US 2015.0017238A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0017238 A1 Kidron (43) Pub. Date: Jan. 15, 2015

(54) METHODS AND COMPOSITIONS FOR Publication Classification TREATING DABETES (51) Int. Cl. (71) Applicant: Oramed Ltd., Jerusalem (IL) A638/28 (2006.01) A638/57 (2006.01) (72) Inventor: Miriam Kidron, Jerusalem (IL) A638/55 (2006.01) A638/56 (2006.01) (73) Assignee: Oramed, Ltd., Jerusalem (IL) 469/48 (2006.01) A638/26 (2006.01) (21) Appl. No.: 14/370,452 (52) U.S. Cl. CPC ...... A61K 38/28 (2013.01); A61 K9/4808 (22) PCT Filed: Jan. 3, 2013 (2013.01); A61 K38/26 (2013.01); A61K38/55 (2013.01); A61 K38/56 (2013.01); A61K38/57 S371 (c)(1), USPC ...... 424/455; 514/6.5

Related U.S. Application Data d herein a methods and compositions for treating iabetes mellitus, concerning oral pharmaceutical composi (60) Provisional application No. 61/631,339, filed on Jan. tions comprising insulin in combination with a GLP-1 ana 3, 2012. logue. Patent Application Publication Jan. 15, 2015 Sheet 1 of 5 US 2015/001 7238A1

Patent Application Publication Jan. 15, 2015 Sheet 2 of 5 US 2015/0017238A1

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Patent Application Publication Jan. 15, 2015 Sheet 3 of 5 US 201S/0017238A1

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Figure 3 US 2015/001 7238 A1 Jan. 15, 2015

METHODS AND COMPOSITIONS FOR mimics physiological first-pass metabolism of insulin and TREATING DABETES GLP-1. These results provide a route to an entirely new class 0001 Benefit is claimed to U.S. Provisional Application of therapeutic modalities. 61/631,339, filed Jan. 3, 2012, which is incorporated herein 0009. The terms “protein’ and “peptide' are used inter by reference in its entirety. changeably herein. Neither term is intended to confer a limi tation of the number of amino acids present, except where a FIELD limitation is explicitly indicated. 0002. Described herein are methods and compositions for BRIEF DESCRIPTION OF THE DRAWINGS treating diabetes mellitus. 0010. The following figures are by way of illustrative BACKGROUND example and are not meant to be taken as limiting the claimed invention. 0003 Diabetes, specifically the Type II (NIDDM) variety, 0011 FIG. 1. Testing of various emulsifier formulations. has emerged in the twenty-first century as an epidemic of Foam buildup score was from 1-5, where 1 indicates no foam, global proportions. Numerous long-term complications, and 5 indicates no liquid visible because of the foam. For the including those affecting the kidneys, legs, feet, eyes, heart, Suspension test, the numbers 1-5 indicate full phase separa nerves, and blood circulation, may result from uncontrolled tion; partial phase separation with Some larger oil bubbles; diabetes. Prevention of these conditions requires comprehen small oil bubble, milky consistency; no bubbles initially, with sive treatment, requiring life style modification and medica later phase separation; and stable emulsion, respectively. tion. A number of effective anti-diabetic drugs are available and are generally safe and well tolerated. All the medications 0012 FIGS. 2A and 2B. Blood glucose profiles following become less effective as the disease progresses, and most administration of oral insulin formulations containing vari patients eventually require insulin. Most of the medications ous emulsifiers. 2A: Formulations A (upper left), B (lower are associated with risks of hypoglycemia and weight gain, left), C (upper right), and D (lower right).2B: Formulations E yet do not alter the inexorable progression of diabetes. (left) and F (right). 0004 Orally-delivered formulations for protein-based 0013 FIG. 3. Blood glucose profiles following oral drugs such as insulin are being developed by the present exenatide and oral insulin administration to pigs. Fasting, inventor (Ziv et al 1994: Nissan et al 2000, Kidronetal 2004, commercial pigs were treated with 150 mcg.eXenatide, 8 mg. Eldor et al 2010B, Eldor etal 2010C). One such oral insulin insulin, or the combination thereof, 30 minutes before caloric product is scheduled to be tested in Phase II trials and is intake. 1-mL blood samples were periodically drawn currently being reviewed for IND status. throughout the 180-minute observation period to determine 0005. The incretin hormone Glucagon-like Peptide 1 glucose concentrations. (GLP-1), Secreted within minutes of food ingestion, is asso ciated with induction of insulin release. Therapies based on DETAILED DESCRIPTION OF THE GLP-1 are treatment options for Type 2 Diabetes Mellitus EMBODIMENTS (T2DM) that act through a variety of complementary mecha 0014 Provided herein is a pharmaceutical composition for nisms. The most intriguing aspect of the incretins is the fact oral delivery, comprising an oil-based liquid formulation, the that they cause insulin release in a glucose-dependent manner oil-based liquid formulation comprising an insulin, a GLP-1 and are thought to have a low risk of inducing hypoglycemia. analogue, a trypsin inhibitor, and a chelator of divalent cat Furthermore, the incretins seem to be weight-neutral (or ions, wherein the oil-based liquid formulation is surrounded weight-reducing), preserve beta-cell mass, and possibly also by a coating or capsule that resists degradation in the stom induce neogenesis of insulin-secreting cells. ach. 0006. However, clinical use of the native GLP-1 is limited 0015. Another embodiment provides a multi-component due to its rapid enzymatic inactivation, resulting in a half-life oral pharmaceutical composition, comprising: (a) a first oil of 2-3 minutes. To overcome this obstacle, long-acting deg based liquid formulation, the first oil-based liquid formula radation-resistant peptides, both natural and synthetic, tion comprising an insulin, a trypsin inhibitor, and a chelator referred to as GLP-1 mimetic agents or analogues, have been of divalent cations; and (b) a second oil-based liquid formu designed and tested. lation, the second oil-based liquid formulation comprising a 0007 To date, GLP-1 analogues are only available as GLP-1 analogue, a trypsin inhibitor, and a chelator of divalent injectable dosage forms. The present inventor is developing cations; wherein each of the first oil-based liquid formulation an oral exenatide GLP-1 analogue capsule. A first-in-humans and the second oil-based liquid formulation is surrounded by trial (n=4) testing its safety in healthy humans demonstrated a coating or capsule that resists degradation in the stomach. In retained biological functionality of orally delivered exenatide Some embodiments, the two liquid formulations can be in (Eldor et al 2010A). separate dosage forms. In other embodiments, the two liquid formulations are in the same dosage form; for example, in SUMMARY separate encased compartments within the same pill. 0008 To the inventor's knowledge, oral insulin formula 0016 “Liquid” as used herein refers to a phase that flows tions have not been tested in combination with oral GLP-1 freely and has a constant Volume under ambient conditions. analogue formulations. The data provided herein illustrate a Fish oil, for instance, is a liquid under ambient conditions. previously-unrecognized, strong cooperative interaction The term includes oil-based solutions, Suspensions, and com between these components when formulated as described binations thereof. In alternative embodiments, the term may herein. This enables a potent anti-diabetes effect in a conve refers to a composition that has a viscosity within the range of nient form that both facilitates patent compliance and also 1-1000 millipascal seconds, inclusive, at 20°C. US 2015/001 7238 A1 Jan. 15, 2015

0017. In certain embodiments, the different components no. 141732-76-5: SEQ ID NO: 4), lixisenatide (CAS no. of a multi-component pharmaceutical composition are indi 320367-13-3), liraglutide (CAS no. 204656-20-2), exendin-9 cated for co-administration together. "Co-administration', in (CAS no. 133514-43-9), AC3174 (Leuc14)lexendin-4, this regard, may refer either to simultaneous administration Amylin Pharmaceuticals, Inc.), taspoglutide (CAS no. or, in another embodiment, to administration within 30 min 275371-94-3), albiglutide (CAS no. 782500-75-8), sema utes of each other. In still other embodiments, the different glutide (CAS no.910463-68-2), LY2189265 (DulaglutideTM: components are indicated for administration in a particular CAS no. 923950-08-7), and CJC-1 134-PC (a modified Exen order, separated by a set time interval that will typically be 30 din-4 analogue conjugated to recombinant human albumin minutes or less. For example, the insulin-containing dosage manufactured by ConjuChemTM). All CAS records were form may be indicated for administration 2-10 minutes after accessed on Dec. 19, 2011. Thus, in certain embodiments, the the exenatide-containing dosage form; in other embodiments, described method or composition utilizes any of the above 10-20 minutes after the exenatide-containing dosage form; in listed GLP-1 analogues. In other embodiments, one of the other embodiments, 20-30 minutes after the exenatide-con above-listed GLP-1 analogues is selected. Those of skill in taining dosage form; and in other embodiments, 30-60 min the art will appreciate in light of the findings of described utes after the exenatide-containing dosage form. Oral dosage herein that other GLP-1 analogues can also be utilized. forms such as those provided herein lend themselves to 0027. Therapeutic insulin and GLP-1 proteins suitable for sequential administration more than injected dosage forms, use in the present invention include derivatives that are modi since regimens requiring repeated injections are likely to be fied (i.e., by the covalent attachment of a non-amino acid associated with low rates of compliance. residue to the protein). For example, but not by way of limi 0018. According to other aspects, a combination medica tation, the protein includes proteins that have been modified, ment for treatment of type 2 diabetes is provided, said com e.g., by glycosylation, acetylation, PEGylation, phosphory bination medicament comprising lation, amidation, or derivatization by known protecting/ 0019 insulin, at least one trypsin inhibitor, and a chela blocking groups. High-MW PEG can be attached to thera tor of divalent cations, and peutic proteins with or without a multifunctional linker either 0020 a GLP-1 analogue from the group consisting of through site-specific conjugation of the PEG to the N- or exenatide, liraglutide, AC3174, taspoglutide, lixisen C-terminus thereof or via epsilon-amino groups present on atide, semaglutide, albiglutide, exendin-9, LY2189265, lysine residues. Additionally, the derivative may contain one and CJC-1 134-PC, or more non-classical amino acids, for example D-isomers of 0021 all comprised jointly in an oil-based liquid for the common amino acids, 2,4-diaminobutyric acid, C-amino mulation in a dosage form for oral delivery. isobutyric acid, A-aminobutyric acid, Abu, 2-aminobutyric acid, Y-Abu, e-Ahk, 6-amino hexanoic acid, Aib, 2-amino Insulin Proteins and GLP-1 Analogues isobutyric acid, 3-amino propionic acid, ornithine, norleu 0022. Insulin proteins and GLP-1 analogues for use as cine, norvaline, hydroxyproline, sarcosine, citruline, homoc described herein are in some embodiments isolated prior to itrulline, cysteic acid, t-butylglycine, t-butylalanine, phe their introduction into the described pharmaceutical compo nylglycine, cyclohexylalanine, B-alanine, fluoro-amino sitions. “Isolated in this regard excludes provision of the acids, designer amino acids such as B-methyl amino acids, insulin and/or GLP-1 analogue as a homogenized tissue CO-methylamino acids, and NC.-methyl amino acids. preparation or other form containing Substantial amounts of contaminating proteins. An example of an isolated protein or Emulsifiers peptide is a recombinant protein or peptide. An alternative 0028. In certain embodiments, an oil-based liquid formu embodiment is a synthetic protein or peptide. lation utilized in the described methods and pharmaceutical 0023. A person skilled in the art will appreciate in light of compositions, or in other embodiments, each of the oil-based the present disclosure that various types of insulinare Suitable liquid formulation that is present, further comprises a com for the described methods and compositions. Exemplary ponent provided as a mixture of (a) a monoacylglycerol insulin proteins include but are not limited to both wild-type (monoglyceride), a diacylglycerol (diglyceride), a triacylg and mutated insulin proteins, including synthetic human lycerol (triglyceride), or a mixture thereof; and (b) a polyeth insulin, synthetic bovine insulin, synthetic porcine insulin, ylene glycol (PEG) ester of a fatty acid. In this regard, each of synthetic whale insulin, and metal complexes of insulin, Such the terms “monoacylglycerol”, “diacylglycerol, and “tria as Zinc complexes of insulin, protamine Zinc insulin, and cylglycerol’ need not refer to a single compound, but rather globin Zinc. can include mixtures of compounds, for example mixtures of 0024. Various classes of insulin may also be utilized, for monoacylglycerols, diacylglycerols, or triacylglycerols hav example fast-acting insulin, lente insulin, semilente insulin, ing fatty acids of varying lengths. In certain preferred ultralente insulin, NPH insulin, glargine insulin, lispro insu embodiments, monoacylglycerols, diacylglycerols, or tria lin, aspart insulin, or combinations of two or more of the cylglycerols utilized in the described methods and composi above types of insulin. tions, for example those used to general PEG esters, are from 0025. In certain embodiments, the insulin of the described an oil source that is Generally Recognized As Safe (GRAS). methods and compositions is wild-type human insulin (Uni 0029. Examples of GRAS oil sources are coconut oil, corn prot ID PO1308). In some embodiments, human insulin is oil, peanut oil, soybean oil, My vacet 9-45 (Diacetylated produced as a recombinant protein in bacterial cells. In other monoglycerides of C-18 fatty acids). embodiments, human insulin is produced synthetically. 0030. A more specific embodiment of (a) is a mixture of 0026 GLP-1 analogues are also referred to in the art as Cs-Cs monoacylglycerols, diacylglycerols, and triacylglyc GLP-1 mimetics. A person of skill in the art will appreciate erols. A more specific embodiment of component (b) is a that the described compositions may include at least one of mixture of PEG monoesters and diesters of a mixture of the following GLP-1 analogues: exenatide (ByettaTM; CAS Cs-C1s fatty acids. US 2015/001 7238 A1 Jan. 15, 2015

