BR112015016111B1 - DELAYED RELEASE PHARMACEUTICAL COMPOSITION FOR DELIVERY OF A BIGUANIDE COMPOUND AND USE OF A BIGUANIDE COMPOUND OR SALT THEREOF - Google Patents

Abstract

DELAYED RELEASE PHARMACEUTICAL COMPOSITION FOR DELIVERY OF A BIGUANIDE COMPOUND AND USE OF A BIGUANIDE COMPOUND OR SALT THEREOF Compositions and methods are provided for improving the pharmacokinetics and reducing the risk of adverse events resulting from the administration of the Biguanide Compound, comprising the administration of delayed release formulas of said compounds having a latency phase release.

Description

FIELD OF THE INVENTION

[0001] The present invention generally relates to the treatment of metabolic disorders with biguanide compounds and to improving the pharmacokinetic and gastrointestinal tolerability of such compounds by administering the biguanide compounds to patients using sustained-release formulations.

FUNDAMENTALS OF THE INVENTION

[0002] Hyperglycemia or high blood sugar is a condition in which an excessive amount of glucose, i.e. greater than approximately 125 mg/dL, circulates in the blood plasma. Chronic hyperglycemia at levels that are more than a little above normal can produce a wide variety of serious complications over a period of years, including kidney damage, neurological damage, cardiovascular damage, retinal damage, or damage to the feet and legs. Diabetic neuropathy can be a result of long-term hyperglycemia.

[0003] Hyperglycemia can be caused by or associated with dysfunction of the thyroid, adrenal and pituitary glands, diseases of the pancreas, severe sepsis and intracranial diseases such as encephalitis, brain tumors and meningitis. By far the most common cause of chronic hyperglycemia is diabetes mellitus, which is widely considered by many to be a looming health care epidemic. In diabetes mellitus, hyperglycemia usually results from low insulin levels (type I diabetes) and/or insulin resistance at the cellular level (type II diabetes).

[0004] Many type II diabetes medications are designed to lower blood glucose levels. A first-line drug of choice for the treatment of type II diabetes and the most commonly prescribed antidiabetic drug in the world is metformin. In contrast to most diabetes medications, hypoglycemia with metformin is rare; it is also weight neutral and is associated with reduced cardiovascular events and reduced mortality.

[0005] Metformin (dimethylbiguanide) belongs to a class of biguanide drugs developed on the basis of a glucose-reducing extract containing guanidines from the Galega officinalis plant. (Bailey & Turner Metformin. N Engl J Med. Feb 29, 1996; 334(9):574-9; Bailey et al. Metformin: its botanical background. Practical Diabetes Int. 2004;21(3): 115-7 ). Originally synthesized as an independent product in 1921, (Werner E, Bell J. The preparation of methylguanidine, and of ββ-dimethylguanidine by the interaction of dicyanodiamide, and methylammonium and dimethylammonium chlorides respectively. J Chem Soc, Transactions. 1921;121:1790- 5), metformin and other biguanides have been found to lower blood glucose in animals. Studies of the glucose-lowering effects of metformin, phenformin, and buformin in humans were published in the 1950s. First, the greater potency of phenformin and buformin resulted in their more widespread use; however, its association with lactic acidosis eventually led to discontinuation in most countries in the late 1970s.

[0006] Metformin improves glucose tolerance in patients by lowering both basal and postprandial plasma glucoses. Metformin alone usually decreases fasting blood glucose by 20% and HbA1c levels by about 1.5%. (Bailey & Turner, supra', DeFronzo & Goodman Efficacy of metformin in patients with non-insulin-dependent diabetes mellitus. The Multicenter Metformin Study Group. N Engl J Med. Aug 31, 1995; 333(9):541-9 ). Metformin has also been shown to improve serum lipids, lower triglycerides, free fatty acids, and LDL cholesterol, and modestly increase HDL cholesterol (Bailey & Turner, supra).

[0007] Metformin's antihyperglycemic effects have been postulated to result from a wide variety of systemic biochemical interactions, including, for example, suppressing glucose production by the liver, increasing insulin sensitivity, elevating peripheral glucose uptake (by phosphorylate transcription factor GLUT-4), increase fatty acid oxidation, and/or decrease glucose absorption from the gastrointestinal tract. (Hundal & Inzucchi Metformin: new understandings, new uses. Drugs. 2003;63(18): 1879-94). More recently, investigators have focused on its apparent impact on glucagon-like peptide 1 (GLP-1) secretion, apparently determining that metformin does not act directly on L cells in the gut to induce GLP-1 secretion or elevate GLP-1 secretion. L cell sensitivity to several known secretagogues. (Mulherin et al., Mechanisms underlying metformin-induced secretion of glucagon-like peptide-1 from the intestinal L cell. Endocrinology 152:4610-19 (December 2011)). These investigators suggested that metformin stimulates GLP-1 release through an indirect mechanism involving both muscarinic (M3) receptor-dependent and gastrin-releasing peptide (GRP) pathways independent of intestinal L cells, such that systemic bioavailability of metformin is essential for therapeutic efficacy.

[0008] Unfortunately, however, systemic exposure to metformin still constitutes a serious risk of lactic acidosis for various patient populations. Lactic acidosis is a potentially fatal metabolic complication that occurs when lactic acid levels increase in the bloodstream. Consequently, metformin is contraindicated in people with any condition that may increase the risk of lactic acidosis, including kidney disorders, lung disease and liver disease. According to the prescribing information, heart failure, in particular, unstable or acute congestive heart failure, also increases the risk of lactic acidosis with metformin. Thus, metformin remains unavailable to treat hyperglycemia in patients with these contraindications.

[0009] Furthermore, conventional metformin formulations often produce adverse gastrointestinal complications of limited doses (Gl) including diarrhea, nausea, vomiting, dizziness, headaches and dyspepsia. Consequently, patient administration is generally titrated upwards over a period of time to a maximum tolerated dose based in no insignificant part on any resulting patient-specific adverse GI effects. Extended-release formulations have been developed in hopes of solving this, but they have not adequately addressed these problems.

[0010] Certainly, there continues to be a need for better and safer compositions and methods for delivering biguanide compounds that address these tolerability and safety concerns. Ideally, these would also provide more effective treatment options for metabolic disorders in patients with contraindications to metformin and/or other biguanides.

SUMMARY OF THE INVENTION

[0011] The present invention solves these longstanding problems with conventional delivery of biguanide compounds by delaying the release of the compounds in the upper gastrointestinal tract and ensuring passage into and preferably through the duodenum prior to dissolution. As demonstrated herein, short-term fluctuations in stomach pH due to meals and other factors can lead to aberrant release patterns and produce spikes in systemic exposure to the biguanide compound, thereby increasing the risk of adverse events, including gastrointestinal complications. and, more dangerously, lactic acidosis in other contraindicated patients.

[0012] In particular, the invention provides compositions and pharmaceutical formulations that deliver biguanide compounds to specific segments of the intestine, while significantly preventing the absorption of biguanide in the stomach and duodenum. In preferred embodiments, the subject formulations are adapted to include both delayed delivery (DR) for delivery at a desired pH, for example, delivery at the pH of the distal small intestine as well as a lag phase (LR) after contact with the desired pH during which drug release from the object formula is minimized in order to accommodate temporary pH fluctuations in the stomach and ensure delayed delivery beyond the duodenum.

[0013] In one aspect, the formula comprises an oral dosage form comprising a biguanide compound, wherein the oral dosage form is adapted to minimize the release of the biguanide compound for a lag phase of at least about 5 to 10 minutes after contact with pH 6.0, 6.5, 6.8 and 7.0, more preferably for a lag phase of at least about 15 to 20 minutes after contact with the desired pH, even more preferably for a latency phase of at least about 25 to 30 minutes after contact with the desired pH in the distal small intestine. In a specifically preferred embodiment, the formula ensures passage of the biguanide through the stomach and release beyond the duodenum, with full release after the lag phase begins at a pH of at least about 6.0, more preferably at least about 6.5 and even more preferably at 6.8 or 7.0.

[0014] In one embodiment, an enteric coating is applied to the object compositions at a weight gain of at least about 4.5 mg/cm2, 5 mg/cm2, 5.5 mg/cm2, 6 mg/cm2 , 6.5 mg/cm2, 7 mg/cm2, 7.5 mg/cm2, 8 mg/cm2, 9 mg/cm2, 10 mg/cm2, 11 mg/cm2, 12 mg/cm2, or 15 mg/cm2 . In a further embodiment, the enteric coating comprises an outer layer of said formula of at least about 5 mg/cm2 to 7 mg/cm2, more preferably at least about 5.5 mg/cm2 to at least about 6.5 mg/cm2. As demonstrated herein, this ensures adequate delivery at the desired intestinal site in order to avoid abnormal spikes in systemic exposure of the biguanide compound.

[0015] In alternative embodiments, an enteric coating of at least about 2.5% or 3% (w/w) weight gain, even more preferably at least about 3.5% or 4% (w/w) p) in weight gain is applied to the composition of the object to produce the desired latency phase. For tablets and capsules, the enteric coating can be applied to achieve about 2.5% to about 5%, 6%, 7%, 8%, 9% or 10% (w/w) weight gain, plus preferably about 3% to at least about 6% (w/w) weight gain. For granules and other multiparticulate dosage forms up to 20 or 30% (w/w) weight gain or more may be applied, preferably from about 20% - 50% (w/w), more preferably from about 30% - 50% (w/w) carcass.

[0016] In another aspect, pharmaceutical compositions are provided comprising an enteric coated oral dosage form comprising a biguanide compound, wherein the dosage form is adapted to minimize biguanide release for a latency phase of at least about from 5 to 10 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for a lag phase of at least about 15 or 20 minutes after contact with the desired pH , even more preferably for a lag phase of at least about 25 or 30 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0. In one embodiment, an enteric coating is applied to the pharmaceutical composition at a weight gain of at least about 4.5 mg/cm2, 5.5 mg/cm2, 6 mg/cm2, 6.5 mg/cm2, cm2, 7 mg/cm2, 7.5 mg/cm2, 8 mg/cm2, 9 mg/cm2, 10 mg/cm2, 11 mg/cm2, 12 mg/cm2, or 15 mg/cm2. In another embodiment, the enteric coating is applied at a weight gain of at least about 5 mg/cm 2 to 9.5 mg/cm 2 , more preferably at about 5.5 mg/cm 2 to about at least 7. 6 mg/cm2. In alternative embodiments, an enteric coating is applied to the pharmaceutical composition to achieve at least about 3.0% to at least about 7.0% (w/w) weight gain, more preferably at least about 4% to at least about 4% least about 6% (w/w) in weight gain.

[0017] In specific embodiments, pharmaceutical compositions are provided comprising an enteric coated oral dosage form comprising a biguanide compound, wherein the dosage form is adapted to release less than about 10%, 5%, 4%, 3 %, 2%, and preferably less than 1% of the biguanide compound after contact with an aqueous medium (e.g., submersion) at a pH of less than about 2 for about an hour or so followed by contact with an aqueous medium at a pH equal to or less than about 5.5 for at least 30 or 45 minutes. In a preferred embodiment, the enteric coated dosage form releases less than about 5%, 2% or 1% of the biguanide compound in a 0.1 N HCI aqueous medium per two hours and less than about 5%, 2 % or 1% when transferred to an aqueous medium at pH 5.5 for at least 30 to 45 minutes.

[0018] In additional embodiments, the enteric coated dosage form releases less than 15%, 10%, 5%, 3%, 2% or less than 1% of the biguanide compound during the lag phase after the dosage form enters in contact with an aqueous medium at a pH of about 6.5 or 6.8, wherein the lag phase is at least ten, fifteen or twenty minutes. In a preferred embodiment, the enteric coated dosage form releases less than about 15% of the biguanide compound when the dosage form is contacted with an aqueous medium at a pH of 6.5 to 6.8 for a phase. of at least ten minutes lag and releases from about 75% to about 100%, and more preferably greater than 90%, 95%, 98% or 99% of the biguanide compound after contacting an aqueous medium in a Carcass pH 6.5 or 6.8 for a total of 90 to 120 minutes.

[0019] In two-stage dissolution embodiments, pharmaceutical compositions are provided comprising an enteric-coated oral dosage comprising a biguanide compound, wherein the dosage form is adapted to release less than 5%, 2% or 1% of the biguanide compound in an aqueous medium of 0.1 N HCI for two hours. In these embodiments, less than 15%, 10%, 5%, 3%, 2% or preferably 1% of the biguanide compound is released after contacting an aqueous medium of 0.1 N HCI for two hours and subsequently transferred to an aqueous medium at a pH of about 6.8 for a lag phase of at least ten, fifteen or twenty minutes. In preferred embodiments, less than 15% of the biguanide compound is released after two hours at acidic pH and a lag phase of at least ten or fifteen minutes at pH 6.8, and at least 60% of the biguanide compound is released after lag phase, and within 60 minutes at pH 6.8, and at least 90% of the biguanide compound is released within 90 to 120 minutes at pH 6.8.

[0020] In three-stage dissolution embodiments, pharmaceutical compositions are provided comprising an enteric-coated oral dosage comprising a biguanide compound, wherein the dosage form is adapted to release less than 5%, 2% or 1% of the biguanide compound in an aqueous medium of 0.1 N HCI for two hours and less than 5%, 2%, or 1% when transferred to an aqueous medium at pH 5.5 for at least one hour. In these embodiments, less than 25%, 20%, 15%, 10%, or 5% of the biguanide compound is released after two hours in an aqueous medium of 0.1 N HCl, 30 minutes in an aqueous medium at pH 5.5, and during a lag phase of at least ten or fifteen minutes at pH 6.8. In preferred embodiments, less than 15%, 10% or 5% of the biguanide compound is released after two hours at acidic pH, 30 minutes at pH 5.5 and a lag phase of at least ten or fifteen minutes at pH 6, 8, and at least 60% of the biguanide compound is released after the lag phase, and within 60 minutes at pH 6.8, and at least 90% of the biguanide compound is released within 90 to 120 minutes at pH 6.8 .

[0021] In some embodiments, the enteric coating comprises a first polymer that minimizes the release of the biguanide compound for at least about 5 to 10 minutes after coming into contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for at least about 15 or 20 minutes, even more preferably for at least about 25 or 30 minutes after coming into contact with a pH of 6.0, 6.5, 6.8 or 7. ,0. In preferred embodiments, the polymer is insoluble in an acid medium, but dissolves upon salt formation or the like above pH 7.0. In an exemplary preferred embodiment, the polymer is selected from the group consisting of Eudragit FS, Eugragit S, shellac, and/or combinations thereof.

[0022] In additional embodiments, the enteric coating comprises a second polymer that dissolves at a lower pH than the first polymer. In preferred embodiments, the second polymer is insoluble at pH 5.5 and below, but dissolves by salt formation or the like above pH 5.5. In an exemplary preferred embodiment, the second polymer is selected from the group consisting of Eudragit L, cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate (hypromellose acetate succinate), polyvinyl acetate phthalate ( PVAP) and sodium alginate, stearic acid, and/or combinations thereof.

[0023] In some embodiments, the enteric coating comprises about 90% Eudragit FS and about 10% Eudragit L, about 80% Eudragit FS and about 20% Eudragit L, about 70% Eudragit FS and about 30% Eudragit L, about 60% Eudragit FS and about 40% Eudragit L, about 50% Eudragit FS and about 50% Eudragit L, about 40% Eudragit FS and about about 60% Eudragit L, about 30% Eudragit FS and about 70% Eudragit L, about 20% Eudragit FS and about 80% Eudragit L, or about 10% Eudragit FS and about 90% Eudragit L. In preferred embodiments, Eudragit FS and said Eudragit L are present in about 7:5 to about 5:7 ratios, and more preferably, from 6:4 to about 4:6 ratios . In an exemplary preferred embodiment, the enteric coating comprises about 60% Eudragit FS and about 40% Eudragit L.

[0024] In some embodiments, the sealing coating may be added between the core comprising the biguanide compound and the enteric coating. The seal liner material can be selected such that it has no effect on drug release. Suitable materials include, for example, hydroxypropylmethyl cellulose (HPMC). In other embodiments, the sealing coating material can be selected to extend the lag phase in order to promote slow drug release after the enteric coating ruptures. Suitable materials include, for example, Eudragit E which dissolves in acid but swells at higher pH and can be used to extend the lag phase after enteric coating rupture.

[0025] In an exemplary and preferred embodiment, the sealing coating comprises a mixture of hypromellose, titanium dioxide, polyethylene glycol 400 (macrogol), and polysorbate 80, for example, Opadry® White YS-1-7003 available from Colorcon Inc. In an alternative preferred and exemplary embodiment, the sealing coating comprises hypromellose, triacetin, and talc, for example, Opadry® 03K19229 Clear also available from Colorcon Inc.

[0026] Accordingly, the object formulas and compositions may further comprise a sealing coating between the biguanide compound and the enteric coating, providing a total coating thickness corresponding to at least about 4% to 8% (w/w) in weight gain, more preferably at least about 4.5% to 6.0% (w/w) in weight gain. In some embodiments, the combination of the outer enteric coating and inner coating comprises at least about 6.9 mg/cm 2 to 13.3 mg/cm 2 , more preferably at least about 7.8 mg/cm 2 to at least about 11.4 mg/cm 2 . mg/cm2.

[0027] In alternative embodiments, the subject formulas and compositions further comprise one or more disintegrants to accelerate dissolution of the core upon disruption of the enteric coating. In preferred embodiments, the disintegrant comprises croscarmellose sodium, sodium starch glycollate or combinations thereof.

[0028] In one embodiment, the biguanide compound is selected from the group consisting of metformin, phenformin, buformin and imeglimine. In a preferred embodiment, the biguanide compound comprises metformin or a salt thereof, preferably metformin hydrochloride.

[0029] In additional embodiments, the oral dosage forms disclosed herein further comprise a DPP-IV inhibitor. In another embodiment, the oral dosage forms disclosed herein further comprise an additional anti-obesity and/or anti-diabetes agent.

[0030] The oral dosage forms disclosed herein may preferably take the form of a tablet, capsule or microsphere, which is preferably enteric coated. Preferably, the tablet, capsule or microsphere is smooth and does not comprise an embossed surface. However, tablets, capsules or microspheres comprising an embossed surface are alternative embodiments embraced, wherein the coating thickness of the enteric coating and/or sealing coating are adjusted accordingly to provide delayed delivery and release phase profiles. latency described in this document.

[0031] Accordingly, methods and compositions are provided for treating metabolic disorders in patients, including otherwise contraindicated patient populations, by administering phase-enhanced delayed-release formulas having the requisite coating to ensure targeted delivery of the compound of biguanide to the patient's small intestine, and preferably the distal small intestine, thereby minimizing systemic bioavailability. The biguanide compounds of the disclosure can be administered to an object in need thereof to treat various metabolic disorders, including obesity, dyslipidemia or other disorders of lipid metabolism, as well as hyperglycemic conditions and histopathological diseases associated with hyperglycemia, including type II diabetes, pre- diabetes, gestational diabetes and polycystic ovary syndrome. The effective use of biguanide compounds for the prophylaxis and prevention of such diseases and disorders, as well as for more general weight loss purposes in overweight or mildly to severely obese individuals, is also explicitly contemplated.

[0032] In one aspect, methods of reducing the risk of an adverse event from administration of biguanide are provided, comprising administering a therapeutically effective amount of a biguanide compound to a subject in need thereof in a delivery formula delayed 5 or 10 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for a lag phase of at least about 15 or 20 minutes, even more preferably for a lag phase latency of at least about 25 or 30 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0. In one embodiment, the adverse event is lactic acidosis. In another event, the adverse event is a gastrointestinal complication selected from the group comprising nausea, diarrhea, dyspepsia and vomiting.

