US 2011 O1428.00A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0142800 A1 Kidron et al. (43) Pub. Date: Jun. 16, 2011

(54) METHODS AND COMPOSITIONS FOR ORAL Publication Classification ADMINISTRATION OF PROTEINS (51) Int. Cl. A638/2 (2006.01) (75) Inventors: Miriam Kidron, Jerusalem (IL); A638/02 (2006.01) Ehud Arbit, Englewood, NJ (US) A638/28 (2006.01) A638/26 (2006.01) (73) Assignee: Oramed Ltd., Jerusalem (IL) A638/27 (2006.01) Appl. No.: 13/058,259 A638/18 (2006.01) (21) A638/43 (2006.01) (22) PCT Fled: Aug. 11, 2009 A6IP3/10 (2006.01) (52) U.S. Cl...... 424/85.5; 514/1.1 : 514/6.5: 514/11.7: (86) PCT NO.: PCT/IL2O09/OOO786 424/85.7; 514/11.3; 514/7.7: 514/7.6: 424/94.1 S371 (c)(1), (57) ABSTRACT (2), (4) Date: Feb. 9, 2011 This invention provides compositions comprising a protein, an absorption enhancer, a protease inhibitor, methods for Related U.S. Application Data treating diabetes mellitus, comprising administering same, (60) Provisional application No. 61/089,812, filed on Aug. and methods for oral administration of a protein with an 18, 2008. enzymatic activity, comprising orally administering same. Patent Application Publication Jun. 16, 2011 US 2011/0142800 A1

Figure 1 Average reduction in blood glucose levels 100

-- Glucose 1 i 50 -- Glucose 2 3:S 40 - -T-T O 20 40 60 8O 100 12O Time (min.)

Figure 2 — Average reduction in blood C-peptide

--Glucose 1 --Glucose 2 -A-Glucose 3 -ee-Glucose 4 O 1 O 20 30 40 50 60 7O 8O 90 100 110 120 Time (min.) US 2011/0142800 A1 Jun. 16, 2011

METHODS AND COMPOSITIONS FOR ORAL absorption enhancer, wherein an absorption enhancer ADMINISTRATION OF PROTEINS enhances the absorption of a protein through an intestinal mucosal barrier. FIELD OF INVENTION 0010. In another embodiment, the present invention pro vides a method for treating diabetes mellitus in a Subject, 0001. This invention provides oral compositions compris comprising administering orally to a Subject a pharmaceutical ing a protein, an absorption enhancer, and a protease inhibitor composition comprising insulin, Exenatide, or a combination and a method for administering same. thereof a protease inhibitor, and a absorption enhancer, wherein an absorption enhancer enhances the absorption of a BACKGROUND OF THE INVENTION protein through an intestinal mucosalbarrier, thereby treating diabetes mellitus. 0002. Due to improved biotechnology, the accessibility of biologically active peptides to the pharmaceutical industry BRIEF DESCRIPTION OF THE DRAWINGS has increased considerably. However, a limiting factor in the development of peptide drugs is the relative ineffectiveness 0011 FIG. 1 is a graph showing the changes in blood when given perorally. Almost all peptide drugs are parenter glucose levels after treatment with the formulations of the ally administered, although parenterally administered pep invention. tide drugs are often connected with low patient compliance. 0012 FIG. 2 is a graph showing the changes in blood 0003 Insulin is a medicament used to treat patients suf C-peptide levels after treatment with the formulations of the fering from diabetes, and is the only treatment for insulin invention. dependent diabetes mellitus. Diabetes Mellitus is character ized by a pathological condition of absolute or relative insulin DETAILED DESCRIPTION OF THE INVENTION deficiency, leading to hyperglycemia, and is one of the main 0013 This invention provides compositions and methods threats to human health in the 21st century. The global figure comprising a protein, a protease inhibitor, and a absorption of people with diabetes is set to rise to 220 million in 2010, enhancer. In another embodiment, the present invention pro and 300 million in 2025. Type I diabetes is caused primarily vides compositions and methods comprising a protein having by the failure of the pancreas to produce insulin. Type II a molecular weight of up to 100,000 Daltons, a protease diabetes, involves a lack of responsiveness of the body to the inhibitor, and aabsorption enhancer. In another, embodiment, action of insulin. the absorption enhancer enhances the absorption of the pro 0004. Approximately 20%-30% of all diabetics use daily tein through an intestinal mucosalbarrier. In another, embodi insulin injections to maintain their glucose levels. An esti ment, the absorption enhancer enhances the absorption of a mated 10% of all diabetics are totally dependent on insulin protein having a molecular weight of up to 100,000 Daltons injections. through an intestinal mucosal barrier. 0005. Currently, the only route of insulin administration is 0014. In another embodiment, the compositions of the injection. Daily injection of insulin is causes considerable present invention include an active agent, a protease inhibitor, suffering for patients. Side effects such as lipodystrophy at and a carrier. In another embodiment, the compositions of the the site of the injection, lipatrophy, lilpohypertrophy, and present invention are used to deliver various proteins through occasional hypoglycemia are known to occur. In addition, various biological, chemical, and physical barriers and are Subcutaneous administration of insulin does not typically particularly suited for delivering proteins which are subject to provide the fine continuous regulation of metabolism that environmental degradation. In another embodiment, the com occurs normally with insulin secreted from the pancreas positions of the present invention are used to deliver active directly into the liver via the portal vein. proteins through various biological, chemical, and physical 0006. The present invention addresses the need for an barriers and are particularly Suited for delivering active pro alternate solution for administration of insulin and peptides teins which are Subject to environmental degradation. In Such as insulin. another embodiment, the compositions comprise an active protein, SNAC, a protease inhibitor, and EDTA or Na-EDTA. SUMMARY OF THE INVENTION In another embodiment, the compositions comprise an active protein, SNAC, a protease inhibitor, and an omega-3 fatty 0007. This invention provides, in one embodiment, a com acid. In another embodiment, the compositions comprise an position comprising a protein having a molecular weight of active protein, SNAC, a protease inhibitor, EDTA or Na up to 100,000 Daltons, a protease inhibitor, and an absorption EDTA, and an omega-3 fatty acid. In another embodiment, enhancer, wherein the absorption enhancer enhances the the composition as described herein is an oral pharmaceutical absorption of a protein through an intestinal mucosal barrier. composition. In another embodiment, the composition as 0008. In another embodiment, the present invention pro described herein is an encapsulated oral pharmaceutical com vides a composition comprising a protein having a molecular position. In another embodiment, the composition as weight of up to 100,000 Daltons, a protease inhibitor, and described herein is in a liquid oral dosage form. In another SNAC or SNAD. embodiment, the composition as described herein is in a dry 0009. In another embodiment, the present invention pro oral dosage form (tablet etc.). vides a method for oral administration of a protein having a 0015. In another, embodiment, the protein of the present molecular weight up to 100,000 Daltons to a subject, whereby invention has a molecular weight of 1,000-5,000 Daltons. In a Substantial fraction of said protein retains its activity after another, embodiment, the protein of the present invention has absorption, through an intestinal mucosalbarrier of a subject, a molecular weight of 5,000-10,000 Daltons. In another, comprising administering orally to a Subject a pharmaceutical embodiment, the protein of the present invention has a composition comprising a protein, a protease inhibitor, and an molecular weight of 10,000-20,000 Daltons. In another, US 2011/0142800 A1 Jun. 16, 2011

embodiment, the protein of the present invention has a In another embodiment, the insulin is whale insulin. In molecular weight of 20,000-30,000 Daltons. In another, another embodiment, the insulin is a metal complex of insulin embodiment, the protein of the present invention has a (e.g. a Zinc complex of insulin, protamine Zinc insulin, or molecular weight of 40,000-50,000 Daltons. In another, globin Zinc). embodiment, the protein of the present invention has a 0019. In another embodiment, the insulin is endogenous molecular weight of 50,000-60,000 Daltons. In another, insulin. In another embodiment, the insulin is fast-acting embodiment, the protein of the present invention has a insulin. In another embodiment, the insulin is lente insulin. In molecular weight of 60,000-70,000 Daltons. In another, another embodiment, the insulin is semilente insulin. In embodiment, the protein of the present invention has a another embodiment, the insulin is ultralente insulin. In molecular weight of 70,000-80,000 Daltons. In another, another embodiment, the insulin is NPH insulin. In another embodiment, the protein of the present invention has a embodiment, the insulin is glargine insulin. In another molecular weight of 80,000-90,000 Daltons. In another, embodiment, the insulin is lispro insulin. In another embodi embodiment, the protein of the present invention has a ment, the insulin is aspart insulin. In another embodiment, the molecular weight of 90,000-100,000 Daltons. In another, insulin is a combination of two or more of any of the above embodiment, the protein of the present invention has a types of insulin. In another embodiment, the insulin is any molecular weight of 100,000-150,000 Daltons. other type of insulin known in the art. Each possibility rep 0016. In another embodiment, the protein has a molecular resents a separate embodiment of the present invention. weight (MW) of 1-50 kilodalton (kDa). In another embodi 0020. In one embodiment, the amount of insulinutilized in ment, the MW is 1-45 kDa. In another embodiment, the MW the methods and compositions of the present invention is is 1-40 kDa. In another embodiment, the MW is 1-35kDa. In 0.5-3 units (u)/kg in humans. In one embodiment, the units another embodiment, the MW is 1-30 kDa. In another used to measure insulin in methods and compositions of the embodiment, the MW is 1-25 kDa. In another embodiment, present invention are USP Insulin Units. In one embodiment, the MW is 1-20 kDa. In another embodiment, the MW is the units used to measure insulin are milligrams. In another 10-50 kDa. In another embodiment, the MW is 15-50kDa. In embodiment, one international Unit (IU) of Insulin is equiva another embodiment, the MW is 20-50 kDa. In another lent to 45.5 mg insulin. embodiment, the MW is 25-50 kDa. In another embodiment, 0021. In another embodiment, the amount of insulin is the MW is 30-50 kDa. In another embodiment, the MW is 0.1-1 u/kg. In another embodiment, the amount is 0.2-1 u/kg. 35-50kDa. In another embodiment, the MW is 1-100kDa. In In another embodiment, the amount is 0.3-1 u/kg. In another another embodiment, the MW is 1-90 kDa. In another embodiment, the amount is 0.5-1 u/kg. In another embodi embodiment, the MW is 1-80 kDa. In another embodiment, ment, the amount is 0.1-2 u?kg. In another embodiment, the the MW is 1-70kDa. In another embodiment, the MW is 1-60 amount is 0.2-2 u?kg. In another embodiment, the amount is kDa. In another embodiment, the MW is 10-100 kDa. In 0.3-2 u/kg. In another embodiment, the amount is 0.5-2 u/kg. another embodiment, the MW is 15-100 kDa. In another In another embodiment, the amount is 0.7-2 u/kg. In another embodiment, the MW is 20-100kDa. In another embodiment, embodiment, the amount is 1-2 u?kg. the MW is 25-100 kDa. In another embodiment, the MW is 0022. In another embodiment, the amount is 1.2-2 u/kg. In 30-100 kDa. In another embodiment, the MW is 10-80 kDa. another embodiment, the amount is 1-1.2 u/kg. In another In another embodiment, the MW is 15-80 kDa. In another embodiment, the amount is 1-1.5 u?kg. In another embodi embodiment, the MW is 20-80 kDa. In another embodiment, ment, the amount is 1-2.5 u?kg. In another embodiment, the the MW is 25-80 kDa. In another embodiment, the MW is amount is 1-3 u?kg. In another embodiment, the amount is 2-3 30-80 kDa. Each possibility represents a separate embodi u/kg. In another embodiment, the amount is 1-5 u/kg. In ment of the present invention. another embodiment, the amount is 2-5 u/kg. In another 0017. In another embodiment, the MW is less than 20 kDa. embodiment, the amount is 3-5 u/kg. In another embodiment, the MW is less than 25 kDa. In 0023. In another embodiment, the amount of insulin is 0.1 another embodiment, the MW is less than 30 kDa. In another u/kg. In another embodiment, the amount is 0.2 u/kg. In embodiment, the MW is less than 35kDa. In another embodi another embodiment, the amount is 0.3 u?kg. In another ment, the MW is less than 40 kDa. In another embodiment, embodiment, the amount is 0.4 u/kg. In another embodiment, the MW is less than 45kDa. In another embodiment, the MW the amount is 0.5u?kg. In another embodiment, the amount is is less than 50 kDa. In another embodiment, the MW is less 0.6 u/kg. In another embodiment, the amount is 0.8 u/kg. In than 55 kDa. In another embodiment, the MW is less than 60 another embodiment, the amount is 1 u?kg. In another kDa. In another embodiment, the MW is less than 65 kDa. In embodiment, the amount is 1.2 u/kg. In another embodiment, another embodiment, the MW is less than 70 kDa. In another the amount is 1.4 u/kg. In another embodiment, the amount is embodiment, the MW is less than 75 kDa. In another embodi 1.6 u/kg. In another embodiment, the amount is 1.8 u?kg. In ment, the MW is less than 80 kDa. In another embodiment, another embodiment, the amount is 2 u?kg. In another the MW is less than 85 kDa. In another embodiment, the MW embodiment, the amount is 2.2 u/kg. In another embodiment, is less than 90 kDa. In another embodiment, the MW is less the amount is 2.5u?kg. In another embodiment, the amount is than 95 kDa. In another embodiment, the MW is less than 100 3 u?kg. kDa. 0024. In another embodiment, the amount of insulin is 0018. In another, embodiment, the protein of the present 1-10 u. In another embodiment, the amount is 2-10 u. In invention is insulin. In another embodiment, the insulin of another embodiment, the amount is 3-10 u. In another methods and compositions of the present invention is human embodiment, the amount is 5-10 u. In another embodiment, insulin. In another embodiment, the insulin is recombinant the amount is 1-20 u. In another embodiment, the amount is insulin. In another embodiment, the insulin is recombinant 2-20 u. In another embodiment, the amount is 3-20 u. In human insulin. In another embodiment, the insulin is bovine another embodiment, the amount is 5-20 u. In another insulin. In another embodiment, the insulin is porcine insulin. embodiment, the amount is 7-20 u. In another embodiment, US 2011/0142800 A1 Jun. 16, 2011