0031. In other, more specific embodiments, the liquid for free PEG. In other embodiments, the self-emulsifying com mulation further comprises a free PEG. ponent has an HLB of 14 and comprises medium and long 0032. In alternate embodiments, an oil-based liquid for chain triacylglycerols conjugated to PEG, free triacylglycer mulation utilized in the described methods and pharmaceuti ols, and free PEG. In other embodiments, the self-emulsify cal compositions, or in other embodiments, each of the oil ing component has an HLB of 14 and consists of a mixture of based liquid formulation that is present, further comprises a medium and long chain triacylglycerols conjugated to PEG, PEG ester of a monoacylglycerol, a diacylglycerol, a triacylg free triacylglycerols, and free PEG. lycerol, or a mixture thereof. In this regard, each of the terms “monoacylglycerol”, “diacylglycerol, and “triacylglycerol 0036 Certain, more specific embodiments utilize self need not refer to a single compound, but rather can include emulsifying components that comprise (a) a monoacylglyc mixtures of compounds, for example mixtures of monoacylg erol, a diacylglycerol, a triacylglycerol, or a mixture thereof. lycerols, diacylglycerols, or triacylglycerols having fatty and (b) a polyethylene glycol (PEG) ester of a fatty acid. In acids of varying lengths. In more specific embodiments, an this regard, each of the terms "monoacylglycerol', 'diacylg additional non-ionic detergent, for example a polysorbate lycerol, and “triacylglycerol’ need not refer to a single com based detergent, is present in addition to the PEG ester. In pound, but rather can include mixtures of compounds, for other, more specific embodiments, a free PEG is also present. example mixtures of monoacylglycerols, diacylglycerols, or In still more specific embodiments, both an additional non triacylglycerols having fatty acids of varying lengths. A more ionic detergent and a free PEG are also present. specific embodiment is a mixture of C-C monoacylglyc 0033. In a still more specific embodiment of the described erols, diacylglycerols, and triacylglycerols. methods and compositions, a liquid formulation used therein 0037. A more specific embodiment of component (b) is a comprises: (a) a mixture of Cs-Cs monoacylglycerols, dia mixture of PEG monoesters and diesters of a mixture of cylglycerols, and triacylglycerols; (b) PEG-32 monoesters Cs-C1s fatty acids. and diesters of a mixture of Cs-Cs fatty acids; and (c) free PEG-32. Even more specifically, the weight/weight ratio of 0038. In other, more specific embodiments, the self-emul component (a) to components (b)+(c) is between 10:90-30:70 sifying component further comprises molecules of free PEG. inclusive; more specifically between 15:85-25:75 inclusive: 0039. Preferred lengths of PEG moieties for use in the more specifically 20:80. In certain embodiments, compo described compositions and methods contain between 5-100 nents (a)-(c) together constitute 8-1.6% weight/weight inclu monomers. In more specific embodiments, the PEG may sive of the oil-based liquid formulation. In more specific contain between 15-50 monomers. In still more specific embodiments, the amount is 9-15% inclusive. In more spe embodiments, the PEG may contain between 25-40 mono cific embodiments, the amount is 10-14% inclusive. In more mers. In more specific embodiments, the PEG may contain 32 specific embodiments, the amount is 11-13% inclusive. In OOCS. more specific embodiments, the amount is 12%. 0034. In other embodiments, an oil-based liquid formula 0040. In a still more specific embodiment of the described tion utilized in the described methods and pharmaceutical methods and compositions, a self-emulsifying component compositions further comprises a self-emulsifying compo used therein comprises: (a) a mixture of Cs-Cs monoacylg nent, which may or may not be one of the mixtures of com lycerols, diacylglycerols, and triacylglycerols; (b) PEG-32 ponents described in the preceding paragraphs. “Self-emul monoesters and diesters of a mixture of Cs-Cs fatty acids; Sifying component' in some embodiments refers to a and (c) free PEG-32; and the weight/weight ratio of compo component that spontaneously forms an emulsion. Typically, nent (a) to components (b)+(c) is 20:80. In certain embodi Such components will form an emulsion under on contact ments, such a component constitutes 8-1.6% weight/weight with aqueous media, forming a fine dispersion i.e. a micro inclusive of the oil-based liquid formulation. In more specific emulsion (SMEDDS). Certain embodiments of such compo embodiments, the amount is 9-15% inclusive. In more spe nents comprise a mixture of triacylglycerols and a high cific embodiments, the amount is 10-14% inclusive. In more hydrophile/lipophile balance (HLB: see Griffin W C: “Cal specific embodiments, the amount is 11-13% inclusive. In culation of HLB Values of Non-Ionic Surfactants. J Soc more specific embodiments, the amount is 12%. Cosmetic Chemists 5:259 (1954)) surfactant. Other embodi 0041 Examples of self-emulsifying components meeting ments of the self-emulsifying component have a waxy, semi the above specifications are GelucireTM 44/14, GelucireTM Solid consistency. 53/10, and GelucireTM 50/13. A particularly preferred 0035. Preferably, the HLB of a self-emulsifying compo example is GelucireTM 44/14. The suffixes 44 and 14 refer nent utilized in the described methods and compositions is 10 respectively to its melting point and its hydrophilic/lypophilic or greater. In other embodiments, it is between 11-19 inclu balance (HLB). GelucireTM 44/14 (Gattefossé SAS, Saint sive. In other embodiments, it is between 12-18 inclusive. In Priest, France) is obtained by polyglycolysis of hydrogenated other embodiments, it is between 12-17 inclusive. In other coconut oil (medium and long chain triacylglycerols with embodiments, it is between 12-16 inclusive, which is indica PEG-32. It has a hydrophile/lipophile balance of 14. It is tive of an oil-in-water (O/W) emulsifier. In other embodi composed of a defined admixture of Cs-C mono-, di- and ments, it is between 13-15 inclusive. In other embodiments, it triacylglycerols (20% w/w); PEG-32 mono- and diesters and is 14. Still more specific embodiments of self-emulsifying free PEG-32 (80% w/w). The main fatty acid present is lauric components have an HLB of 12-16 inclusive and comprise acid, accounting for 45% on average of the total fatty acid medium and long chain triacylglycerols conjugated to PEG, content. It is a solid dispersion composed of a PEG ester free triacylglycerols, and free PEG. In other embodiments, fraction under a lamellar phase of 120 A with a helical con the self-emulsifying component has an HLB of 12-16 inclu formation and an acylglycerol fraction under a hexagonal sive and consists of a mixture of medium and long chain packing. The main products of simulated gastrointestinal triacylglycerols conjugated to PEG, free triacylglycerols, and lipolysis of GelucireTM 44/14 are PEG-32 mono and diesters. US 2015/001 7238 A1 Jan. 15, 2015

Non-Ionic Detergents those described herein that contain insulin but lack a GLP-1 0042. In certain embodiments, the oil-based liquid formu analogue; or, in a more specific embodiment, a formulation lation utilized in the described methods and pharmaceutical identical except for the absence of the GLP-1 analogue. Simi compositions further comprises a non-ionic detergent in addi larly, a subclinical dose of a GLP-1 analogue would be less tion to the self-emulsifying component. In certain embodi than that required using formulations similar to those ments, the non-ionic detergent is selected from the group described herein that contain a GLP-1 analogue but lack consisting of polysorbate-20, polysorbate-40, polysorbate insulin; or, in a more specific embodiment, a formulation 80, lauromacrogol 400, polyoxyl 40 stearate, polyoxyethyl identical except for the absence of the insulin. ene hydrogenated castor oil 10, 50 and 60, glycerol 0048 Those skilled in the art will appreciate, in light of the monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty present disclosure, that characterization of a dose as Subclini acid ester, methyl cellulose, carboxymethyl cellulose, n-oc cal will depend on the weight and health status (including tylglucoside, n-dodecylglucoside, n-dodecylmaltoside, insulin resistance, if relevant) of the patient, the circum octanoyl-N-methylglucamide, decanoyl-N-methylglucam stances of the administration, co-administration of other dia ide, TritonTM-X-100, TritonTM-X-114, ThesitTM, Isotridecy betes medications, the robustness of the active ingredient and poly(ethylene glycol ether), 3-(3-cholamidopropyl)dim the excipients, and the desired physiological effect. For ethylamminio-1-propane sulfonate (CHAPS), 3-(3- example, studies to date of oral formulations similar to those cholamidopropyl)dimethylamminio-2-hydroxy-1-propane described herein, but containing insulin only, have shown that sulfonate (CHAPSO), and N-dodecyl-N,N-dimethyl-3-am 8 mg of an encapsulated oral formulation in combination with monio-1-propane Sulfonate. In other embodiments, one of the a protease inhibitor and EDTA, in fish oil (similar to the one above-listed non-ionic detergents is selected. described herein but lacking exenatide) is a Subclinical dose 0043. In certain, more specific embodiments, a non-ionic for fasting, adult, human Type 2 diabetic patients, if the goal detergent used in the described methods and compositions is is a robust change in blood glucose levels; while 16 mg. is a a polysorbate-based detergent. Examples of polysorbate clinical dose under the same circumstances. Doses of the based detergent are detergents derived by covalently bonding same formulation necessary to achieve modulation of post polyethoxylated sorbitan to a fatty acid. More specific prandial glucose excursions in the same patients have not embodiments of polysorbate-based detergents are polysor been determined, but are likely to be slightly higher. How bate-20, polysorbate-40, and polysorbate-80. ever, doses such as these also depend on the potency of the 0044) For example, polysorbate 80 (Tween-80) is a mild, formulation, and thus the clinical dose threshold may be non-ionic detergent derived from polyethoxylated sorbitan slightly lower if a more potent protease inhibitor is used, for and oleic acid and having the following structure: example. Determination of a Subclinical dose for a particular

HO(CH2CH2O). (OCH2CH3),OH

O CH(OCH2CH2),OH O

Sum of w + x + y + z = 20

0045. In the case of polysorbate 80, the moiety shown on set of circumstances, for example via empirical testing, is the right side is a mixture of fatty acids, containing 60-70% well within the ability of one skilled in the art. oleic acid (as depicted), with the balance being primarily 0049. In more specific embodiments, the subclinical linoleic, palmitic, and Stearic acids. amount of insulin of the described methods and compositions 0046. In a more specific embodiment, the polysorbate 80 is between 6-16 mg inclusive for an adult patent having dia constitutes 3-10% weight/weight inclusive of an oil-based betes mellitus, for exampleType 2 diabetes mellitus (T2DM), liquid formulation used in the described methods and com for example for preventing post-prandial glucose excursions positions. In a more specific embodiment, the percentage is when administered between 30-60 minutes (min) before a 4-8% inclusive. In a more specific embodiment, the percent meal, in more specific embodiments 30 min, 45 min, or 60 age is 4.5-6% inclusive. In a more specific embodiment, the min before a meal. In other embodiments, the subclinical percentage is 5%. amount is between 6-14 mg inclusive. In other embodiments, Dosages the subclinical amount is between 6-12 mg inclusive. In other embodiments, the subclinical amount is between 6-10 mg 0047 Alternatively or in addition, the insulin and/or inclusive. In other embodiments, the subclinical amount is 8 GLP-1 analogue present in the described compositions or mg. In other embodiments, the Subclinical amount is 12 mg. used in the described methods is present in a subclinical In other embodiments, the subclinical amount is between amount. The term "subclinical amount” in this context refers 8-16 mg inclusive. In other embodiments, the subclinical to an amount less than that required to elicit a complete amount is between 8-14 mg inclusive. In other embodiments, desired physiological effect, for example control of post the subclinical amount is between 8-12 mg inclusive. In other prandial blood glucose levels, in the context of its formulation embodiments, the subclinical amount is between 8-10 mg and the patient. Accordingly, a Subclinical dose of insulin inclusive. In other embodiments, the subclinical amount is 16 would be less than that required using formulations similar to mg. In other embodiments, the Subclinical amount is between US 2015/001 7238 A1 Jan. 15, 2015