[0033] In another aspect, methods of reducing the risk of an adverse event from administration of biguanide are provided, comprising administering an enteric coated oral dosage form comprising a biguanide compound to a patient in need thereof, in that the dosage form is adapted to release less than 10%, 5%, 4%, 3%, 2% and preferably less than 1% of the biguanide compound upon contact with an aqueous medium at a pH of less than about 2 for about two hours followed by contact with an aqueous medium at a pH equal to or less than about 5.5 for less than about 30 to 45 minutes. In a preferred embodiment, the enteric coated dosage form releases less than about 5%, 2%, or 1% of the biguanide compound in a 0.1 N HCl aqueous medium per two hours and less than about 5%, 2 % or 1% when transferred to an aqueous medium at pH 5.5 for at least 30 to 45 minutes.

[0034] In additional embodiments, the enteric coated dosage form releases less than 15%, 10% or 5% of the biguanide compound during the lag phase after the dosage form comes into contact with an aqueous medium at a pH of about 6.5 or 6.8, where the latency phase is at least ten, fifteen or twenty minutes. In a preferred embodiment, the enteric coated dosage form releases less than about 15% of the biguanide compound when the dosage form is contacted with an aqueous medium at a pH of 6.5 to 6.8 for a phase. of at least ten minutes lag and releases from about 75% to about 100%, and more preferably greater than 90%, 95%, 98% or 99% of the biguanide compound after contacting an aqueous medium in a Carcass pH 6.5 or 6.8 for a total of 90 to 120 minutes.

[0035] Also provided herein are methods of treating metabolic disorders in a patient in need thereof, comprising administering a therapeutically effective amount of a biguanide compound to said patient in a delayed-release formula that minimizes the release of the compound of biguanide for a lag phase of at least about 5 or 10 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for a lag phase of at least about 15 or 20 minutes, even more preferably for a lag phase of at least about 25 or 30 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0. In some embodiments, the subject methods comprise administering an enteric coated oral dosage comprising a biguanide compound to a patient in need thereof, where the dosage form is adapted to deliver less than about 10%, 5%, 4% , 3%, 2%, or 1% of the biguanide compound at a pH of less than 2 for at least 2 hours, followed by a pH equal to or less than about 5.5 for at least 30 to 45 minutes. In further embodiments, the dosage form is adapted to release less than 15%, 10% or 5% of the biguanide compound when the dosage form is contacted with an aqueous medium at a pH of about 6.8 or less than at least least about 10 or 15 minutes.

[0036] Also provided herein are methods of reducing the onset of diabetes in a subject with pre-diabetes, comprising administering a therapeutically effective amount of a biguanide compound to said subject in a delayed-release formula that minimizes the release of the biguanide compound for a lag phase of at least about 5 or 10 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for a lag phase of at least about 15 or 20 minutes, even more preferably for a lag phase of at least about 25 or 30 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0. In some embodiments, the subject methods comprise administering an enteric coated oral dosage comprising a biguanide compound to a patient in need thereof, where the dosage form is adapted to deliver less than about 10%, 5%, 4% , 3%, 2%, or 1% of the biguanide compound at a pH of less than 2 for at least 2 hours, followed by a pH equal to or less than about 5.5 for at least 30 to 45 minutes. In further embodiments, the dosage form is adapted to release less than 15%, 10% or 5% of the biguanide compound when the dosage form is contacted with an aqueous medium at a pH of about 6.8 or less than at least least about 10 or 15 minutes.

[0037] Also provided herein are methods of inducing weight loss in a patient in need thereof, comprising administering a therapeutically effective amount of a biguanide compound to said patient in a delayed-release formula that minimizes the release of the biguanide compound for a lag phase of at least about 5 or 10 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for a lag phase of at least about of 15 or 20 minutes, even more preferably for a lag phase of at least about 25 or 30 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0. In some embodiments, the subject methods comprise administering an enteric coated oral dosage comprising a biguanide compound to a patient in need thereof, where the dosage form is adapted to deliver less than about 10%, 5%, 4% , 3%, 2%, or 1% of the biguanide compound at a pH of less than 2 for at least 2 hours, followed by a pH equal to or less than about 5.5 for at least 30 to 45 minutes. In preferred embodiments, the dosage form is adapted to release less than 15%, 10% or 5% of the biguanide compound when the dosage form is contacted with an aqueous medium at a pH of about 6.8 or less than at least least about 10 or 15 minutes.

[0038] In some embodiments, clinically induced weight loss results in greater than 2.27 kilograms (5 pounds) lost in the subject, for example, greater than 4.55 kilograms (10 pounds) lost, preferably greater than 11.36 kilos (25 pounds) lost, and even more preferably more than 22.73 kilos (50 pounds) lost. In other embodiments, the induced weight loss results in the subject having a body mass index between 18.5 and 24.9. In another embodiment, the induced weight loss results in at least a loss of at least 0.5 inches in the waist circumference.

[0039] The methods and compositions disclosed herein are also suitable for patients having a contraindication to the biguanide compound, for example metformin, phenformin or buformin. Such a contraindication may be a condition of hypoxia, impaired lactate clearance, and/or impaired clearance of the biguanide compound, for example, impaired metformin clearance.

[0040] For example, in one embodiment, the methods disclosed herein can be used to treat a patient who may have a hypoxic condition, such as, but not limited to, respiratory failure and heart failure. In another embodiment, the patient may have impaired lactate elimination. In another embodiment, the patient may suffer from liver failure, which may result in impaired lactate elimination. In another embodiment, the patient may have impaired elimination of the biguanide compound, which may be caused, for example, by renal dysfunction and/or renal disease. Likewise, in one embodiment the patient may have renal dysfunction. Such renal dysfunction may be moderate or severe renal dysfunction, or end-stage renal disease. In another embodiment, the patient may have kidney disease, which may be chronic. In another embodiment, the patient may have hyperglycemia, which may be chronic, and which may be caused by type II diabetes.

[0041] Biguanide compounds suitable for use in the subject methods include, for example, metformin, phenformin, buformin or imeglimin, including the analogues, salts, solvates, polymorphs, hydrates, N-oxides and prodrugs of such compounds.

[0042] In preferred embodiments, the biguanide compound has a relative bioavailability reduced by 70%, 60%, 50%, 40%, 30%, 20% or 10% in the subject for the delayed release formulation compared to an immediate release conventional (IR) or the extended release (XR) composition having the same amount of the biguanide compound. Likewise, in specific embodiments, administration of the subject's delayed-release formulation minimizes the mean plasma AUG, mean plasma Cmax, and/or circulating plasma concentration of the biguanide compound in said patient compared to an identical protocol administering a IR or XR formulation having the same amount of the biguanide compound. In preferred embodiments, the biguanide compound is metformin, the IR composition is Glucophage® and the XR composition is Glucophage® XR.

[0043] In one embodiment, the plasma AUCQ-36 mean of the biguanide compound is less than about 15,000 ng*hr/ml_ or 14,000 ng*hr/ml_, preferably less than about 12,000 ng*hr/ml_, more preferably less than about 11,000 ng*hr/ml_ and most preferably less than about 10,000 ng*hr/ml_ when administered at a total daily dose (TDD) of 2000 mg or 1000 mg twice daily (b/s en die, abbreviated as "b.i.d" or "BID"). In another embodiment, the mean plasma AUCo-36 of the biguanide compound is less than about 10,000 ng*hr/ml_, preferably less than about 9,000 ng*hr/ml_, more preferably less than about 8,000 ng*hr/ml_ ml_ or 7,000 ng*hr/ml_ and most preferably less than about 6,000 ng*hr/ml_ or 5,000 ng*hr/ml_ when administered at 1000 mg TDD, 500 mg BID or lower effective doses.

[0044] In one embodiment, the mean plasma Cmax of the biguanide compound is less than about 1100 ng/ml or 14,000 ng*hr/ml_, preferably less than about 1000 ng/ml, more preferably less than about 950 ng /mL and more preferably less than about 900 ng/mL when administered as 2000 mg TDD or 1000 mg BID. In another embodiment, the average plasma Cmax of the biguanide compound is less than about 800 ng/mL, preferably less than about 700 ng/mL, more preferably less than about 600 ng*hr/mL, and most preferably less than about of 600 ng/mL or 500 ng/mL when administered at 2000 mg TDD, 500 mg BID or lower effective doses.

[0045] In one embodiment, the resulting circulating plasma concentration of the biguanide compound is below about 5 µg/ml or 4 µg/ml, preferably below about 3 µg/ml or 2.5 µg/ml, more preferably below about 2 µg/ml, 1 µg/ml, 0.5 µg/ml or 0.25 µg/ml in the patient.

[0046] Administration of the subject's formulations may be twice a day in the morning and evening, or once a day (omni in die, abbreviated as "OD"). In certain preferred embodiments, administration may be once daily, in the morning, for example, before 1:00 pm, preferably before 12:00 or 11:00 am, more preferably before 10:00 am or 9:00 am, or with the morning meal. In other preferred embodiments, administration may be once daily in the evening, for example, after 5 pm, more preferably after 6 or 7 pm, or with the evening meal. In another preferred embodiment, administration can be once daily before bedtime.

[0047] The subject methods administer therapeutically effective amounts of the biguanide compound(s). Notably, however, the inventive methods provided herein advantageously allow for lower therapeutic doses than prior art formulations, on a per unit dose basis and/or on a daily basis. In certain embodiments of the methods disclosed herein, the biguanide compound is administered twice daily in an oral dosage form at a unit dose greater than 500 mg BID, for example, 600 or 800 mg BID. In certain preferred embodiments of the methods disclosed herein, the twice-daily oral dosage is less than 500 mg BID, e.g., less than 400 mg BID, e.g., less than 300 mg BID, e.g., about 150, 200 or 250 mg BID. In alternative preferred embodiments, the biguanide compound is administered once daily at a per unit dose of 75 mg OD, 125 mg OD, 250 mg OD, 300 mg OD, 500 mg OD, 600 mg OD, 750 mg OD, 800 mg OD, or 1000 mg OD. In further embodiments, the total daily dose of the biguanide compound is less than 2000 mg/day, preferably less than 1500 mg/day, more preferably less than 1000 or 750 mg/day, more preferably less than 500, 400, 300 or 200 mg/day.

[0048] In another embodiment, the oral dosage form can additionally include an extended-release formula for the biguanide compound. In preferred embodiments, the biguanide compound is targeted for delivery to the distal small intestine.

[0049] In the methods disclosed herein, the biguanide compound may be or comprise metformin, a metformin salt, solvate, polymorphine, hydrate, N-oxide or prodrug. In preferred embodiments, the biguanide compound is a metformin salt selected from the group consisting of hydrochloride, phosphate, sulfate, hydrobromide, salicylate, maleate, hemi-maleate, benzoate, succinate, hemisuccinate, ethanesulfonate, fumarate, hemi-fumarate , glycolate, palmoate, oratate, acetate, isobutyrate, acetylsalicylate, nicotinic acid, adamanthoate, chlorophyllin including zinc-chlorophyllin, carboxylic acid, benzoic acid, dichloroacetic acid, theophylline-7-acetate, clofibrate, tartate, oxalate, hemi-oxlate, tannate and hydroxyl acid. In a particularly preferred embodiment, the biguanide compound is metformin hydrochloride.

[0050] The methods disclosed herein may also further comprise administering an immediate-release, extended-release, or delayed-release form of one or more additional therapeutic agents, e.g., a DPP-IV inhibitor (e.g., sitagliptin , saxagliptin, berberine, vildagliptin, linagliptin, alogliptin, and the like), a chemosensory receptor ligand (e.g., a sweet receptor ligand, bitter receptor ligand, umami receptor ligand, sour receptor ligand, fat receptor or bile acid receptor ligand), an anti-obesity or anti-diabetes agent, or a chemosensory receptor antagonist, e.g., lactisole. Non-limiting examples include modalities further comprising administering 100 mg sitagliptin OD, or 50 mg sitagliptin BID. The delayed-release form can be a double-layer tablet, or a capsule with the two components as encapsulated mini-tablets. The delayed release form can also additionally include an immediate release component that is enteric coated at pH 5.0 for the additional therapeutic agent.

BRIEF DESCRIPTION OF THE FIGURES

[0051] FIG. 1 shows the study design described in Example 1.

[0052] FIG. 2 shows the events during the study treatment period described in Example 1.

[0053] FIG. 3 shows the plasma concentration of metformin immediate release (Metformin IR) (•) and metformin delayed release (Metformin DR) (■) (x-axis; ng/mL) as a function of time (y-axis; min) after ingestion at t = -240 and after a meal at t = 0 min.

[0054] FIG. 4A shows plasma concentration of PYY (x-axis; pg/mL) versus time (y-axis; min) in subjects at baseline (n,o) OR after ingestion of Metformin IR (•) or Metformin DR (■) and after a meal at t = 0 min. Figure 4B shows plasma concentration of active GLP-1 (x-axis; GLP-1 A pmol/L) as a function of time (y-axis; min) in subjects at baseline (□, o) or after ingestion of Metformin IR (•) or Metformin DR (■) and after a meal at t = 0 min. FIG. 4C shows total GLP-1 plasma concentration (x-axis; GLP-1 T pmol/L) versus time (y-axis; min) in subjects at baseline (□, o) or after ingestion of either Metformin IR (•) or Metformin DR (■) and after a meal at t = 0 min. For figures 4A to 4C, percentage increase in Abs AUG is compared to baseline values.

[0055] FIG. 5A shows plasma glucose concentration (x-axis; mg/dL) versus time (y-axis; min) in subjects at baseline (□, o) or after ingestion of either Metformin IR (•) or Metformin DR (■) and after a meal at t = 0 min. FIG. 5B shows plasma insulin concentration (x-axis; pmol/L) versus time (y-axis; min) in subjects at baseline (□, o) or after ingestion of either Metformin IR (•) or Metformin DR (■) and after a meal at t = 0 min. For figures 5A through 5B, percentage increase in Abs AUG is compared to baseline values.

[0056] FIG. 6 is a graph showing the area under the PYY curve (x-axis; log-transformed) versus the metformin area under the curve (ng/mL*min) after ingestion of Metformin IR (•) and Metformin DR (■)

[0057] FIG. 7A shows the plasma concentration of Metformin IR (•) and Metformin DR (■) (x-axis; ng/mL) as a function of time (y-axis; min) after ingestion at t =-240 and after a meal at t = 0 min. FIG. 7B shows plasma PYY concentration (x-axis; mg/dL) versus time (y-axis; min) in subjects at baseline (□, o) or after ingestion of Metformin IR (•) or Metformin DR (■) and after a meal at t = 0 min.

[0058] FIG. 8 shows the mean plasma concentrations of metformin (x-axis; ng/mL) on day 5 of 500mg (♦) and 1000 mg (■) Metformin DR, 1000 mg Metformin IR (o) and 500 mg Metformin Metformin IR + 1000 mg of Metformin DR (▲) as a function of time (y-axis; min). The dose was administered at t = -1 minute.

[0059] FIG. 9 shows the relative steady-state bioavailability in subjects with type 2 diabetes of Metformin DR 500 mg and 1000 mg BID compared to Metformin IR 1000 mg BID based on the 11-hour AUG of metformin plasma on day 5 (y-axis; % AUC(oa n hr)). These levels constitute a 45% and 57% reduction in the total extent of plasma metformin exposure for Metformin DR 500 mg BID and Metformin 1000 mg BID compared to Metformin IR 1000 mg BID.

[0060] FIG. 10 shows mean total plasma PYY concentrations (x-axis; pg/mL) as a function of time (y-axis; min) in subjects at baseline (o) or day 5 of the designated treatment (•).

[0061] FIG. 11 shows mean active plasma GLP-1 concentration (x-axis; pg/mL) as a function of time (y-axis; min) in subjects at baseline (o) or day 5 of the designated treatment (•). Breakfast was given at t = 0 min, dose was given at t = -1 minute, and lunch was given at t = 300 min.

[0062] FIG. 12 shows mean plasma glucose concentration (x-axis; mg/dL) as a function of time (y-axis; min) in subjects at baseline (o) or day 5 of the designated treatment (•).

[0063] FIG. 13 shows the individual change in fasting plasma glucose concentrations (x-axis; mg/dL) versus time (y-axis; min) from baseline to day 5 by scatter plot in subjects treated with 500mg (♦ ) and 1000mg (■) Metformin DR, 1000 mg Metformin IR (•) and 500mg Metformin IR + 1000mg Metformin DR (▲) (y-axis). The line on the panel marks the mean LS change in glucose (mg/dL) for each treatment.

[0064] FIG. 14 shows the mean plasma concentration of metformin (x-axis; ng/mL) of 500mg (♦) and 1000 mg (■) Metformin DR, 1000 mg Metformin IR (o) and 2000 mg extended-release (Metformin XR) ) as a function of time (y-axis; hours). The dose was administered at t = 0 hours. The second dose was administered by BID regimens at t = 12 hours. Meals/snacks were provided at t = -.42; 2.08; 11.5; 18 and 24 hours.

[0065] FIG. 15 shows the Cmax (left panel) and AUCo-36 (right panel) of one-day dosing of metformin 1000 mg BID IR, metformin 500 mg BID and 1000 mg BID DR, and metformin 2000 mg QD XR. The * means a statistically significant reduction in exposure compared to metformin IR and metformin XR (all p < 0.0001)

[0066] FIG. 16 shows the relative bioavailability of Metformin DR 500 and 1000 mg BID one-day dosing compared to Metformin IR 1000 mg BID (left panel) and the relative bioavailability of Metformin RD 500 and 1000 mg BID one-day dosing compared to 2000 mg Metformin XR QD (right panel)

[0067] FIG. 17 shows individual patient pharmacokinetic profiles on Day 5 of dosing with the anticipated delay in metformin release following a Metformin DR dose at t=-240 shown in the left panel and no delay shown in the right panel.

[0068] FIG. 18 shows the dissolution performance of exemplary delayed release formulas having nominal enteric coatings of 2.4% (Batch No. K111511-89A) and 3.8% (Batch No. K260512-127).

[0069] FIG. 19 shows the two-stage dissolution performance of exemplary delayed release formulas having 3.0% to 3.9% nominal enteric coatings. The first stage is 2 hours at pH < 2, and the second stage is 2 hours at pH 6.8.

[0070] FIG. 20 shows the three-stage dissolution performance of exemplary delayed release formulas having 3.0% to 3.9% nominal enteric coatings. The first stage is 2 hours at pH < 2, the second stage is 1 hour at pH 5.5 and the third stage is 3 hours at pH 6.8.

DETAILED DESCRIPTION

[0071] Contemplated herein are methods and compositions that consistently minimize the systemic bioavailability of biguanide compounds, such as metformin, in subjects, but still provide significant salutary metabolic effects, eg, reduction of hyperglycemia. Contrary to conventional understanding (see, e.g., Mulherin et al, supra), the biguanide compounds of the disclosure actually cause GLP-1 release through a mechanism of action that may include interaction with the luminal or epithelial aspect. (i.e., gastrointestinal tract side) of enteroendocrine cells, and systemic bioavailability can therefore be minimized and still achieve significant therapeutic efficacy. Advantageously, the object's methods and compositions significantly improve the pharmacokinetics of drug release and also dramatically reduce the possibility of adverse effects such as lactic acidosis and gastrointestinal complications.

[0072] Accordingly, methods of reducing the risk of adverse events from administration of biguanide are provided, comprising administering a therapeutically effective amount of a biguanide compound in a delayed-release formula to an object in need thereof; wherein said delayed-release formula comprises a lag phase profile that minimizes dissolution of the formula for at least about 5, 10, 12, 15 or 20 minutes after reaching the distal small intestine. Also provided herein are methods of treating metabolic disorders in subjects, including subjects having a contraindication to a biguanide compound(s), comprising administering a therapeutically effective amount of a biguanide compound in a delayed-release formula to a object in need of this; wherein said delayed formula comprises a lag phase profile that minimizes dissolution of the formula for at least about 5, 10, 12, 15 or 20 minutes after reaching the distal small intestine. In preferred embodiments, the biguanide compound is selected from the group consisting of metformin, buformin, phenoformin and imeglymine and is administered at lower doses and/or with lower bioavailability than currently indicated and still achieves the desired metabolic improvements.

[0073] Definitions

[0074] The terms "gastrointestinal tract" and "intestine", as used herein, refer to the stomach and intestines. The "small" or "upper" intestine includes the duodenum, jejunum and ileum and the "large" or "lower" intestine includes the cecum, colon and rectum. The "distal" small intestine includes the jejunum and ileum.