the amount is 10-20 u. In another embodiment, the amount is ment, the amount of Exenatide in a formulation as described 12-20 u. In another embodiment, the amount is 10-12 u. In herein is 50 mcg to 60 mcg. In another embodiment, the another embodiment, the amount is 10-15 u. In another amount of Exenatide in a formulation as described herein is embodiment, the amount is 10-25 u. In another embodiment, 60 mcg to 70 mcg. In another embodiment, the amount of the amount is 10-30 u. In another embodiment, the amount is Exenatide in a formulation as described herein is 70mcg to 80 20-30 u. In another embodiment, the amount is 10-50 u. In mcg. In another embodiment, the amount of Exenatide in a another embodiment, the amount is 20-50 u. In another formulation as described herein is 80 mcg to 90 mcg. In embodiment, the amount is 30-50 u. In another embodiment, another embodiment, the amount of Exenatide in a formula the amount is 20-100 u. In another embodiment, the amount tion as described herein is 90 mcg to 100 mcg. In another is 30-100u. In another embodiment, the amount is 100-150 u. embodiment, the amount of Exenatide in a formulation as In another embodiment, the amount is 100-250 u. In another described herein is 100 mcg to 110 mcg. In another embodi embodiment, the amount is 100-300 u. In another embodi ment, the amount of Exenatide in a formulation as described ment, the amount is 200-300 u. In another embodiment, the herein is 110 mcg to 125 mcg. In another embodiment, the amount is 100-500 u. In another embodiment, the amount is amount of Exenatide in a formulation as described herein is 200-500 u. In another embodiment, the amount is 300-500 u. 125 mcg to 150 mcg. In another embodiment, the amount of In another embodiment, the amount is 200-1000 u. In another Exenatide in a formulation as described herein is 150 mcg to embodiment, the amount is 300-1000 u. 175 meg. In another embodiment, the amount of Exenatide in 0.025 Inanother embodiment, the amount of insulin is 1 u. a formulation as described herein is 175 mcg to 200 mcg. In In another embodiment, the amount is 2 u. In another embodi another embodiment, the amount of Exenatide in a formula ment, the amount is 3 u. In another embodiment, the amount tion as described herein is 200 mcg to220 mcg. In another is 4 u. In another embodiment, the amount is 5 u. In another embodiment, the amount of Exenatide in a formulation as embodiment, the amount is 6 u. In another embodiment, the described herein is 220 mcg to 240 mcg. In another embodi amount is 8 u. In another embodiment, the amount is 10 u. In ment, the amount of Exenatide in a formulation as described another embodiment, the amount is 12 u. In another embodi herein is 240 mcg to 260 mcg. In another embodiment, the ment, the amount is 14 u. In another embodiment, the amount amount of Exenatide in a formulation as described herein is is 16 u. In another embodiment, the amount is 18 u. In another 260 mcg to 300 mcg. embodiment, the amount is 20 u. In another embodiment, the 0029. In another embodiment, the amount of Exenatide in amount is 22 u. In another embodiment, the amount is 25u. In a formulation as described herein is 300 mcg to 350 mcg. In another embodiment, the amount is 30 u. In another embodi another embodiment, the amount of Exenatide in a formula ment, the amount is 50 u. In another embodiment, the amount tion as described herein is 350 mcg to 400 mcg. In another is 80 u. In another embodiment, the amount is 100 u. In embodiment, the amount of Exenatide in a formulation as another embodiment, the amount is 120 u. In another embodi described herein is 400 mcg to 450 mcg. In another embodi ment, the amount is 140 u. In another embodiment, the ment, the amount of Exenatide in a formulation as described amount is 160 u. In another embodiment, the amount is 180 u. herein is 450 mcg to 500 mcg. In another embodiment, the In another embodiment, the amount is 200 u. In another amount of Exenatide in a formulation as described herein is embodiment, the amount is 300 u. In another embodiment, 550 mcg to 600 mcg. In another embodiment, the amount of the amount is 500 u. Exenatide in a formulation as described herein is 600 mcg to 0026. In another embodiment, the protein is an insulin 700 mcg. In another embodiment, the amount of Exenatide in secretagogue. In another embodiment, the protein is GLP-1. a formulation as described herein is 700 mcg to 800 mcg. In In another embodiment, the protein is a GLP-1 analogue. In another embodiment, the amount of Exenatide in a formula another embodiment, the protein is a GLP-1 mimetic. In tion as described herein is 800 mcg to 900 mcg. In another another embodiment, the protein is an incretin mimetic. In embodiment, the amount of Exenatide in a formulation as another embodiment, the protein mimics the GLP-1 incretin. described herein is 900 mcg to 1 mg. In another embodiment, the protein is GLP-2. In another 0030. In another embodiment, the Exenatide formulation embodiment, the protein is a GLP-2 analogue. In another as described herein is taken twice a week. In another embodi embodiment, the protein is a GLP-2 mimetic. ment, the Exenatide formulation as described herein is taken 0027. In another embodiment, the protein is Exenatide. In once every two days. In another embodiment, the Exenatide another embodiment, the oral formulations of the present formulation as described hereinistaken once a day. In another invention protect Exenatide breakdown in the stomach. In embodiment, the Exenatide formulation as described herein another embodiment, Exenatide formulation of the invention is taken twice a day. In another embodiment, the Exenatide controls blood Sugar levels. In another embodiment, formulation as described herein is taken three times a day. In Exenatide formulation of the invention helps control blood another embodiment, the Exenatide formulation as described Sugar levels. In another embodiment, EXenatide formulation herein is taken four times a day. In another embodiment, the of the invention induces pancreatic production of insulin. In Exenatide formulation as described herein is taken five times another embodiment, Exenatide formulation of the invention a day. In another embodiment, one of skill in the art deter is used to treat type-2 (non-insulin dependent) diabetes. In mines the dosage of a Exenatide formulation as described another embodiment, Exenatide formulation of the invention herein. In another embodiment, one of skill in the art deter is used in conjunction with other diabetes medicines. mines the daily dose of a Exenatide formulation as described 0028. In another embodiment, the amount of Exenatide in herein. In another embodiment, one of skill in the art deter a formulation as described herein is 10 mcg to 1 mg. In mines the daily dosing regimen of a Exenatide formulation as another embodiment, the amount of Exenatide in a formula described herein. tion as described herein is 10 mcg to 25 mcg. In another 0031. In another embodiment, the Exenatide formulation embodiment, the amount of Exenatide in a formulation as as described herein is taken at least 15 minutes before a meal. described herein is 25 mcg to 50 mcg. In another embodi In another embodiment, the Exenatide formulation as US 2011/0142800 A1 Jun. 16, 2011