10-16 mg inclusive. In other embodiments, the subclinical 0054. In other embodiments, the subclinical amount of amount is between 10-14 mg inclusive. In other embodi GLP-1 analogue of the described methods and compositions ments, the subclinical amount is between 10-18 mg inclusive. is between 0.100-0.400 mcg/kg inclusive for an adult patent 0050. In other embodiments, the subclinical amount of having T2D, for example for preventing post-prandial glu insulin of the described methods and compositions is between cose excursions when administered before a meal. In other 0.06-0.16 mg/kg (milligrams per kilogram body weight) embodiments, the subclinical amount is between 0.100-0.300 inclusive for an adult patent having T2DM, for example for mcg/kg inclusive. In other embodiments, the Subclinical preventing post-prandial glucose excursions when adminis amount is between 0.100-0.250 mcg/kg inclusive. In other tered before a meal. In other embodiments, the subclinical embodiments, the subclinical amount is between 0.100-0.200 amount is between 0.06-0.14 mg/kg inclusive. In other mcg/kg inclusive. In other embodiments, the Subclinical embodiments, the subclinical amount is between 0.06-0.12 amount is between 0.100-0.150 mcg/kg inclusive. In other mg/kg inclusive. In other embodiments, the Subclinical embodiments, the Subclinical amount is 0.100 mcg/kg. In amount is between 0.06-0.10 mg/kg inclusive. In other other embodiments, the subclinical amount is 0.150 mcg/kg. embodiments, the subclinical amount is 0.08 mg/kg. In other In other embodiments, the subclinical amount is 0.200 mcg/ embodiments, the Subclinical amount is 0.12 mg/kg. In other kg. In other embodiments, the subclinical amount is 0.250 embodiments, the Subclinical amount is 0.16 mg/kg. In other mcg/kg. In other embodiments, the Subclinical amount is embodiments, the subclinical amount is between 0.08–0.16 0.300 mcg/kg. In other embodiments, the subclinical amount mg/kg inclusive. In other embodiments, the Subclinical is between 0.150-0.400 mcg/kg inclusive. In other embodi amount is between 0.08-0.14 mg/kg inclusive. In other ments, the subclinical amount is between 0.150-0.300 mcg/ embodiments, the subclinical amount is between 0.08-0.12 kg inclusive. In other embodiments, the Subclinical amount is mg/kg inclusive. In other embodiments, the Subclinical between 0.150-0.250 mcg/kg inclusive. In other embodi amount is between 0.08–0.10 mg/kg inclusive. In other ments, the subclinical amount is between 0.100-0.200 mcg/ embodiments, the subclinical amount is between 0.10-0.16 kg inclusive. mg/kg inclusive. In other embodiments, the Subclinical 0055. In other embodiments, the subclinical amount of amount is between 0.10-0.18 mg/kg inclusive. In other GLP-1 analogue is an amount corresponding to one of the embodiments, the subclinical amount is between 0.10-0.14 above amounts or ranges for an adult, adjusted per body mg/kg inclusive. weight for a pediatric patient. In other embodiments, the 0051. In still other embodiments, the subclinical amount GLP-1 analogue is present in a Subclinical amount adjusted of insulin is an amount corresponding to one of the above for a pediatric patient, and the insulin is also present in a amounts or ranges for an adult, adjusted per body weight for Subclinical amount adjusted for a pediatric patient, for a pediatric patient. In other embodiments, the insulin is example an amount corresponding to 4-12 mg inclusive for an present in a Subclinical amount adjusted for a pediatric adult patent, adjusted for the weight of the a pediatric patient. patient, and the GLP-1 analogue is also present in a subclini 0056. The above amounts may be for exenatide, or in cal amount adjusted for a pediatric patient. another embodiment, for one of the other GLP-1 analogues 0052. The above amounts may be for wild-type human known in the art. insulin, or in another embodiment, for one of the other types 0057. In other embodiments, the described dosage form of insulin known in the art. contains exenatide in an amount of between 100-600 mcg 0053. In other, more specific embodiments, a subclinical inclusive, 100-500 mcg inclusive, 100-400 mcg inclusive, amount of a GLP-1 analogue is presentina dosage form of the 100-300mcg inclusive, 200-600mcg inclusive, 200-500mcg described methods and compositions. In some embodiments, inclusive, 200-400 mcg inclusive, 200-300 mcg inclusive, 150 micrograms (mcg), 200 mcg, 250 mcg, or 300 mcg is 150-300 mcg inclusive, or 150-250 mcg inclusive; together considered a Subclinical dose for adult, human Subjects with with 8-16 mg insulin inclusive. In other embodiments, the T2DM for example for preventing post-prandial glucose described dosage form contains exenatide in an amount of excursions when administered between 30-60 min before a between 100-600mcg inclusive, 100-500mcg inclusive, 100 meal, in more specific embodiments 30 min, 45 min, or 60 400 mcg inclusive, 100-300 mcg inclusive, 200-600 mcg min before a meal. In other embodiments, the subclinical inclusive, 200-500 mcg inclusive, 200-400 mcg inclusive, amount of GLP-1 analogue is between 100-400mcg inclusive 200-300 mcg inclusive, 150-300 mcg inclusive, or 150-250 for an adult patent having T2DM. In other embodiments, the mcg inclusive; together with 8-12 mg insulin inclusive. In subclinical amount is between 100-300 mcg. inclusive. In other embodiments, the described dosage form contains other embodiments, the subclinical amount is between 100 exenatide in an amount of between 100-600 mcg inclusive, 250 mcg. inclusive. In other embodiments, the subclinical 100-500mcg inclusive, 100-400mcg inclusive, 100-300mcg amount is between 100-200 mcg. inclusive. In other embodi inclusive, 200-600 mcg inclusive, 200-500 mcg inclusive, ments, the subclinical amount is between 100-150 mcg. 200-400mcg inclusive, 200-300mcg inclusive, 150-300mcg inclusive. In other embodiments, the Subclinical amount is inclusive, or 150-250 mcg inclusive; together with 12-16 mg 100 mcg. In other embodiments, the subclinical amount is insulin inclusive. In other embodiments, the described dosage 150 mcg. In other embodiments, the subclinical amount is form contains exenatide in an amount of between 100-600 200 mcg. In other embodiments, the subclinical amount is mcg inclusive, 100-500 mcg inclusive, 100-400 mcg inclu 250 mcg. In other embodiments, the subclinical amount is sive, 100-300mcg inclusive, 200-600mcg inclusive, 200-500 300 mcg. In other embodiments, the subclinical amount is mcg inclusive, 200-400 mcg inclusive, 200-300 mcg inclu between 150-400mcg. In other embodiments, the subclinical sive, 150-300 mcg inclusive, or 150-250 mcg inclusive: amount is between 150-300 mcg. inclusive. In other embodi together with 16-24 mg insulin inclusive. In other embodi ments, the subclinical amount is between 150-250 mcg. ments, the described dosage form contains exenatide in an inclusive. In other embodiments, the Subclinical amount is amount of between 100-600 mcg inclusive, 100-500 mcg between 150-200 mcg. inclusive. inclusive, 100-400 mcg inclusive, 100-300 mcg inclusive, US 2015/001 7238 A1 Jan. 15, 2015

200-600mcg inclusive, 200-500mcg inclusive, 200-400mcg mg inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 inclusive, 200-300 mcg inclusive, 150-300 mcg inclusive, or mg inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 150-250 mcg inclusive; together with 24-32 mg insulin inclu mg inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; sive. In other embodiments, the described dosage form con together with 300-450 mcg exenatide inclusive. In other tains exenatide in an amount of between 100-600mcg inclu embodiments, the described dosage form contains insulin in sive, 100-500mcg inclusive, 100-400mcg inclusive, 100-300 an amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg mcg inclusive, 200-600 mcg inclusive, 200-500 mcg inclu inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg sive, 200-400mcg inclusive, 200-300mcg inclusive, 150-300 inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg mcg inclusive, or 150-250 mcg inclusive; together with 12-16 inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg mg insulin inclusive. In other embodiments, the described inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; dosage form contains exenatide in an amount of between together with 450-600 mcg exenatide inclusive. In other 100-600mcg inclusive, 100-500mcg inclusive, 100-400mcg embodiments, the described dosage form contains insulin in inclusive, 100-300 mcg inclusive, 200-600 mcg inclusive, an amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg 200-500mcg inclusive, 200-400mcg inclusive, 200-300mcg inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg inclusive, 150-300 mcg inclusive, or 150-250 mcg inclusive: inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg together with 8 mg insulin inclusive. In other embodiments, inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg the described dosage form contains exenatide in an amount of inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; between 100-600mcg inclusive, 100-500mcg inclusive, 100 together with 100-150 mcg exenatide inclusive. In other 400 mcg inclusive, 100-300 mcg inclusive, 200-600 mcg embodiments, the described dosage form contains insulin in inclusive, 200-500 mcg inclusive, 200-400 mcg inclusive, an amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg 200-300 mcg inclusive, 150-300 mcg inclusive, or 150-250 inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg mcg inclusive; together with 12 mg insulin inclusive. In other inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg embodiments, the described dosage form contains exenatide inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg in an amount of between 100-600 mcg inclusive, 100-500 inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; mcg inclusive, 100-400 mcg inclusive, 100-300 mcg inclu together with 150-200 mcg exenatide inclusive. In other sive, 200-600mcg inclusive, 200-500mcg inclusive, 200-400 embodiments, the described dosage form contains insulin in mcg inclusive, 200-300 mcg inclusive, 150-300 mcg inclu an amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg sive, or 150-250 mcg inclusive; together with 16 mg insulin inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg inclusive. In other embodiments, the described dosage form inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg contains exenatide in an amount of between 100-600 mcg inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg inclusive, 100-500 mcg inclusive, 100-400 mcg inclusive, inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; 100-300mcg inclusive, 200-600mcg inclusive, 200-500mcg together with 200-250 mcg exenatide inclusive. In other inclusive, 200-400 mcg inclusive, 200-300 mcg inclusive, embodiments, the described dosage form contains insulin in 150-300 mcg inclusive, or 150-250 mcg inclusive; together an amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg with 20 mg insulin inclusive. In other embodiments, the inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg described dosage form contains exenatide in an amount of inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg between 100-600mcg inclusive, 100-500mcg inclusive, 100 inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg 400 mcg inclusive, 100-300 mcg inclusive, 200-600 mcg inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; inclusive, 200-500 mcg inclusive, 200-400 mcg inclusive, together with 250-300 mcg exenatide inclusive. 200-300 mcg inclusive, 150-300 mcg inclusive, or 150-250 0059. In other embodiments, the described dosage form mcg inclusive; together with 24 mg insulin inclusive. In other contains insulin in an amount of 8-32 mg inclusive, 8-28 mg embodiments, the described dosage form contains exenatide inclusive, 8-24 mg inclusive, 8-20 mg inclusive, 8-16 mg in an amount of between 100-600 mcg inclusive, 100-500 inclusive, 8-12 mg inclusive, 12-32 mg inclusive, 16-32 mg mcg inclusive, 100-400 mcg inclusive, 100-300 mcg inclu inclusive, 20-32 mg inclusive, 24-32 mg inclusive, 12-24 mg sive, 200-600mcg inclusive, 200-500mcg inclusive, 200-400 inclusive, 16-24 mg inclusive, 12-20 mg inclusive, or 16-20 mcg inclusive, 200-300 mcg inclusive, 150-300 mcg inclu mg inclusive; together with 100 mcg exenatide inclusive. In sive, or 150-250 mcg inclusive; together with 28 mg insulin other embodiments, the described dosage form contains insu inclusive. In other embodiments, the described dosage form lininanamount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 contains exenatide in an amount of between 100-600 mcg mg inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg inclusive, 100-500 mcg inclusive, 100-400 mcg inclusive, inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg 100-300mcg inclusive, 200-600mcg inclusive, 200-500mcg inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg inclusive, 200-400 mcg inclusive, 200-300 mcg inclusive, inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; 150-300 mcg inclusive, or 150-250 mcg inclusive; together together with 200 mcg exenatide inclusive. In other embodi with 32 mg insulin inclusive. ments, the described dosage form contains insulin in an 0058. In other embodiments, the described dosage form amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg contains insulin in an amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg inclusive, 8-24 mg inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg inclusive, 8-12 mg inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg inclusive, 20-32 mg inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; inclusive, 16-24 mg inclusive, 12-20 mg inclusive, or 16-20 together with 250 mcg exenatide inclusive. In other embodi mg inclusive; together with 150-300mcg exenatide inclusive. ments, the described dosage form contains insulin in an In other embodiments, the described dosage form contains amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg insulin in an amount of 8-32 mg inclusive, 8-28 mg inclusive, inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg 8-24 mg inclusive,8-20 mg inclusive, 8-16 mg inclusive, 8-12 inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg US 2015/001 7238 A1 Jan. 15, 2015

inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg 0065. In some embodiments, the patient receiving the inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; described pharmaceutical composition is receiving a small together with 300 mcg exenatide inclusive. In other embodi molecule DM therapeutic agent such as Metformin and/or a ments, the described dosage form contains insulin in an thiazolidinedione (TZD). In other embodiments, the patient is amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg not receiving a small-molecule DM therapeutic agent. The inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg described compositions are believed to be effective in either inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg instance. inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; Protease Inhibitors together with 400 mcg exenatide inclusive. In other embodi 0066. As used herein, the term “trypsin inhibitor” refers to ments, the described dosage form contains insulin in an any agent capable of inhibiting the action of trypsin on a amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg substrate. The ability of an agent to inhibit trypsin can be inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg measured using assays well known in the art. For example, in inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg a typical assay, one unit corresponds to the amount of inhibi inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg torthat reduces the trypsinactivity by one benzoyl-L-arginine inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; ethyl ester unit (BAEE-U). One BAEE-U is the amount of together with 500mcg exenatide inclusive. In other embodi enzyme that increases the absorbance at 253 nm by 0.001 per ments, the described dosage form contains insulin in an minute at pH 7.6 and 25°C. See, for example, K. Ozawa, M. amount of 8-32 mg inclusive, 8-28 mg inclusive, 8-24 mg Laskowski, 1966, J. Biol. Chem. 241:3955; andY. Birk, 1976, inclusive, 8-20 mg inclusive, 8-16 mg inclusive, 8-12 mg Meth. Enzymol. 45:700. inclusive, 12-32 mg inclusive, 16-32 mg inclusive, 20-32 mg 0067. Some trypsin inhibitors known in the art are specific inclusive, 24-32 mg inclusive, 12-24 mg inclusive, 16-24 mg to trypsin, while others inhibit trypsin and other proteases inclusive, 12-20 mg inclusive, or 16-20 mg inclusive; such as chymotrypsin. Trypsin inhibitors can be derived from together with 600 mcg exenatide inclusive. animal or vegetable sources: for example, soybean, corn, lima 0060. In other embodiments, the described dosage form and other beans, squash, Sunflower, bovine and other animal contains 8-16 mg insulin inclusive and 150-300 mcg. pancreas and lung, chicken and turkey egg white, Soy-based exenatide. In other embodiments, the described dosage form infant formula, and mammalian blood. Trypsin inhibitors can contains 8-12 mg insulin inclusive and 150-300 mcg. also be of microbial origin: for example, antipain; see, for exenatide. In other embodiments, the described dosage form example, H. Umezawa, 1976, Meth. Enzymol. 45, 678. A contains 12-16 mg insulin inclusive and 150-300 mcg. trypsin inhibitor can also be an arginine or lysine mimic or exenatide. In other embodiments, the described dosage form other synthetic compound: for example arylguanidine, ben contains 6-16 mg insulin inclusive and 150-300 mcg. Zamidine, 3,4-dichloroisocoumarin, diisopropylfluorophos exenatide. phate, gabexate mesylate, or phenylmethanesulfonyl fluo 0061. In other embodiments, the described dosage form ride. As used herein, an arginine or lysine mimic is a contains 8-16 mg insulin inclusive and 100-400 mcg. compound that is capable of binding to the P' pocket of exenatide. In other embodiments, the described dosage form trypsin and/or interfering with trypsin active site function. contains 8-12 mg insulin inclusive and 100-400 mcg. 0068. In certain embodiments, the trypsin inhibitor uti exenatide. In other embodiments, the described dosage form lized in methods and compositions of the present invention is contains 12-16 mg insulin inclusive and 100-400 mcg. selected from the group consisting of lima bean trypsin exenatide. In other embodiments, the described dosage form inhibitor, aprotinin, (a.k.a. pancreatic trypsin inhibitor or contains 6-16 mg insulin inclusive and 100-400 mcg. basic pancreatic trypsin inhibitor BPTI; Uniprot No. exenatide. P00974 database accessed on Jan. 2, 2013), Kazal inhibitor 0062. In other embodiments, the described dosage form (pancreatic secretory trypsin inhibitor), ovomucoid, Alpha contains 8-16 mg insulin inclusive and 100-200 mcg. 1-antitrypsin, Cortisol binding globulin, Centerin (SER exenatide. In other embodiments, the described dosage form PINA9/GCET1 (germinal centre B-cell-expressed transcript contains 8-12 mg insulin inclusive and 100-200 mcg. 1), PI-6 (Sun et al 1995), PI-8 (Sprecher et al 1995), exenatide. In other embodiments, the described dosage form Bomapin, a Glade A serpin for example Serpina3 (NCBI contains 12-16 mg insulin inclusive and 100-200 mcg. Gene ID: 12; database accessed on Dec. 27, 2012), Serpinao exenatide. In other embodiments, the described dosage form (NCBI Gene ID: 866; database accessed on Dec. 27, 2012), contains 6-16 mg insulin inclusive and 100-200 mcg. Serpinal 2 (NCBI Gene ID: 145264; database accessed on exenatide. Dec. 27, 2012); Serpinal 0 (NCBI Gene ID: 51156; database accessed on Dec. 27, 2012); Serpina7 (NCBI Gene ID: 6906; 0063. In other embodiments, the described dosage form database accessed on Dec. 27, 2012); Serpina'9 (NCBIGene contains 8-16 mg insulin inclusive and 200-400 mcg. ID: 327657; database accessed on Dec. 27, 2012); Serpinal 1 exenatide. In other embodiments, the described dosage form (NCBI Gene ID: 256394; database accessed on Dec. 27, contains 8-12 mg insulin inclusive and 200-400 mcg. 2012); Serpinal 3 (NCBI Gene ID: 388007; database exenatide. In other embodiments, the described dosage form accessed on Dec. 27, 2012); Serpina2 (NCBI Gene ID: contains 12-16 mg insulin inclusive and 200-400 mcg. 390502; database accessed on Dec. 27, 2012); and Serpina4 exenatide. In other embodiments, the described dosage form (NCBI Gene ID: 5104; database accessed on Dec. 27, 2012) contains 6-16 mg insulin inclusive and 200-400 mcg. Yukopin (SerpinB12: Gene ID: 89777; database accessed on exenatide. Dec. 27, 2012), antipain, benzamidine, 3,4-dichloroisocou 0064. In other embodiments, the described dosage form marin, diisopropylfluorophosphate, and gabexate mesylate. contains 8-16 mg insulin inclusive and 150-300 mcg. In other embodiments, more than one, for example 2, 3, or 4. exenatide. of the above inhibitors is selected. US 2015/001 7238 A1 Jan. 15, 2015

0069. A representative precursor sequence of aprotinin is: inhibitor, another trypsin inhibitor is also present. Non-lim iting examples of a trypsin inhibitor and a trypsin/chymot rypsin inhibitor are isolated KTI, which inhibits trypsin, and (SEQ ID NO: 1) isolated BBI (Bowman-Birkinhibitor), which inhibits trypsin MKMSRLCLSW ALLWLLGTLA ASTPGCDTSN OAKAQRPDFC and chymotrypsin; these are present together in the compo LEPPYTGPCK ARIIRYFYNA KAGLCOTFWY GGCRAKRNNF sition, in some embodiments. KSAEDCMRTC GGAIGPWENL. Chelators of Divalent Cations 0070. Of these 100 residues, residues 1-21 are the signal 0078. The chelator of divalent cations utilized in the peptide, 22-35 and 94-100 are propeptides, and the mature described methods and compositions is, in one embodiment, chain BBI chain is composed of residues 36-93 (58 AA). any physiologically acceptable compound having a high 0071. In other embodiments, the trypsin inhibitor is affinity for at least one of calcium, magnesium, and manga derived from soybean. Trypsin inhibitors derived from soy nese ions. In another embodiment, the chelator is selected bean (Glycine max) are readily available and are considered from the group consisting of citrate or a salt thereof ethyl to be safe for human consumption. They include, but are not enediamine tetracetic acid (EDTA) or a salt thereof (for limited to, SBTI, KTI (Kuntz, Trypsin Inhibitor), for example example disodium EDTA and calcium disodium EDTA); KTI3, and BBI (Bowman-Birk inhibitor: Uniprot number EGTA (ethylene glycol tetraacetic acid) or a salt thereof; P01055 database accessed on Jan. 3, 2013). SBTI is com diethylene triamine pentaacetic acid (DTPA) or a salt thereof; posed of KTI, which inhibits trypsin, and BBI, which inhibits and BAPTA (1.2-bis(o-aminophenoxy)ethane-N,N,N',N'- trypsin and chymotrypsin. Such trypsin inhibitors are avail tetraacetic acid) or a salt thereof. In other embodiments, one able for example from Sigma-Aldrich, St. Louis, Mo., USA. of the above-listed chelators is utilized. In more specific 0072 A representative precursor sequence of BBI is: embodiments, the chelator is EDTA. Oils (SEQ ID NO: 2) MWWLKWCLVL LFLWGGTTSA NLRLSKLGLL MKSDHOHSND 0079 Pharmaceutical compositions and methods described herein utilize one or more oils as the basis of their DESSKPCCDO CACTKSNPPO CRCSDMRLNS CHSACKSCIC liquid phase. In certain embodiments, the oil may be any physiologically acceptable oil that is liquid at ambient tem ALSYPAQCFC VDITDFCYEP CKPSEDDKEN. perature. 0073. Of these 110 residues, residues 1-19 are the signal 0080. In more specific embodiments, the oil comprises an peptide, 20-39 are a propeptide, and the mature chain BBI omega-3 fatty acid. In other embodiments, the omega-3 fatty chain is composed of residues 40-110 (71 AA). acid is an omega-3 polyunsaturated fatty acid. In another 0074 KTI3 has Uniprot number P01070 (database embodiment, the omega-3 fatty acid is DHA, an omega-3, accessed on Jan. 3, 2013). A representative precursor polyunsaturated, 22-carbon fatty acid also referred to as 4, 7. sequence of KTI3 is: 10, 13, 16, 19-docosahexaenoic acid. In another embodiment, the omega-3 fatty acid is -linolenic acid (9, 12, 15-octadec atrienoic acid). In another embodiment, the omega-3 fatty SEO ID NO : 3) acid is Stearidonic acid (6, 9, 12, 15-octadecatetraenoic acid). MKSTIFFLFL FCAFTTSYLP SAIADFWLDN EGNPLENGGT In another embodiment, the omega-3 fatty acid is eicosa YYILSDITAF GGIRAAPTGN ERCPLTWVOS RNELDKGIGT trienoic acid (ETA; 11, 14, 17-eicosatrienoic acid). In another embodiment, the omega-3 fatty acid is eicSoatetraenoic acid IISSPYRIRF IAEGHPLSLK FDSFAWIMLC WGIPTEWSWW (8, 11, 14, 17-eicosatetraenoic acid). In one embodiment, the omega-3 fatty acid is eicosapentaenoic acid (EPA; 5, 8, 11, EDLPEGPAWK IGENKDAMDG WFRLERWSDD EFNNYKLWFC 14, 17-eicosapentaenoic acid). In another embodiment, the POOAEDDKCG DIGISIDHDD GTRRLVVSKN KPLVVOFOKL omega-3 fatty acid is eicosahexaenoic acid (also referred to as 5, 7, 9, 11, 14, 17-eicosahexaenoic acid). In another embodi DKESLAKKNH GLSRSE ment, the omega-3 fatty acid is docosapentaenoic acid (DPA; 0075 Of the above sequence, residues 1-24 are the signal 7, 10, 13, 16, 19-docosapenatenoic acid). In another embodi peptide, 206-216 are a propeptide, and the mature KTI chain ment, the omega-3 fatty acid is tetracosahexaenoic acid (6,9. is composed of residues 25-205 (181 AA). 12, 15, 18, 21-tetracosahexaenoic acid). 0076. In other embodiments, a method or oral pharmaceu I0081. In other embodiments, the oil is a naturally-occur tical composition described herein utilizes two trypsin inhibi ring oil comprising an omega-3 fatty acid. In more specific tors. In other embodiments, more than two trypsin inhibitors embodiments, the oil is selected from the group consisting of are present. In other embodiments, three trypsin inhibitors are a fish oil, canola oil, flaxseed oil, algal oil and hemp seed oil. present. In other embodiments, four trypsin inhibitors are In more specific embodiments, the oil is a fish oil. Several present. In other embodiments, two of the trypsin inhibitors types of fish oil have been tested in the compositions are SBTI and aprotinin. In yet other embodiments, the only described herein and have all been found to work equally two trypsin inhibitors are SBTI and aprotinin. In still other well. embodiments, the only two trypsin inhibitors are isolated BBI and isolated aprotinin. Representative Specific Formulations 0077. In other embodiments, a chymotrypsin inhibitor is I0082 In still more specific embodiments, a liquid formu present together with a trypsin inhibitor. In other embodi lation utilized in the described method or composition com ments, when the chymotrypsin inhibitor is also a trypsin prises insulin, exenatide, Gelucire 44/14, EDTA, SBTI, apro US 2015/001 7238 A1 Jan. 15, 2015