[0075] As used herein, the term "lag phase" as applied to the release of biguanide from object formulations refers to the period of time that begins when the enteric-coated formula comes into contact with the pH at which the enteric coating is designed to dissolve and during which there is a minimum initial rate of release, for example, less than about 20% or 15%, more preferably less than about 10% or 5%, even more preferably less than about of 3%, 2% or 1% for at least the first 5, 10, 15 or 20 minutes at the desired pH.

[0076] "Treating" or "the treatment" of any condition, disease, or disorder refers, in some embodiments, to ameliorating the disease, disorder, or condition (i.e., arresting or slowing the development of the disease, disorder, or condition, or at least one of its clinical symptoms). In other embodiments, "treat" or "treatment" refers to mitigating at least one physical parameter, which may or may not be perceptible to the subject, including physical parameters that are undesirable but not clinically significant. In still other embodiments, "treat" or "treatment" refers to inhibiting the disease, disorder, or condition, either physically, (e.g., stabilization of an apparent symptom), physiologically, (e.g., stabilization of a parameter physical) is both. In still other embodiments, "treat" or "treatment" refers to preventing or delaying the onset of the disease, disorder or condition.

[0077] "Therapeutically effective amount" or "effective amount" means the amount of a composition, compound, therapy or course of treatment that, when administered to a subject to treat a disease, disorder or condition, is sufficient to effect such treatment to the disease, disorder or condition. The "therapeutically effective amount" will vary depending on the composition, compound, therapy, course of treatment, disease, disorder or condition and its severity, and the age, weight, etc., of the subject being treated.

[0078] As used herein, the term "hyperglycemic" or "hyperglycemia", when used in reference to a condition of a patient, means an abnormally high transient or chronic level of glucose present in the blood of a patient. The condition may be caused by a delay in glucose metabolism or absorption, such that the patient exhibits glucose intolerance or an elevated glucose state typically not found in normal patients (e.g., in glucose-intolerant subdiabetic patients at risk of developing diabetes or in diabetic patients). Fasting plasma glucose (FPG) levels for normoglycemia are less than about 110 mg/dl, for impaired glucose metabolism, between about 110 and 126 mg/dl, and for diabetics greater than about 126 mg/dl .

[0079] When the biguanide compounds described herein include one or more chiral centers, the stereochemistry of such chiral centers can independently be in the R or S configuration, or a mixture of the two. The chiral centers may additionally be designated as R or S or R, S or d, D, 1, L or d, I, D, L. Correspondingly, the biguanide compounds of the invention, if they can be present in form optically active, may actually be present in the form of a racemic mixture of enantiomers or in the form of any one of the enantiomers separated in substantially isolated and purified form, or as a mixture comprising any relative proportions of the enantiomers.

[0080] When the biguanide compounds described herein contain two or more chiral centers, diastereoisomers are then possible. Such diastereoisomers may be present as pure diastereoisomeric enantiomers, pure racemic mixtures of diastereoisomeric enantiomers, mixtures of diastereoisomers which may be racemic or may have optical activity in their own right, due to the complex permutations of enantiomeric diastereoisomers in the equilibrium of mixtures.

[0081] When the biguanide compounds of the invention can be present in geometrically isomeric forms around, for example, the guanide bond, then they can even be present in the form of a mixture of geometric isomers comprising any relative proportions of the isomers, or in some cases in the form of any of the geometric isomers separated in substantially isolated and purified form.

[0082] When the biguanide compounds described herein include one or more linearly conjugated or isolated double bonds, the geometry around such double bonds may independently be a cis/trans, E/Z mixture or a respective geometric E or Z isomer.

[0083] "Alkyl" means a straight or branched chain saturated monovalent hydrocarbon radical. By way of example, the hydrocarbon chain may have from one to twenty carbons, one to sixteen carbons, one to fourteen carbons, one to twelve carbons, one to ten carbons, one to eight carbons, one to six carbons, one to four carbons, etc. "Lower alkyl" can refer to alkyls having, for example, one to six carbons, one to four carbons, and the like. In certain examples, a straight chain alkyl can have from one to six carbon atoms and a branched alkyl from three to six carbon atoms, for example, methyl, ethyl, propyl, 2-propyl, butyl (including all isomeric forms ), pentyl (including all isomeric forms), and the like. "Me" means methyl, "Et" means ethyl, and "iPr" means isopropyl.

[0084] "Aryl" means a monocyclic or bicyclic monovalent aromatic hydrocarbon radical, for example, having from 6 to 20 or 6 to 10 atom rings, for example, phenyl or naphthyl.

[0085] "Alkylaryl" means an (alkylene)-R radical, where R is aryl as defined above.

[0086] "Cycloalkyl" means a saturated or partially saturated cyclic monovalent hydrocarbon radical (or an alicyclic radical). By way of example, cycloalkyl may have from three to twenty carbon atoms, from three to sixteen carbon atoms, from three to fourteen carbon atoms, from three to twelve carbon atoms, from three to ten carbon atoms, from three to eight carbon atoms, three to six carbon atoms, etc., where one or two carbon atoms may be replaced by an oxo group, for example, admantanil, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, indanyl and others.

[0087] "Alkylcycloalkyl" means an (alkylene)-R radical, where R is cycloalkyl as defined above; for example, cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl, or cyclohexylmethyl and the like.

[0088] "Heterocyclyl" or "heterocycloalkyl" means a saturated or unsaturated monovalent monocyclic group in which one or two atom rings are heteroatoms selected from N, O or S, the remaining atom rings being C. heterocyclyl is optionally fused to a (single) aryl or heteroaryl ring as defined herein. The heterocyclyl ring fused to the monocyclic aryl or heteroaryl ring is also referred to in this Application as a "bicyclic heterocyclyl" ring. Additionally, one or two ring carbon atoms in the heterocyclyl ring may optionally be replaced with a -CO- group. More specifically, the term heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2-oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, tetrahydropyranyl, thiomorpholino and the like. When the heterocyclyl ring is unsaturated it may contain one or two ring double bonds. When the heterocyclyl group contains at least one nitrogen atom, it is also referred to herein as heterocycloamino and is a subset of the heterocyclyl group. When the heterocyclyl group is a saturated ring and is not fused to the aryl or heteroaryl ring, as stated above, it is also referred to herein as saturated monocyclic heterocyclyl.

[0089] "Alkylheterocycloalkyl" means a -(alkylene)-R radical, where R is the heterocyclyl ring as defined above, for example, tetrahydrofuranylmethyl, piperazinylmethyl, morpholinylethyl and the like.

[0090] "Heteroaryl" means a monovalent monocyclic or bicyclic aromatic radical, where one or more, preferably one, two or three ring atoms are heteroatoms selected from N, O or S, the remaining ring atoms being carbon. Representative examples include, but are not limited to, pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, diazolyl, pyrazolyl, triazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, tetrazolyl and others.

[0091] "Oxo" or "carbonyl" means =(O) group or C=O group, respectively.

[0092] The term "substituted" means that the referenced group is replaced with one or more additional groups individually and independently selected from groups described herein. In some embodiments, an optional substituent is selected from oxo, halogen, -CN, -NH2, -OH, -NH(CHs), -N CH3)2, alkyl (including straight chain, branched, and/or unsaturated alkyl ), substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, fluoroalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxy, fluoroalkoxy, -S-alkyl, -S(O)2-alkyl, -CONH((substituted alkyl or unsubstituted) or (substituted or unsubstituted phenyl)), -CON(H or unsubstituted alkyl)2, -OCON(substituted or unsubstituted alkyl)2, -NHCONH((substituted or unsubstituted alkyl) or (substituted or unsubstituted phenyl) ), -NHCOalkyl, -N (substituted or unsubstituted alkyl)CO(substituted or unsubstituted alkyl), -NHCOO(substituted or unsubstituted alkyl), -C(OH)(substituted or unsubstituted alkyl^ and -C(NH2 )(substituted or unsubstituted alkyl)2- In some embodiments, by way of example, an optional substituent is selected from oxo, fluorine, chlorine, bromine, iodine, -CN, -NH2, -OH, -NH (CH3 ), -N (CH3)2, -CH31- CH2CH3, -CH(CH3)2, -CF3, -CH2CF3, -OCH3, -OCH2CH3I - OCH(CH3)2I -OCF3I -OCH2CF3I -S(O)2I - CH3 , -CONH2I-CONHCH3I-NHCONHCH31 -COCH3r -COOH and the like. In some embodiments, substituted groups are replaced with one, two or three of the previous groups. In some embodiments, substituted groups are replaced with one or two of the preceding groups. In some embodiments, substituted groups are replaced with one of the preceding groups. Additionally, unless otherwise noted, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates every possible isomer, for example, every enantiomer and diastereomer and a mixture of isomers, such as racemic or scalemic mixtures.

[0093] In some embodiments, a biguanide compound of the disclosure is present in a composition as a salt. In some embodiments, salts are obtained by reacting a compound of the disclosure with acids. In certain other embodiments, pharmaceutically acceptable salts are obtained by reacting a compound of the disclosure with a base. In other embodiments, the compounds are used as the free acid or free base form in the manufacture of the compositions described herein. Types of salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with a pharmaceutically acceptable: inorganic acid, such as, for example, hydrochloric acid, hydrobromic acid sulfuric acid, phosphoric acid, metaphosphoric acid and the like; or with an organic acid, such as, for example, acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, acid 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, butyric acid, phenylacetic acid, phenylbutyric acid, valproic acid and the like; (2) salts formed when an acidic proton present in the parent compound is replaced by a metal ion, e.g. an alkali metal ion (e.g. lithium, sodium, potassium), an alkaline earth ion (e.g. magnesium or calcium) or an aluminum ion. In some cases, the biguanide compound described herein is reacted with an organic base, such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. In other instances, the compounds described herein form salts with amino acids such as, but not limited to, arginine, lysine and the like. Acceptable inorganic bases used to form salts with compounds that include an acidic proton include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide and the like.

[0094] The term "amino acid" includes any of the twenty naturally occurring amino acids or the D form of any of the naturally occurring amino acids. Furthermore, the term "amino acid" also includes non-naturally occurring amino acids other than D-amino acids, which are functional equivalents of naturally occurring amino acids. Such non-naturally occurring amino acids include, for example, norleucine ("Nle"), norvaline ("Nva"), L- or D-naphthalanine, ornithine ("Orn"), homoarginine (homoArg) and others known in the peptide art. , such as those described in M. Bodanzsky, “Principles of Peptide Synthesis,” 1st and 2nd Ed. Revised, Springer-Verlag, New York, N.Y., 1984 and 1993, and Stewart and Young, "Solid Phase Peptide Synthesis," 2nd Ed., Pierce Chemical Co., Rockford, III., 1984, both of which are incorporated herein by reference.

[0095] Amino acids and amino acid analogues can be purchased commercially (Sigma Chemical Co.; Advanced Chemtech) or synthesized using methods known in the art.

[0096] Within the scope of embodiments, the biguanide compounds described herein additionally include forms of the compounds such as pharmaceutically acceptable salts, solvates (including hydrates), amorphous phases, partially crystalline and crystalline forms (including all polymorphs), pro -drugs, metabolites, N-oxides, isotopically labeled, epimers, pure epimers, mixtures of epimers, enantiomers including but not limited to single enantiomers and enantiomeric diastereoisomers, meso compounds, stereoisomers, racemic mixtures and diastereoisomeric mixtures. Biguanide compounds described herein having one or more double bonds include cis/trans isomers, E/Z isomers and geometric isomers. Biguanide compounds described herein can be prepared as pharmaceutically acceptable salts formed when an acidic proton present in the parent compound is replaced by a metal ion, for example an alkali metal ion, an alkaline earth ion or an aluminum ion; or coordinates with an organic basis. Furthermore, salt forms of the disclosed compounds can be prepared using raw material salts or intermediates.

[0097] In some embodiments, the biguanide compounds described herein include solvent addition forms or crystal forms thereof, particularly solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric amounts of a solvent and can be formed during the crystallization process with pharmaceutically acceptable solvents such as water, ethanol and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol.

[0098] As noted above, in some embodiments, the biguanide compounds disclosed herein possess one or more stereocenters, and each center exists independently in the R or S configuration. The biguanide compounds disclosed herein include all diastereomeric, enantiomeric, and epimeric forms , as well as their appropriate mixtures.

[0100] In some embodiments, sites on the biguanide compounds disclosed herein are susceptible to various metabolic reactions. Therefore, incorporation of the appropriate substituents into metabolic reaction sites will reduce, minimize or eliminate metabolic pathways. In specific embodiments, the substituent suitable for decreasing or eliminating the susceptibility of the aromatic ring to metabolic reactions is, by way of example only, a halogen, deuterium or an alkyl group.

[0101] In some embodiments, the biguanide compounds described in this document are isotopically labeled, which are identical to those recited in the various formulas and structures presented in this document, except that one or more atoms are replaced by an atom with a mass atomic mass or mass number different from the atomic mass or mass number normally found in nature. In some embodiments, one or more hydrogen atoms are replaced with deuterium. In some embodiments, metabolic sites on the compounds described herein are deuterated. In some embodiments, deuterium replacement offers certain therapeutic advantages resulting from greater metabolic stability, such as, for example, prolonged half-life in vivo or reduced dosage requirements. Throughout the specification, groups and their substituents can be chosen by one skilled in the art to provide stable moieties and compounds.

[0102] Biguanides

[0103] The compositions and methods disclosed in this document relate to metformin and other biguanides. By way of background, metformin is one of the simplest structural variants of a class of compounds known as the biguanides. From a structural perspective, metformin resembles a pharmacophore or fragment of a larger biologically active chemical structure.

em que: Ri, R2, R3, R4, Rs, Re e R7 são selecionados independentemente de: H, OH, O-Rx, em que Rx é alquil, cicloalquil, alquilcicloalquil, acil, éster, tioéster; alquil opcionalmente substituído (por exemplo, um Ci a C12 alquil de reta cadeia ou de cadeia ramificada, opcionalmente, substituído com oxigênio, silício, enxofre ou opcionalmente substituído com OH, O-alquil, SH, S-alquil, NH2, NH- alquil); cicloalquil (por exemplo, cicloalquil C3 para C7); alquilcicloalquil (por exemplo, alquilcicloalquil C4 para C12 ); heterocicloalquil (por exemplo, onde 0 heterociclo compreende um ou dois heteroátomos selecionados de O, S ou N, incluindo um heterocicloalquil C2 a Ce); alquilheterocicloalquil (por exemplo, onde 0 heterociclo compreende um ou dois heteroátomos selecionados de O, S ou N, incluindo um alquilheterocicloalquil C3 a Cu, e incluindo em que quando N está presente no anel heterocíclico, 0 átomo de nitrogênio pode ser na forma de uma amida, carbamato ou ureia); alquenil opcionalmente substituído (por exemplo, alquenil de cadeia reta ou de cadeia ramificada Ci para C12, opcionalmente, substituído com oxigênio, silício, enxofre ou opcionalmente substituído com OH, O-alquil, SH, S-alquil, NH2, NH-alquil); alquinil opcionalmente substituído (por exemplo, alquinil de cadeia ramificada ou de cadeia reta Ci a C12 opcionalmente substituído com oxigênio, silício, enxofre ou opcionalmente substituído com OH, O-alquil, SH, S-alquil, NH2, NH-alquil); aril opcionalmente substituído (por exemplo, fenil, fenil substituído, naftil, naftil substituído); alquilaril opcionalmente substituído (por exemplo, alquilfenil, fenil com alquil substituído, alquilnaftil, naftil com alquil substituído); heteroaril opcionalmente substituído (por exemplo, piridil, furanil, tiofenil, pirrolil, oxazolil, isoxazolil, tiazolil, diazolil, pyrazolil, triazolil, os quais são todos substituídos opcionalmente,); alquilheteroaril opcionalmente substituído; e ou Re e R? podem se juntar para formar uma ligação, juntos formando um anel incluindo os átomos de nitrogênio aos quais estão ligados; ou Ri e R2 juntos podem formar um anel heterocíclico de 3 a 8 membros, incluindo os átomos de nitrogênio aos quais estão ligados; ou R4 e Rs podem, juntos, formar um anel selecionado a partir do grupo aziridina, pirrolil, imidazolil, pirazolil, indolil, indolinil, pirrolidinil, piperazinil e piperidil, incluindo os átomos de nitrogênio ao quais estão ligados.[0104] In one embodiment, the biguanide compounds of the subject's invention include the following:

wherein: R1, R2, R3, R4, Rs, Re and R7 are independently selected from: H, OH, O-Rx, wherein Rx is alkyl, cycloalkyl, alkylcycloalkyl, acyl, ester, thioester; optionally substituted alkyl (e.g. a C1 to C12 straight chain or branched chain alkyl optionally substituted with oxygen, silicon, sulfur or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl ); cycloalkyl (e.g. C3 to C7 cycloalkyl); alkylcycloalkyl (for example, C4 to C12 alkylcycloalkyl); heterocycloalkyl (for example, where the heterocycle comprises one or two heteroatoms selected from O, S or N, including a C2 to Ce heterocycloalkyl); alkylheterocycloalkyl (for example, where the heterocycle comprises one or two heteroatoms selected from O, S or N, including a C3 to Cu alkylheterocycloalkyl, and including where when N is present on the heterocyclic ring, the nitrogen atom may be in the form of a amide, carbamate or urea); optionally substituted alkenyl (e.g. straight-chain or branched-chain C1 to C12 alkenyl optionally substituted with oxygen, silicon, sulfur or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl) ; optionally substituted alkynyl (e.g. branched chain or straight chain C1 to C12 alkynyl optionally substituted with oxygen, silicon, sulfur or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl); optionally substituted aryl (e.g. phenyl, substituted phenyl, naphthyl, substituted naphthyl); optionally substituted alkylaryl (for example, alkylphenyl, alkyl-substituted phenyl, alkylnaphthyl, alkyl-substituted naphthyl); optionally substituted heteroaryl (for example, pyridyl, furanyl, thiophenyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, diazolyl, pyrazolyl, triazolyl, which are all optionally substituted); optionally substituted alkylheteroaryl; and or Re and R? can come together to form a bond, together forming a ring including the nitrogen atoms to which they are bonded; or R1 and R2 together can form a 3- to 8-membered heterocyclic ring, including the nitrogen atoms to which they are attached; or R4 and Rs may together form a ring selected from the group aziridine, pyrrolyl, imidazolyl, pyrazolyl, indolyl, indolinyl, pyrrolidinyl, piperazinyl and piperidyl, including the nitrogen atoms to which they are attached.

[0105] In certain embodiments, O-Rx may be selected from: straight or branched chain alkyl O-C1 for Cs; O-C3 to C7 cycloalkyl; O-C4 alkylcycloalkyl to Cs; O-acyl; O-esters; and O-thioesters.

[0106] In other embodiments, optional substitutions can include, for example, OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl. Additionally, an alkyl, alkenyl, alkynyl, etc. can be substituted for an oxygen, silicon, sulfur, etc., to form a heteroalkyl, heteroalkenyl, heteroalkynyl, etc.