described herein is taken at least 30 minutes before a meal. In tion of molecular oxygen, on paired donors, on Superoxide as another embodiment, the Exenatide formulation as described acceptor, oxidizing metal ions, on CH or CH(2) groups, on herein is taken at least 45 minutes before a meal. In another iron-sulfur proteins as donors, on reduced flavodoxin as embodiment, the Exenatide formulation as described herein donor, on phosphorus or arsenic in donors, or on X-Handy-H is taken at least 60 minutes before a meal. In another embodi to form an X-y bond. ment, the Exenatide formulation as described herein is taken 0038. In some embodiments, transferases are acyltrans at least 75 minutes before a meal. In another embodiment, the ferases or glycosyltransferases. In some embodiments, trans Exenatide formulation as described herein is taken at least 90 ferases transferaldehyde or ketone residues. In some embodi minutes before a meal. In another embodiment, the Exenatide ments, transferases transfer alkyl or aryl groups, other than formulation as described herein is taken at least 100 minutes methyl groups. In some embodiments, transferases transfer before a meal. In another embodiment, the Exenatide formu nitrogenous, phosphorous, Sulfur or selenium containing lation as described herein is taken at least 120 minutes before groups. a meal. In another embodiment, the Exenatide formulation as 0039. In some embodiments, hydrolases are glycosylases described herein is taken at least 150 minutes before a meal. or act on ether bonds, on peptide bonds, on carbon-nitrogen In another embodiment, the Exenatide formulation as bonds, other than peptide bonds, on acid anhydrides, on car described herein is taken at least 180 minutes before a meal. bon-carbon bonds, on halide bonds, on phosphorus-nitrogen 0032. In another embodiment, the Exenatide formulation bonds, on Sulfur-nitrogen bonds, on carbon-phosphorus as described herein reduces the side effects associated with an bonds, on sulfur-sulfur bonds, or on carbon-sulfur bonds. injectable dosage form comprising Exenatide. In another 0040. In some embodiments, lyases are carbon-carbon embodiment, the Exenatide formulation as described herein lyases, carbon-oxygen lyases, carbon-nitrogen lyases, car reduces nausea as a side effect which is associated with an bon-sulfur lyases, carbon-halide lyases, phosphorus-oxygen injectable dosage form comprising Exenatide. In another lyases, or other lyases. embodiment, the Exenatide formulation as described herein 0041. In some embodiments, isomerases are racemases or does not induce nausea as a side effect which is associated epimerases, cis-trans-isomerases, intramolecular oxi with an injectable dosage form comprising Exenatide. doreductases, intramolecular transferases, intramolecular 0033. In another embodiment, the protein is glucagon. In lyases, or other isomerases. another embodiment, the protein is interferon gamma. In another embodiment, the protein is interferon alpha. In 0042. In some embodiments, ligases form carbon-sulfur another embodiment, the protein is a growth hormone. In bonds, carbon-nitrogen bonds, carbon-carbon bonds, phos another embodiment, the protein is erythropoietin. In another phoric ester bonds, or nitrogen-metal bonds. embodiment, the protein is granulocyte colony stimulating 0043. In some embodiments, transporter proteins are factor (G-CSF). In another embodiment, the protein is omen annexins, ATP-binding cassette transporters, hemoglobin, tin. In another embodiment, the protein is calcitonin. In ATPases, calcium channels, potassium channels, sodium another embodiment, the protein is PTH. In another embodi channels, or solute carriers. ment, the protein is PYY. 0044. In some embodiments, storage proteins comprise 0034. In another embodiment, the protein is any other albumins, lactoglobulins, casein ovomucin, ferritin, phos protein known in the art. In another embodiment, the protein vitin, lactoferrin, or Vitellogenin. In one embodiment, albu is a growth hormone. In one embodiment, the growth hor mins comprise avidin, ovalbumin, serum albumin, parvalbu mone is somatotropin. In another embodiment, the growth min, c-reactive proteinprealbumin, conalbumin, ricin, hormone is Insulin Growth Factor-I (IGF-I). In another lactalbumin, methemalbumin, or transthyretin. embodiment, the growth hormone is any other growth hor 0045. In some embodiments, structural proteins comprise mone known in the art. amyloid, collagen elastin, or fibrillin. 0035. In another embodiment, methods and compositions 0046. In some embodiments, the protein is a viral protein, of the present invention are used to administer a human serum bacterial protein, invertebrate protein, or vertebrate protein. albumin. Human serum albumin is not, in one embodiment, In some embodiments, the protein is a recombinant protein. considered to be a pharmaceutically-active component; how In one embodiment, the protein is a recombinant protein. In ever, it can be used in the context of the present invention as one embodiment, the recombinant protein is a recombinant a therapeutically-beneficial carrier for an active component. human protein. Each type of protein represents a separate embodiment of the 0047. The molecular weights of some of the proteins men present invention. tioned above are as follows: insulin—6 kilodalton (kDa); 0036. In one embodiment, the protein is an enzyme. In glucagon—3.5 kDa, interferon, 28 kDa, growth hormone— Some embodiments, the protein is a receptor ligand, trans 21.5-47 kDa; human serum albumin 69 kDa; erythropoi porter, or a storage protein. In one embodiment, the protein is etin 34 kDa; G-CSF 30-34 kDa. Thus, in one embodi a structural protein. ment, the molecular weight of these proteins is appropriate 0037. In some embodiments, the enzyme is an oxi for administration by methods of the present invention. doreductase, transferase, hydrolase, lyase, isomerase, or 0048. As provided herein, a protease inhibitor protects the ligase. In some embodiments, oxidoreductases act on the protein of the present invention from cleavage. In another aldehyde or oxo group of donors, on the CH-CH group of embodiment, the present invention provides that a protease donors, on the CH-NH(2) group of donors, on the CH NH inhibitor protects insulin of the present invention from cleav group of donors, on NADH or NADPH, on the CH-OH age. In another, embodiment, the present invention provides group of donors, on nitrogenous compounds as donors, on a that a protease inhibitor facilitates the protein absorption in Sulfur group of donors, on a heme group of donors, on diphe the intestine of a subject. In another, embodiment, the present nols and related Substances as donors, on a peroxide as accep invention provides that a protease inhibitor facilitates the tor, on hydrogen as donor, on single donors with incorpora absorption of insulin in the intestine of a Subject. US 2011/0142800 A1 Jun. 16, 2011

0049. In another, embodiment, a serpin is: Alpha 1-antit phosphate), leupeptin, PEFABLOC(R) SC (4-(2-Aminoethyl)- rypsin, Antitrypsin-related protein, Alpha 1-antichymot benzenesulfonyl fluoride hydrochloride), PMSF (phenylm rypsin, Kallistatin, Protein C inhibitor, Cortisol binding ethyl sulfonyl fluoride), TLCK (1-Chloro-3-tosylamido-7- globulin, Thyroxine-binding globulin, Angiotensinogen, amino-2-heptanone HC1), TPCK (1-Chloro-3- to sylamido Centerin, Protein Z-related protease inhibitor, Vaspin, Mono 4-phenyl-2-butanone), pentamidine isethionate, pepstatin, cyte neutrophil elastase inhibitor, Plasminogen activator guanidium, alpha2-macroglobulin, a chelating agent of zinc, inhibitor-2, Squamous cell carcinoma antigen-1 (SCCA-1), or iodoacetate, zinc. Each possibility represents a separate Squamous cell carcinoma antigen-2 (SCCA-2), Maspin, embodiment of the present invention. PI-6, Megsin, PI-8, PI-9, Bomapin, Yukopin, Hurpin/Head 0056. In another embodiment, the amount of a protease pin, Antithrombin, Heparin cofactor II, Plasminogen activa inhibitor utilized in methods and compositions of the present tor inhibitor 1, Glia derived nexin/Protease nexin I, Pigment invention is 0.1 mg/dosage unit. In another embodiment, the epithelium derived factor, Alpha2-antiplasmin, Complement amount of a protease inhibitor is 0.2 mg/dosage unit. In 1-inhibitor, 47 kDa Heat shock protein (HSP47), Neuroser another embodiment, the amount is 0.3 mg/dosage unit. In pin, or Pancpin. another embodiment, the amount is 0.4 mg/dosage unit. In 0050. In another embodiment, the present invention pro another embodiment, the amount is 0.6 mg/dosage unit. In vides that the protease inhibitor is a trypsin inhibitor such as another embodiment, the amount is 0.8 mg/dosage unit. In but not limited to: Lima bean trypsin inhibitor, Aprotinin, Soy another embodiment, the amount is 1 mg/dosage unit. In bean trypsin inhibitor (SBTI), or Ovomucoid. another embodiment, the amount is 1.5 mg/dosage unit. In 0051. In another embodiment, the present invention pro another embodiment, the amount is 2 mg/dosage unit. In vides that the protease inhibitor is a Cysteine protease inhibi another embodiment, the amount is 2.5 mg/dosage unit. In tor. In another, embodiment, the present invention provides another embodiment, the amount is 3 mg/dosage unit. In that Cysteine protease inhibitors of the invention comprise: another embodiment, the amount is 5 mg/dosage unit. In cyStatin, type 1 cystatins (or Stefins), CyStatins of type 2.hu another embodiment, the amount is 7 mg/dosage unit. In man cystatins C, D, S, SN, and SA, cystatin E/M, cystatin F, another embodiment, the amount is 10 mg/dosage unit. In type 3 cystatins, or kininogens. another embodiment, the amount is 12 mg/dosage unit. In 0052. In another embodiment, the present invention pro another embodiment, the amount is 15 mg/dosage unit. In vides that the protease inhibitor is a Threonine protease another embodiment, the amount is 20 mg/dosage unit. In inhibitor. In another, embodiment, the present invention pro another embodiment, the amount is 30 mg/dosage unit. In vides that Threonine protease inhibitors of the invention com another embodiment, the amount is 50 mg/dosage unit. In prise: Bortezomib, MLN-519, ER-807446, TMC-95A. another embodiment, the amount is 70 mg/dosage unit. In 0053. In another embodiment, the present invention pro another embodiment, the amount is 100 mg/dosage unit. vides that the protease inhibitor is an Aspartic protease inhibi 0057. In another embodiment, the amount of a protease tor. In another, embodiment, the present invention provides inhibitor is 0.1-1 mg/dosage unit. In another embodiment, the that Aspartic protease inhibitors of the invention comprise: amount of a protease inhibitor is 0.2-1 mg/dosage unit. In C-Macroglobulin, Pepstatin A, Aspartic protease inhibitor another embodiment, the amount is 0.3-1 mg/dosage unit. In 11, Aspartic protease inhibitor 1, Aspartic protease inhibitor another embodiment, the amount is 0.5-1 mg/dosage unit. In 2, Aspartic protease inhibitor 3, Aspartic 20 protease inhibi another embodiment, the amount is 0.1-2 mg/dosage unit. In tor 4. Aspartic protease inhibitor 5. Aspartic protease inhibi another embodiment, the amount is 0.2-2 mg/dosage unit. In tor 6, Aspartic protease inhibitor 7. Aspartic protease inhibi another embodiment, the amount is 0.3-2 mg/dosage unit. In tor 8, Aspartic protease inhibitor 9, Pepsin inhibitor Dit33, another embodiment, the amount is 0.5-2 mg/dosage unit. In Aspartyl protease inhibitor, or Protease A inhibitor 3. another embodiment, the amount is 1-2 mg/dosage unit. In 0054. In another embodiment, the present invention pro another embodiment, the amount is 1-10 mg/dosage unit. In vides that the protease inhibitorisa Metalloprotease inhibitor. another embodiment, the amount is 2-10 mg/dosage unit. In In another, embodiment, the present invention provides that another embodiment, the amount is 3-10 mg/dosage unit. In Metalloprotease inhibitors of the invention comprise: Angio another embodiment, the amount is 5-10 mg/dosage unit. In tensin-1 -converting enzyme inhibitory peptide, Antihemor another embodiment, the amount is 1-20 mg/dosage unit. In ragic factor BJ46a, Beta-casein, Proteinase inhibitor CeKI, another embodiment, the amount is 2-20 mg/dosage unit. In Venom metalloproteinase inhibitor DM43, Carboxypepti another embodiment, the amount is 3-20 mg/dosage unit. In dase A inhibitor, Smpl, IMPI, Alkaline proteinase, inh, another embodiment, the amount is 5-20 mg/dosage unit. In Latexin, Carboxypeptidase inhibitor, Antihemorragic factor another embodiment, the amount is 10-20 mg/dosage unit. In HSF, Testican-3, SPOCK3, TIMP1, Metalloproteinase another embodiment, the amount is 10-100 mg/dosage unit. inhibitor 1, Metalloproteinase inhibitor 2, TIMP2, Metallo In another embodiment, the amount is 20-100 mg/dosage proteinase inhibitor 3, TIMP3, Metalloproteinase inhibitor 4. unit. In another embodiment, the amount is 30-100 mg/dos TIMP4, Putative metalloproteinase inhibitor tag-225, Tissue age unit. In another embodiment, the amount is 50-100 inhibitor of metalloprotease, WAP. kazal, immunoglobulin, mg/dosage unit. In another embodiment, the amount is or kunitz and NTR domain-containing protein 1. 10-200 mg/dosage unit. In another embodiment, the amount 0055. In some embodiments, a protease inhibitor is a sui is 20-200 mg/dosage unit. In another embodiment, the cide inhibitor, a transition state inhibitor, or a chelating agent. amount is 30-200 mg/dosage unit. In another embodiment, In some embodiments, the protease inhibitor of the present the amount is 50-200 mg/dosage unit. In another embodi invention is: AEBSF-HC1 (epsilon)-aminocaproic acid,(al ment, the amount is 100-200 mg/dosage unit. pha) 1-antichymotypsin, antipain, antithrombin III.(alpha) 0058. In another embodiment, the amount of a protease 1-antitrypsin (alpha 1-proteinase inhibitor). APMSF-HC1 inhibitor utilized in the methods and compositions of the (4-amidinophenyl-methane Sulfonyl-fluoride), sprotinin, present invention is 1000 ki.u. (kallikrein inactivator units)/ benzamidine-HC1.chymostatin, DFP (diisopropylfluoro dosage unit. In another embodiment, the amount is 10 k.i.u./ US 2011/0142800 A1 Jun. 16, 2011