tinin, and fish oil. In other embodiments, the liquid water-free. “Water-free” refers, in certain embodiments, to a formulation consists essentially of insulin, eXenatide, Gelu formulation into which no aqueous components have been cire 44/14, EDTA, SBTI, aprotinin, and fish oil. “Consists intentionally added. It does not preclude the presence of trace essentially of for these purposes indicates that the liquid amounts of water that have been absorbed from the atmo formulation does not contain any other components that sphere into the components thereof. In another embodiment, appreciably affect its physiological characteristics. In other the liquid formulation is free of aqueous components. If more embodiments, the liquid formulation consists of insulin, than one liquid formulation is present, for example in a multi exenatide, Gelucire 44/14, EDTA, SBTI, aprotinin, and fish component composition, each liquid formulation may be free oil. In other, even more specific embodiments, the amounts of of aqueous components. In yet other embodiments, one or insulin, exenatide, EDTA, SBTI, aprotinin, and fish oil per more oils are the only liquid components of each of the one or dosage form are 8-16 mg, 150-300mcg, 100-200 mg, 50-100 more liquid formulations. In yet another embodiment, fish oil mg, 20-30 mg, and 0.4-0.7 ml, respectively, and the amount of is the only liquid component of each of the one or more liquid Gelucire 44/14 is 8-1.6%. In still more specific embodiments, formulations. the amounts of insulin, exenatide. EDTA, SBTI, aprotinin, and fish oil per dosage form are 8-16 mg, 150-300 mcg, 150 Coatings mg, 75 mg, 24 mg, and 0.5-0.7 ml, respectively, and the I0086 Those of skill in the art will appreciate, given the amount of Gelucire 44/14 is 8-1.6%. In other embodiments, present disclosure, that various pH-sensitive coatings may be the above composition further comprises a non-ionic deter utilized in the described methods and compositions. In certain gent. In more specific embodiments, the non-ionic detergent embodiments, any coating that inhibits digestion of the com is a polysorbate-based detergent. In even more specific position in the stomach of a Subject may be utilized. Typically, embodiments, the polysorbate-based detergent is polysorbate Such coatings will not dissolve in human gastric juices within 80. Preferably, the polysorbate 80 constitutes 3-10% weight/ 2 hours, and will dissolve within 30 minutes in duodenal fluid. weight inclusive of the oil-based liquid formulation. In other I0087. In other embodiments, the coating comprises a bio embodiments, the above composition is coated by a coating degradable polysaccharide. In other embodiments, a hydro that resists degradation in the stomach. gel is utilized. In other embodiments, the coating comprises 0083. In still more specific embodiments, a liquid formu one of the following excipients: chitosan, an aquacoat ECD lation utilized in the described method or composition com coating, an azo-crosslinked polymer, cellulose acetate phtha prises insulin, exenatide, a self-emulsifying component, late, cellulose acetate trimellitate (CAT), cellulose acetate EDTA, SBTI, aprotinin, and fish oil. In other embodiments, butyrate, hydroxypropylmethyl cellulose phthalate, or poly the liquid formulation consists essentially of insulin, vinyl acetate phthalate. exenatide, a self-emulsifying component, EDTA, SBTI, apro I0088. In other embodiments, a timed-release system such tinin, and fish oil. “Consists essentially of for these purposes as PulsincapTM is utilized. indicates that the liquid formulation does not contain any I0089. In preferred embodiments, the coated dosage forms other components that appreciably affect its physiological described herein release the core (containing the oil-based characteristics. In other embodiments, the liquid formulation formulation) when pH reaches the range found in the intes consists of insulin, exenatide, a self-emulsifying component, tines, which is alkaline relative to that in the stomach. In more EDTA, SBTI, aprotinin, and fish oil. In other, even more specific embodiments, the coating comprises a pH-sensitive specific embodiments, the amounts of insulin, eXenatide, EDTA, SBTI, aprotinin, and fish oil per dosage form are 8-16 polymer. In various embodiments, either mono-layer or mg, 150-300 mcg, 100-200 mg, 50-100 mg, 20-30 mg, and multi-layer coatings may be utilized. 0.4-0.7 ml, respectively, and the amount of self-emulsifying 0090. In one embodiment, the coating is an enteric coat component is 8-16%. In still more specific embodiments, the ing. Methods for enteric coating are well known in the art amounts of insulin, exenatide, EDTA, SBTI, aprotinin, and (see, for example, Siepmann Fetal 2005). In more specific fish oil per dosage form are 8-16 mg, 150-300 mcg, 150 mg. embodiments, a EudragitTM coating is utilized as the enteric 75 mg, 24 mg, and 0.5-0.7 ml, respectively, and the amount of coating. EudragitTM coatings are acrylic polymers, the use of self-emulsifying component is 8-16%. In other embodi which is well known in the art. ments, the above composition further comprises a non-ionic 0091. In another embodiment, microencapsulation is used detergent. In more specific embodiments, the non-ionic deter as a stomach-resistant coating in the compositions described gent is a polysorbate-based detergent. In even more specific herein. Methods for microencapsulation are well known in embodiments, the polysorbate-based detergent is polysorbate the art, and are described inter alia in United States Patent 80. Preferably, the polysorbate 80 constitutes 3-10% weight/ Application Publication No. 2011/0305768, which is incor weight inclusive of the oil-based liquid formulation. In other porated by reference herein. embodiments, the above composition is coated by a coating 0092. In other embodiments, the coating is a capsule. that resists degradation in the stomach. Gelatin capsules are most preferred. Methods for inserting an 0084 “Weight/weight percentages referred to herein uti oil-based formulation into a gelatin capsule are well known in lize the amount of oil base in the formulation, for example fish the art. oil, as the denominator; thus, 60 mg of Gelucire in 500 mg fish oil is considered as 12% w/w, regardless of the weight of the Pharmaceutical Compositions and Methods of Making Same other components. Similarly, 50 mg. Tween-80 mixed with 0093. In another aspect a pharmaceutical composition 500 mg fish oil is considered as 10% Tween-80. described herein is provided for treating diabetes mellitus, for 0085. In other embodiments, a liquid formulation utilized example Type 2 diabetes mellitus, in a human. in the described method or composition is water-free. If more 0094. In yet another aspect, a use of a combination of than one liquid formulation is present, for example in a multi ingredients described herein is provided in the preparation of component composition, each liquid formulation may be a medicament for treating diabetes mellitus in a human. US 2015/001 7238 A1 Jan. 15, 2015

0095 Still another aspect provides a method for treating tering to a Subject in need of such treatment a pharmaceutical diabetes mellitus in a human, the method comprising the composition described hereinabove, thereby treating hyper optional step of selecting a subject by diagnosing diabetes triglyceridemia. mellitus, followed by the step of administering to a subject in 0103) Another aspect provides a method of treating need of Such treatment a pharmaceutical composition elevated serum apolipoprotein B (ApoB), the method com described herein, thereby treating diabetes mellitus in a prising the step of administering to a subject in need of Such human. treatment a pharmaceutical composition described herein 0096. Still another aspect provides a method of manufac above. turing a pharmaceutical composition, comprising the steps of 0104. Another aspect provides a method of treating meltinga waxy, self-emulsifying component; adding the mol elevated total cholesterol/HDL ratio, the method comprising ten component to fish oil, optionally cooling the resulting the step of administering to a Subject in need of such treatment mixture; adding to the oil EDTA, SBTI in powder form, a pharmaceutical composition described hereinabove. aprotinin in powderform, crystalline insulin, and exenatide in 0105. Another aspect provides a method of treating an powder form, in some embodiments in the order listed; and elevated apolipoprotein B/apolipoprotein A1 ratio, the mixing and homogenizing the resulting liquid, in some method comprising the step of administering to a Subject in embodiments on a roller mill. need of Such treatment a pharmaceutical composition 0097. Yet another aspect provides a method of manufac described hereinabove, thereby treating an elevated apolipo turing a pharmaceutical composition, comprising the steps of protein B/apolipoprotein A1 ratio. meltinga waxy, self-emulsifying component; adding the mol 0106 Methods for measuring each of the aforementioned ten component to fish oil, optionally cooling the resulting lipid parameters are well known to those skilled in the art. mixture; adding to the oil EDTA, isolated BBI in powder Exemplary methods are described interalia in Chiquette E et form, isolated KTI3 in powder form, crystalline insulin, and al and Martinez-Colubi Metal. exenatide in powder form, in Some embodiments in the order 0107 Another aspect provides a method of treating an listed; and mixing and homogenizing the resulting liquid, in impaired insulin-induced enhancement of vasodilator some embodiments on a roller mill. responses in a Subject with metabolic syndrome, the method 0098. Yet another aspect provides a method of manufac comprising the step of administering to a Subject in need of turing a pharmaceutical composition, comprising the steps of Such treatment a pharmaceutical composition described here meltinga waxy, self-emulsifying component; adding the mol inabove, thereby treating an impaired insulin-induced ten component to fish oil, optionally cooling the resulting enhancement of vasodilator responses in a subject with meta mixture; adding to the oil EDTA, isolated KTI3 in powder bolic syndrome. Methods for measuring insulin-induced form, aprotinin in powder form, crystalline insulin, and vasodilator responses are known in the art, and include, for exenatide in powder form, in Some embodiments in the order example, measuring blood flow responses (for example in the listed; and mixing and homogenizing the resulting liquid, in forearm) to acetylcholine (ACh) and sodium nitroprus side some embodiments on a roller mill. (SNP) (Tesauro et al). 0099. Yet another aspect provides a use of a combination 0.108 Metabolic syndrome is considered to be presentifat of ingredients described hereinabove in the preparation of a least three of the following factors are present: medicament for treating unstable diabetes, also known as 0.109 1. A large waistline (abdominal obesity). glycemic lability (Ryan et al., 2004) in a human. Yet another 0110 2. A high triglyceride level (or a high triglyceride aspect provides a method for treating unstable diabetes, the level in the absence of medicine to treat high triglycer method comprising the step of administering to a Subject in ides). 0111. 3. A low HDL cholesterol level (or low HDL need of Such treatment a pharmaceutical composition cholesterol in the absence of medicine to treat low HDL described hereinabove, thereby treating unstable diabetes. cholesterol). 0100 Provided in another embodiment is a use of a com 0112 4. Hypertension (or hypertension in the absence bination of ingredients described hereinabove in the prepara of medicine to treat hypertension. tion of a medicament for treating an elevated fasting blood 0113 5. High fasting blood sugar (or high fasting blood glucose level in a human. Provided in another embodiment is Sugar in the absence of medicine to treat high fasting a method for treating an elevated fasting blood glucose level. blood Sugar). the method comprising the step of administering to a subject 0114. Another aspect provides a method of treating non in need of Such treatment a pharmaceutical composition alcoholic steatohepatitis in a Subject with metabolic Syn described hereinabove, thereby treating an elevated fasting drome, the method comprising the step of administering to a blood glucose. In certain preferred embodiments, the subject Subject in need of such treatment a pharmaceutical composi is a human Subject. In various embodiments, elevated fasting tion described herein, thereby treating non-alcoholic Steato blood glucose is considered to exist in a subject having a hepatitis in a subject with metabolic syndrome. Methods of glycated hemoglobin HgA1c level of 8-10%, or a fasting diagnosing and measuring non-alcoholic Steatohepatitis are plasma sugar level from 100 to 125 mg/dL, or 5.6 to 6.9 well known in the art, and include, for example, measuring mmol/L. plasma aspartate transaminase (AST) levels, plasma alanine 0101 Provided herein, in another embodiment, is a transaminase (ALT) levels, hepatic mRNA levels of genes method of treating elevated total cholesterol, the method involved in lipogenesis, and diacylglycerol acyltransferase-2 comprising the step of administering to a subject in need of (DGAT2) levels in the liver (Miyashita Tet al). Such treatment a pharmaceutical composition described here 0.115. Another aspect provides a method of treating inabove, thereby treating elevated total cholesterol. elevated total cholesterol, the method comprising the step of 0102) Another aspect provides a method of treating hyper administering to a Subject in need of Such treatment an oral, triglyceridemia, the method comprising the step of adminis oil-based liquid formulation, the oil-based liquid formulation US 2015/001 7238 A1 Jan. 15, 2015