[0107] In certain embodiments, each of: R3, RΘ and R7, or R3, R4, Rs and R7, or R3, R4, Rs and R7 or R3, R4, Rs, Re and R7, or R2, R3, R4, Rs, Re and R7 are independently selected from: H, methyl, ethyl, propyl or isopropyl; and each of the remaining substituent groups: Ri, R2, R* and Rs, or Ri, R2θ Re, or Ri, R∑ and Re or Ri and R2 or Ri, respectively, are independently selected from: H; optionally substituted alkyl (e.g. C1 to C12 straight-chain or branched-chain alkyl optionally hetero-substituted with oxygen, silicon, sulfur or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl) ; optionally substituted alkenyl (e.g. C1 to C12 straight chain or branched chain alkenyl optionally hetero-substituted with oxygen, silicon, sulfur or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl) ; optionally substituted alkynyl (for example, C1 to C12 straight chain or branched chain alkynyl optionally hetero substituted with oxygen, silicon, sulfur or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH2, NH-alkyl); cycloalkyl (e.g. C3 to C7 cycloalkyl); alkylcycloalkyl (e.g. C4 to C12 alkylcycloalkyl); heterocycloalkyl (for example, where the heterocycle comprises one or two heteroatoms, selected from O, S or N, including C2 to C6 heterocycloalkyl); alkylheterocycloalkyl (e.g., where the heterocycle comprises one or two heteroatoms, selected from O, S or N, including C3 to Cu alkylheterocycloalkyl, and including which when N is present on the heterocyclic ring, the nitrogen atom may be in the form amide, carbamate or urea); aryl (e.g., phenyl, substituted phenyl, naphthyl, substituted naphthyl); alkylaryl (e.g., alkylphenyl, alkyl-substituted phenyl, alkylnaphthyl, alkyl-substituted naphthyl); heteroaryl (for example, pyridyl, furanyl, thiophenyl, pyrrolyl, oxazolyl, isoxazolyl, thiazoyl, diazolyl, pyrazolyl, triazolyl, all of which are optionally substituted); alkylheteroarill; or R1 and R2 together can form a 3- to 8-membered heterocyclic ring, including the nitrogen atoms to which they are attached; or R4 and Rs may together form a ring selected from the group aziridine, pyrrolyl, imidazolyl, pyrazolyl, indolyl, indolinyl, pyrrolidinyl, piperazinyl and piperidyl, including the nitrogen atoms to which they are attached.

[0108] Exemplary compounds and substituents of Ri, R2, R3, R4, Rs, Re and R7 of Formula I are shown below. Additional combinations of R1, R2, R3, R4, Rs, Re, and R substituent selections? are envisioned and disclosed in co-pending US Patent Application Serial No. 3/547,022, which disclosure is expressly incorporated by reference herein.

[0109] In certain embodiments, the biguanide compounds of Formula I may include an asymmetric center or centers, and may be in the form of a composition of a racemic mixture, a diastereoisomeric mixture, a single enantiomer, an enantiomeric diastereoisomer, a meso compound , a pure epimer or a mixture of its epimers, etc. Additionally, the biguanide compounds can have one or more double bonds and can be in a form of a cis/trans, E/Z mixture or a respective E or Z geometric isomer.

[0110] The biguanide compounds of Formula I may also be prepared as a salt form, for example, pharmaceutically acceptable salts, including suitable acid forms, for example, selected salt forms of hydrochloride, hydrobromide, acetate, propionate, butyrate , sulfate, hydrogen sulfate, sulfite, carbonate, hydrogen carbonate, phosphate, phosphinate, oxalate, hemi-oxalate, malonate, hemi-malonate, fumarate, hemi-fumarate, maleate, hemi-maleate, citrate, hemi-citrate, tartrate , hemi-tartrate, aspartate, glutamate, etc.

Alternative embodiments of biguanide compounds specifically contemplated for use in the subject invention include the related heterocyclic compounds described in copending US Patent Application Serial No. 13/547,022, the disclosure of which is expressly incorporated herein by reference. The phrase "biguanide compound" as used herein includes these related heterocyclic compounds, exemplary embodiments of which include the following:

[0112] In one embodiment, the compounds of the disclosure may be prepared as a three component salt form including components A, B and C, wherein: A is the protonated form of a natural or artificial amino acid; B is the dianion of an acid; and C is the protonated form of a Compound of Formula I.

[0113] In certain aspects, stoichiometric amounts of A, B, and C may be included, where: A is the protonated form of a natural amino acid selected from alanine, aspartic acid, asparagine, arginine, glycine, glutamine, lysine, acid glutamic, phenylalanine, tyrosine, serine, threonine, tryptophan, leucine, isoleucine, histidine, methionine, proline, cysteine or cystine; B is the dianion of an acid selected from oxalic, malonic, citric, maleic, fumaric, tartaric, aspartic, glutamic and the like; and C is the protonated form of a Compound of Formula I.

[0114] Contraindications for biguanide compounds, including metformin

[0115] Since systemic biguanides, including metformin, are reported to be substantially excreted by the kidneys, the risk of accumulation of biguanide compounds and lactic acidosis increases with the degree of impaired renal function. Other contraindications for biguanide compounds such as metformin include impaired lactate elimination and a hypoxic condition. Likewise, patients with these contraindications are currently not treatable with conventional biguanide compounds.

[0116] However, as demonstrated herein, the therapeutic efficacy of metformin and other biguanide compounds does not require an increase in the systemic level of metformin that poses an increased risk of lactic acidosis. As such, the risk of metformin accumulation and lactic acidosis is drastically lower, and the methods provided herein can therefore be used to treat a condition in a patient in need thereof, even where the patient has a contraindication to metformin. . For example, the methods provided herein can be used to treat a patient in need thereof, where the patient has a hypoxic condition (e.g. respiratory failure and/or heart failure), impaired lactate elimination (e.g. , due to liver failure), metformin extraction deficiency, and/or renal dysfunction, which may be moderate, severe, or terminal dysfunction and may be the result of chronic kidney disease.

[0117] Metabolic disorders

[0118] The compositions and methods of the present invention find advantageous use in the treatment and/or prophylaxis of metabolic disorders, including overweight, obesity, pre-diabetes, polycystic ovary syndrome, dyslipidemia or lipid metabolism disorders, as well as hyperglycemic conditions, such as insulin-dependent (type 1) or -independent (type 2) diabetes, as well as the physiological conditions or disorders associated with or resulting from the hyperglycemic condition. Thus, hyperglycemic conditions treatable via a method of the invention also include a histopathologic change associated with chronic or acute hyperglycemia (e.g., diabetes). Particular examples include degeneration of the pancreas (destruction of β cells), calcification of the renal tubule, degeneration of the liver, eye damage (diabetic retinopathy), diabetic foot, mucosal ulcerations such as mouth and gums, excessive bleeding, delayed blood clotting or wound healing and increased risk of coronary heart disease, stroke, peripheral vascular disease, dyslipidemia, hypertension and obesity.

[0119] As used herein, the term "hyperglycemic" or "hyperglycemia", when used in reference to a condition of a patient, means an abnormally high transient or chronic level of glucose present in the blood of a patient. The condition may be caused by a delay in glucose metabolism or absorption, such that the patient exhibits glucose intolerance or an elevated glucose state typically not found in normal patients (e.g., in glucose-intolerant subdiabetic patients at risk of developing diabetes or in diabetic patients). Fasting plasma glucose (FPG) levels for normoglycemia are less than about 110 mg/dl, for impaired glucose metabolism, between about 110 and 126 mg/dl, and for diabetics greater than about 126 mg/dl .

[0120] Metabolic disorders also include obesity or an undesirable body mass. Leptin, cholecystokinin, PYY and GLP-1 decrease hunger, increase energy expenditure, induce weight loss or provide normal glucose homeostasis. Thus, in various embodiments, a method of the invention for treating obesity or undesirable body mass or hyperglycemia involves the local administration of metformin to activate enteroendocrine cell production of cholecystokinin, oxyntomodulin, GIP, GLP-2, PYY, or GLP-1 . Treatable disorders also include those typically associated with obesity, e.g. abnormally elevated serum/plasma LDL, VLDL, triglycerides, cholesterol, plaque formation leading to narrowing or blockage of blood vessels, increased risk of hypertension/stroke, coronary heart disease , etc.

[0121] Synthesis of Compounds

[0122] Compounds described herein can be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. Additionally, solvents, temperatures and other reaction conditions presented herein may vary according to the practice and knowledge of those skilled in the art.

[0123] The raw material used for the synthesis of compounds described in this document can be obtained from commercial sources, such as Aldrich Chemical Co. (Milwaukee, Wis.), Sigma Chemical Co. (St Louis, Mo.) or the raw materials can be synthesized. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and materials known to those skilled in the art, as described, for example, in March, ADVANCED ORGANIC CHEMISTRY 4th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANICCHEMISTRY 4th Ed., Vols. A and B (Plenum 2000, 2001), and Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3rd Ed., (Wiley 1999) (all of which are incorporated by reference in their entirety). General methods for preparing the compounds as disclosed herein can be derived from reactions known in the field and the reactions can be modified by using appropriate reagents and conditions, as would be recognized by one skilled in the art, for introducing the various fractions found in formulas as provided in this document.

[0124] Additional biguanide synthesis methods and schemes for the compounds described herein can be found in U.S. Application with Serial No. 12/593,479 (issued as U.S. 2010/0130498); US order with Serial No. 12/593,398 (issued as U.S. 2010/0184796); Pat. U.S. No. 7,829,299; US order with Serial No. 11/578,013 (issued as U.S. 2010/0056621); U.S. patent At the. 7,416,867; US order with Serial No. 11/455,693 (issued as U.S. 2007/0037212); US order with Serial No. 13/059,730 (issued as U.S. 2011/0143376), U.S. with Serial No. 12/996,670 (issued as U.S. 2011/0311991), Pat. U.S. At the. 7,811,788; US order with Serial No. 11/182,942 (issued as U.S. 2006/0019346); US order with Serial No. 12/993,542 (issued as U.S. 2011/0086138), U.S. with Serial No. 12/373,235 (issued as U.S. 2010/0055209); International Application Serial No. PCT/IL2007/000454 (published as WO 2007/116404); US order with Serial No. 10/472,056 (issued as U.S. 2004/0138189); Pat. U.S. At the. 5,891,919; Pat. U.S. At the. 6,376,657; US order with Serial No. 11/554,982 (issued as U.S. 2007/0104805); US order with Serial No. 11/926,745 (issued as U.S. 2008/0108604); International Application Serial No. PCT/CA2009/001688 (published as WO 2010/060198); US order with Serial No. 12/735,557 (issued as U.S. 2010/0330205); International Application Serial No. PCT/CA2007/001066 (published as WO 2008/000063); US order with Serial No. 11/438,204 (issued as U.S. 2006/0269617); US order with Serial No. 10/563,713 (issued as U.S. 2006/0172020); US order with Serial No. 10/902,352 (issued as U.S. 2006/0024335); US order with Serial No. 10/538,038 (issued as U.S. 2006/0275765), U.S. with Serial No. 11/555,617 (issued as U.S. 2008/0187936); US order with Serial No. 12/739,264 (issued as U.S. 2010/0316736); US order with Serial No. 12/215,609 (issued as U.S. 2009/0042813); US order with Serial No. 11/893,088 (issued as U.S. 2008/0050499); Pat. U.S. At the. 7,807,204; US order with Serial No. 11/811,166 (issued as U.S. 2008/0003268); Pat. U.S. At the. 6,376,657; International Application Serial No. PCT/US2011/041183 (published as WO 2011/163183); International Application Serial No. PCT/EP2011/059814 (published as WO 2011/157692); US order with Serial No. 12/790,292 (issued as U.S. 2011/0293753); International Application Serial No. PCT/JP2009/071700 (published as WO 2010/076879); US order with Serial No. 13/032,530 (issued as U.S. 2011/0217394); International Application Serial No. PCT/EP2011/000110 (published as WO 2011/085979); International Application Serial No. PCT/US2010/058467 (published as WO 2011/068814); US order with Serial No. 13/060,996 (issued as U.S. 2011/0152361); US order with Serial No. 12/09,253 (issued as U.S. 2011/0124609); US order with Serial No. 12/687,962 (issued as U.S. 2011/0119499); and International Application Serial No. PCT/EP2010/004623 (published as WO 2011/012298); each of which is incorporated by reference in its entirety.

[0125] Administration and Methods

[0126] The biguanide compounds of the disclosure, including analogues, salts, solvates, polymorphs, hydrates, N-oxides and prodrugs of such compounds can be administered to a subject in need thereof to treat various metabolic disorders, including obesity, dyslipidemia or other disorders of lipid metabolism, as well as hyperglycemic conditions and histopathological diseases associated with hyperglycemia, including type II diabetes. Particularly in view of the surprising and unexpected dissociation of systemic bioavailability and therapeutic efficacy achieved herein and consequent improvement in toxicity and safety, the effective use of such compounds for prophylaxis and prevention of such diseases and disorders, as well as use for weight loss purposes More general weight is also explicitly contemplated in this document.

[0127] In preferred embodiments, the compound is metformin. Previous metformin formulations are reported to have an average systemic bioavailability of 30% to 60%, while many comparatively small molecules have bioavailability in excess of 60%. See, for example, Tucker et al., “Metformin kinetics in healthy subjects and in patients with diabetes mellitus” Br. J.Clin. Pharmacol. 1981, 12(2) 235-246. Notably, metformin administration increases plasma GLP-1 concentrations in normal, diabetic, and DPP-IV deficient rodents, as well as in humans with and without type II diabetes, but has been reported to do so indirectly and independently of a direct impact on intestinal L cells. Mulherin et al., supra.

[0128] As demonstrated herein, however, and contrary to well-established convention in the art, enteroendocrine activation by metformin can be triggered by luminal signals in the epithelial aspect of the intestine, and therefore, improvement of the systemic bioavailability of metformin is actually unnecessary after oral ingestion in order to stimulate the release of gastrointestinal hormones such as GLP-1. Likewise, the effective treatment of otherwise contraindicated patients is now made possible through the administration of compositions comprising biguanide compounds (including analogues, salts, solvates, polymorphs, hydrates, N-oxides and prodrugs thereof) tailored to minimize the systemic bioavailability of the compound. In preferred embodiments, the subject compositions and methods are formulated to minimize and preferably avoid an initial release in the stomach and/or proximal small intestine (areas of greatest absorption) in order to reduce systemic bioavailability after oral administration.

[0129] Delivery to Specific Intestinal Locations

[0130] The embodiments described in this document provide a method of treatment comprising administering a delayed-release composition comprising a biguanide compound (including any analogues, salts, solvates, polymorphs, hydrates, N-oxides or prodrugs thereof) formulated to be delivered to one or more sites in the small intestine and/or large intestine, and preferably the distal small intestine, in order to minimize systemic bioavailability by avoiding absorption in the stomach and proximal small intestine and corresponding rapid increase in Cmax-

[0131] Biguanide compounds target beyond the stomach to one or more regions of the small intestine and preferably target downstream or distal to the duodenum. In preferred embodiments, the compounds are delivered to the jejunum, ileum, cecum and colon or a combination thereof. In preferred embodiments, the compounds are delivered to the jejunum, ileum and cecum or a combination thereof. In preferred embodiments, the compounds are preferably targeted to the ileum. In further embodiments, the compound is delivered downstream or distally to the jejunum or to the large intestine alone.

[0132] In still other embodiments, the biguanide compound (including a respective analogue, salt, solvate, polymorph, hydrate, N-oxide or prodrug) is delivered to one or more regions of the upper intestine and one or more regions of the lower intestine. For example, the compound can be delivered to the duodenum and the colon. In another non-limiting example, the compound can be delivered to the duodenum, jejunum, ileum and colon.

[0133] The administration of biguanides such as metformin to the preferred regions or sites of the intestine can be accomplished by any known method. In preferred embodiments, the biguanide compound is formulated into a delayed-release composition for oral delivery that delivers the compound to targeted regions or sites of the intestine. When delivery of the biguanide compound is intended for two or more regions of the gastrointestinal tract, the compound can be delivered in any proportion and manner.

[0134] Minimizing systemic exposure

[0135] As described above, the methods disclosed herein minimize the systemic bioavailability of the biguanide compound in contraindicated patients. In some embodiments, biguanide compounds have reduced mean systemic bioavailability. In some embodiments, reduced mean systemic bioavailability is lower mean systemic bioavailability compared to an immediate-release or extended-release formulation having an equivalent amount of the biguanide compound. In other embodiments, reduced mean systemic bioavailability is when the mean systemic bioavailability is less than 30%, less than 25%, less than 15%, less than 10% and less than 5% compared to an immediate or extended release formulation having a equivalent amount of the biguanide compound. In certain cases, the mean systemic bioavailability is less than 15%.

[0136] In some embodiments, the subject methods minimize mean plasma Cmax and/or mean AUC levels of the biguanide compound in contraindicated patients. In some embodiments, the methods of administration result in minimal plasma absorption, mean Cmax and/or mean AUC levels of the biguanide compounds in the patient. In other embodiments, mean plasma Cmax, and/or mean AUC levels of the biguanide compound are considered subtherapeutic for the described compositions compared to the reported Cmax and/or AUC levels of conventional immediate-release and extended-release formulations. with identical amounts of metformin. For example, subtherapeutic or irrelevant metformin plasma Cmax and/or AUC levels include 75%, 60%, 50%, 40%, and 30% of the reported Cmax or AUC levels of known metformin formulations (e.g., GLUMETZA® , GLUCOPHAGE®, GLUCOPHAGE® XR, RIOMET®, FORTAMET®, OBIMET®, GLUFORMIN®, DIANBEN®, DIABEX®, DIAFORMIN®, Metformin IR®, Metformin SR® and the like).

[0137] In specific embodiments, the compositions and inventive methods directed to metformin produce a Cmax that is not more than 75% or 85%, preferably not more than 50% or 60%, more preferably not more than 25% or 30% or 40% of the same dose of an immediate-release metformin formulation (eg GLUCOPHAGE®) after oral ingestion. In other embodiments, the inventive methods provide a Cmax that is no more than 3x, more preferably no more than 2.5x, 2x, even more preferably no more than 1.8x or 1.5x the initial minimum plasma concentration. to 12 hours after the last oral intake of metformin. In other embodiments, the inventive compositions and methods provide for an average plasma AUC over the dosage range that is no more than 75% or 80%, preferably no more than 50% or 60%, most preferably no more than 25% , 30% or 40% of the same dose of an immediate-release formulation (eg, GLUCOPHAGE®) after oral ingestion.

[0138] Likewise, in specific embodiments, administration of the subject's delayed-release formulation minimizes mean plasma AUG, mean plasma Cmax, and/or circulating plasma concentration of the biguanide compound in contraindicated patients compared to a protocol identical by administering an IR or XR formulation having the same amount of the biguanide compound. In one embodiment, the average plasma AUCo20 of the biguanide compound resulting from administration is less than about 15,000 ng*hr/ml_ or 14,000 ng*hr/ml_, preferably less than about 12,000 ng*hr/ml_, 11,000 ng*hr/ml_ or 10,000 ng*hr/ml_, more preferably less than about 9,000 ng*hr/ml_, 8,000 ng*hr/ml_ or 7,000 ng*hr/ml_. In one embodiment, the resulting plasma mean Cmax of the biguanide compound is less than about 1000 ng/mL, preferably less than about 900 ng/mL or 800 ng/mL, more preferably less than about 700 ng/mL , 600 ng/ml or 500 ng/ml. In one embodiment, the resulting circulating plasma concentration of the biguanide compound is below about 5 µg/ml or 4 µg/ml, preferably below about 3 µg/ml or 2.5 µg/ml, most preferably below about about 2 μg/ml, 1 μg/ml, 0.5 μg/ml or 0.25 μg/ml in the patient. In preferred embodiments, the biguanide compound is metformin, the IR composition is Glucophage® and the XR composition is Glucophage® XR.

[0139] Formulations

[0140] To limit their systemic bioavailability, compositions comprising the biguanide compound are adapted for delayed release in order to minimize plasma absorption. Delivery of biguanide compounds such as metformin to enteroendocrine cells is by a known method, including, for example, oral, rectal, nasogastric tube injection, parenteral injection such as intraluminal intestinal injection. In preferred embodiments, oral dosage forms are administered. Oral delivery of biguanide compounds is described in the section on delayed release formulations and includes timed release systems, enteric coatings and pH dependent systems and the like. In some embodiments, compositions comprising compounds described herein utilize a multi-component system where the biguanide compound is delivered to various sites in the gastrointestinal tract such as the duodenum, jejunum, ileum, large intestine or combinations thereof after administration. For example, a delayed release formulation comprising the biguanide compound can be delivered to the lower intestine by use of timed or delayed (enteric) release components. Multi-component systems of such compounds can be in unit dosage forms such as double, triple or multi-layer slugs or multi-particle forms such as encapsulated micro-pellets, granules or as separate dosage forms, e.g. separate lozenges taken together or at periodic intervals.