dosage unit. In another embodiment, the amount is 12 k.i.u./ embodiment, the enhancer is n-dodecyl-B-D-maltopyrano dosage unit. In another embodiment, the amount is 15 ki.u./ side. In some embodiments, Surfactants serve as absorption dosage unit. In another embodiment, the amount is 20 k.i.u./ enhancer. In one embodiment, the enhancer is chitisan Such as dosage unit. In another embodiment, the amount is 30 ki.u./ N.N.N-trimethyl chitosan chloride (TMC). dosage unit. In another embodiment, the amount is 40 ki.u./ 0062. In one embodiment, NO donors of the present inven dosage unit. In another embodiment, the amount is 50 ki.u./ tion comprise 3-(2-Hydroxy-1-(1-methylethyl)-2-nitrosohy dosage unit. In another embodiment, the amount is 70 k.i.u./ drazino)-1-propanamine, N-ethyl-2-(1-ethyl-hydroxy-2-ni dosage unit. In another embodiment, the amount is 100 ki. troSohydrazino)-ethanamine, O S-Nitroso-N- u./dosage unit. In another embodiment, the amount is 150 acetylpenicillamine ki.u./dosage unit. In another embodiment, the amount is 200 0063. In another embodiment, the bile acid is cholic acid. ki.u./dosage unit. In another embodiment, the amount is 300 In another embodiment, the bile acid is chenodeoxycholic ki.u./dosage unit. In another embodiment, the amount is 500 acid. In another embodiment, the bile acid is taurocholic acid. k.i.u./dosage unit. In another embodiment, the amount is 700 In another embodiment, the bile acid is taurochenodeoxy ki.u./dosage unit. In another embodiment, the amount is cholic acid. In another embodiment, the bile acid is glyco 1500 ki.u./dosage unit. In another embodiment, the amount cholic acid. In another embodiment, the bile acid is glyco is 3000 ki.u./dosage unit. In another embodiment, the chenocholic acid. In another embodiment, the bile acid is 3 amount is 4000 ki.u./dosage unit. In another embodiment, beta-monohydroxychloric acid. In another embodiment, the the amount is 5000 k.i.u./dosage unit. Each amount of a first bile acid is lithocholic acid. In another embodiment, the bile or a second protease inhibitor represents a separate embodi acid is 5 beta-cholanic acid. In another embodiment, the bile ment of the present invention. acid is 3,12-diol-7-one-5 beta-cholanic acid. In another 0059. In another embodiment, the amount is 50-5000 ki. embodiment, the bile acid is 3 alpha-hydroxy-12-ketocholic u./dosage unit. In another embodiment, the amount is 500 acid. In another embodiment, the bile acid is 3 beta-hydroxy 4000 ki.u./dosage unit. In another embodiment, the amount 12-ketocholic acid. In another embodiment, the bile acid is 12 is 100-1000 ki.u./dosage unit. In another embodiment, the alpha-3 beta-dihydrocholic acid. In another embodiment, the amount is 200-1500 ki.u./dosage unit. bile acid is ursodesoxycholic acid. 0060. In another embodiment, compositions of the present 0064. In one embodiment, the enhancer is a nonionic sur invention comprise a Substance that enhances absorption of a factant. In one embodiment, the enhancer is a nonionic poly protein of the invention through an intestinal mucosal barrier. oxyethylene ether Surface active agent (e.g. one having an In another embodiment, compositions of the present inven HLB value of 6 to 19, wherein the average number of poly tion further comprise a Substance that enhances absorption of oxyethylene units is 4 to 30). In another embodiment, the insulin through an intestinal mucosal barrier. In another enhancer is an anionic Surface active agents. In another embodiment, compositions of the present invention further embodiment, the enhancer is a cationic Surface active agent. comprise a substance that enhances absorption of Exenatide In another embodiment, the enhancer is an ampholytic Sur through an intestinal mucosal barrier. In another embodi face active agent. In one embodiment, Zwitteruionic Surfac ment, compositions of the present invention further comprise tants such as acylcarnitines serve as absorption enhancers. a Substance that reduces the degradation of Exenatide in the 0065. In another embodiment, the absorption enhancer is digestive system.In another embodiment, compositions of the an effective oral absorption enhancer for macromolecular present invention further comprise a Substance that reduces drugs. In another embodiment, the absorption enhancer is the degradation of Exenatide in the stomach. In another very water soluble. In another embodiment, the absorption embodiment, compositions of the present invention further enhancer is fully, i. e. greater than 85%, absorbed by the comprise a Substance that reduces the degradation of gastro-intestinal tract. In another embodiment, the absorption Exenatide in the intestine. Such a substance is referred to enhancer is a coarse form. In another embodiment, the hereinas an "enhancer. As provided herein, enhancers, when absorption enhancer is micronized. In another embodiment, used together with omega-3 fatty acids or a protease inhibitor, the absorption enhancer is amorphous. In another embodi enhance the ability of a proitein to be absorbed in the intes ment, the absorption enhancer is N-(5-chlorosalicyloyl)-8- tine. As provided herein, enhancers, when used together with aminocaprylic acid (5-CNAC). In another embodiment, the omega-3 fatty acids and a protease inhibitor, enhance the absorption enhancer is N-(102-hydroxybenzoylamino) ability of insulin to be absorbed in the intestine. As provided decanoic acid (SNAD). In another embodiment, the absorp herein, enhancers, when used together with omega-3 fatty tion enhancer is N-(8- 2-hydroxybenzoylamino) caprylic acids and a protease inhibitor, enhance the ability of acid (SNAC). In another embodiment, the absorption Exenatide to be absorbed in the intestine. enhancer is CNAC, SNAD, SNAC, a monosodium and/or 0061. In one embodiment, the enhancer is dide disodium salts thereof, ethanol solvates of sodium salts canoylphosphatidylcholine (DDPC). In one embodiment, the thereof and the monohydrates of sodium salts thereof and any enhancer is a chelating agent such as ethylenediaminetet combinations thereof. In another embodiment, the absorption raacetic acid (EDTA) or egtazic acid EGTA. In another enhancer is 8-(N-2-hydroxy-4-methoxybenzoyl)-aminoca embodiment, EDTA is sodium-EDTA. In some embodi prylic acid (4-MOAC) and pharmaceutically acceptable salts ments, the enhancer is NO donor. In some embodiments, the thereof and/or amorphous and polymorphic forms of enhancer is abile acid, glycine-conjugated form of a bile acid, 4-MOAC. In another embodiment, the absorption enhancer is or an alkali metal salt. In one embodiment, absorption N-(8-2-hydroxy-5-chlorobenzoyl-amino)octanoic acid enhancement is achieved through utilization of a combination (also known as 8-(N-2-hydroxy-5-chlorobenzoyl)aminoca of a-galactosidase and B-mannanase. In some embodiments, prylic acid)) (5-CNAC) and pharmaceutically acceptable the enhancer is a fatty acid such as sodium caprate. In one salts thereof and/or amorphous and polymorphic forms of embodiment, the enhancer is sodium glycocholate. In one 5-CNAC. In another embodiment, the absorption enhancer is embodiment, the enhancer is sodium salicylate. In one 4-(2-hydroxy-4- chlorobenzoyl)aminobutanoate (also US 2011/0142800 A1 Jun. 16, 2011

known as 4-(4-chloro-2-hydroxy-benzoyl)aminobutanoic amount is 7 mg/dosage unit. In another embodiment, the acid) (4-CNAB) and pharmaceutically acceptable salts amount is 10 mg/dosage unit. In another embodiment, the thereof, including its monosodium salt and/or amorphousand amount is 12 mg/dosage unit. In another embodiment, the polymorphic forms of 4-CNAB. amount is 15 mg/dosage unit. In another embodiment, the 0066. In another embodiment, there is a synergistic effect amount is 20 mg/dosage unit. In another embodiment, the in lowering blood glucose level of SNAD and a protease amount is 30 mg/dosage unit. In another embodiment, the inhibitor of the invention. In another embodiment, there is a amount is 50 mg/dosage unit. In another embodiment, the synergistic effect in lowering blood glucose level of SNAC amount is 70 mg/dosage unit. In another embodiment, the and a protease inhibitor of the invention. amount is 100 mg/dosage unit. 0067. In another embodiment, the pharmaceutical compo (0071. In another embodiment, the amount of enhancer is sitions of the present invention comprise a delivery effective 0.1-1 mg/dosage unit. In another embodiment, the amount of amount of one or more of the absorption enhancers.In another enhancer is 0.2-1 mg/dosage unit. In another embodiment, the embodiment, the pharmaceutical compositions of the present amount is 0.3-1 mg/dosage unit. In another embodiment, the invention comprise an amount Sufficient to deliver the active amount is 0.5-1 mg/dosage unit. In another embodiment, the agent for the desired effect. amount is 0.1-2 mg/dosage unit. In another embodiment, the 0068. In another embodiment, the pharmaceutical compo amount is 0.2-2 mg/dosage unit. In another embodiment, the sitions of the present invention comprise an amount of 2.5% amount is 0.3-2 mg/dosage unit. In another embodiment, the to 99.4% by weight of an absorption enhancer. In another amount is 0.5-2 mg/dosage unit. In another embodiment, the embodiment, the pharmaceutical compositions of the present amount is 1-2 mg/dosage unit. In another embodiment, the invention comprise an amount of 2.5% to 10% by weight of a amount is 1-10 mg/dosage unit. In another embodiment, the absorption enhancer. In another embodiment, the pharmaceu amount is 2-10 mg/dosage unit. In another embodiment, the tical compositions of the present invention comprise an amount is 3-10 mg/dosage unit. In another embodiment, the amount of 8% to 15% by weight of a absorption enhancer. In amount is 5-10 mg/dosage unit. In another embodiment, the another embodiment, the pharmaceutical compositions of the amount is 1-20 mg/dosage unit. In another embodiment, the present invention comprise an amount of 10% to 20% by amount is 2-20 mg/dosage unit. In another embodiment, the weight of a absorption enhancer. In another embodiment, the amount is 3-20 mg/dosage unit. In another embodiment, the pharmaceutical compositions of the present invention com amount is 5-20 mg/dosage unit. In another embodiment, the prise an amount of 15% to 30% by weight of an absorption amount is 10-20 mg/dosage unit. In another embodiment, the enhancer. In another embodiment, the pharmaceutical com amount is 10-100 mg/dosage unit. In another embodiment, positions of the present invention comprise an amount of 20% the amount is 20-100 mg/dosage unit. In another embodi to 40% by weight of an absorption enhancer. In another ment, the amount is 30-100 mg/dosage unit. In another embodiment, the pharmaceutical compositions of the present embodiment, the amount is 50-100 mg/dosage unit. In invention comprise an amount of 30% to 50% by weight of an another embodiment, the amount is 10-200 mg/dosage unit. absorption enhancer. In another embodiment, the pharmaceu In another embodiment, the amount is 20-200 mg/dosage tical compositions of the present invention comprise an unit. In another embodiment, the amount is 30-200 mg/dos amount of 40% to 60% by weight of an absorption enhancer. age unit. In another embodiment, the amount is 50-200 In another embodiment, the pharmaceutical compositions of mg/dosage unit. In another embodiment, the amount is 100 the present invention comprise an amount of 50% to 70% by 200 mg/dosage unit. Each type and amount of enhancer rep weight of an absorption enhancer. In another embodiment, resents a separate embodiment of the present invention. the pharmaceutical compositions of the present invention 0072. In some embodiments, omega-3 fatty acid can be comprise an amount of 70% to 99.4% by weight of an absorp found in vegetable sources such as the seeds of chia, perilla, tion enhancer. In another embodiment, the amount of an flax, walnuts, purslane, lingonberry, Seabuckthorn, and hemp. absorption enhancer in the present composition is a delivery In some embodiments, omega-3 fatty acids can also be found effective amount and can be determined for any particular in the fruit of the acai palm. In another embodiment, the carrier or biologically or chemically active agent by methods omega-3 fatty acid has been provided in the form of a syn known to those skilled in the art. thetic omega-3 fatty acid. In one embodiment, the omega-3 0069. In another embodiment, an absorption enhancer is a fatty acid of methods and compositions of the present inven peptide. In another embodiment, an absorption enhancer is an tion has been provided to the composition in the form of fish amino acid. In another embodiment, a absorption enhancer is oil. In another embodiment, the omega-3 fatty acid has been derived from amino acids. provided in the form of canola oil. In another embodiment, 0070. In another embodiment, the amount of enhancer the omega-3 fatty acid has been provided in the form of utilized in methods and compositions of the present invention flaxseed oil. In another embodiment, the omega-3 fatty acid is 0.1 mg/dosage unit. In another embodiment, the amount of has been provided in the form of any other omega-3 fatty enhancer is 0.2 mg/dosage unit. In another embodiment, the acid-rich source known in the art. In another embodiment, the amount is 0.3 mg/dosage unit. In another embodiment, the omega-3 fatty acid has been provided in the form of a syn amount is 0.4 mg/dosage unit. In another embodiment, the thetic omega-3 fatty acid. Each form of omega-3 fatty acids amount is 0.6 mg/dosage unit. In another embodiment, the represents a separate embodiment of the present invention. amount is 0.8 mg/dosage unit. In another embodiment, the 0073. In another embodiment, the omega-3 fatty acid of amount is 1 mg/dosage unit. In another embodiment, the methods and compositions of the present invention is an amount is 1.5 mg/dosage unit. In another embodiment, the omega-3 polyunsaturated fatty acid. In another embodiment, amount is 2 mg/dosage unit. In another embodiment, the the omega-3 fatty acid is DHA, an omega-3, polyunsaturated, amount is 2.5 mg/dosage unit. In another embodiment, the 22-carbon fatty acid also referred to as 4, 7, 10, 13, 16, amount is 3 mg/dosage unit. In another embodiment, the 19-docosahexaenoic acid. In another embodiment, the amount is 5 mg/dosage unit. In another embodiment, the omega-3 fatty acid is -linolenic acid (9, 12, 15-octadecatri US 2011/0142800 A1 Jun. 16, 2011