comprising an insulin, one or, in another embodiment more ment more than one, trypsin inhibitor, and a chelator of diva than one, trypsin inhibitor, and a chelator of divalent cations, lent cations, wherein the oil-based liquid formulation is Sur wherein the oil-based liquid formulation is surrounded by a rounded by a coating or capsule that resists degradation in the coating or capsule that resists degradation in the stomach. stomach. Another aspect provides a method of treating an Another aspect provides a method of treating elevated total elevated total cholesterol/HDL ratio, the method comprising cholesterol, the method comprising the step of administering the step of administering to a Subject in need of such treatment to a subject in need of such treatment an oral, oil-based liquid an oral, oil-based liquid formulation, the oil-based liquid formulation, the oil-based liquid formulation comprising a formulation comprising a GLP-1 analogue, one or, in another GLP-1 analogue, one or, in another embodiment more than embodiment more than one, trypsin inhibitor, and a chelator one, trypsin inhibitor, and a chelator of divalent cations, of divalent cations, wherein the oil-based liquid formulation wherein the oil-based liquid formulation is surrounded by a is Surrounded by a coating or capsule that resists degradation coating or capsule that resists degradation in the stomach. The in the stomach. The oil, insulin, GLP-1 analogue, trypsin oil, insulin, GLP-1 analogue, trypsin inhibitor(s), chelator, inhibitor(s), chelator, coating, and other optional ingredients coating, and other optional ingredients of the above compo of the above compositions may be any described herein; each sitions may be any described herein; each alternative may be alternative may be combined freely to form discrete embodi combined freely to form discrete embodiments of the inven ments of the invention disclosed herein. tion disclosed herein. 0119) Another aspect provides a method of treating an 0116. Another aspect provides a method of treating hyper elevated apolipoprotein B/apolipoprotein A1 ratio, the triglyceridemia, the method comprising the step of adminis method comprising the step of administering to a Subject in tering to a subject in need of Such treatment an oral, oil-based need of Such treatment an oral, oil-based liquid formulation, liquid formulation, the oil-based liquid formulation compris the oil-based liquid formulation comprising an insulin, one ing an insulin, one or, in another embodiment more than one, or, in another embodiment more than one, trypsin inhibitor, trypsin inhibitor, and a chelator of divalent cations, wherein and a chelator of divalent cations, wherein the oil-based liquid the oil-based liquid formulation is Surrounded by a coating or formulation is surrounded by a coating or capsule that resists capsule that resists degradation in the stomach. Another degradation in the stomach. Another aspect provides a aspect provides a method of treating hypertriglyceridemia, method of treating an elevated apolipoprotein B/apolipopro the method comprising the step of administering to a subject tein A1 ratio, the method comprising the step of administering in need of Such treatment an oral, oil-based liquid formula to a Subject in need of such treatment an oral, oil-based liquid tion, the oil-based liquid formulation comprising a GLP-1 formulation, the oil-based liquid formulation comprising a analogue, one or, in another embodiment more than one, GLP-1 analogue, one or, in another embodiment more than trypsin inhibitor, and a chelator of divalent cations, wherein one, trypsin inhibitor, and a chelator of divalent cations, the oil-based liquid formulation is Surrounded by a coating or wherein the oil-based liquid formulation is surrounded by a capsule that resists degradation in the stomach. The oil, insu coating or capsule that resists degradation in the stomach. The lin, GLP-1 analogue, trypsin inhibitor(s), chelator, coating, oil, insulin, GLP-1 analogue, trypsin inhibitor(s), chelator, and other optional ingredients of the above compositions may coating, and other optional ingredients of the above compo be any described herein; each alternative may be combined sitions may be any described herein; each alternative may be freely to form discrete embodiments of the invention dis combined freely to form discrete embodiments of the inven closed herein. tion disclosed herein. 0117. Another aspect provides a method of treating I0120 Another aspect provides a method of treating an elevated serum ApoB, the method comprising the step of impaired insulin-induced enhancement of vasodilator administering to a Subject in need of Such treatment an oral, responses in a Subject with metabolic syndrome, the method oil-based liquid formulation, the oil-based liquid formulation comprising the step of administering to a Subject in need of comprising an insulin, one or, in another embodiment more Such treatment an oral, oil-based liquid formulation, the oil than one, trypsin inhibitor, and a chelator of divalent cations, based liquid formulation comprising an insulin, one or, in wherein the oil-based liquid formulation is surrounded by a another embodiment more than one, trypsin inhibitor, and a coating or capsule that resists degradation in the stomach. chelator of divalent cations, wherein the oil-based liquid for Another aspect provides a method of treating elevated serum mulation is surrounded by a coating or capsule that resists ApoB, the method comprising the step of administering to a degradation in the stomach. Subject in need of Such treatment an oral, oil-based liquid I0121 Another aspect provides a method of treating an formulation, the oil-based liquid formulation comprising a impaired insulin-induced enhancement of vasodilator GLP-1 analogue, one or, in another embodiment more than responses in a Subject with metabolic syndrome, the method one, trypsin inhibitor, and a chelator of divalent cations, comprising the step of administering to a Subject in need of wherein the oil-based liquid formulation is surrounded by a Such treatment an oral, oil-based liquid formulation, the oil coating or capsule that resists degradation in the stomach. The based liquid formulation comprising a GLP-1 analogue, one oil, insulin, GLP-1 analogue, trypsin inhibitor(s), chelator, or, in another embodiment more than one, trypsin inhibitor, coating, and other optional ingredients of the above compo and a chelator of divalent cations, wherein the oil-based liquid sitions may be any described herein; each alternative may be formulation is surrounded by a coating or capsule that resists combined freely to form discrete embodiments of the inven degradation in the stomach. The oil, insulin, GLP-1 analogue, tion disclosed herein. trypsin inhibitor(s), chelator, coating, and other optional 0118. Another aspect provides a method of treating an ingredients of the above compositions may be any described elevated total cholesterol/HDL ratio, the method comprising herein; each alternative may be combined freely to form the step of administering to a subject in need of such treatment discrete embodiments of the invention disclosed herein. an oral, oil-based liquid formulation, the oil-based liquid I0122) Another aspect provides a method of treating non formulation comprising an insulin, one or, in another embodi alcoholic steatohepatitis in a Subject with metabolic Syn US 2015/001 7238 A1 Jan. 15, 2015

drome, the method comprising the step of administering to a TABLE 1 Subject in need of Such treatment an oral, oil-based liquid formulation, the oil-based liquid formulation comprising an Formula insulin, one or, in another embodiment more than one, trypsin tion name Emulsifiers Other ingredients inhibitor, and a chelator of divalent cations, wherein the oil Experiment 2A based liquid formulation is Surrounded by a coating or cap Sule that resists degradation in the stomach. Another aspect GMS 290 GMS 2% only 3.375 g. SBTI, provides a method of treating non-alcoholic steatohepatitis in 6.75 mg EDTA, 1.08 mg aprotinin, a Subject with metabolic syndrome, the method comprising 0.39 mg. human the step of administering to a subject in need of such treatment insulin, an oral, oil-based liquid formulation, the oil-based liquid 22.5 g. of fish oil formulation comprising a GLP-1 analogue, one or, in another 2%-296 2% GMS, 2% lecithin Same as above. embodiment more than one, trypsin inhibitor, and a chelator 2%-10% 2% GMS, 10% lecithin Same as above. of divalent cations, wherein the oil-based liquid formulation 10% lec. 10% lecithin only Same as above. is Surrounded by a coating or capsule that resists degradation Experiment 2B in the stomach. The oil, insulin, GLP-1 analogue, trypsin A. 5% lecithin, 2% GMS Same as above. inhibitor(s), chelator, coating, and other optional ingredients B 3% lecithin, 12% Gelucire 44/14 Same as above. of the above compositions may be any described herein; each C 6% lecithin, 12% Gelucire 44/14 Same as above. alternative may be combined freely to form discrete embodi D 5% Tween-80, 12% Gelucire 44/14 Same as above. ments of the invention disclosed herein. E 10% Tween-80, 12% Gelucire 44f14 Same as above. 0123. Wherever alternatives for single separable features F 12% Gelucire 44f14 only Same as above. Such as, for example, a insulin protein or dosage thereof, a GLP-1 analogue of dosage thereof, a protease inhibitor, a chelator, an emulsifier, or a coating are laid out herein as Husbandry "embodiments', it is to be understood that such alternatives may be combined freely to form discrete embodiments of the I0128 Animal health: Only healthy pigs, as certified by a invention disclosed herein. clinical veterinarian, were used for the study. Housing: Soli 0.124 With respect to the jurisdictions allowing it, all pat tary when with CVC and grouped at other times. Bedding: ents, patent applications, and publications mentioned herein, Concrete--woodchips. Illumination: 12-12 hlight cycle. Tem both Supra and infra, are incorporated herein by reference. perature: 19-25° C. 0125. The invention is further illustrated by the following examples and the figures, from which further embodiments and advantages can be drawn. These examples are meant to Identification illustrate the invention but not to limit its scope. I0129. Each animal was uniquely identified via ear tags. EXPERIMENTAL DETAILS SECTION Example 1 Experimental Design Identification of Effective Emulsifiers for 0.130 Animals were deprived of food 24-36 hours prior to Homogenous Therapeutic Protein/Fish Oil testing. Access to water was ad libitum. Preparations I0131 Animals were anesthetized with 20 mg/kg ket 0126 Previous formulations of insulin in fish oil were amine--2 mg/kg Xylazine. Fasting and anesthetized pigs were found to exhibit precipitation with the passage of time; they positioned on their left side before liquid formulations were were thus inconvenient for large-scale pharmaceutical dosage administered under endoscopic guidance, directly to the form preparation. New formulations containing 3.375 g. duodenum. After injection of the formulation, 1 mL fish oil SBTI per 22.5 g. of fish oil and containing the following was injected, followed by 10 mL air, to flush the apparatus, emulsifiers were tested: lecithin (trial sequence 1), Polysor thereby ensuring administration of the entire formulation. bate 80 (Tween-80) (sequence 2), or Gelucire R 44/14 (se Pigs were then returned to their pens to allow for full recovery quence 3), alone or in combination with each other or glycerol from the anesthetic treatment, which required 10-15 min. monostearate (GMS) (FIG. 1). Subsequently, the most prom Blood samples (0.5 mL of which were analyzed) were peri ising formulations (indicated with an asterisk) were produced odically drawn from the central line catheter (CVC) over the again by melting the Gelucire R (which was a waxy Solid as ensuing 240-min monitoring period. Blood glucose concen ambient temperature), then adding it to the fish oil. After trations were determined from each sample, at each time cooling this mixture, the Solid components were added in point. Piglets were intravenously treated with gentamycin powder form in the following order: EDTA, SBTI, aprotinin, (100 mg/10 kg) after every experiment day to avoid infection. and insulin; and the resulting liquid was mixed and homog In cases where glucose concentrations dropped below 30 enized on a roller mill. mg/dL, piglets were served commercial pig chow, and glu cose concentrations were monitored for an additional 30 min Example 2 utes thereafter. In Vivo Testing of Various Emulsifier Formulations 0.132. A washout period of at least 2 days was enforced Materials and Experimental Methods between test days. Formulations Results 0127. The formulations tested in Experiments 2A and 2B are shown below in Table 1. Percentages of emulsifiers are 0.133 10 insulin-fish oil formulations with different emul expressed as weight/weight with respect to the weight of the sifiers were tested for in-vivo activity on blood glucose levels liquids present. in 2 separate experiments. Results are described below. US 2015/001 7238 A1 Jan. 15, 2015