[0141] In some embodiments, the delayed release formulation releases the biguanide compound after the emergence of a desired pH, due to the enteric coating. Contemplated pHs include about pH 6.0, more preferably about pH 6.5 and about pH 7.0. Once a desired pH is achieved, the compound begins to release. Such compositions may release the biguanide compound within about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, or about 30 minutes after the desired pH has been achieved, and/or may have timed, sustained, or slow release aspects that release the biguanide compound over a longer period, such as about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours or about 8 hours. An exemplary two-component delivery system may be, in some embodiments, a double layer lozenge. Three, four and additional components are contemplated within the embodiments.

[0142] For sustained release formulations comprising the biguanide compound, dosages of the compound can range from about 1 mg to about 2000 mg, about 10 mg to about 1500 mg, about 50 mg to about 1000 mg or about 100 mg or about 500 mg per day. In some cases, the dose of the compound is about 2000 mg, about 1500 about 1000 mg, about 800 mg, about 600 mg, about 500 mg, about 400 mg, about 300 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, about 75 mg, about 50 mg, about 25 mg, about 10 mg, or about 1 mg per day. In some embodiments, the dose of the compound is less than 400 mg. In some embodiments, the dose of the compound is 250 mg.

[0143] Biguanide compound salts include, but are not limited to hydrochloride, phosphate, sulfate, hydrobromide, salicylate, maleate, benzoate, succinate, ethanesulfonate, fumarate, glycolate, pyrantel, oratate, acetate, isobutyrate, acetylsalicylate, nicotinic acid, adamanthoate, zinc-chlorophyllin, carboxylic acid, benzoic acid, dichloroacetic acid, theophylline-7-acetate, clofibrate, tartate, oxalate, tannate and hydroxyl acid salts. In the preferred embodiment, the salt is metformin hydrochloride.

[0144] The biguanide compounds of the subject invention can advantageously be administered or combined with additional therapeutic agents, such as anti-obesity and/or anti-diabetic agents described herein. Notable agents for combinations with the metformin compositions described herein include DPP-IV inhibitors (e.g., sitagliptin, saxagliptin, berberine, vildagliptin, linagliptin, alogliptin, and the like), SGLT-2 and/or SGLT-1 inhibitors (e.g., , dapafloglizine, LX4211), agonists of GPR40, GPR120, GPR119, GPR41, GPR43, etc., thiazolidinedione (e.g., pioglitazone, rivoglitazone, rosiglitazone, troglitazone, and the like), sulfonylureas (e.g., glipzide, glibenclamide (glyburide), gliquidone , gliclopyramide, glimepiride, gliclazide, acetohexamide, carbutamide, chlorpropamide, tolbutamide, tolazamide and the like), dual PPAR modulators (aleglitazar, muraglitazar, tesaglitazar and the like), lipid lowering agents (eg, statins) and antihypertensive agents.

[0145] Formulations for the compositions provided herein include those suitable for oral or rectal administration, and administration, although the most suitable route may depend, for example, on the condition and disorder of the recipient. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods known in the art of pharmacy. All methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients.

[0146] Formulations suitable for oral administration may be presented as discrete units such as capsules, pills or lozenges, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.

[0147] Preparations of compositions that can be used orally include lozenges, hard capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Pastilles can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient into a free-flowing form, such as a powder or granules, optionally mixed with binders (e.g. povidone, gelatin, hydroxypropylmethyl cellulose), inert diluents, preservative , disintegrant (eg sodium starch glycollate, cross-linked povidone, cross-linked sodium carboxymethylcellulose) or lubricating agents, surface active or dispersing. Molded tablets can be made by molding, in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. The lozenges, optionally, can be coated or marked and can be formulated so as to provide a slow or controlled release of the active ingredient therein. Pellets may optionally be provided with an enteric coating, to provide for release in parts of the intestine other than the stomach. All formulations for oral administration must be in dosages suitable for such administration. The hard capsules can contain the active ingredients in admixture with the filler, such as lactose, binders, such as starches, and/or lubricants, such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycols. Also, stabilizers can be added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide and lacquer solutions, and suitable organic solvents or solvent mixtures. . Dyes or pigments can be added to tablet or tablet coatings for identification or to characterize different combinations of active compound doses.

[0148] It should be understood that, in addition to the ingredients particularly mentioned above, the compounds and compositions described herein may include other agents conventional in the art, taking into account the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.

[0149] The compositions described in this document may also contain the biguanide compound in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from, by way of non-limiting example, sweetening agents, flavoring agents , coloring agents and preserving agents, in order to provide pharmaceutically elegant and palatable preparations.

[0150] Delayed Release Formulations

[0151] Many strategies can be pursued to achieve delayed release in which the location of release is controlled in order to minimize systemic absorption. For example, delayed release can be achieved by appropriate selection of formulation parameters and ingredients (eg, appropriate controlled release compositions and coatings). Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles and liposomes. The release mechanism can be controlled such that the biguanide compounds are released at timed intervals or the release is controlled, the release of combined agents can be simultaneous, or a delayed release of the biguanide compound in a combination can be affected. when early release of a particular agent is preferred over another. Different delivery systems described herein may also be combined to deliver on a burst of various time intervals (e.g., about 30 minutes, about 120 minutes, about 180 minutes, and about 240 minutes after oral administration) or at different sites (e.g. release in the lower intestine, upper intestine, jejunum, ileum, cecum, colon and/or rectum) or a combination thereof. For example, a pH dependent system can be combined with a timed release system or any other system described in this document to achieve a desired release profile.

[00147] In certain preferred embodiments, the formula comprises an oral dosage form comprising a biguanide compound and having a delayed release system designed to include a lag phase of at least about 3, 5, 7 or 10 minutes prior to drug release at the desired pH and/or intestinal site, more preferably at least about 12, 15 or 18 minutes before drug delivery, even more preferably at least about 20, 30 or 60 minutes before drug delivery at the desired pH and/or intestinal site, for example, the small intestine, and preferably, the distal small intestine.

[00148] In certain embodiments, the biguanide compounds are provided in the form of a sustained release formulation together with a sustained release component of the biguanide compound and/or an additional therapeutic agent in a unit dosage form. The sustained release component can be formulated by any known method such as a layer surrounding a portion of the sustained release component or the like. Exemplary extended release ratios of an additional therapeutic agent to immediate release of a biguanide compound are about 10% XR to about 90% DR, about 15% XR to about 85% DR, about 20% XR to about about 80% DR, about 25% XR to about 75% DR, about 30% XR to about 70% DR, about 35% XR to about 65% DR, about 40% XR to about 60 %DR, about 45% XR to about 55% DR, or about 50% XR to about 50% DR. In certain embodiments, the extended release of an active agent to modified release of an active agent is about 25% XR to about 75% DR. In certain embodiments, the sustained release of an active agent to modified release of an active agent is from about 20% XR to about 80% DR. Unit dosage forms with an XR and DR component include any known formulation, including bilayer tablets, coated pellets and the like.

[0149] In certain embodiments, the biguanide compounds are provided in the form of a delayed-release formulation together with an immediate-release component of an additional therapeutic agent in a unit dosage form. The immediate release component can be formulated by any known method, such as a layer surrounding the delayed release component or the like. Exemplary immediate release ratios of an additional therapeutic agent to delayed release of a biguanide compound are from about 10% IR to about 90% DR, 15% IR to about 85% DR, from about 20% IR to about from 80% DR, about 25% RI to about 75% DR, about 30% RI to about 70% DR, about 35% RI to about 65% DR, about 40% RI to about 60 % DR, about 45% RI for about 55% DR, or about 50% RI for about 50% DR. In certain embodiments, the immediate release of an active agent to the delayed release of an active agent is from about 25% IR to about 75% DR. In certain embodiments, the immediate release of an active agent to the delayed release of an active agent is from about 20% IR to about 80% DR. Unit dosage forms with an IR and DR component include any known formulation, including bilayer tablets, coated pellets and the like.

[0150] Enteric Coatings and pH Dependent Systems

[0151] The formulation may also be coated with an enteric coating, which protects an active agent, for example a biguanide compound, from degradation in an acidic environment, such as the stomach, and allows for a delayed release to a target area , for example, the ileum, for absorption.

[0152] The enteric coating may be, as a non-limiting example, wax or wax-like substance, such as carnauba wax, fatty alcohols, hydrogenated vegetable oils, zein, shellac, sucrose, gum arabic, gelatin, dextrin , psyllium husk powder, polymethacrylates, anionic polymethacrylates, blends of poly(methacrylic acid, methyl methacrylate), polymers or copolymers derived from acrylic and/or esters of methacrylic acid, cellulose acetate phthalate, cellulose acetate trimelliate, phthalate hydroxypropyl methylcellulose (HPMCP), cellulose propionate phthalate, cellulose acetate maleate, polyvinyl alcohol phthalate, hydroxypropyl methylcellulose acetate succinate (HPMCAS), hydroxypropyl methylcellulose hexahydrophthalate, polyvinyl acetate phthalate, poly(acid) blends methacrylic, ethyl acrylate), ethylcellulose, methylcellulose, propylcellulose, chitosan succinate, polyvinyl acetate phthalate (PVAP), carboxymethylethyl cellulose of polyvinyl acetate polymers and compatible mixtures thereof. Furthermore, an inactive intermediate film can be provided between the biguanide compound, for example, and the enteric coating to prevent interaction of the biguanide compound with the enteric coating.

[0153] In a non-limiting example, silicone microspheres for pH-controlled delivery of gastrointestinal drugs have been described by Carelli et al., int. J. Pharmaceutics 179: 73-83, 1999. The microspheres are semi-interpenetrating, pH-sensitive polymer hydrogels made of varying proportions of poly(methylmethacrylate-co-methacrylic acid) (EUDRAGIT® L100 or EUDRAGIT® S100) and cross-linked polyethylene glycol 8000 that are encapsulated in silicone microspheres. The EUDRAGIT® series of methacrylic acid copolymers is commercially available from the Evonik Industries factory in Darmstadt, Germany.

[0154] Enteric coatings can be formulated to release a biguanide compound at a desired pH using combinations of enteric polymers. It is known that different sites in the gastrointestinal system have specific pHs. For example, the duodenum may correspond to a pH 5.5 environment and the jejunum may correspond to a pH 6.0 environment. In preferred embodiments, the enteric coatings are formulated to release the compound upon emergence of a desired pH, for example, in the distal small intestine and lower intestine, i.e., at pH 6, at pH 6.5, or at a pH 7. In multi-release modalities, enteric coatings are formulated to release on a surge of two or more pH values. In certain embodiments, enteric coatings are formulated to release at an onset of pH 5.5, 6.0, 6.5 and 7.0. In certain embodiments, the enteric coatings are formulated to release at a pH range of 6.5 and 7.0. In certain embodiments, enteric coatings are formulated to release in the jejunum, ileum and lower intestine. In still other embodiments, enteric coatings are used in combination with other alternative delivery systems, such as a timed release system.

[00155] In certain embodiments, the enteric coating is applied for oral dosing from a thickness of at least about 4.5 mg/cm2, 5 mg/cm2, 5.5 mg/cm2, 6 mg/cm2, 6.5 mg/cm2, 7 mg/cm2, 7.5 mg/cm2, 8 mg/cm2, 9 mg/cm2, 10mg/cm2, 11 mg/cm2, 15 mg/cm2, or 20 mg/ cm2. For tablets and capsules, enteric coating can be applied to achieve about 2.5% to about 5%, 6%, 7%, 8%, 9%, 10% or 12% (w/w) gain in weight, more preferably about 3.0% to about 6% (w/w) weight gain, even more preferably at least about 3.5% or 4% (w/w) weight gain. For granules and other multiparticulate dosage forms up to 20 or 30% (w/w) weight gain or more may apply, preferably from about 20% - 50% (w/w), more preferably from of carcass from 30% - 50% (w/w). As demonstrated herein, this ensures adequate delivery at the desired intestinal site in order to avoid abnormal spikes in systemic exposure of the biguanide compound.

[00156] In other embodiments, pharmaceutical compositions are provided comprising an enteric coated oral dosage form comprising a biguanide compound, wherein the dosage form is adapted to minimize biguanide release for a latency phase of at least about 5 to 10 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for a lag phase of at least about 15 or 20 minutes after contact with the desired pH, even more preferably for a lag phase of at least about 25 or 30 minutes after contact with a pH of 6.0, 6.5, 6.8 or 7.0. In one embodiment, an enteric coating is applied to the pharmaceutical composition at a weight gain of at least about 4.5 mg/cm2, 5.5 mg/cm2, 6 mg/cm2, 6.5 mg/cm2, cm2, 7 mg/cm2, 7.5 mg/cm2, 8 mg/cm2, 9 mg/cm2, 10 mg/cm2, 11 mg/cm2, 12 mg/cm2, or 15 mg/cm2. In another embodiment, the enteric coating is applied at a weight gain of at least about 5 mg/cm 2 to 9.5 mg/cm 2 , more preferably at about 5.5 mg/cm 2 to about at least 7. 6 mg/cm2. In alternative embodiments, an enteric coating is applied to the pharmaceutical composition to achieve at least about 3.0% to at least about 7.0% (w/w) weight gain, more preferably at least about 4% to at least about 4% least about 6% (w/w) in weight gain.

[00157] In specific embodiments, pharmaceutical compositions are provided comprising an enteric coated oral dosage form comprising a biguanide compound, wherein the dosage form is adapted to release less than about 10%, 5%, 4%, 3 %, 2%, and preferably less than 1% of the biguanide compound after contact with an aqueous medium (e.g., submersion) at a pH of less than about 2 for about one hours followed by contact with an aqueous medium at a pH equal to or less than about 5.5 for at least 30 or 45 minutes. In a preferred embodiment, the enteric coated dosage form releases less than about 5%, 2% or 1% of the biguanide compound in a 0.1 N HCI aqueous medium per two hours and less than about 5%, 2 % or 1% when transferred to an aqueous medium at pH 5.5 for at least 30 to 45 minutes.

[00158] In additional embodiments, the enteric coated dosage form releases less than 15%, 10%, 5%, 3%, 2% or less than 1% of the biguanide compound during the lag phase after the dosage form enters in contact with an aqueous medium at a pH of about 6.5 or 6.8, wherein the lag phase is at least ten, fifteen or twenty minutes. In a preferred embodiment, the enteric coated dosage form releases less than about 15% of the biguanide compound when the dosage form is contacted with an aqueous medium at a pH of 6.5 to 6.8 for a phase. of at least ten minutes lag and releases from about 75% to about 100%, and more preferably greater than 90%, 95%, 98% or 99% of the biguanide compound after contacting an aqueous medium in a Carcass pH 6.5 or 6.8 for a total of 90 to 120 minutes.

[00159] In two-stage dissolution embodiments, pharmaceutical compositions are provided comprising an enteric-coated oral dosage comprising a biguanide compound, wherein the dosage form is adapted to release less than 5%, 2% or 1% of the biguanide compound in an aqueous medium of 0.1 N HCI for two hours. In these embodiments, less than 15%, 10%, 5%, 3%, 2% or preferably 1% of the biguanide compound is released after contacting an aqueous medium of 0.1 N HCI for two hours and subsequently transferred to an aqueous medium at a pH of about 6.8 for a lag phase of at least ten, fifteen or twenty minutes. In preferred embodiments, less than 15% of the biguanide compound is released after two hours at acidic pH and a lag phase of at least ten or fifteen minutes at pH 6.8, and at least 60% of the biguanide compound is released after lag phase, and within 60 minutes at pH 6.8, and at least 90% of the biguanide compound is released within 90 to 120 minutes at pH 6.8.

[00160] In three-stage dissolution embodiments, pharmaceutical compositions are provided comprising an enteric-coated oral dosage comprising a biguanide compound, wherein the dosage form is adapted to release less than 5%, 2% or 1% of the biguanide compound in an aqueous medium of 0.1 N HCl for two hours and less than 5%, 2%, or 1% when transferred to an aqueous medium at pH 5.5 for at least one hour. In these embodiments, less than 25%, 20%, 15%, 10%, or 5% of the biguanide compound is released after two hours in an aqueous medium of 0.1 N HCl, 30 minutes in an aqueous medium at pH 5.5, and during a lag phase of at least ten or fifteen minutes at pH 6.8. In preferred embodiments, less than 15%, 10% or 5% of the biguanide compound is released after two hours at acidic pH, 30 minutes at pH 5.5 and a lag phase of at least ten or fifteen minutes at pH 6, 8, and at least 60% of the biguanide compound is released after the lag phase, and within 60 minutes at pH 6.8, and at least 90% of the biguanide compound is released within 90 to 120 minutes at pH 6.8 .

[00161] In alternative embodiments, the subject formulas and compositions further comprise one or more disintegrants to accelerate dissolution of the core upon disruption of the enteric coating. In preferred embodiments, the disintegrant comprises croscarmellose sodium, sodium starch glycollate or combinations thereof.

[00162] In additional embodiments, the formulas and compositions of the object may further comprise a sealing coating between the biguanide compound and the enteric coating, providing a total coating thickness corresponding to at least about 4% to 8% (w/ p) in weight gain, more preferably at least about 4.5% to 6.0% (w/w) in weight gain. In some embodiments, the combination of the outer enteric coating and inner coating comprises at least about 6.9 mg/cm 2 to 13.3 mg/cm 2 , more preferably at least about 7.8 mg/cm 2 to at least about 11.4 mg/cm 2 . mg/cm2.

[00163] In some embodiments, the enteric coating comprises a first polymer that minimizes the release of the biguanide compound for at least about 5 to 10 minutes after coming into contact with a pH of 6.0, 6.5, 6.8 or 7.0, more preferably for at least about 15 or 20 minutes, even more preferably for at least about 25, 30, 45 or 60 minutes after coming into contact with a pH of 6.0, 6.5, 6, 8 or 7.0. In preferred embodiments, the polymer is insoluble in an acid medium, but dissolves upon salt formation or the like above pH 7.0. In a preferred embodiment, the polymer is Eudragit FS or Eugragit S.

[00164] In additional embodiments, the enteric coating comprises a second polymer that dissolves at a lower pH than the first polymer. In preferred embodiments, the second polymer is insoluble at pH 5.5 and below, but dissolves by salt formation or the like above pH 5.5. In a preferred embodiment, the second polymer is Efragit L. In some embodiments, Eudragit L is replaced with cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate (hypromellose acetate succinate), hydroxypropyl methylcellulose acetate phthalate (PVAP) and sodium alginate, stearic acid, and/or combinations thereof.

[00165] In some embodiments, the enteric coating comprises about 90% Eudragit FS and about 10% Eudragit L, about 80% Eudragit FS and about 20% Eudragit L, about 70% Eudragit FS and about 30% Eudragit L, about 60% Eudragit FS and about 40% Eudragit L, about 50% Eudragit FS and about 50% Eudragit L, about 40% Eudragit FS and about about 60% Eudragit L, about 30% Eudragit FS and about 70% Eudragit L, about 20% Eudragit FS and about 80% Eudragit L, or about 10% Eudragit FS and about 90% Eudragit L. In preferred embodiments, Eudragit FS and said Eudragit L are present in about 7:5 to about 5:7 ratios, and more preferably, from 6:4 to about 4:6 ratios . In an exemplary preferred embodiment, the enteric coating comprises about 60% Eudragit FS and about 40% Eudragit L.

[00166] In some embodiments, the sealing coating can be added between the biguanide compound and the enteric coating. The sealing liner material can be selected such that it has no effect on drug release. Suitable materials include, for example, hydroxypropylmethylcellulose (HPMC). In other embodiments, the sealing coating material can be selected to extend the lag phase, slowing drug release after the enteric coating ruptures. Suitable materials include, for example, Eudragit E which dissolves in acid but swells at higher pH and can be used to extend the lag phase after enteric coating rupture.