enoic acid). In another embodiment, the omega-3 fatty acid is another embodiment, the coating is a water-based dispersion Stearidonic acid (6, 9, 12, 15-octadecatetraenoic acid). In of ethylcellulose, e.g. hydroxypropylmethylcelullose another embodiment, the omega-3 fatty acid is eicosatrienoic (HPMC) E15. In another embodiment, the coating is a gastro acid (ETA: 11, 14, 17-eicosatrienoic acid). In another resistant coating, e.g. a polymer containing carboxylic acid embodiment, the omega-3 fatty acid is eicSoatetraenoic acid groups as a functional moiety. In another embodiment, the (8, 11, 14, 17-eicosatetraenoic acid). In one embodiment, the coating is a monolithic matrix. In another embodiment, the omega-3 fatty acid is eicosapentaenoic acid (EPA; 5, 8, 11, coating is cellulose ether (e.g. hypromellose (HPMC). Each 14, 17-eicosapentaenoic acid). In another embodiment, the type of coating represents a separate embodiment of the omega-3 fatty acid is eicosahexaenoic acid (also referred to as present invention. “EPA'; 5, 7, 9, 11, 14, 17-eicosahexaenoic acid). In another embodiment, the omega-3 fatty acid is docosapentaenoic acid 0079. In another embodiment, the present invention pro (DPA; 7, 10, 13, 16, 19-docosapenatenoic acid). In another vides a composition comprising an active protein of the embodiment, the omega-3 fatty acid is tetracosahexaenoic invention, a protease inhibitor, and an absorption enhancer. In acid (6, 9, 12, 15, 18, 21-tetracosahexaenoic acid). In another another embodiment, the present invention provides a com embodiment, the omega-3 fatty acid is any other omega-3 position comprising an active protein of the invention, a pro fatty acid known in the art. Each omega-3 fatty acid repre tease inhibitor, an omega-3 fatty acid, and an absorption sents a separate embodiment of the present invention. enhancer. In another embodiment, the present invention pro 0074. In another embodiment, compositions of the present vides a composition comprising an active protein of the invention further comprise a coating that inhibits digestion of invention, a protease inhibitor, an omega-3 fatty acid, EDTA the composition in a subject's stomach. In one embodiment, or Na-EDTA, and an absorption enhancer. coating inhibits digestion of the composition in a subject's 0080. In another embodiment, the present invention pro stomach. In one embodiment, the coated dosage forms of the vides that the use of at least one protease inhibitor and an present invention release drug when pH move towards alka absorption enhancer in a single oral composition dramati line range. In one embodiment, coating is a monolayer, cally, unexpectedly, increase the bioavailability of a protein of wherein in other embodiments coating applied in multilayers. the invention. In another embodiment, the present invention In one embodiment, coating is a bioadhesive polymer that provides that the use of at least one protease inhibitor and an selectively binds the intestinal mucosa and thus enables drug absorption enhancer in a single oral composition dramati release in the attachment site. In one embodiment, the enteric cally, unexpectedly, increase the bioavailability of insulin.In coating is an enteric film coating. In some embodiment, coat another embodiment, the present invention provides that the ing comprises biodegradable polysaccharide, chitosan, aqua use of at least one protease inhibitor and an absorption teric aqueous, aquacoat ECD, azo polymer, cellulose acetate enhancer in a single oral composition dramatically, unexpect phthalate, cellulose acetate trimelliate, hydroxypropylmethyl edly, increase the bioavailability of Exenatide. cellulose phthalate, gelatin, poly vinyl acetate phthalate, I0081. In another embodiment, the present invention pro hydrogel, pulsincap, or a combination thereof. In one vides that the use of at least one protease inhibitor and an embodiment, pH sensitive coating will be used according to absorption enhancer in a single oral composition dramati the desired release site and/or profile as known to one skilled cally, unexpectedly, increase the bioavailability of a protein of in the art. the invention. In another embodiment, the present invention 0075. In one embodiment, the coating is an enteric coat provides that the use of at least one protease inhibitor and an ing. Methods for enteric coating are well known in the art, and absorption enhancer in a single oral composition dramati are described, for example, in Siepmann F. Siepmann Jet al. cally, unexpectedly, increase the bioavailability of insulin.In Blends of aqueous polymer dispersions used for pellet coat another embodiment, the present invention provides that the ing: importance of the particle size. J Control Release 2005; use of at least one protease inhibitor and an absorption 105(3): 226-39; and Huyghebaert N. Vermeire A, Remon JP. enhancer in a single oral composition dramatically, unexpect In vitro evaluation of coating polymers for enteric coating and edly, increase the bioavailability of Exenatide. human ileal targeting. Int J Pharm 2005; 298(1): 26-37. Each I0082 In another embodiment, the present invention pro method represents a separate embodiment of the present vides that the use of at least one protease inhibitor and SNAC invention. in a single oral composition dramatically, unexpectedly, 0076. In another embodiment, EudragitR), an acrylic poly increase the bioavailability of a protein of the invention.In mer, is used as the enteric coating. The use of acrylic poly another embodiment, the present invention provides that the mers for the coating of pharmaceutical preparations is well use of at least one protease inhibitor and SNAC in a single oral known in the art. Eudragit Acrylic Polymers have been shown composition dramatically, unexpectedly, increase the bio to be safe, and are neither absorbed nor metabolized by the availability of insulin. In another embodiment, the present body, but rather are eliminated. invention provides that the use of at least one protease inhibi 0077. In another embodiment, the coating is a gelatin coat tor and SNAC in a single oral composition dramatically, ing. In another embodiment, microencapsulation is used to unexpectedly, increase the bioavailability of Exenatide. protect the insulin against decomposition in the stomach. In I0083. In another embodiment, the present invention pro another embodiment, the coating is a gelatin coating. In vides that the use of at least one protease inhibitor and SNAD another embodiment, microencapsulation is used to protect in a single oral composition dramatically, unexpectedly, Exenatide against decomposition in the stomach. Methods for increase the bioavailability of a protein of the invention.In applying a gelatin coating and for microencapsulation are another embodiment, the present invention provides that the well known in the art. Each method represents a separate use of at least one protease inhibitor and SNAD in a single embodiment of the present invention. oral composition dramatically, unexpectedly, increase the 0078. In another embodiment, the coating is a film-coat bioavailability of insulin. In another embodiment, the present ing. In another embodiment, the coating is ethylcellulose. In invention provides that the use of at least one protease inhibi US 2011/0142800 A1 Jun. 16, 2011 tor and SNAD in a single oral composition dramatically, SNAC or SNAD in a single oral composition dramatically, unexpectedly, increase the bioavailability of Exenatide. unexpectedly, increase the bioavailability of insulin. In 0084. In another embodiment, the present invention pro another embodiment, the present invention provides that the vides that the use of a serpin and SNAC or SNAD in a single use of a Metalloprotease protease inhibitor and SNAC or oral composition dramatically, unexpectedly, increase the SNAD in a single oral composition dramatically, unexpect bioavailability of a protein of the invention. In another edly, increase the bioavailability of Exenatide. embodiment, the present invention provides that the use of a I0089. In another embodiment, the present invention pro serpin and SNAC or SNAD in a single oral composition vides that the use of an Aspartic protease inhibitor and SNAC dramatically, unexpectedly, increase the bioavailability of or SNAD in a single oral composition dramatically, unexpect insulin.In another embodiment, the present invention pro edly, increase the bioavailability of a protein of the invention. vides that the use of a serpin and SNAC or SNAD in a single In another embodiment, the present invention provides that oral composition dramatically, unexpectedly, increase the the use of an Aspartic protease inhibitor and SNAC or SNAD bioavailability of Exenatide. In another embodiment, the in a single oral composition dramatically, unexpectedly, present invention provides that the use of a serpin and SNAC increase the bioavailability of insulin. In another embodi or SNAD in a single oral composition dramatically, unexpect ment, the present invention provides that the use of an Aspar edly, increase the bioavailability of a protein of the invention. tic protease inhibitor and SNAC or SNAD in a single oral 0085. In another embodiment, the present invention pro composition dramatically, unexpectedly, increase the bio vides that the use of a trypsin inhibitor and SNAC or SNAD in availability of Exenatide. a single oral composition dramatically, unexpectedly, 0090. In another embodiment, the present invention pro increase the bioavailability of insulin. In another embodi vides that the use of a protease inhibitor and SNAC or SNAD ment, the present invention provides that the use of a trypsin in a single oral composition dramatically, unexpectedly, inhibitor and SNAC or SNAD in a single oral composition increase the bioavailability of insulin in a human subject by at dramatically, unexpectedly, increase the bioavailability of least 10%. In another embodiment, the present invention pro Exenatide. In another embodiment, the present invention pro vides that the use of a protease inhibitor and SNAC or SNAD vides that the use of SBTI or Aprotinin and SNAC or SNAD in a single oral composition dramatically, unexpectedly, in a single oral composition dramatically, unexpectedly, increase the bioavailability of Exenatide in a human subject increase the bioavailability of a protein of the invention. In by at least 10%. In another embodiment, the present invention another embodiment, the present invention provides that the provides that the use of a protease inhibitor and SNAC or use of SBTI or Aprotinin and SNAC or SNAD in a single oral SNAD in a single oral composition dramatically, unexpect composition dramatically, unexpectedly, increase the bio edly, increase the bioavailability of insulin in a human subject availability of insulin. In another embodiment, the present by at least 20%. In another embodiment, the present invention invention provides that the use of SBTI or Aprotinin and provides that the use of a protease inhibitor and SNAC or SNAC or SNAD in a single oral composition dramatically, SNAD in a single oral composition dramatically, unexpect unexpectedly, increase the bioavailability of Exenatide. edly, increase the bioavailability of Exenatide in a human I0086. In another embodiment, the present invention pro subject by at least 20%. In another embodiment, the present vides that the use of a Cysteine protease inhibitor and SNAC invention provides that the use of a protease inhibitor and or SNAD in a single oral composition dramatically, unexpect SNAC or SNAD in a single oral composition dramatically, edly, increase the bioavailability of a protein of the invention. unexpectedly, increase the bioavailability of insulin in a In another embodiment, the present invention provides that human subject by at least 30%. In another embodiment, the the use of a a Cysteine protease inhibitor and SNAC or SNAD present invention provides that the use of a protease inhibitor in a single oral composition dramatically, unexpectedly, and SNAC or SNAD in a single oral composition dramati increase the bioavailability of insulin. In another embodi cally, unexpectedly, increase the bioavailability of Exenatide ment, the present invention provides that the use of a Cysteine in a human subject by at least 30%. In another embodiment, protease inhibitor and SNAC or SNAD in a single oral com the present invention provides that the use of a protease position dramatically, unexpectedly, increase the bioavail inhibitor and SNAC or SNAD in a single oral composition ability of Exenatide. dramatically, unexpectedly, increase the bioavailability of 0087. In another embodiment, the present invention pro insulin in a human subject by at least 40%. In another embodi vides that the use of a Threonine protease inhibitor and SNAC ment, the present invention provides that the use of a protease or SNAD in a single oral composition dramatically, unexpect inhibitor and SNAC or SNAD in a single oral composition edly, increase the bioavailability of a protein of the invention. dramatically, unexpectedly, increase the bioavailability of In another embodiment, the present invention provides that Exenatide in a human subject by at least 40%. In another the use of a Threonine protease inhibitor and SNAC or SNAD embodiment, the present invention provides that the use of a in a single oral composition dramatically, unexpectedly, protease inhibitor and SNAC or SNAD in a single oral com increase the bioavailability of insulin. In another embodi position dramatically, unexpectedly, increase the bioavail ment, the present invention provides that the use of a Threo ability of insulin in a human subject by at least 50%. In nine protease inhibitor and SNAC or SNAD in a single oral another embodiment, the present invention provides that the composition dramatically, unexpectedly, increase the bio use of a protease inhibitor and SNAC or SNAD in a single oral availability of Exenatide. composition dramatically, unexpectedly, increase the bio 0088. In another embodiment, the present invention pro availability of Exenatide in a human subject by at least 50%. vides that the use of a Metalloprotease protease inhibitor and In another embodiment, the present invention provides that SNAC or SNAD in a single oral composition dramatically, the use of a protease inhibitor and SNAC or SNAD in a single unexpectedly, increase the bioavailability of a protein of the oral composition dramatically, unexpectedly, increase the invention. In another embodiment, the present invention pro bioavailability of insulin in a human subject by at least 60%. vides that the use of a Metalloprotease protease inhibitor and In another embodiment, the present invention provides that US 2011/0142800 A1 Jun. 16, 2011

