LOCATION: 32 <223> OTHER INFORMATION: Amidation

<4 OOs, SEQUENCE: 24 Cys Ser Asn Lieu. Ser Thr Cys Val Lieu. Gly Lys Lieu. Ser Glin Glu Lieu. 1. 5 1O 15 His Llys Lieu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Asn Ala Tyr 2O 25

45 What is claimed is: SuccCSNLSTCVLGKLSQELHKLQTYPRTDV 1. A method for treating type I diabetes, type II diabetes, GANAY-NH2 SEQ ID NO: 17. metabolic syndrome, or obesity, or appetite Suppression, or 4. The method of claim 1, wherein the peptide is formu for mitigating insulin resistance, or for improving glycemic lated for enteral administration. control, or for improving serum insulin level control in a 50 5. The method of claim 1, wherein the peptide is formu patient in need of Such treatment, comprising administering lated for parenteral administration. to the patient as a combination therapy an effective amount 6. The method of claim 1, wherein the peptide is formu of an insulin sensitizer and of a peptide selected from the lated with a carrier for oral administration. group consisting of SEQ ID NO: 12, SEQ ID NO: 15, SEQ 7. The method of claim 6, wherein the carrier comprises ID NO: 17, SEQID NO: 19, SEQ ID NO: 22, and SEQ ID 55 N-(5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), NO: 24. sodium salt of 10-(2-Hydroxybenzamido)decanoic acid 2. The method of claim 1, wherein the peptide is selected (SNAD), or sodium salt of N-(8-2-hydroxybenzoylamino) from the group consisting of caprylic acid (SNAC). R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANAP 8. The method of claim 1, wherein the peptide is formu NH2 SEQID NO: 19, and 60 lated in a pharmaceutical composition for oral administra R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANAY tion comprising coated citric acid particles, and wherein the NH2 SEQ ID NO: 24; coated citric acid particles increase the oral bioavailability of wherein each peptide R is absent or is an acylation moiety. the peptide. 3. The method of claim 1, wherein the peptide is selected 9. The method of claim 1, wherein said treatment is for from the group consisting of 65 effecting a weight reduction of the patient. AcCSNLSTCVLGKLSQELHKLQTYPRTDVGANAP 10. The method of claim 1, wherein the treatment is to NH2 SEQID NO: 15, and improve postprandial glycemic control in the patient. US 9,533,022 B2 39 40 11. The method of claim 1, wherein the insulin sensitiser propamide, gliclazide, tolbutamide, tolaZamide, glipizide, is at least one selected from the group consisting of a glibenclamide, glyburide, gliquidone, glyclopyramide and biguanide, a thiazolidinedione, a dual peroxisome prolifera glimeprinide. tor-activated receptor (PPAR) agonist, a Sulphonylurea, a 16. The method of claim 11, wherein the meglitinide/ glinide is selected from the group consisting of nateglinide, meglitinide/glinide, a glucagon-like peptide-1 (GLP-1) ana repaglinide and mitiglinide. log, a dipetidyl peptidase 4 (DPP-4) inhibitor, an insulin 17. The method of claim 11, wherein the glucagon-like analog, an alpha-glucosidase inhibitor, an amylin analog, a peptide-1 (GLP-1) analog is selected from the group con sodium/glucose cotransporter 2 (SGLT2) inhibitor, benfluo sisting of exenatide, liraglutide and albiglutide. rex, and tolrestat. 18. The method of claim 11, wherein the dipetidyl pep 12. The method of claim 11, wherein the biguanide is 10 tidase 4 (DPP-4) inhibitor is selected from the group con selected from the group consisting of metformin, buformin sisting of alogliptin, linagliptin, saxagliptin, Sitagliptin and and phenformin. Vildagliptin. 19. The method of claim 11, wherein the alpha-glucosi 13. The method of claim 11, wherein the thiazolidin dase inhibitor is selected from the group consisting of edione is selected from the group consisting of balaglita 15 acarbose, miglitol and Voglibose. Zone, pioglitaZone, rivoglitaZone, rosiglitaZone and troglita 20. The method of claim 11, wherein the amylin analog is ZO. pramlintide. 14. The method of claim 11, wherein the dual peroxisome 21. The method of claim 11, wherein the sodium/glucose proliferator-activated receptor (PPAR) agonist is selected cotransporter 2 (SGLT2) inhibitor is selected from the group from the group consisting of aleglitazar, muraglitazar and consisting of dapagliflozin, remogliflozin and Sergliflozin. tesaglitazar. 22. The method of claim 1, wherein the insulin sensitizer 15. The method of claim 11, wherein the sulphonylurea is is metformin. selected from the group consisting of carbutamide, chloro