[0168] In some embodiments, the sealing coating is a mixture of hypromellose, titanium dioxide, polyethylene glycol 400 (macrogol) and polysorbate 80, where hypromellose is the polymeric coating, titanium dioxide is a coloring agent, polyethylene glycol 400 serves as an anti-caking agent and polysorbate 80 is present as a dispersant (in aqueous suspension) and plasticizer. In other embodiments, the sealing coating is a mixture of hypromellose, triacetin and talc, where hypromellose is the polymeric coating, triacetin is present as a plasticizer and talc is present as an anti-tack agent. In some embodiments, the sealing coating is Opadry® White YS-1-7003 (Colorcon). In other embodiments, the sealing coating is Opadry® 03K19229 Clear.

[0169] Microcapsule gastroretentive systems described in Pat. U.S. Us. 6,022,562, 5,846,566 and 5,603,957 may be used in the delayed release delivery methods described herein. Microparticles of an active agent or drug are coated by spraying with a material consisting of a mixture of a film-forming polymeric derivative, a hydrophobic plasticizer, a functional agent and a nitrogen-containing polymer. The resulting microcapsules are less than or equal to 1000 microns (gm) in size and in certain cases such microcapsules are between 100 and 500 microns. These microcapsules remain in the small intestine for at least 5 hours.

[0170] Film-forming polymer derivatives used in such microcapsules include, but are not limited to, ethylcellulose derivatives, cellulose acetate, and non-water soluble cellulose. Nitrogen-containing polymers include, but are not limited to, polyacrylamide, poly-N-vinylamide, poly-N-vinyl-lactam and polyvinylpyrrolidone. The plasticizer used in such a microcapsule includes, but is not limited to, glycerol esters, italates, citrates, sebacates, cetyl alcohol esters, castor oil and cutin. The lubricating or surface active agent used in such a lubricating microcapsule includes, but is not limited to, anionic surfactants, such as, by way of example, the alkali metal or alkaline earth metal salts of fatty acids, stearic acid and/or acid. oleic, nonionic surfactants, such as, by way of example, polyoxyethylenated sorbitan esters and/or polyoxyethylenated sorbitan esters and/or polyoxyethylenated castor oil derivatives; and/or lubricants such as stearates, such as, by way of example, calcium, magnesium, aluminum stearate, zinc stearate, stearyl fumarate, sodium stearyl fumarate and glyceryl behenate.

[0171] A non-limiting example of a lower Gl delivery formulation comprises a tablet for lower Gl delivery. The internal composition of the lozenge comprises about 0.01% by weight of about 10.0% by weight of a suitable active ingredient; about 50% by weight to about 98% by weight of a hydrocolloid gum obtainable from higher plants; and about 2% by weight to about 50% by weight of a pharmaceutically acceptable excipient, such as a binder. Other optional materials may be present that will help establish the desired characteristics of the pharmaceutical composition. These include materials that can enhance absorption of the active ingredient in the lower Gl, can protect the active ingredient from degradation, can prevent dissolution, and the like. Optionally surrounding the inner composition of the tablet is a coating which is preferably an enteric polymeric material.

[0172] In one embodiment, the formula comprises an excipient from the group consisting of sodium starch gluconate, povidone, corn starch, colloidal silicon dioxide, magnesium stearate, hypromellose, polyethylene glycol and combinations thereof. In one embodiment, the formula comprises sodium starch gluconate, povidone, corn starch, colloidal silicon dioxide and magnesium stearate as excipients. In another embodiment, the formula comprises povidone, magnesium stearate, hypromellose and polyethylene glycol as excipients.

[0173] The formulation is designed to take advantage of (1) the protective characteristics of the hydrocolloid obtainable from higher plants in the upper Gl and (2) the disintegrative characteristics of the hydrocolloid in the lower Gl. Thus, the internal composition of the lozenge can be one of several designs: (a) it can be a matrix of a therapeutically effective amount of the active ingredient uniformly dispersed in combination with a high percentage of the hydrocolloid and a generally smaller amount of other excipients; (b) it may have a core, in which the active principle is concentrated, surrounded by a layer of material which is free of the active principle and which has a high percentage of the hydrocolloid and a generally smaller amount of other excipients; (c) may have a concentration gradient of the active ingredient such that there is a greater amount in the core of the tablet with smaller amounts in multiple layers surrounding the core and very little or no active ingredient in the outer layer. If the insert design is (a), (b) or (c) above, specificity for regional delivery to the lower Gl is enhanced by enteric coating the insert with an appropriate enteric coating material.

[0174] Suitable hydrocolloids are well known in the art. See, for example, "The Chemistry of Plant Gums and Mucilages" by Smith and Montgomery from the A.C.S. Monograph series, #141, 1959, Reinhold Publishing Co. and the 18th Edition of The Merck Index. In general, the amount of hydrocolloid that will be used is an amount that allows the composition to travel through the upper GI tract without significant disintegration and without releasing a significant amount of active principle in the upper GI tract, i.e., providing a delayed release profile. Generally, this amount of hydrocolloid will be greater than about 50% but less than about 98%. Depending on individual variability, whether a patient ate or fasted, and other factors, a lozenge will travel through the stomach and upper intestinal tract in about 3 to 6 hours. During this time, little active principle (less than 20%, preferably less than 10%) is released from the tablet of this invention. Once the tablet reaches the lower Gl, the release of the active principle is triggered by enzymatic degradation of the galactomannan gum.

[0175] Timed Release Systems

[0176] In one embodiment, the delayed-release mechanism is a "timed" release or time-release ("LT") system that releases an active agent, e.g., a biguanide compound, at certain times after administration. Timed release systems are well known in the art and suitable timed release systems can include any known excipient and/or coating. For example, excipients in a matrix, layer, or coating can delay the release of an active agent by slowing the diffusion of the active agent into an environment. Suitable timed release excipients include, but are not limited to, acacia (gum arabic), agar, magnesium aluminum silicate, alginates (sodium alginate), sodium stearate, sea bean, bentonite, carbomer, carrageenan, carbopol, cellulose, microcrystalline cellulose, keratonia, chondrus, dextrose, furcellaran, gelatin, ghatti gum, guar gum, galactomannan, hectorite, lactose, sucrose, maltodextrin, mannitol, sorbitol, honey, corn starch, wheat starch, starch rice, potato starch, gelatin, sterculia gum, xanthan gum, glyceryl behenate (e.g. Compritol 888 ato), glyceryl distearate (e.g. Precirol act 5), polyethylene glycol (e.g. PEG 200-4500), oxide polyethylene, adipic acid, gum tragacanth, ethylcellulose (e.g. ethylcellulose 100), ethylhydroxyethyl cellulose, ethylmethyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxyethylmethyl cellulose (e.g. KIOOLV, K4M, KI5M), hydroxypropyl cellulose, poly(methacrylate) hydroxyethyl), cellulose acetate (e.g., cellulose acetate CA-398-10 NF), cellulose acetate phthalate, cellulose acetate propionate, cellulose acetate butyrate, methyl cellulose acetate hydroxypropyl succinate, hydroxypropyl phthalate methyl cellulose, cellulose butyrate, cellulose nitrate, oxypolygelatin, pectin, polygeline, povidone, propylene carbonate, polyanhydrides, methyl vinyl ether/maleic anhydride copolymer (PVM/MA), polymethoxyethyl methacrylate), poly(methoxyethoxyethyl methacrylate) ), hydroxypropyl cellulose, hydroxypropylmethyl cellulose, sodium carboxymethylcellulose (CMC), silicon dioxide, vinyl polymers, e.g. polyvinylpyrrolidones (PVP: povidone), polyvinyl acetates or polyvinyl acetate phthalates and mixtures, Kollidon SR, acrylic derivatives (e.g. polyacrylates, e.g. cross-linked polyacrylates, methicrylic acid copolymers), Splenda® (dextrose, maltodextrin and sucralose) or combinations thereof. The timed-release excipient can be in a matrix with the active agent, in another compartment or layer of the formulation, as part of the coating, or any combination thereof. Varying amounts of one or more timed release excipients can be used to achieve a designated release time.

[0177] A non-limiting example includes TIMERx® system formulations. This controlled release formulation system provides altered time release (SyncroDoseTM) as well as biphasic release (Geminex®). (See, for example, Staniforth & Baichwal, TIMERx®: novel polysaccharide composites for controlled/programmed release of active ingredients in the gastrointestinal tract, Expert Opin. Drug Deliv., 2(3): 587-89 (2005)). Using formulations such as these for the invention described herein, compositions can be created that target the upper gastrointestinal tract, lower gastrointestinal tract, or both, in addition to temporarily controlling the release of such compounds at either location.

[0178] In some embodiments, timed release systems are formulated to release the compound in a burst of about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 60 minutes, about 70 minutes, about 80 minutes, about 90 minutes, about 100 minutes, about 110 minutes, about 120 minutes, about 130 minutes, about 140 minutes, about 150 minutes, about 160 minutes, about 170 minutes, about 180 minutes, about 190 minutes, about 200 minutes, about 210 minutes, about 220 minutes, about 230 minutes, about 240 minutes, about 250 minutes, about 260 minutes, about 270 minutes, about 280 minutes, about 290 minutes, about 300 minutes, about 310 minutes, about 320 minutes, about 330 minutes, about 340 minutes, about 350 minutes, about 360 minutes, about 370 minutes, about 380 minutes, about 390 minutes, about 400, about 400, about 410, or about 420 minutes after administration. In multiple release modalities, timed delivery systems are formulated to deliver at more than one time point. In certain embodiments, timed release systems are formulated to release in a burst of about 10 minutes, about 30 minutes, and about 120 minutes, about 180 minutes, and about 240 minutes after administration. In certain embodiments, timed release systems are formulated to release in a burst of about 5 to about 45 minutes, about 105 to about 135 minutes, about 165 to about 195 minutes, about 225 to about 255 minutes or a combination of the respective times after administration to a patient.

[0179] Modified Release Formulations

[0180] In a further embodiment, methods and compositions directed at delivering the biguanide compound may further employ controlled, sustained, or extended release formulations known collectively as "modified release" formulations. Compositions can be administered via modified release systems or by delivery devices that are well known to those of ordinary skill in the art. Examples include, but are not limited to, those described in U.S. Pat. U.S. Us. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; and 5,733,566. These dosage forms can be used to provide modified release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres or a combination thereof. to provide the desired release profile, in varying proportions. Suitable modified release formulations known to those of ordinary skill in the art, including those described herein, can be easily selected for use with the active ingredients of the invention. The invention thus encompasses single unit dosage forms suitable for oral administration such as, but not limited to, tablets, capsules, gelcaps and capsules that are further adapted for modified release.

[0181] In some embodiments, modified release systems are formulated to release the compound in a duration of about 30 minutes, about 40 minutes, about 50 minutes, about 60 minutes, about 70 minutes, about 80 minutes, about 90 minutes, about 100 minutes, about 110 minutes, about 120 minutes, about 130 minutes, about 140 minutes, about 150 minutes, about 160 minutes, about 170 minutes, about 180 minutes , about 190 minutes, about 200 minutes, about 210 minutes, about 220 minutes, about 230 minutes, about 240 minutes, about 250 minutes, about 260 minutes, about 270 minutes, about 280 minutes , about 290 minutes, about 300 minutes, about 310 minutes, about 320 minutes, about 330 minutes, about 340 minutes, about 350 minutes, about 350 minutes, about 360 minutes, about 370 minutes , about 380 minutes, about 390 minutes, about 400, about 400, about 410, or about 420 minutes right after the release started. In multiple-release embodiments, modified release systems are formulated to deliver over more than one time duration at different time points.

[0182] In a non-limiting example, chitosan and mixtures of chitosan with carboxymethylcellulose sodium (CMC-Na) have been used as vehicles for the sustained release of active ingredients, as described by Inouye et al, Drug Design and Delivery 1: 297 -305, 1987. Mixtures of these compounds and agents from the combinations of the invention, when compressed below 200 kg/cm 2 , form a slug from which the active agent is slowly released after administration to a patient. The release profile can be altered by varying the proportions of chitosan, CMC-Na and active agent(s). Lozenges may also contain other additives, including lactose, CaHPO4 dihydrate, sucrose, crystalline cellulose or croscarmellose sodium.

[0183] In another non-limiting example, Baichwal, in U.S. Pat. U.S. No. 6,245,356 describes solid, oral, sustained release dosage forms that include agglomerated particles of a therapeutically active drug in amorphous form, a gelling agent, an ionizable gel strength enhancing agent, and an inert diluent. The gelling agent may be a mixture of a xanthan gum and locust bean gum capable of cross-linking with the xanthan gum when the gums are exposed to an ambient fluid. Preferably, the ionizable gel reinforcing agent acts to reinforce the strength of the hinged bond between the xanthan gum and the locust bean gum and thereby prolong the release of the drug component from the formulation. In addition to xanthan gum and locust bean gum, acceptable gelling agents that may also be used include those gelling agents well known in the art. Examples include natural or modified natural gums such as alginates, carrageenan, pectin, guar gum, modified starch, hydroxypropylmethylcellulose, methylcellulose and other cellulosic materials or polymers, such as, for example, sodium carboxymethylcellulose and hydroxypropyl cellulose and mixtures thereof.

[0184] In another non-limiting formulation useful for the combinations of the invention, Baichwal and Staniforth in U.S. Pat. U.S. No. 5,135,757 describes a free-flowing slow release granulation for use as a pharmaceutical excipient that includes from about 20 to about 70 percent or more by weight of a hydrophilic material that includes a heteropolysaccharide (such as, for example, xanthan gum or a derivative thereof) and a polysaccharide material capable of cross-linking the heteropolysaccharide (such as, for example, galactomannans and more preferably locust bean gum) in the presence of aqueous solutions, and from about 30 to about 80 percent by weight of an inert pharmaceutical filler (such as, for example, lactose, dextrose, sucrose, sorbitol, xylitol, fructose or mixtures thereof). After mixing the excipient with a tricyclic compound/corticosteroid combination or combination agent of the invention, the mixture is directly compressed into solid dosage forms, such as tablets. The lozenges thus formed slowly release the drug when swallowed and exposed to gastric fluids. By varying the amount of excipient in relation to the drug, a slow release profile can be achieved.

[0185] Slow-release formulations may also include a coating that is not readily soluble in water, but which is slowly attacked and removed by water, or through which water can slowly permeate. Thus, for example, the combinations of the invention can be spray coated with a solution of a binder under continuously fluidizing conditions, such as described by Kitamori et al., U.S. Pat. U.S. At the. 4,036,948. Examples of water soluble binders include pregelatinized starch (e.g. pregelatinized corn starch, pregelatinized white potato starch), pregelatinized modified starch, water soluble celluloses (e.g. hydroxypropyl cellulose, hydroxymethyl -cellulose, hydroxypropylmethylcellulose, carboxymethylcellulose), polyvinylpyrrolidone, polyvinyl alcohol, dextrin, gum arabic and gelatin, binders soluble in organic solvents such as cellulose derivatives (e.g. cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, ethylcellulose).

[0186] In another non-limiting example, Villa et al., in U.S. Pat. No. 6,773,720 describes a modified release system containing an inner lipophilic matrix where an active ingredient is engulfed and an outer hydrophilic matrix in which the lipophilic matrix is dispersed. An active ingredient, such as a biguanide or related heterocyclic compound, is first encapsulated in a low-melt lipophilic excipient or excipient mixture while undergoing heating to soften and/or melt the excipient itself, thereby incorporating the active ingredient. by simple dispersion. Upon cooling to room temperature, an inert matrix forms, which can be reduced in size to obtain matrix granules containing the active ingredient particles. The inert matrix granules are further blended together with one or more hydrophilic water-swellable excipients. In this regard, when the composition is in contact with biological fluids, a swollen layer of high viscosity is formed, which coordinates the solvent molecules and acts as a barrier to the penetration of the aqueous fluid itself within the new structure. Said barrier antagonizes the "disruption effect" caused by the dissolution of the active ingredient engulfed within the inert matrix, which is in turn within the hydrophilic matrix. A commercially available system of this type is from Cosmo Technologies Limited (Italy), under the trade name MMX® technology. Lipophilic/hydrophilic matrices can additionally be enteric coated for specific pH delivery.

[0187] Formulations for upper intestinal delivery, lower intestinal delivery or both are known in the art. Destination of active ingredients to various regions of the gut is described, for example, in The Encyclopedia of Pharmaceutical Technology, by James Swarbrick and James Boylan, Informa Health Care, 1999, on pgs. 287-308. Any formulation suitable for gastrointestinal delivery for site-specific delivery and/or time-specific delivery (i.e., delayed, controlled, extended, or sustained release) can be used with the invention and is contemplated herein.

[0188] Any of the delivery systems described in this document can be used in combination with others to achieve multiple releases and/or specific release profiles. In some embodiments, the biguanide compound is in a formulation that achieves multiple releases at gastrointestinal sites after administration. In certain embodiments, the biguanide compound is in a multiple-release formulation that releases at an onset of about 10 minutes, about 30 minutes, about 120 minutes, about 180 minutes, about 240 minutes, or combinations of the respective times. after administration. In certain embodiments, the biguanide compound is in a multiple-release formulation that releases at an onset of about 5 to about 45 minutes, about 105 to about 135 minutes, about 165 to about 195 minutes, about 225 to about 255 minutes or a combination of the respective times after administration.

[0189] In certain embodiments, the biguanide compound is in a multiple-release formulation that releases from the jejunum, ileum, lower intestine, or combinations thereof, after administration. In still other embodiments, the biguanide compound is in a multiple-release formulation that releases at an onset of about pH 6.0, about pH 6.5, about pH 7.0, or combinations thereof, after administration. In still other embodiments, the biguanide compound is in a multiple-release formulation that releases at ranges from about pH 6.0 to about pH 7.0, about pH 7.0 to about pH 8.0, or combinations of the respective times after administration.

[0190] Oral Dosage Forms

[0191] Oral dosage forms suitable for use in the subject methods and compositions include tablets, hard capsules, push-fit capsules made of gelatin, as well as sealed capsules made of gelatin and a plasticizer, such as glycerol and sorbitol, as well as troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, syrups or elixirs. Appropriate oral dosage forms may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from, by way of non-limiting example, sweetening agents, flavoring agents, colorants and preserving agents, in order to provide pharmaceutically elegant and palatable preparations.

[0192] Tablets contain the active ingredient in admixture with pharmaceutically acceptable excipients, which are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binding agents, for example starch, gelatin, polyvinyl pyrrolidone or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. Pastilles can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form, such as a powder or granules, optionally mixed with binders (e.g. povidone, gelatin, hydroxypropylmethyl cellulose), inert diluents, preservative, disintegrants (eg, sodium starch glycollate, cross-linked povidone, cross-linked sodium carboxymethylcellulose) or lubricating, surface active or dispersing agents. Molded tablets can be made by molding, in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. Tablets are coated using known techniques in order to delay disintegration and absorption in the gastrointestinal tract and thus minimize systemic bioavailability, as described more fully herein.

[0193] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or soft gelatin capsules, in that the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oil medium, for example peanut oil, liquid paraffin or olive oil. Alternatively, push-fit capsules can contain the active ingredients in admixture with the fill, such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycols. Also, stabilizers can be added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide and lacquer solutions, and suitable organic solvents or solvent mixtures. . Dyes or pigments can be added to tablet or tablet coatings for identification or to characterize different combinations of active compound doses.

[0194] It should be understood that, in addition to the ingredients particularly mentioned above, the compounds and compositions described herein may include other agents conventional in the art, taking into account the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.

[0195] In various embodiments, the compositions provided in this document are in liquid form. Liquid forms include, by way of example and not limitation, neat liquids, solutions, suspensions, dispersions, colloids, foams and the like. In certain cases, liquid forms also contain a nutritional component or base (eg dairy, yogurt, juice or shake). In some aspects, the compounds are micronized or as nanoparticles in liquid form. In certain cases, compounds may be coated to mask taste properties. In other examples, the compounds are coated to modify delivery to the distal small intestine and colon.

[0196] Aqueous solutions or suspensions contain the active ingredient(s) in admixture with suitable excipients for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacane and gum acacia; dispersing or wetting agents can be a naturally occurring phosphatide, e.g. lecithin, or condensation products of an alkylene oxide with fatty acids, e.g. polyoxyethylene stearate or condensation products of ethylene oxide with chain aliphatic alcohols long, for example heptadecaethylene-oxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and anhydrides of hexitol, for example polyethylene sorbitan monooleate. The aqueous solutions or suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame. In certain cases, the flavoring agents are the compounds.