the use of a protease inhibitor and SNAC or SNAD in a single least 70%. In another embodiment, the present invention pro oral composition dramatically, unexpectedly, increase the vides that the use of Aprotinin or SBTI and SNAG or SNAD bioavailability of Exenatide in a human subject by at least in a single oral composition dramatically, unexpectedly, 60%. In another embodiment, the present invention provides increase the bioavailability of insulin in a human subject by at that the use of a protease inhibitor and SNAC or SNAD in a least 80%. In another embodiment, the present invention pro single oral composition dramatically, unexpectedly, increase vides that the use of Aprotinin or SBTI and SNAC or SNAD the bioavailability of insulin in a human subject by at least in a single oral composition dramatically, unexpectedly, 70%. In another embodiment, the present invention provides increase the bioavailability of insulin in a human subject by at that the use of a protease inhibitor and SNAG or SNAD in a least 90%. In another embodiment, the present invention pro single oral composition dramatically, unexpectedly, increase vides that the use of Aprotinin or SBTI and SNAC or SNAD the bioavailability of Exenatide in a human subject by at least in a single oral composition dramatically, unexpectedly, 70%. In another embodiment, the present invention provides increase the bioavailability of insulin in a human subject by at that the use of a protease inhibitor and SNAC or SNAD in a least 100%. single oral composition dramatically, unexpectedly, increase 0092. In another embodiment, this invention further pro the bioavailability of insulin in a human subject by at least vides the use of sustained release dosage forms (e.g. Sustained 80%. In another embodiment, the present invention provides release microencapsulation) that enable the treatment fre that the use of a protease inhibitor and SNAC or SNAD in a quency to be reduced to once or twice a day. In another single oral composition dramatically, unexpectedly, increase embodiment, the insulin dosage is increased correspondingly the bioavailability of Exenatide in a human subject by at least with decreasing frequency of administration. In another 80%. In another embodiment, the present invention provides embodiment, the Exenatide dosage is increased correspond that the use of a protease inhibitor and SNAC or SNAD in a ingly with decreasing frequency of administration. Each type single oral composition dramatically, unexpectedly, increase of coating, absorption enhancer, dosage form, etc., that inhib the bioavailability of insulin in a human subject by at least its digestion of the composition in the stomach represents a 90%. In another embodiment, the present invention provides separate embodiment of the present invention. that the use of a protease inhibitor and SNAC or SNAD in a 0093 Methods of measuring insulin levels are well known single oral composition dramatically, unexpectedly, increase in the art. In one embodiment, levels of recombinant insulin the bioavailability of Exenatide in a human subject by at least are measuring using a human insulin radio-immunoassay 90%. In another embodiment, the present invention provides (RIA) kit, e.g. the kit manufactured by Linco Research Inc. that the use of a protease inhibitor and SNAC or SNAD in a (St. Charles, Mo.). In another embodiment, levels of C pep single oral composition dramatically, unexpectedly, increase tide are measured as well, to determine the relative contribu the bioavailability of insulin in a human subject by at least tions of endogenous and exogenous insulin to observed rises 100%. In another embodiment, the present invention provides in insulin levels. In another embodiment, insulin ELISA kits that the use of a protease inhibitor and SNAC or SNAD in a are used. In another embodiment, insulin levels are measured single oral composition dramatically, unexpectedly, increase by any other method known in the art. In another embodi the bioavailability of Exenatide in a human subject by at least ment, Exenatide levels are measured by a method known in 100%. the art. Each possibility represents a separate embodiment of 0091. In another embodiment, the present invention pro the present invention. vides that the use of Aprotinin or SBTI and SNAC or SNAD 0094. In another embodiment, a multiparticulate dosage in a single oral composition dramatically, unexpectedly, forms is used to inhibit digestion of the composition in the increase the bioavailability of insulin in a human subject by at stomach. In another embodiment, a multiparticulate dosage least 10%. In another embodiment, the present invention pro forms is used to inhibit digestion of the composition in the vides that the use of Aprotinin or SBTI and SNAC or SNAD stomach. in a single oral composition dramatically, unexpectedly, 0095. In another embodiment, the present invention pro increase the bioavailability of insulin in a human subject by at vides a method for oral administration of a protein with an least 20%. In another embodiment, the present invention pro enzymatic activity to a Subject, whereby a Substantial fraction vides that the use of Aprotinin or SBTI and SNAC or SNAD of the protein retains the enzymatic activity after absorption in a single oral composition dramatically, unexpectedly, through an intestinal mucosal barrier of the Subject, compris increase the bioavailability of insulin in a human subject by at ing administering orally to the Subject a pharmaceutical com least 30%.In another embodiment, the present invention pro position comprising the protein, a protease inhibitor, and an vides that the use of Aprotinin or SBTI and SNAC or SNAD absorption enhancer, thereby orally administering a protein in a single oral composition dramatically, unexpectedly, with an enzymatic activity to a Subject. increase the bioavailability of insulin in a human subject by at 0096. In another embodiment, the present invention pro least 40%. In another embodiment, the present invention pro vides a method for oral administration of a protein with an vides that the use of Aprotinin or SBTI and SNAC or SNAD enzymatic activity to a Subject, whereby a Substantial fraction in a single oral composition dramatically, unexpectedly, of the protein retains the enzymatic activity after absorption increase the bioavailability of insulin in a human subject by at through an intestinal mucosal barrier of the Subject, compris least 50%. In another embodiment, the present invention pro ing administering orally to the Subject a pharmaceutical com vides that the use of Aprotinin or SBTI and SNAC or SNAD position comprising the protein, a protease inhibitor, and in a single oral composition dramatically, unexpectedly, SNAC or SNAD, thereby orally administering a protein with increase the bioavailability of insulin in a human subject by at an enzymatic activity to a Subject. least 60%. In another embodiment, the present invention pro 0097. In another embodiment, the present invention pro vides that the use of Aprotinin or SBTI and SNAC or SNAD vides a method for oral administration of insulinto a Subject, in a single oral composition dramatically, unexpectedly, whereby a substantial fraction of insulin retains its activity increase the bioavailability of insulin in a human subject by at after absorption through an intestinal mucosal barrier of the US 2011/0142800 A1 Jun. 16, 2011