TABLE 2 Results of Experiment 2A. The three numbers in each box indicate baseline value, lowest value, and end (20 mg/dL). LOW indicates a value of less than 20 mg/dL. Pig 5 Formulation Pig 1 Pig2 Pig3 Pig4 (stoma) Score GMS 290 78 -> 48-> 72->low 63 -23 - 67 - 32 SS-e 21 3 65 -> low 28 -> 60 -e S8 2% GMS, 82 -> Low 78 -eAS - 85 -30 - 78 - 23 2 2% lecithin -> 65 76 68 -> 61 2% GMS, 76 -> Low 76 ->low 76 ->low 74 -> low 4 10% lecithin -> 77 -> 21 -> low - 70 10% lecithin 51 -> 63 -> 50 ->low 57->low 61 -> low 5 71 -> low -> low -> low

0134. The results of Experiment 2B are indicated in FIG. aprotinin in 0.5-0.7 mL fish oil. The liquid was coated by a 2. soft-gel, enteric-coated capsule. The dosage form was manu factured by Swiss Caps AG. Example 3 0.138. The oral exenatide formulation contained 150 microgram (mcg) exenatide, 150 mg EDTA, 75 mg SBTI, and An Oral Insulin/Oral Exenatide Combination 24 mg aprotinin in 0.5-0.7 mL fish oil. The liquid was coated Sharply Reduces Blood Glucose Levels and Prevents by a soft-gel, enteric-coated capsule. The dosage form was Postprandial Glucose Excursions manufactured by Swiss Caps AG. Materials and Experimental Methods Statistical Analysis 0.139 p-values were calculated using a paired, one-tailed Animals t-teSt. 0135 Healthy 25-30 kg, 3-4 month-old pigs were used. Results Experimental Design 0140 AS expected, placebo-treated pigs experienced glu cose excursions following caloric intake. Pigs treated with 0136. Pigs were deprived of food for 24-36 hours before oral exenatide 30 minutes before caloric intake completely the start of the study. CVCs were replaced every five days, avoided the glucose excursions throughout the 150 minutes unless circumstances required earlier replacement. Animals following caloric intake (FIG. 3). Oral insulin also curbed were anesthetized by isoflurane administration through a glucose excursions, but did not fully prevent a rise in blood mask before tracheal intubation and respirator hook-up (2 L glucose concentrations throughout the entire monitoring O per minute and 3-5% isoflurane, when required), then period. A sharply greater effect was observed upon combined were positioned on their left side and capsules were admin treatment with oral exenatide and oral insulin. In addition to istered directly into the duodenum, using an endoscopic bas fully preventing glucose excursions, the combined treatment ket under endoscopic guidance. When both oral insulin and reduced glucose concentrations to a low of more than 50% oral exenatide were administered, oral exenatide was deliv below baseline values during the entire period from the ered first, followed by oral insulin within 2-10 minutes. Pigs 70-minute timepoint until the end of the 180-minute moni were returned to their pens to allow for full recovery from the toring session. The difference in blood glucose levels was anesthetic treatment and 30 minutes after capsule administra statistically significant compared to no treatment or treatment tion were given 10 g/kg piglet commercial milk powder (Den with insulin or exenatide alone (p-values: combination vs. kapig Premium (Denkavit) prepared in an equal Volume of control: 5.3x10; combination vs. oral insulin: 0.00015, and water. Meals were typically consumed within 7-15 minutes. Blood samples were periodically drawn from the CVC for combination vs. oral exenatide: 1.6x10). glucose concentration testing over the ensuing 240-min Example 4 monitoring period. Piglets were intravenously treated with gentamycin (100 mg/10 kg) after every experiment day to Testing of an Oral Insulin/Oral Exenatide avoid infection. In cases where glucose concentrations Combination in Human Subjects dropped below 30 mg/dL, piglets were served commercial pig chowder and glucose concentrations were monitored for 0.141 Volunteers, either diabetic or non-diabetic, are an additional 30 minutes thereafter. Control pigs were administered prior to a meal one or more dosage forms having untreated. Three pigs were used, each of whom received each a pH-sensitive coating or capsule containing a liquid formu formulation a number of times. The number of tests were n=5 lation containing one or more protease inhibitors, EDTA, for oral insulin, n=5 for oral exenatide, n=7 for the combina insulin and exenatide. One representative formulation is tion, and n=6 for the control. 50-200 mg. per capsule SBTI; 20-30 mg. per capsule Apro tinin: 75-200 mg EDTA; 8-32 mg. per capsule insulin; 150 Formulations 600 mcg. per capsule exenatide; and 0.5-0.7 ml fish oil, optionally with an emulsifier (e.g. 8-20% Gelucire R 44/14). 0.137 The oral insulin formulation contained 8 mg insulin, In other experiments, subjects are administered two encapsu 12% Gelucire 44/14, 150 mg EDTA, 75 mg SBTI, and 24 mg lated liquid formulations, one containing protease inhibitor US 2015/001 7238 A1 Jan. 15, 2015

(S), EDTA, and insulin, and the other containing protease Procedures inhibitor(s), EDTA, and exenatide. One representative formu lation is a first capsule containing one or more protease inhibi (O153 Stage I: tors, EDTA, and insulin in a liquid formulation, optionally 0154 Segment I of the treatment phase will consist of two with an emulsifier (e.g. Gelucire R 44/14), together with a visits, evaluating 150 lug and 300 ug oral exenatide versus second capsule containing protease inhibitors, emulsifier, placebo treatment in healthy Subjects, administered in the EDTA, and exenatide in a liquid formulation, optionally with morning after an 8-hr fast. Further dose escalation to Segment II will be authorized if no serious side effects and safety and an emulsifier (e.g. GelucireR 44/14). tolerability are demonstrated. In Segment II, fasting Subjects 0142. Subjects are challenged with a 75 g oral glucose will be administered 450 and/or 600 ug oral exenatide and/or load after capsule administration (for example 30-60 minutes placebo to fasting individuals (Appendix 1). The highest tol later) and relevant physiological parameters are Subsequently erable dose will then be delivered 60 minutes before intake of monitored by collection of blood samples. a standard meal. 0143. The following protocol is an exemplary protocol (O155 Each dosing will be followed by at least a 72-hour that may be used. Those skilled in the art will appreciate that washout period. non-essential details of the protocol may be modified without compromising its ability to provide the desired information. Screening Phase: Overview 0156 The following evaluations may be performed: (O157 Medical history 0144 Stage I: 0158 Physical examination 0145 Stage I consists of two segments. Segment 1 will 0159 Medication history assess the safety, tolerability and the pharmacokinetics/phar (0160 ECG macodynamics (PK/PD) of escalating doses of oral exenatide 0.161 Vital signs (blood pressure, heart rate). Vital signs in eight (8) healthy volunteers. In the first segment, the two will be measured in the sitting position after at least 5 lower doses of oral exenatide (150 and 300 ug exenatide) will minutes of rest. be randomly tested among all healthy, fasting Subjects. If 0162 Clinical laboratory evaluations (chemistry, deemed safe, further dose escalation (450 and 600 ug hematology, HgA1C) exenatide) will be authorized in Segment 2. The highest tol erable dose will then be administered to the subjects 60 min Treatment Phase: utes before a standard meal (Visit 5). 0146 Stage II: 0163 1. Blood samples will be obtained at -45, —30, 0147 This stage will assess the T2DM patient response to - 15, 0, 15, 30, 45, 60, 75, 90, 105, 120, 135, 150 min, escalating doses of exenatide when delivered 60 min before a where time=0 is the time at which the capsule is ingested, standard meal. Placebo controls will be included in the study, and tested for glucose, insulin, c-peptide, as well as GLP-1 as will treatment with an active control of Byetta R (5ug) analogue content. subcutaneously delivered 30 min before a standard meal. In (0164. 2. A standard meal will be served 60 minutes after addition, the T2DM subjects will be treated with an oral oral exenatide administration to healthy patients at visit 5 insulin capsule containing 16 mg of insulin and with a com only. bination of oral insulin/oral exenatide, at two independent 0.165 3. Gastrointestinal symptoms may be assessed. study visits, 60 minutes before a standard meal. (0166 Stage II: 0.167 300 and 600 micrograms exenatide, or the two high 0148. In addition to exenatide and/or insulin, all formula est tolerable doses determined in Stage I, will be administered tions contain 75 mg SBTI, 24 mg aprotinin, and 150 mg in parallel and in combination with 16 mg insulin, for EDTA, all in fish oil, in enteric-coated capsules. example as 16 mg combined with 300 microgram, 60 minutes before a standard meal. On study visits where an injection of Interpretation: Byetta R (5 lug) is administered, a meal will be served 30 014.9 The AUC of glucose reductions and insulin excur minutes after dosing. sion will be calculated and compared between the different treatmentS. Screening Phase: 0168 Same as for Stage I. Study Population: Treatment Phase: 0150. Healthy: Male, healthy individuals. 0151 T2DM: (0169. 1. Blood samples will be drawn at -15, 0, 15, 30, 45, 0152 The presence of T2DM is determined by WHO cri 60, 75,90, 105, 120, 135, 150, and 240 min, where time=0 is teria. Subjects may be of either gender, between the ages of 18 the time at which the capsule is ingested, and tested for and 60, with stable glycemic control, optionally on oral glucose, insulin, c-peptide, as well as GLP-1 analogue levels antidiabetes medication (e.g. metformin and/or TZDS), and (C.T. and/or AUC) not pregnant or breastfeeding. Subjects will continue their 2. A standard meal will be served 60 minutes after oral dosing regular medications and dosing regimen up to the night prior or 30 minutes after subcutaneous administration of Byetta.R. to the study. Patients will resume their regular medication 3. Gastrointestinal symptoms may be assessed. after completing each study session (same day). Other than 0170 Treatments (delivered in a randomized order): diabetes, Subjects should be in general good health, with 0171 a. 300 or 600 mcg oral exenatide, or placebo, or 5 stable liver enzyme levels (below 2x the upper normal range). mcg Subcutaneous exenatide US 2015/001 7238 A1 Jan. 15, 2015