[0197] Oily suspensions can be formulated by suspending the active ingredient(s) in a vegetable oil, for example, arachis oil, olive oil, sesame oil or coconut oil, or in mineral oil such like liquid paraffin. Oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those defined above and flavoring agents can be added to provide a palatable oral preparation. These compositions can be preserved by the addition of an antioxidant such as butylated hydroxyanisole or alpha-tocopherol.

[0198] Dispersible powders and granules suitable for preparing an aqueous solution or suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already cited above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

[0199] Compositions may also be in the form of an oil-in-water emulsion. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifying agents may be naturally occurring phosphatides, for example soy lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate and condensation products of said partial esters with ethylene oxide, for example for example, polyoxyethylene sorbitan monooleate. Emulsions can also contain sweetening agents, flavoring agents, preservatives and antioxidants.

[0200] Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, flavoring agents, coloring agents and antioxidants.

[0201] The compositions may also be formulated into rectal compositions, such as suppositories or retention enemas, for example, containing conventional suppository bases, such as cocoa butter, polyethylene glycol, or other glycerides. These compositions can also be prepared by mixing the inhibitors with a suitable non-irritating excipient which is solid at normal temperatures but liquid at the rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, glycerin gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and polyethylene glycol fatty acid esters.

[0202] Accordingly, pharmaceutical compositions comprising the biguanide compound in a sustained-release formulation suitable for oral administration, such as a tablet, capsule, wafer, lozenge, powder or granule, solution, liquid or suspension, are also provided. The pharmaceutical composition is preferably in unit dosage form suitable for single administration of precise dosages, for example 100mg, 200mg, 250mg, 300mg, 400mg, 500mg, 600mg, 750mg, 800 mg or 1000 mg of the desired biguanide compound, particularly metformin, phenformin, buformin or imeglimine or a salt thereof. The pharmaceutical composition may comprise a conventional pharmaceutical carrier or excipient and the compound according to the invention as an active ingredient. They may further comprise other medical or pharmaceutical agents, carriers, adjutants, etc.

[0203] Suitable carriers include inert diluents or fillers, water and various organic solvents. The compositions can, if desired, contain additional ingredients such as flavors, binders, excipients and the like. Thus, for oral administration, tablets containing various excipients, such as citric acid, may be employed together with various disintegrants such as starch or other cellulosic material, alginic acid and certain complex silicates, and with binding agents such as sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, talc and sodium lauryl sulfate are often useful for tabletting purposes. Other reagents such as an inhibitor, surfactant or solubilizer, plasticizer, stabilizer, viscosity increasing agent or film forming agent can also be added. Solid compositions of a similar type can also be employed in filled soft and hard gelatine capsules. Materials include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the active compound can be combined with various sweetening or flavoring agents, coloring matter and dyes and, if desired, emulsifying or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin or combinations thereof.

[0204] Excipients

[0205] Any of the compositions or formulations described herein include any excipients used in pharmacy and are selected based on compatibility with the active agent(s) and release profile properties of the desired dosage form. Excipients include, but are not limited to, binders, fillers, flow aids, disintegrants, lubricants, stabilizers, surfactants and the like. A summary of the excipients described herein can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteeth Ed (Easton, PA: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, (Easton, PA: Mack Publishing Co 1975); Libertyman, H.A. and Lachman, L, Eds., Pharmaceutical Dosage Forms (New York, NY: Marcel Decker 1980); and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed (Lippincott Williams & Wilkins 1999), incorporated herein by reference in their entirety.

[0206] Binders impart cohesive qualities and include, for example, alginic acid and salts thereof; cellulose derivatives such as carboxymethylcellulose, methylcellulose (e.g. Methocel®), hydroxypropylmethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose (e.g. Klucel®), ethylcellulose (e.g. Ethocel®) and microcrystalline cellulose (e.g. Avicel®); microcrystalline dextrose; amylose; magnesium aluminum silicate; polysaccharide acids; bentonites; gelatin; polyvinylpyrrolidone/vinyl acetate copolymer; crospovidone; povidone; starch; pregelatinized starch; tragacanth, dextrin, a sugar such as sucrose (eg Dipac®), glucose, dextrose, molasses, mannitol, sorbitol, xylitol (eg Xylitab®) and lactose; a natural or synthetic gum such as acacia, tragacanth, ghatti gum, isapol bark mucilage, polyvinylpyrrolidone (e.g. Polyvidone® CL, Kollidon® CL, Polyplasdone® XL-10), larch arabogalactan, Veegum®, polyethylene glycol, waxes, alginate sodium and the like.

[0207] Disintegrants facilitate the dissolution or disintegration of solid oral dosage forms after administration. Examples of disintegrants include a starch, for example, a natural starch such as corn starch or potato starch, a pregelatinized starch such as National 1551 or Amijel®, or sodium starch glycollate, such as Promogel® or Explotab®; a cellulose as a wood product, methylcrystalline cellulose, for example Avicel®, Avicel® PH101, Avicel® PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, Ming Tia® and Zolka-Flo ®, methylcellulose, croscarmellose or a crosslinked cellulose, such as crosslinked sodium carboxymethylcellulose (Ac-Di-Solt), crosslinked carboxymethylcellulose, or crosslinked croscarmellose; a cross-linked starch such as sodium starch glycollate; a cross-linked polymer such as crospovidone; a cross-linked polyvinylpyrrolidone; alginate such as alginic acid or a salt of alginic acid such as sodium alginate; a clay such as Veegum® HV (magnesium aluminum silicate); a gum such as agar, guar, locust bean, karaya, pectin, or tragacanth; sodium starch glycollate; bentonite; a natural sponge; a resin such as a cation exchange resin; citrus pulp; Sodium lauryl sulfate; combination starch sodium lauryl sulfate; and others.

[0208] Lubricants are compounds that prevent, reduce or inhibit the adhesion or friction of materials. Exemplary lubricants include, for example, stearic acid; calcium hydroxide; baby powder; stearyl sodium fumarate; a hydrocarbon such as mineral oil, hydrogenated castor oil or hydrogenated vegetable oil, such as hydrogenated soybean oil (Sterotex®); higher fatty acids and their alkali metal and alkaline earth metal salts, such as aluminum, calcium, magnesium, zinc; stearic acid, sodium stearates, magnesium stearates, glycerol, talc, waxes, Stearowet® boric acid, sodium benzoate, sodium acetate, sodium chloride, leucine, a polyethylene glycol or a methoxypolyethylene glycol such as Carbowax™, oxide polymers ethylene, sodium oleate, glyceryl behenate (eg Compritol 888 Ato), glyceryl distearate (Precirol Ato 5), polyethylene glycol, magnesium or sodium lauryl sulfate, colloidal silica such as Syloid™, Carb-O-Sil®, DL -leucine, a starch such as corn starch, silicone oil, a surfactant, and the like.

[0209] Flow aids or glidants improve the flow characteristics of powder mixtures. Such compounds include, for example, colloidal silicon dioxide such as Cab-o-sil®; tribasic calcium phosphate, talc, corn starch, DL-leucine, sodium lauryl sulfate, magnesium stearate, calcium stearate, sodium stearate, kaolin, micronized amorphous silicon dioxide (Syloid®) and the like.

[0210] Plasticizers help in coating solid oral dosage forms. Exemplary plasticizers include, but are not limited to, triethyl citrate, triacetin (glyceryl triacetate), triethyl acetyl citrate, polyethylene glycols (PEG 4000, PEG 6000, PEG 8000), Carbowax 400 (polyethylene glycol 400), diethyl phthalate, diethyl sebacate , acetyltriethylcitrate, oleic acid, glyceralmonosterate, tributyl citrate, acetylated monoglycerides, glycerol, fatty acid esters, propylene glycol and dibutylphthalate and the like.

[0211] The above mentioned excipients are given as examples only and are not intended to include all possible options. Other suitable excipient classes include coloring agents, granulating agents, preservatives, antifoaming agents, solubilizers and the like. Furthermore, many excipients may have more than one role or function, or may be classified into more than one group; classifications are descriptive only and are not intended to limit any use of a particular excipient.

[0212] Combination therapies

[0213] The compositions of the embodiments described herein can be co-administered with known therapies for the treatment of any of the conditions described herein. Coadministration can also account for additive or synergistic effects, resulting in the need for lower doses of a known therapy, the compositions described herein, or both. Additional co-administration benefits include reduction in toxicities associated with any of the known therapies.

[0214] Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which both agents are present. Thus, in some embodiments, a known therapy and compositions described herein are administered in a single treatment. In some embodiments, the compositions described herein and a known therapy are blended into a resulting composition. In some embodiments, known therapy and compositions described herein are administered in separate compositions or administrations.

[0215] Administration of the compositions described herein and known therapies described herein can be by any feasible means. Administration of a composition described herein and a second compound (eg, diabetes or obesity drug) can be by any suitable means. If the compositions described herein and a second compound are administered as separate compositions, they may be administered by the same or different routes. If the compositions described herein and a second compound are administered in a single composition, they can be administered by any suitable route such as, for example, oral administration. In certain embodiments, compositions of metformin an analogue thereof (including salts, solvates, polymorphs, hydrates, N-oxides or prodrugs thereof) and second compounds can be administered to the same or different regions of the gastrointestinal tract. For example, metformin or an analogue thereof (including salts, solvates, polymorphs, hydrates, N-oxides or prodrugs thereof) can be administered in combination with an antidiabetic drug to be delivered to the duodenum, jejunum, ileum or colon.

[0216] Therapies, drugs and compounds useful for the treatment of hyperglycemia and/or associated diseases or conditions, for example, diabetes, can be administered with the compositions disclosed herein. Diabetic therapy drugs and compounds include, but are not limited to those that lower triglyceride concentrations, lower glucose concentrations, and/or modulate insulin (e.g., stimulate insulin production, mimic insulin, increase glucose-dependent insulin secretion, suppress glucagon secretion or action, enhance insulin action or insulin sensitizers or are exogenous forms of insulin).

[0217] Drugs that lower the level of triglycerides include, but are not limited to, ascorbic acid, asparaginase, clofibrate, colestipol, mevastatin fenofibrate, pravastatin, simvastatin, fluvastatin or omega-3 fatty acid. Drugs that lower the LDL cholesterol level include, but are not limited to, clofibrate, gemfibrozil and fenofibrate, nicotinic acid, mevinoline, mevastatin, pravastatin, simvastatin, fluvastatin, lovastatin, cholestyrine, colestipol or probucol.

[0218] In another aspect, compositions of the embodiments described herein can be administered in combination with glucose-lowering compounds.

[0219] The drug classes of thiazolidinediones (also called glitazones), sulfonylureas, meglitinides, biguanides, alpha-glucosidase inhibitors, DPP-IV inhibitors, and incretin mimetics have been used as pharmacological therapies for hyperglycemia and diabetes mellitus (type 2 ) and related diseases.

[0220] Drugs that lower blood glucose include but are not limited to glipizides, glyburides, exenatide (Byetta®), incretins, sitagliptin (Januvia®), pioglitazone, glimepiride, rosiglitazone, metformin, vildagliptin, saxagliptin (OnglyzaTM), sulfonylureas , meglitinide glycosidase inhibitor (e.g., Prandin®), biguanides (e.g., Glucophage®), repaglinide, acarbose, troglitazone, nateglinide, synthetic or natural recombinant insulin and its derivatives, and amylin and amylin derivatives.

[0221] When administered sequentially, the combination may be administered in two or more administrations. In an alternative embodiment, it is possible to administer one or more of the biguanide compounds and one or more additional active ingredients via different routes. One skilled in the art will also recognize that a variety of active ingredients can be administered in combination with one or more biguanide compounds that can act to enhance or synergistically improve the control, prevention, amelioration, alleviation or treatment of obesity or eating disorders or conditions. .

[0222] According to the methods provided herein, when co-administered with at least one other obesity (or anti-obesity) or weight reducing drug, the compounds of the disclosure can be: (1) co-formulated and administered or delivered simultaneously in a combined formulation; (2) delivered alternately or in parallel as separate formulations; or (3) by any other combination therapy regimen known in the art. When provided in alternation therapy, the methods provided may include administering or delivering the active ingredients sequentially, for example, in a separate solution, emulsion, suspension, tablets, pills or capsules, or by different injections in separate syringes. In general, during alternation therapy, an effective dosage of each active ingredient is administered sequentially, i.e. serially, while in simultaneous therapy, effective doses of two or more active ingredients are administered together. Various sequences of intermittent combination therapy can also be used.

[0223] In certain embodiments, compositions provided herein may be used with other commercially available dietary aids or other weight loss or anti-obesity agents, such as, by way of example, PYY and PYY agonists, GLP-1 agonists and GLP-1, a DPP-IV inhibitor, CCK and CCK agonists, exendin and exendin agonists, GIP and GIP agonists, amylin and amylin agonists, ghrelin modulators (e.g., inhibitors) and leptin and leptin agonists. In certain cases, biguanide or compositions comprising the biguanide compound provided herein are used in combination with amylin, amylin agonists or mimetics. Exemplary amylin agonists or mimetics include pramlintide and related compounds. In certain instances, the compounds and compositions provided herein are used in combination with leptin, leptin agonists or mimetics. Additional leptin agonists or mimetics can be identified using methods described by U.S. Pat. U.S. At the. 7,247,427, which is incorporated herein by reference. In other cases, the compounds and compositions provided herein increase leptin sensitivity and enhance the effectiveness of leptin, leptin agonists or mimetics.

[0224] Additional anti-obesity agents suitable for use in the methods of the invention include those currently under development. Other anti-obesity agents include phentermine, fenfluramine, sibutramine, rimonabant, topiramate, zonisamide, bupropion, naltrexone, lorcaserin or related sympathomimetics, and orlistat or other intestinal lipase inhibitors, alone or in combination. Therapies, drugs and compounds useful for treating weight loss, binge eating, addictions and food cravings can be administered with the compositions described herein. For example, the subject may still be administered at least one other drug that is known to suppress hunger or control appetite. Such drugs and therapeutic compounds include, but are not limited to, phenteramines such as Meridia® and Xenical®. Additional compounds, drugs and therapies are known in the art and contemplated herein.

[0225] As such, in one aspect, the compound can be used as part of a combination therapy for the control, prevention or treatment of obesity or eating disorders or conditions. Compounds used as part of a combination therapy to treat obesity or reduce weight include, but are not limited to, central nervous system agents that affect neurotransmitters or neural ion channels, including antidepressants (bupropion), noradrenaline reuptake inhibitors ( GW320659), selective serotonin 2C receptor agonists, selective 5HT 2c receptor agonists, anticonvulsant agents (topiramate, zonisamide), certain dopamine antagonists and cannabinoid-1 receptor antagonists (CB-1 receptor antagonists) (rimonabant) ; central nervous system/leptin/insulin pathway agents, including leptin analogues, leptin transport and/or leptin receptor promoters, ciliary neurotrophic factor (Axokine), neuropeptide Y and agouti-related peptide antagonists, regulated transcription factors of pro-opiomelanocortin and cocaine and amphetamine, alpha-melanocyte-stimulating hormone analogues, melanocoritin-4 receptor agonists, and agents that affect insulin metabolism/activity, which include protein-tyrosine phosphatase-1B inhibitors, receptor proliferator-activated peroxisome, gamma-receptor antagonists, short-acting bromocriptine (ergoset), somatostatin agonists (Octreotide) and adiponectin/Acrp30 (Famoxin or fatty acid metabolic oxidation inducer); neural-gastrointestinal pathway agents, including those that increase cholecystokinin (CCK) activity, PYY activity, NPY activity, and PP activity, glucagon-like peptide-1-increasing activity, activity (exendin 4, liraglutide, dipeptidyl peptidase IV) and those that decrease ghrelin activity, as well as amylin analogues (pramlintide); agents that may increase resting metabolic rate (selective β-3 stimulants/agonists, uncoupling protein homologues, and thyroid receptor agonists); other more diverse agents, including melanin concentrating hormone antagonists, phytostanol analogues, functional oils, P57, amylase inhibitors, growth hormone fragments, synthetic analogues of dehydroepiandrosterone sulfate, antagonists of adipocyte 11 B-hydroxysteroid dehydrogenase type 1 activity , corticotropin-releasing hormone agonists, fatty acid synthesis inhibitors (cerulenin and C75), carboxy peptidase inhibitors, indanone/indanols, aminosterols (trodusquemin/trodulamine) and other gastrointestinal lipase inhibitors (ATL962); amphetamines, such as dextroamphetamine; other sympathomimetic adrenergic agents, including phentermine, benzphetamine, phendimetrazine, mazindol and diethylpropion.