Subject, comprising administering orally to the Subject a phar tor, and SNAC or SNAD, EDTA (or a salt thereof), and maceutical composition comprising insulin, a protease omega-3 fatty acid thereby treating diabetes mellitus. inhibitor, and SNAC or SNAD, thereby orally administering 0101. In another embodiment, the present invention pro a protein with an enzymatic activity to a Subject. In another vides a method for treating diabetes mellitus in a human embodiment, the present invention provides a method for oral Subject, comprising administering orally to the Subject a phar administration of Exenatide to a subject, whereby a Substan tial fraction of Exenatide retains its activity after absorption maceutical composition comprising an Exenatide, at least one through an intestinal mucosal barrier of the Subject, compris protease inhibitor, and SNAC or SNAD, thereby treating dia ing administering orally to the Subject a pharmaceutical com betes mellitus. In another embodiment, the present invention position comprising Exenatide at least one protease inhibitor, provides a method for treating diabetes mellitus in a human and SNAC or SNAD, thereby orally administering a protein Subject, comprising administering orally to the Subject a phar with an enzymatic activity to a Subject. maceutical composition comprising an Exenatide and at least 0098. In another embodiment, the present invention pro one protease inhibitor, and SNAC or SNAD, and omega-3 vides a method for oral administration of a protein with an fatty acid thereby treating diabetes mellitus. In another enzymatic activity to a subject, whereby a Substantial fraction embodiment, the present invention provides a method for of the protein retains the enzymatic activity after absorption treating diabetes mellitus in a human Subject, comprising through an intestinal mucosal barrier of the Subject, compris administering orally to the Subject a pharmaceutical compo ing administering orally to the Subject a pharmaceutical com sition comprising an Exenatide and at least one protease position comprising the protein, at least one protease inhibi inhibitor, and SNAC or SNAD, EDTA (or a salt thereof), and tor, SNAC or SNAD, and an omega-3 fatty acid, thereby omega-3 fatty acid thereby treating diabetes mellitus. orally administering a protein with an enzymatic activity to a 0102. In one embodiment, the diabetes mellitus is Type I Subject. In another embodiment, the present invention pro diabetes. In another embodiment, the diabetes mellitus is vides a method for oral administration of a protein with an Type II diabetes. In another embodiment, the diabetes melli enzymatic activity to a subject, whereby a Substantial fraction tus is insulin-dependent diabetes. In another embodiment, the of the protein retains the enzymatic activity after absorption diabetes mellitus is non-insulin-dependent diabetes. In through an intestinal mucosal barrier of the Subject, compris ing administering orally to the Subject a pharmaceutical com another embodiment, the diabetes mellitus is any other type position comprising the protein, at least one protease inhibi of diabetes known in the art. Each possibility represents a tor, and SNAC or SNAD, EDTA (or a salt thereof), and an separate embodiment of the present invention. omega-3 fatty acid, thereby orally administering a protein 0103) In one embodiment, three treatments a day of the with an enzymatic activity to a human Subject. insulin composition are administered. In another embodi 0099. In another embodiment, the present invention pro ment, two treatments a day are administered. In another vides a method for oral administration of insulin to a Subject, embodiment, four treatments a day are administered. In whereby a substantial fraction of insulin retains its activity another embodiment, one treatment a day is administered. In after absorption through an intestinal mucosal barrier of the another embodiment, more than four treatments a day are Subject, comprising administering orally to the Subject a phar administered. Each possibility represents a separate embodi maceutical composition comprising insulin, at least one pro ment of the present invention. tease inhibitor, SNAC or SNAD, an omega-3 fatty acid, and 0104 Any of the methods of the present invention may Na-EDTA, thereby orally administering a protein with an utilize, in various embodiments, any of the compositions of enzymatic activity to a subject. In another embodiment, the the present invention. present invention provides a method for oral administration 0105. In another embodiment, the present invention pro of Exenatide to a subject, whereby a substantial fraction of vides a composition for oral administration of insulin, com Exenatide retains its activity after absorption through an prising an insulin protein at least one protease inhibitor, and intestinal mucosal barrier of the Subject, comprising admin SNAG or SNAD, whereby a substantial fraction of the insulin istering orally to the Subject a pharmaceutical composition retains the enzymatic activity after absorption through an comprising Exenatide, at least one protease inhibitor, SNAC intestinal mucosal barrier of a human Subject. or SNAD.an omega-3 fatty acid, and Na-EDTA, thereby 0106. In one embodiment, the present invention provides orally administering a protein with an enzymatic activity to a the use of a protein, at least one protease inhibitor, and SNAC Subject. or SNAD in the manufacture of a medicament for oral admin 0100. In another embodiment, the present invention pro istration of a protein with an enzymatic activity to a Subject, vides a method for treating diabetes mellitus in a human whereby a substantial fraction of the protein retains the enzy Subject, comprising administering orally to the Subject a phar matic activity after absorption through an intestinal mucosal maceutical composition comprising an insulin, at least one barrier of the subject. In one embodiment, the present inven protease inhibitor, and SNAC or SNAD, thereby treating dia tion provides the use of a protein, at least one protease inhibi betes mellitus. In another embodiment, the present invention tor, SNAC or SNAD, and an omega-3 fatty acid in the manu provides a method for treating diabetes mellitus in a human facture of a medicament for oral administration of a protein Subject, comprising administering orally to the Subject a phar with an enzymatic activity to a Subject, whereby a substantial maceutical composition comprising an insulin and at least fraction of the protein retains the enzymatic activity after one protease inhibitor, and SNAC or SNAD, and omega-3 absorption through an intestinal mucosal barrier of the Sub fatty acid thereby treating diabetes mellitus. In another ject. In one embodiment, the present invention provides the embodiment, the present invention provides a method for use of a protein, at least one protease inhibitor, SNAC or treating diabetes mellitus in a human Subject, comprising SNAD, Na-EDTA, and an omega-3 fatty acid in the manu administering orally to the Subject a pharmaceutical compo facture of a medicament for oral administration of a protein sition comprising an insulin and at least one protease inhibi with an enzymatic activity to a Subject, whereby a substantial US 2011/0142800 A1 Jun. 16, 2011 12 fraction of the protein retains the enzymatic activity after 0113. In another embodiment, solid carriers/diluents for absorption through an intestinal mucosal barrier of the Sub use in methods and compositions of the present invention ject. include, but are not limited to, a gum, a starch (e.g. corn 0107. In one embodiment, the present invention provides starch, pregeletanized starch), a Sugar (e.g., lactose, mannitol, the use of an insulin protein, at least one protease inhibitor, Sucrose, dextrose), a cellulosic material (e.g. microcrystalline and SNAC or SNAD in the manufacture of a medicament for cellulose), an acrylate (e.g. polymethylacrylate), calcium car treating diabetes mellitus in a subject. In one embodiment, the bonate, magnesium oxide, talc, or mixtures thereof. present invention provides the use of an insulin protein, at 0114. In another embodiment, the compositions further least one protease inhibitor, SNAC or SNAD, and an omega-3 comprise binders (e.g. acacia, cornstarch, gelatin, carbomer, fatty acid in the manufacture of a medicament for treating ethyl cellulose, guar gum, hydroxypropyl cellulose, hydrox diabetes mellitus in a subject. In one embodiment, the present ypropyl methyl cellulose, povidone), disintegrating agents invention provides the use of an insulin protein, at least one (e.g. cornstarch, potato starch, alginic acid, silicon dioxide, protease inhibitor, SNAC or SNAD, Na-EDTA, and an croScarmelose sodium, crospovidone, guar gum, Sodium omega-3 fatty acid in the manufacture of a medicament for starch glycolate), buffers (e.g., Tris-HC1... acetate, phosphate) treating diabetes mellitus in a subject. of various pH and ionic strength, additives such as albuminor 0108. In one embodiment, the present invention provides gelatin to prevent absorption to Surfaces, detergents (e.g., the use of an Exenatide protein, at least one protease inhibitor, Tween 20, Tween 80, Pluronic F68, bile acid salts), surfac and SNAC or SNAD in the manufacture of a medicament for tants (e.g. sodium lauryl Sulfate), permeation enhancers, solu treating diabetes mellitus in a subject. In one embodiment, the bilizing agents (e.g., glycerol, polyethylene glycerol), anti present invention provides the use of an Exenatide protein, at oxidants (e.g., ascorbic acid, Sodium metabisulfite, butylated least one protease inhibitor, SNAC or SNAD, and an omega-3 hydroxyanisole), stabilizers (e.g. hydroxypropyl cellulose, fatty acid in the manufacture of a medicament for treating hyroxypropylmethyl cellulose), Viscosity increasing agents diabetes mellitus in a subject. In one embodiment, the present (e.g. carbomer, colloidal silicon dioxide, ethyl cellulose, guar invention provides the use of an Exenatide protein, at least gum), Sweeteners (e.g. aspartame, citric acid), preservatives one protease inhibitor, SNAC or SNAD, Na-EDTA, and an (e.g., Thimerosal, benzyl alcohol, parabens), lubricants (e.g. omega-3 fatty acid in the manufacture of a medicament for Stearic acid, magnesium Stearate, polyethylene glycol, treating diabetes mellitus in a subject. Sodium lauryl Sulfate), flow-aids (e.g. colloidal silicon diox 0109. In one embodiment, methods and compositions of ide), plasticizers (e.g. diethyl phthalate, triethyl citrate), the present invention have the advantage of more closely emulsifiers (e.g. carbomer, hydroxypropyl cellulose, sodium mimicking physiological insulin secretion by the pancreas. lauryl Sulfate), polymer coatings (e.g., poloxamers or poloX When insulin is secreted into the portal vein, the liver is amines), coating and film forming agents (e.g. ethylcellulose, exposed to a greater insulin concentration than peripheral acrylates, polymethacrylates) and/or adjuvants. Each of the tissues. Similarly, insulin administered according to the above excipients represents a separate embodiment of the present invention reaches the intestine and is absorbed in the present invention. body through the intestine and through the portal system to 0.115. In some embodiments, the dosage forms of the the liver. This absorption route thus resembles the physiologi present invention are formulated to achieve an immediate cal secretion of insulin by the pancreas, enabling, in this release profile, an extended release profile, or a delayed embodiment, delicate control of the blood glucose level and release profile. In some embodiments, the release profile of the metabolic activities of the liver and the peripheral organs the composition is determined by using specific excipients controlled by insulin. By contrast, when insulin is adminis that serve for example as binders, disintegrants, fillers, or tered to insulin-deficient diabetic patients via the peripheral coating materials. In one embodiment, the composition will venous system, the concentration of insulin in the portal vein beformulated to achieve a particular release profile as known is similar to that in the peripheral circulation, resulting in to one skilled in the art. hypoinsulinemia in the portal vein and the liver and hyperin 0116. In one embodiment, the composition is formulated Sulinemia in the peripheral venous system. This leads, in one as an oral dosage form. In one embodiment, the composition embodiment, to an abnormal pattern of glucose disposal. is a Solid oral dosage form comprising tablets, chewable 0110. In another embodiment, different constituents of tablets, or capsules. In one embodiment the capsules are soft compositions of the present composition are absorbed at dif gelatin capsules. In another embodiment, capsules as ferent rates from the intestinal lumen into the blood stream. described herein are hard-shelled capsules. In another The absorption of the bile acid, in one embodiment, is sig embodiment, capsules as described herein are soft-shelled nificantly faster than the absorption of insulin. capsules. In another embodiment, capsules as described 0111 For this reason, in another embodiment, a drug regi herein are made from gelatine. In another embodiment, cap men involving ingestion of a pair of pills at spaced intervals, Sules as described herein are made from plant-based gelling e.g., a second pill containing a higher concentration of Substances like carrageenans and modified forms of starch enhancer is taken at a defined interval (e.g. 30 minutes) after and cellulose. the first pill. In another embodiment, certain of the constitu 0117. In other embodiments, controlled- or sustained-re ents are microencapsulated to enhance the absorption of the lease coatings utilized in methods and compositions of the insulin into the system. In another embodiment, certain of the present invention include formulation in lipophilic depots constituents are microencapsulated to enhance the absorption (e.g. fatty acids, waxes, oils). of the Exenatide into the system. 