(0172 b. 300 or 600mcg oral exenatide, or placebo, or 5 changes in several cardiovascular risk markers. Vasc mcg Subcutaneous exenatide Health Risk Manag. 2012: 8:621-9. (0173 c.300 or 600mcg oral exenatide, or placebo, or 5 0188 Eldor R. Kidron M, Arbit E. A single-bling, two mcg Subcutaneous exenatide period study to assess the safety and pharmacodynamics of 0.174 d. 16 mg oral insulin an orally delivered GLP-1 analog (exenatide) in healthy 0.175 e. 300 mcg oral exenatide--16 mg oral insulin. subjects. American Diabetes Association 70' Annual Sci entific Sessions, June. 25-29, 2010A, Orlando, Fla. Safety Evaluations: (0189 Eldor R, Kidron M, Arbit E. Open-label study to 0176 Adverse events will be collected throughout the assess the safety and pharmacodynamics of five oral insu study beginning from the time the Subject signs the lin formulations in healthy subjects. Diabetes Obes Metab. consent form until the end of study evaluations March 2010B; 12(3):219-223. 0177 Concomitant medications/therapies will be (0190. Eldor R, Kidron M, Greenberg-Shushlav Y. Arbit E. recorded throughout the duration of study, beginning Novel glucagon-like peptide-1 analog delivered orally from the time the Subject signs the informed consent. reduces postprandial glucose excursions in porcine and 0.178 Vital signs (blood pressure, heart rate) will be canine models. J Diabetes SciTechnol. 2010C:4(6): 1516 measured in the sitting position after at least 5 minutes of 1523. rest at the following times: (0179 Screening (0191 Kidron M, Dinh S. Menachem Y, et al. A novel 0180 Approximately 20 minutes prior to study drug per-oral insulin formulation: proof of concept study in administration non-diabetic subjects. Diabet Med. April 2004; 21 (4): 354 0181. Approximately 1 and 2.5 hours post study drug 357. administration 0.192 Martinez-Colubi Metal, Switching to darunavir/ 0182 Clinical laboratory evaluations (chemistry, ritonavir monotherapy (DRV/r mx): effect on kidney func hematology) will be performed at Screening and at end tion and lipid profile. J Int AIDS Soc. 2012 Nov. 11; 15(6): of study or early discontinuation. 18348. doi:10.7448/IAS.15.6.18348. 0183 Physical examinations at screening and end of 0193 Miyashita Tetal, Hepatoprotective effect of tamox study or early discontinuation. ifen on Steatosis and non-alcoholic steatohepatitis in 0.184 Electrocardiograms (ECG) at screening. An ECG mouse models. J Toxicol Sci. 2012:37(5):931-42 will be performed at end of study or early discontinua (0194 Ryan EA, Shandro T. Green Ketal. Assessment of tion, only if the investigator deems it necessary. the severity of hypoglycemia and glycemic lability in type 1 diabetic Subjects undergoing islet transplantation. Dia Example 5 betes. 2004 April; 53(4):955-62. Testing of an Oral Insulin/Oral Exenatide 0.195 Siepmann F. Siepmann J et al. Blends of aqueous Combination in Treatment of Unstable Diabetes polymer dispersions used for pellet coating: importance of the particle size. J Control Release 2005: 105(3): 226-39. 0185. Subjects with elevated fasting glucose levels (for example, Subjects having a glycated hemoglobin HgA1c (0196) Sun.J., Rose J. B., Bird P. (1995).J. Biol. Chem. 270, level of 8-10%, or a fasting plasma sugar level from 100 to 16089-16096. 125 mg/dL or 5.6 to 6.9 mmol/L).) are monitored for several 0.197 Nissan A. Ziv E. Kidron M, et al. Intestinal absorp days using a blinded continuous glucose monitor (CGM) to tion of low molecular weight heparin in animals and human establish a baseline. During several Subsequent days, they are subjects. Haemostasis. September-October 2000; 30(5): administered a formulation described herein, optionally prior 225-232. to meals. Blinded CGM is performed to determine the effi (0198 Sprecher C A, Morgenstern K A, Mathewes S, cacy of the formulations. Dahlen J. R. Schrader S K, Foster DC, Kisiel W. J Biol. 0186. In the claims, the word “comprise', and variations Chem. 1995 Dec. 15: 270(50):29854-61. thereof such as “comprises”, “comprising, and the like indi (0199 Tesauro et al. Effects of GLP-1 on Forearm Vasodi cate that the components listed are included, but not generally lator Function and Glucose Disposal During Hyperin to the exclusion of other components. sulinemia in the Metabolic Syndrome. Diabetes Care. 2012 Oct. 15. REFERENCES 0200. Ziv E. Kidron M, Raz I, et al. Oral administration of 0187 Chiquette E et al. Treatment with exenatide once insulin in Solid form to nondiabetic and diabetic dogs. J weekly or twice daily for 30 weeks is associated with Pharm Sci. June 1994; 83(6):792-794.

SEQUENCE LISTING

<16 Os NUMBER OF SEO ID NOS: 4

<21 Os SEQ ID NO 1 &211s LENGTH: 1.OO 212s. TYPE: PRT &213s ORGANISM: Bos taurus

<4 OOs SEQUENCE: 1 US 2015/001 7238 A1 Jan. 15, 2015 16

- Continued

Met Lys Met Ser Arg Lieu. Cys Lieu. Ser Val Ala Lieu. Lieu Val Lieu. Lieu.

Gly Thr Lieu Ala Ala Ser Thr Pro Gly Cys Asp Thr Ser Asn Glin Ala 2O 25 3O Lys Ala Glin Arg Pro Asp Phe Cys Lieu. Glu Pro Pro Tyr Thr Gly Pro 35 4 O 45 Cys Lys Ala Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Lieu. SO 55 6 O Cys Glin Thr Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn. Asn. Phe 65 70 7s 8O Llys Ser Ala Glu Asp Cys Met Arg Thr Cys Gly Gly Ala Ile Gly Pro 85 90 95 Trp Glu Asn Lieu. 1OO

<210s, SEQ ID NO 2 &211s LENGTH: 110 212. TYPE: PRT <213> ORGANISM: Glycine max

<4 OOs, SEQUENCE: 2 Met Val Val Lieu Lys Val Cys Lieu Val Lieu. Lieu. Phe Lieu Val Gly Gly 1. 5 1O 15 Thir Thir Ser Ala Asn Lieu. Arg Lieu Ser Llys Lieu. Gly Lieu Lieu Met Lys 2O 25 3O Ser Asp His Gln His Ser Asn Asp Asp Glu Ser Ser Llys Pro Cys Cys 35 4 O 45 Asp Glin Cys Ala Cys Thr Lys Ser Asn Pro Pro Glin Cys Arg Cys Ser SO 55 6 O Asp Met Arg Lieu. Asn. Ser Cys His Ser Ala Cys Llys Ser Cys Ile Cys 65 70 7s 8O Ala Leu Ser Tyr Pro Ala Glin Cys Phe Cys Val Asp Ile Thr Asp Phe 85 90 95 Cys Tyr Glu Pro Cys Llys Pro Ser Glu Asp Asp Llys Glu Asn 1OO 105 11 O

<210s, SEQ ID NO 3 &211s LENGTH: 216 212. TYPE: PRT <213> ORGANISM: Glycine max <4 OOs, SEQUENCE: 3 Met Lys Ser Thr Ile Phe Phe Leu Phe Leu Phe Cys Ala Phe Thr Thr 1. 5 1O 15

Ser Tyr Lieu Pro Ser Ala Ile Ala Asp Phe Val Lieu. Asp Asn. Glu Gly 2O 25 3O

Asn Pro Leu Glu Asn Gly Gly Thr Tyr Tyr Ile Leu Ser Asp Ile Thr 35 4 O 45

Ala Phe Gly Gly Ile Arg Ala Ala Pro Thr Gly Asn. Glu Arg Cys Pro SO 55 6 O

Lieu. Thr Val Val Glin Ser Arg Asn. Glu Lieu. Asp Llys Gly Ile Gly Thr 65 70 7s 8O

Ile Ile Ser Ser Pro Tyr Arg Ile Arg Phe Ile Ala Glu Gly His Pro 85 90 95 US 2015/001 7238 A1 Jan. 15, 2015

- Continued

Lieu. Ser Lieu Lys Phe Asp Ser Phe Ala Val Ile Met Lieu. CyS Val Gly 1OO 105 11 O Ile Pro Thr Glu Trp Ser Val Val Glu Asp Leu Pro Glu Gly Pro Ala 115 12 O 125 Val Lys Ile Gly Glu Asn Lys Asp Ala Met Asp Gly Trp Phe Arg Lieu 13 O 135 14 O Glu Arg Val Ser Asp Asp Glu Phe Asn. Asn Tyr Llys Lieu Val Phe Cys 145 150 155 160 Pro Glin Glin Ala Glu Asp Asp Llys Cys Gly Asp Ile Gly Ile Ser Ile 1.65 17O 17s Asp His Asp Asp Gly. Thir Arg Arg Lieu Val Val Ser Lys Asn Llys Pro 18O 185 19 O Lieu Val Val Glin Phe Glin Llys Lieu. Asp Llys Glu Ser Lieu Ala Lys Llys 195 2OO 2O5 Asn His Gly Lieu. Ser Arg Ser Glu 21 O 215

<210s, SEQ ID NO 4 &211s LENGTH: 39 212. TYPE: PRT <213> ORGANISM: Heloderma suspectum 22 Os. FEATURE: <221 > NAMEAKEY: misc feature <222s. LOCATION: (39) . . (39) <223 OTHER INFORMATION: residue is amidated on C-terminus

<4 OOs, SEQUENCE: 4 His Gly Glu Gly Thr Phe Thir Ser Asp Lieu. Ser Lys Gln Met Glu Glu 1. 5 1O 15 Glu Ala Val Arg Lieu. Phe Ile Glu Trp Lieu Lys Asn Gly Gly Pro Ser 2O 25 Ser Gly Ala Pro Pro Pro Ser 35

1. An oral pharmaceutical composition, comprising an oil 4. The oral pharmaceutical composition of claim 1, based liquid formulation, said oral pharmaceutical composi wherein said oil-based liquid formulation, or both said first tion comprising an insulin, a GLP-1 analogue, a trypsin oil-based liquid formulation and said second oil-based liquid inhibitor, and a chelator of divalent cations, wherein said formulation, further comprises a self-emulsifying compo oil-based liquid formulation is surrounded by a coating that nent. resists degradation in the stomach. 5. The oral pharmaceutical composition of claim 1, 2. An oral pharmaceutical composition, comprising a com wherein said insulin is present in an amount between 4-12 mg bination of i). a first oil-based liquid formulation, said first inclusive for an adult patent or a corresponding amount per oil-based liquid formulation comprising an insulin, a trypsin body weight for a pediatric patient. inhibitor, and a chelator of divalent cations; and ii). a second 6. The oral pharmaceutical composition of claim 5, oil-based liquid formulation, said second oil-based liquid wherein said GLP-1 analogue is present in an amount formulation comprising a GLP-1 analogue, a trypsin inhibi between 100-300 micrograms inclusive for an adult patent or tor, and a chelator of divalent cations, wherein each of said a corresponding amount per body weight for a pediatric first oil-based liquid formulation and said second oil-based patient. liquid formulation is Surrounded by a coating that resists 7. The oral pharmaceutical composition of claim 1, degradation in the stomach. wherein said GLP-1 analogue is present in an amount 3. The oral pharmaceutical composition of claim 2, between 100-300 micrograms inclusive for an adult patent or wherein said oil-based liquid formulation, or both said first a corresponding amount per body weight for a pediatric oil-based liquid formulation and said second oil-based liquid patient. formulation, further comprises a component provided as a 8. The oral pharmaceutical composition of claim 1, mixture of (a) a monoacylglycerol, a diacylglycerol, a tria wherein said trypsin inhibitor is selected from the group cylglycerol, or a mixture thereof, and (b) a polyethylene consisting of soybean trypsin inhibitor (SBTI), Bowman glycol (PEG) ester of a fatty acid. Birk inhibitor (BBI), and aprotinin. US 2015/001 7238 A1 Jan. 15, 2015

9. The oral pharmaceutical composition of claim 1, further 19. A method for treating a human with Type 2 diabetes comprising a second trypsin inhibitor. mellitus, said method comprising the steps of selecting a 10. The oral pharmaceutical composition of claim 9. Subject diagnosed with Type 2 diabetes mellitus, and admin wherein said first and second trypsin inhibitors are (i) SBTI istering said subject oral pharmaceutical composition of and (ii) aprotinin; or said first and second trypsin inhibitors claim 1, thereby treating a human with Type 2 diabetes mel are (i) isolated KTI3 and (ii) isolated BBI. litus. 11. The oral pharmaceutical composition of claim 1, wherein said chelator is EDTA. 20. A method for treating a non-human animal with diabe 12. The oral pharmaceutical composition of claim 1, tes mellitus, said method comprising the step of administer wherein said oil is fish oil. ing to said non-human animal the oral pharmaceutical com 13. The oral pharmaceutical composition of claim 1, position of claim 1, thereby treating a non-human animal with wherein said oil-based liquid formulation is water-free. diabetes mellitus. 14. The oral pharmaceutical composition of claim 1, 21. A method for reducing preprandial glucose excursion wherein said GLP-1 analogue is exenatide. in a human with Type 2 diabetes mellitus, said method com 15. The oral pharmaceutical composition of claim 1, prising the steps of selecting a subject diagnosed with Type 2 wherein said coating is a pH-sensitive capsule. diabetes mellitus, and administering said subject oral phar 16. The oral pharmaceutical composition of claim 1 for maceutical composition of claim 1, thereby reducing reducing preprandial glucose excursion in a having human preprandial glucose excursion in a human with Type 2 diabe Type 2 diabetes mellitus. tes mellitus. 17. The oral pharmaceutical composition of claim 16, wherein said insulin is present in an amount between 4-12 mg 22. A method for reducing preprandial glucose excursion inclusive, said GLP-1 analogue is exenatide, and said in a non-human animal with diabetes mellitus, said method exenatide is present in an amount between 100-300 micro comprising the step of administering to said non-human ani grams inclusive. mal the oral pharmaceutical composition of claim 1, thereby 18. Use of the oral pharmaceutical composition of claim 1 reducing preprandial glucose excursion in a non-human ani for reducing preprandial glucose excursionina having human mal with diabetes mellitus. Type 2 diabetes mellitus. k k k k k