0118. The compositions also include, in another embodi 0112. In one embodiment, a treatment protocol of the ment, incorporation of the active material into or onto par present invention is therapeutic. In another embodiment, the ticulate preparations of polymeric compounds such as poly protocol is prophylactic. Each possibility represents a sepa lactic acid, polglycolic acid, hydrogels, etc, or onto rate embodiment of the present invention. liposomes, microemulsions, micelles, unilamellar or multila US 2011/0142800 A1 Jun. 16, 2011 mellar vesicles, erythrocyte ghosts, or spheroplasts.) Such by 3 healthy human subjects. As shown in FIG. 1, blood compositions will influence the physical state, solubility, sta glucose levels were significantly reduced and more stable in bility, rate of in vivo release, and rate of in vivo clearance. In human subjects treated with formulations (1) and (3). These another embodiment, particulate compositions of the active results suggest that formulations comprising a combination ingredients are coated with polymers (e.g. poloxamers or of a protease inhibitor and SNAC are superior to formulations poloxamines) comprising only SNAC. Specifically, the combination of 0119. In another embodiment, the compositions contain SNAC and SBTI is superior to any other formulation used. It ing the insulin and omega-3 fatty acid are delivered in a is important to notice that the effect of SNAG and a protease vesicle, e.g. a liposome (see Langer, Science 249:1527-1533 inhibitor is synergistic because formulations consisting insu (1990); Treat et al., in Liposomes in the Therapy of Infectious lin and a single protease inhibitor do not have a blood glucose Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, lowering effect- no differences in the reduction of blood New York, pp. 353–365 (1989); Lopez-Berestein, ibid., pp. glucose levels were observed in previous experiment with the 317-327; see generally ibid). In another embodiment, the same formulations comprising a single protease inhibitor compositions containing the Exenatide and omega-3 fatty (SBTI or Aprotinin). Thus, the results regarding the reduction acid are delivered in a vesicle, e.g. a liposome (see Langer, in blood glucose levels with insulin and a combination of Science 249:1527-1533 (1990); Treat et al., in Liposomes in SNAC and at least one protease inhibitor was unexpected. the Therapy of Infectious Disease and Cancer, Lopez-Ber The formulation comprising SNAC and SBTI had a synergis estein and Fidler (eds.), Liss, New York, pp. 353–365 (1989); tic, utmost effect in lowering blood glucose levels Lopez-Berestein, ibid., pp. 317-327; see generally ibid). 0.125. As shown in FIG. 2, blood C-peptide levels were 0120. The preparation of pharmaceutical compositions significantly reduced in human Subjects treated with formu that contain an active component, for example by mixing, lation (1) and (3). These results also suggest that the combi granulating, or tablet-forming processes, is well understood nation of SNAC and a protease inhibitor such as SBTI had a in the art. The active therapeutic ingredient is often mixed synergistic effect in lowering blood C-peptide levels. with excipients that are pharmaceutically acceptable and I0126. It should be emphasized that no reduction of C-pep compatible with the active ingredient. For oral administra tide levels were observed in a previous experiment wherein tion, the active ingredients of compositions of the present formulations comprising a single protease inhibitor (SBTI or invention are mixed with additives customary for this pur Aprotinin) were used. Thus, the results regarding the reduc pose. Such as vehicles, stabilizers, or inert diluents, and con tion in blood C-peptide levels with insulin and a combination verted by customary methods into suitable forms for admin of a protease inhibitor such as SBTI and SNAG was unex istration, such as tablets, coated tablets, hard or soft gelatin pected. capsules, aqueous, alcoholic or oily Solutions. 0121 Each of the above additives, excipients, formula Example 2 tions and methods of administration represents a separate embodiment of the present invention. Optimization of Source of Omega-3 Fatty Acids 0122. In one embodiment, the term “treating” refers to I0127. Various omega-3 fatty acids or sources of omega-3 curing a disease. In another embodiment, “treating refers to fatty acids (e.g. those listed above in the specification) are preventing a disease. In another embodiment, “treating compared for their ability to preserve insulin following oral refers to reducing the incidence of a disease. In another administration in methods and compositions of the present embodiment, “treating refers to ameliorating symptoms of a invention. Insulin tablets or capsules are formulated as disease. In another embodiment, “treating refers to inducing described in the above Examples, except that the insulin is remission. In another embodiment, “treating refers to slow dissolved in the alternate source instead of in fish oil. The ing the progression of a disease. most effective source of omega-3 fatty acids is used in Sub sequent Examples. EXPERIMENTAL DETAILS SECTION Example 1 Example 3 Insulin, SNAC, AND Protease Inhibitors Oral For Optimization of a Protease Inhibitor mulations I0128 Various protease inhibitors (either non-toxic or hav Materials and Experimental Methods ing an acceptable toxicity profile; e.g. those listed above in the specification) are compared for their ability to preserve insu Formulation lin following oral administration in methods and composi 0123 X please provide days prior to dosing, formulation tions of the present invention. Insulin and/or Exenatide tablets were prepared containing: (1) 6 mg insulin, 250 mg SNAC, or capsules are formulated as described in the above 125 mg SBTI in what is the exact carrier, (2). 6 mg insulin, Examples, except that the alternate protease inhibitors are 250 mg SNAC, 2.5 mg Aprotinin, 125 mg SBTI in what is the substituted for SBTI and/or Aprotinin. Amounts of the pro exact carrier, (3) 6 mg insulin, 250 mg SNAC, 2.5 mg Apro tease inhibitors are also varied, to determine the optimal tinin, 125 mg SBTI in what is the exact carrier. (4) 6 mg amounts. The most effective protease inhibitor/amount is insulin, 250 mg SNAC in what is the exact carrier. The used in Subsequent Examples. formulations were stored in the refrigerator (4°C.) until dos 1ng. Example 4 Results Optimization of an Enhancer 0.124. In the next experiment, the formulations as I0129. Various enhancers (e.g. those listed above in the describes in the “Formulation' section were orally consumed specification) are compared for their ability to facilitate US 2011/0142800 A1 Jun. 16, 2011 absorption of insulin following oral administration in meth consisting of N-(8-2- hydroxybenzoylamino)caprylate ods and compositions of the present invention. Insulin tablets (SNAC), N-(10-2-hydroxybenzoylamino)decanoate or capsules are formulated as described in the above (SNAD), a salt of said SNAC or said SNAD, and a combina Examples, except that the alternate enhancers are substituted tion thereof. for EDTA. Amounts of the enhancers are also varied, to 43. The method of claim 42, wherein said recombinant determine the optimal amounts. The most effective enhancer/ protein is an enzyme. amount is used in Subsequent experiments. 44. The method of claim 42, wherein said recombinant protein is selected from the group consisting of insulin, a Example 5 glucagon, an interferon gamma, an interferon alpha, a growth hormone, an erythropoietin, a GLP-1, a GLP-1 analogue, and Optimization of Type and Amount of Insulin granulocyte colony stimulating factor (G-CSF). 0130 Various types and amounts of insulin e.g. those 45. The method of claim 42, wherein said pharmaceutical listed above in the specification) are compared for their ability composition further comprises an omega-3 fatty acid. to regulate blood Sugar in methods and compositions of the 46. The method of claim 42, wherein said protease inhibi present invention. Insulin tablets or capsules are formulated tor is selected from the group consisting of a serpin, a Suicide as described in the above Examples, except that the type and inhibitor, a transition state inhibitor, a protein protease inhibi amount of insulin is varied. The most effective typefamount tor, a chelating agent, a Cysteine protease inhibitor, a Threo of insulin is used in clinical trials. nine protease inhibitor, an Aspartic protease inhibitor, and a 1-31. (canceled) Metalloprotease inhibitor. 32. An oral pharmaceutical composition comprising a pro 47. The method of claim 46, wherein said protease inhibi tein having a molecular weight of up to 100,000 Daltons; a tor is a serpin, and said serpin is a trypsin inhibitor. protease inhibitor; and a compound selected from the group 48. The method of claim 42, wherein said protease inhibi consisting of N-(8-2-hydroxybenzoylamino)caprylate tor is selected from the group consisting of Lima bean trypsin (SNAC), N-(10-2-hydroxybenzoylamino)decanoate inhibitor, Aprotinin, soybean trypsin inhibitor (SBTI), and (SNAD), a salt of said SNAC or said SNAD, and a combina Ovomucoid. tion thereof. 49. The method of claim 42, wherein said pharmaceutical 33. The composition of claim 32, wherein said protein is a composition further comprises EDTA or a salt thereof. recombinant protein. 50. The method of claim 42, wherein said pharmaceutical 34. The composition of claim 32, wherein said enhancer is composition further comprises a coating that inhibits diges a salt of said SNAC or said SNAD, and said salt is selected tion of said pharmaceutical composition in a stomach of a from the group consisting of a monosodium salt, a disodium Subject; an enteric coating; or a gelatin coating. salt, and a combination thereof. 51. A method for treating diabetes mellitus in a subject, 35. The composition of claim 32, wherein said protein is comprising administering orally to said Subject a pharmaceu selected from the group consisting of insulin, a glucagon, an tical composition comprising insulin, Exenatide, or a combi interferon gamma, an interferon alpha, a growth hormone, an nation thereof; a protease inhibitor, and a compound selected erythropoietin, a GLP-1, a GLP-1 analogue, and granulocyte from the group consisting of N-(8-2-hydroxybenzoyl colony stimulating factor (G-CSF). amino)caprylate (SNAC), N-(10-2-hydroxybenzoylamino) 36. The composition of claim 32, wherein said protease decanoate (SNAD), a salt of said SNAC or said SNAD, and a inhibitor is selected from the group consisting of a serpin, a combination thereof. Suicide inhibitor, a transition state inhibitor, a protein protease 52. The method of claim 51, wherein said insulin or inhibitor, a chelating agent, a Cysteine protease inhibitor, a Exenatide is recombinant insulin or Exenatide. Threonine protease inhibitor, an Aspartic protease inhibitor, 53. The method of claim 51, wherein said pharmaceutical and a Metalloprotease inhibitor. composition further comprises an omega-3 fatty acid. 37. The composition of claim 36, wherein said protease 54. The method of claim 51, wherein said protease inhibi inhibitor is a serpin, and said serpin is a trypsin inhibitor. tor is selected from the group consisting of a serpin, a Suicide 38. The composition of claim 32, wherein said protease inhibitor, a transition state inhibitor, a protein protease inhibi inhibitor is selected from the group consisting of Lima bean tor, a chelating agent, a Cysteine protease inhibitor, a Threo trypsin inhibitor, Aprotinin, soy bean trypsin inhibitor nine protease inhibitor, an Aspartic protease inhibitor, and a (SBTI), and Ovomucoid. Metalloprotease inhibitor. 39. The composition of claim 32, further comprising an 55. The method of claim 51, wherein said protease inhibi omega-3 fatty acid. tor is selected from the group consisting of Lima bean trypsin 40. The composition of claim 32, further comprising inhibitor, Aprotinin, soybean trypsin inhibitor (SBTI), and EDTA or a salt thereof. Ovomucoid. 41. The composition of claim 32, further comprising a 56. The method of claim 51, wherein said pharmaceutical coating that inhibits digestion of said composition in a stom composition further comprises EDTA or a salt thereof. ach of a Subject; an enteric coating; or a gelatin coating. 57. The method of claim 51, wherein said pharmaceutical 42. A method for oral administration of a recombinant composition further comprises a coating that inhibits diges protein having a molecular weight up to 100,000 Daltons to a tion of said pharmaceutical composition in a stomach of a Subject, comprising administering orally to said Subject a Subject; an enteric coating; or a gelatin coating. pharmaceutical composition comprising said protein; a pro tease inhibitor; and a compound selected from the group c c c c c