[0226] Other compounds include ecopipam; oxyntomodulin (OM); glucose-dependent insulinotropic polypeptide (GIP) inhibitors; gastrin-releasing peptide; neuromedin B; enterostatin; amfebutamone, SR-58611; CP-045598; AOD-0604; QC-BT16; rGLP-1; 1426 (HMR-1426); N-5984; ISIS-113715; solabegron; SR-147778; Org-34517; melanotan-ll; cetilistat; c-2735; c-5093; c-2624; APD-356; radafaxine; intranasal peptide YY [3-36]; androgen receptor agonists; PYY 3-36; DOV-102677; tagatose; SLV-319; 1954 (Aventis Pharma AG); oxyntomodulin, Thiakis; bromocriptine, PLIVA; diabetes/hyperlipidemia therapy, Yissum; CKD-502; thyroid receptor beta-agonists; beta-3 adrenoceptor agonist; CDK-A agonists; galanin agonists; dopamine D1/D2 agonists; melanocortin modulators; verongamine; neuropeptide Y antagonists; melanin-concentrating hormone receptor antagonists; dual PPAR alpha/gamma agonists; CGEN-P-4; kinase inhibitors, human MCH receptor antagonists; GHS-R antagonists, ghrelin receptor agonists; DG70 inhibitors; cotinine; CRF-BP inhibitors, urocortin agonists; UCL-2000; impentamine; beta-3 adrenergic receptor; MC4 pentapeptide agonists; trodusquemine; GT-2016; C-75; CPOP; MCH-1 receptor antagonists; RED-103004; aminosterols; orexin-1 antagonists; neuropeptide Y5 receptor antagonists; DRF-4158; DRF-4158; PT-15; PTPase inhibitors; A37215; SA-0204; glycolipid metabolites; MC-4 agonist; produlestan; PTP-1B inhibitors; GT-2394; neuropeptide Y5 antagonists; melanocortin receptor modulators; MLN-4760; PPAR dual gamma/delta agonists; NPY5RA-972; 5-HT2C receptor agonist; neuropeptide Y5 receptor antagonists (phenyl urea analogues); AGRP/MC4 antagonists; neuropeptide Y5 antagonists (benzimidazole); glucocorticoid antagonists; MCHR1 antagonists; acetyl-CoA carboxylase inhibitors; R-1496; HOB1 modulators; NOX-B11; peptide YY 3-36 (eligen); 5-HT 1 modulators; pancreatic lipase inhibitors; GRC-1087; CB-1 antagonists; MCH-1 antagonists; LY-448100; BRS3 bombesin agonists; ghrelin antagonists; MC4 antagonists; stearoyl-CoA desaturase modulators; histamine H3 antagonists; PPARpan agonists; EP-01492; hormone-sensitive lipase inhibitors; fatty acid binding protein 4 inhibitors; thiolactone derivatives; protein tyrosine phosphatase 1B inhibitors; MCH-1 antagonist; P-64; PPAR gamma ligands; hormonal antagonists that concentrate melanin; gastroprokinetic thiazoles; PA-452; T-226296; A-331440; immunodrug vaccines; diabetes/obesity therapies (Bioagency, Biofrontera Discovery GmbH); P-7 (Genfit); DT-011M; PTP1B inhibitor; antidiabetes peptide conjugates; KATP agonists; obesity therapies (Lexicon); 5-HT2 agonists; MCH-1 receptor antagonists; GMAD-1/GMAD-2; STG-to-MD; neuropeptide Y antagonist; angiogenesis inhibitors; G protein-coupled receptor agonists; nicotine therapies (ChemGenex); anti-obesity agents (Abbott); neuropeptide Y modulators; melanin-concentrating hormone; GW-594884A; MC-4R agonist; histamine H3 antagonists; orphan GPCR modulators; MYTH-3108; NLC-002; HE-2300; IGF/IBP-2-13; 5-HT2C agonists; ML-22952; neuropeptide Y receptor antagonists; AZ-40140; anti-obesity therapy (Nisshin Flour); GNTI; melanocortin receptor modulators; alpha-amylase inhibitors; neuropeptide Y1 antagonist; beta-3 adrenoceptor agonists; Ob gene products (Eli Lilly &Co.);SWR-0342-SA; beta-3 adrenoceptor agonist; SWR-0335; SP-18904; oral insulin mimetics; beta-3 adrenoceptor agonists; NPY-1 antagonists; beta-3 agonists; obesity therapies (7TM Pharma); 11-beta-hydroxysteroid dehydrogenase (HSD) 1 inhibitors; QRX-431; E-6776; RI-450; melanocortin-4 antagonists; melanocortin 4 receptor agonists; obesity therapies (CuraGen); leptin mimetics; A-74498; second-generation leptin; NBI-103; CL-314698; CP-114271; beta-3 receptor agonists; NMI-8739; UCL-1283; BMS-192548; CP-94253; PD-160170; nicotinic agonist; LG-100754; SB-226552; LY-355124; CKD-711; L-751250; PPAR inhibitors; protein G therapeutics; obesity therapy (Amylin Pharmaceuticals Inc.); BW-1229; monoclonal antibody (ObeSys/CAT); L-742791; (S)-sibutramine; MBU-23; YM-268; BTS-78050; tube-like protein genes; genomics (eating disorders; Allelix/Lilly); MS-706; GI-264879A; GW-409890; FR-79620 analogues; obesity therapy (Hybrigenics SA); ICI198157; ESP-A; 5-HT2C agonists; PD-170292; AIT-202; LG-100641; Gl-181771; anti-obesity therapies (Genzyme); leptin modulator; GHRH mimetics; obesity therapy (Yamanouchi Pharmaceutical Co. Ltd.); SB-251023; CP-331684; BIBO-3304; cholesten-3-ones; LY-362884; BRL- 48962; NPY-1 antagonists; A-71378; RTM-didesmethylsibutramine; starch derivatives; obesity therapies (Bristol-Myers Squibb Co.); obesity therapeutics (Ligand Pharmaceuticals Inc.); LY-226936; NPY antagonists; CCK-A agonists; FPL-14294; PD-145942; ZA-7114; CL-316243; SR-58878; R-1065; BIBP-3226; HP-228; talibegron; FR-165914; AZM-008; AZM-016; AZM-120; AZM-090; vomeroferin; BMS-187257; D-3800; AZM-131; gene discovery (Axys/Glaxo); BRL-26830A SX-013; ERR modulators; adipsin; AC-253; A-71623; A-68552; BMS-210285; TAK-677; MPV-1743; obesity therapy (Modex); GI-248573; AZM-134; AZM-127; AZM-083; AZM-132; AZM-115; exopipan; SSR-125180; obesity therapy (Melacure Therapeutics AB); BRL-35135; SR-146131; P-57; AZM-140; CGP-71583A; RF-1051; BMS-196085; manifaxin; beta-3 agonists; DMNJ (Korea Research Institute of Bioscience and Biotechnology); BVT-5182; LY-255582; SNX-024; galanin antagonists; neurokinin-3 antagonists; dexfenfluramine; mazindol; diethylpropion; phendimetrazine; benzphetamine; amfebutmon; sertraline; metformin; AOD-9604; ATL-062; BVT-933; GT389-255; SLV319; HE-2500; PEG-axoquine; L-796568; and ABT-239.

[0227] In some embodiments, compounds for use in combination with a composition comprising the biguanide compound provided herein include rimonabant, sibutramine, orlistat, PYY or an analogue thereof, CB-1 antagonist, leptin, phentermine and exendin analogues. Exemplary dosage ranges include phentermine resin (30 mg in the morning), fenfluramine hydrochloride (20 mg three times daily), and a combination of phentermine resin (15 mg in the morning) and lorcaserin (30 mg before the evening meal). and sibutramine (10-20 mg). Weintraub et al. (1984) Arch. Intern. Med. 144:1143-1148.

[0228] In other embodiments, compounds for use in combination with a composition provided herein include GPR119 agonists (eg, anandamide; AR-231, 453; MBX-2982; oleoylethanolamide; PSN-365963; PSN-632408; palmitoylethanolamide) ,GPR120 agonists (e.g., omega-3 fatty acids including, but not limited to, all-linolenic acid, docosapentaenoic acid, docosahexaenoic acid, eicosatrienoic acid, eicosatetraenoic acid, eicosapentaenoic acid, heneicosapentaenoic acid, hexadecatrienoic acid, stearidonic acid, tetracosahexaenoic acid and tetracosapentaenoic acid) and GPR 40, GPR41 and GPR43 agonists (eg free fatty acids including short, medium and long chain unsaturated and saturated fatty acids).

[0229] In some embodiments, a composition provided herein is used as an adjunctive therapy to bariatric surgery. Bariatric surgery is a weight loss procedure and refers to modifications to the gastrointestinal tract and includes procedures such as gastric banding, sleeve gastrectomy, gastrointestinal bypass procedure (e.g., roux en Y, biliary duodenal bypass, gastric loop bypass) , intragastric balloon, vertical bands, gastroplasty, endoluminal sleeve, biliopancreatic diversion and the like. In certain cases, the composition provided herein is attached to the gastric band. In certain instances, a composition is adjunct to gastrointestinal bypass procedures. In still other cases, a composition provided herein is adjunct to the sleeve gastrectomy. In some embodiments, a composition provided herein as adjunctive therapy to bariatric surgery is administered prior to the bariatric procedure. In certain embodiments, a biguanide or related heterocyclic compound composition as adjunctive bariatric surgery therapy is administered prior to the bariatric procedure. In certain cases, when used as an adjunctive therapy, the dosages and amounts of a composition provided herein may be adjusted as needed with respect to the bariatric procedure. For example, amounts of a composition provided herein administered as an adjunctive therapy to a bariatric procedure can be reduced by half normal dosages or as directed by a medical professional.

[0230] Combination therapy can be explored, for example, in modulating the metabolic syndrome (or treating the metabolic syndrome and its related symptoms, complications and disorders), where the compositions of compounds provided herein can be effectively used in combination with , for example, the active agents discussed above for modulating, preventing or treating diabetes, obesity, hyperlipidemia, atherosclerosis, and/or their respective related symptoms, complications and disorders.

[0231] Methods for Evaluating Treatment

[0232] Evaluation of Diabetes Treatment

[0233] The effect of treatment with the biguanide compound of the invention on aspects of diabetic disease can be assessed according to common and art-known methods practiced by physicians for treating diabetic patients.

[0234] The effectiveness of treating metabolic syndrome/diabetes and conditions associated with diabetes with the compositions and methods described herein can be evaluated using methodologies and assays known in the art. By way of example, quantitative assessment of renal function and parameters of renal dysfunction are well known in the art. Examples of assays for determining renal function/dysfunction include serum creatinine level, creatinine clearance rate; cystatin C clearance rate, 24-hour urinary creatinine clearance, 24-hour urinary protein secretion; glomerular filtration rate (GFR); urinary albumin creatinine ratio (ACR); albumin excretion rate (AER); and renal biopsy.

[0235] Quantitative assessment of pancreatic function and parameters of pancreatic dysfunction or insufficiency are well known in the art. Examples of assays for determining pancreas function/dysfunction include evaluating pancreatic functions using biological and/or physiological parameters such as evaluation of size, growth and/or secretory activity of Islets of Langerhans, size, growth and/or secretory activity of cells- beta, circulating blood levels and insulin secretion, blood glucose levels, pancreas imaging, pancreas biopsy, oral glucose challenge glucose uptake studies, evaluation of cytokine profiles, blood gas analysis, extension of blood perfusion of tissues and angiogenesis within tissues.

[0236] Additional assays for treating diabetes and conditions associated with diabetes are known in the art and are contemplated herein.

[0237] Evaluation of Treatment for Weight Loss, Obesity and Eating Disorders

[0238] In the treatment of obesity it is desired that weight and/or fat be reduced in a patient. By weight reduction, it is understood that the subject loses a part of his total body weight over the course of treatment (whether the course of treatment lasts for days, weeks, months or years). Alternatively, weight reduction can be defined as a decrease in the percentage of fat mass to lean mass (in other words, the patient has lost fat mass but maintained or gained lean mass, without necessarily a corresponding loss in total body weight). An effective amount of biguanide compound treatment administered in this embodiment is an amount effective to reduce a patient's body weight over the course of treatment, or, alternatively, an amount effective to reduce the patient's percent fat mass over the course of treatment. In certain embodiments, the patient's body weight is reduced, over the course of treatment, by at least about 1%, at least about 5%, at least about 10%, at least about 15%, or by at least about of 20%. Alternatively, the patient's percentage of body fat is reduced over the course of treatment by at least 1%, at least 5%, at least 10%, at least 15%, at least 20% or at least 25%.

[0239] Fat content and total body weight can be measured at the end of the diet period. In rats, a frequently used method of determining total body fat is to surgically remove and weigh the retroperitoneal fat pad, a body of fat located in the retroperitoneum, the area between the posterior abdominal wall and the posterior parietal peritoneum. Cushion weight is considered directly related to the animal's body fat percentage. Since the relationship between body weight and body fat in rats is linear, obese animals have a correspondingly greater percentage of body fat and retroperitoneal fat pad weight.

[0240] In modalities where methods of treating, reducing or preventing compulsions in a patient are provided, compulsions may be measured by means of a questionnaire, either known in the art or created by the person studying compulsions. Such a questionnaire would rather rate the level of binge eating on a numerical scale, with the patient scoring 0 if they do not have compulsive overeating and scoring (if on a scale of 1-10) 10 if the patient has severe compulsive overeating. The questionnaire would preferably also include questions about what types of foods the patient is craving. Binge eating can be determined or measured using a questionnaire and a Binge Eating Scale (BES). The severity of binge eating can be divided into three categories (mild, moderate and severe) based on the total SWB score (calculated by summing the scores for each individual item). Also provided are methods of reducing a patient's BES score, comprising administering to a patient in need thereof a compound treatment in an amount effective to reduce the patient's BES score. In some embodiments, administration of a composite treatment changes the patient's SWB category, for example, from severe to moderate, from severe to mild, or from moderate to mild.

[0241] Pretreatment Evaluation of the Patient's Hormonal Profile

[0242] In some embodiments, patients are previously evaluated for expression of metabolic hormones using the methods described herein. The therapy provided to the individual, therefore, can be tailored to their specific needs. In embodiments, the patient's hormone profile is pre-assessed and, depending on the changes the physician wishes to effect, a predetermined amount of the metabolite/compound combination is administered. The assessment process can be repeated and treatment adjusted accordingly at any time during or after treatment.

[0243] Hormone Tests

[0244] In embodiments, levels of hormones tested in association with the methods of the invention, including, but not limited to, GLP-1, GLP-2, GIP, oxyntomodulin, PYY, CCK, glycentin, insulin, glucagon, ghrelin , amylin, uroguanylin, C-peptide and/or combinations thereof, are detected according to standard methods described in the literature. For example, proteins can be measured by immunological assays and transcripts by nucleic acid amplification techniques. Functional assays described in the art can also be used as appropriate. In embodiments, test samples comprise cultured cells, patient cells or tissue samples, patient body fluids, e.g., blood or plasma, etc. Likewise, analyte levels (eg, glucose, triglycerides, HDL, LDL, apoB and the like) analyzed in association with the methods of the invention are detected according to any known method.

[0245] For example, immunofluorescence can be used to assay for GLP-1. Cells can be grown on confluent monolayer matrigel-coated coverslips in 12-well plates at 37°C, fixed in 4% paraformaldehyde in phosphate buffered saline (PBS) and incubated with primary antiserum (e.g., gustducin anti- rabbit alpha, 1:150; Santa Cruz Biotechnology and rabbit anti-GLP-1, Phoenix) overnight at 4°C, followed by permeabilization with 0.4% Triton-X in PBS for 10 minutes and blocking. if for 1 hour at room temperature. After three washing steps with blocking buffer, the appropriate secondary antibody is applied (AlexaFluor 488 anti-rabbit immunoglobulin, 1:1000; Molecular Probes) for 1 hour at room temperature. After three washing steps, cells can be fixed in Vectashield medium and immunofluorescence visualized.

[0246] GLP-1 RNA isolated from cells can be tested using RT-PCR. RT-PCR isolation of RNA from cells can be performed using standard methodology. The RT-PCR reaction can be performed in a volume of 50 µl in a Peltier thermal cycler (PTC-225 DNA Engine Tetrad Cycler; MJ Research) using the published primer sequences (Integrated DNA Technologies). Reverse transcription can be performed at 50°C for 30 minutes; after an initial activation step at 95°C for 15 minutes. PCR can be performed by denaturing at 94°C for 1 minute, annealing at 55°C for 1 minute, and extending at 72°C for 1 minute for 40 cycles, followed by a final extension step at 72°C for 10 minutes. Negative controls can be included as appropriate, for example by substituting water for the template or omitted reverse transcriptase. The control can be isolated RNA, for example, rat lingual epithelium. PCR products can be separated on a 2% ethidium bromide agarose gel and visualized under UV light.

[0247] Radioimmunoassay (RIA) for total GLP-1 in patient blood samples can be performed as described in the art, for example, by Laferrere, et al., 2007, “Incretin Levels and Effect are Markedly Enhanced IMonth after Roux- en-Y Gastric Bypass Surgery in Obese Patients with Type 2 Diabetes", Diabetes Care 30(7): 1709-1716 (using commercially available materials obtained from Phoenix Pharmaceutical, Belmont, CA). The authors describe measuring the effect of GIP and GLP-1 on insulin secretion by measuring the difference in insulin secretion (area under the curve, or AUC) in response to an oral glucose tolerance test and an isoglycemic intravenous glucose test.

[0248] Measurement of plasma concentrations of GLP-1, GIP, glucagon, insulin, C-peptide, pancreatic peptide, non-esterified fatty acids, glutamic acid decarboxylase antibodies and islet antigen antibodies, is described, for example, by Toft- Nielsen, et al., 2001, “Determinants of the Impaired Secretion of Glucagon-Like Peptide-1 in Type 2 Diabetic Patients,” J. Clin. End. Met. 86(8): 3717_,3723. The authors describe the use of radioimmunoassay for GLP-1 to measure plasma concentrations of amidated GLP-1-(7-36) using antibody code no. 89390. This assay measures the sum of GLP-1-(7-36) and its metabolite GLP-1-(9-36). The authors describe the measurement of GIP using C-terminally directed antibody code no. R65 (RIA), which reacts 100% with a human GIP, but not with 8-kDA GIP.

[0249] GLP-1 and PYY can be directly assayed in the supernatant of venous effluents as described, for example, by Claustre, et al. (1999, “Stimulatory effect of β-adrenergic agonists on ileal L cell secretion and modulation by α-adrenergic activation, J. Endocrin. 162:271-8). (See also Plaisancie' et al., 1994, “Regulation of glucagon-like peptide-1-(7-36) amide secretion by intestinal neurotransmitters and hormones in the isolated vasculariy perfused rat colon," Endocrinology 135:2398-2403 and Plaisancie ' et al., 1995, “Release of peptide YY by neurotransmitters and gut hormones in the isolated, vasculariy perfused rat colon,” Scandinavian Journal of Gastroenterology 30:568-574) In this method, the anti-GLP-1 antibody 199D is used at a dilution of 1:250,000. This antibody reacts 100% with GLP-1-(7-36) amide, 84% with GLP-1-(1-36) amide, and less than 0.1% with GLP- 1-(1-37), GLP-1-(7-37), GLP-2 and glucagon PYY is analyzed with A4D anti-porcine PYY antiserum at a dilution of 1:800,000.

[0250] Methods for analyzing GLP-1 and GIP are also described elsewhere in the art, for example by Jong, et al., PNAS, 2007.

[0251] PYY can also be analyzed in the blood using a radioimmunoassay as described by, for example, Weickert, et al., 2006, “Soy isoflavones increase preprandial peptide YY (PYY), but have no effect on ghrelin and body weight in healthy postmenopausal women” Journal of Negative Results in BioMedicine, 5:11. Blood is collected into ice-cooled EDTA tubes for glucose, ghrelin, and PYY analysis. After centrifugation at 1600g for 10 minutes at 4°C, aliquots were immediately frozen at -20°C until assayed. All individual patient samples were measured in the same assay. The authors described measuring total immunoreactive ghrelin being measured by a commercially available radioimmunoassay (Phoenix Pharmaceuticals, Mountain View, CA, USA). (See also Weickert, et al., 2006, “Cereal fiber improves whole-body insulin sensitivity in overweight and obese women,” Diabetes Care 29:775-780). Total human immunoreactive PYY is measured by a commercially available radioimmunoassay (LINCO Research, Missouri, USA), using Bioactive 1251-labeled PYY as a tracer and a PYY antiserum to determine the level of active PYY by the dual antibody/PEG technique. The PYY antibody is grown in guinea pigs and recognizes both PYY 1-36 and PYY 3-36 (active) forms of human PYY.

[0252] SGLT-1, sodium-dependent intestinal glucose transporter 1 is a protein involved in the supply of glucose to the body. It has been reported to be expressed in response to sugar in the gut lumen through a pathway involving T1R3 (Margolskee, et al., 2007 “T1R3 and gustducin in gut sense sugars to regulate expression of Na+-glucose cotransporter 1,” Proc Natl Acad Sci USA 104, 15075-15080”). SGLT-1 expression can be detected as described, for example, by Margolskee, et al., for example, using quantitative methods, PCR and Western Blotting, known in the art. Measurement of glucose transport has been described in the literature, for example by Dyer, et al., 1997, Gut 41:56-9 and Dyer, et al., 2003, Eur. J. Biochem 270:3377-88. Measurement of glucose transport in brush border membrane vesicles can be done, for example, by starting D-glucose uptake by adding 100 μl of incubation medium containing 100 mM NaSCN (or KSCN), 100 mM of mannitol, 20 mM Hepes/Tris (pH 7.4), 0.1 mM MgSO4, 0.02% (wt/vol) NaN3 and 0.1 mM D-[U14C] glucose for BBMV (100 µg protein ). The reaction is stopped after 3 sec by adding 1 ml of frozen stop buffer, containing 150 mM KSCN, 20 mM Hepes/Tris (pH 7.4), 0.1 mM MgSO4, 0.02% (wt/ vol) of NaN3 and 0.1 mM of phlorizin. A 0.9 ml portion of the reaction mixture is removed and filtered under vacuum through a 0.22 µm pore cellulose acetate/nitrate filter (GSTF02500; Millipore, Bedford, MA). The filter is washed 5 times with 1 ml of stop buffer, and the radioactivity retained on the filter is measured by liquid scintillation counting.

EXAMPLES

[0253] EXAMPLE 1: ENTEROENDOCRINE PRODUCTION OF PYY, GLP-1 A AND GLP-1 T AND GLUCOSE AND INSULIN REDUCTION IS INDEPENDENT OF METFORMIN PLASMA ABSORPTION

[0254] Example 1.1 Materials and Methods

[0255] Population: Approximately 18 eligible male and female subjects, 18 to 65 years of age, with a BMI of 25.0 to 35.0 kg/m2, were randomized in this study. To be eligible, each subject also met the following criteria: (a) was not breastfeeding; (b) had a negative pregnancy test result (human chorionic gonadotropin, beta subunit); (c) surgically sterile, past menopause or, if of potential pregnancy, practiced appropriate birth control throughout the duration of the study; (d) had a physical examination free of dynamically significant abnormalities, including but not limited to the following conditions: (i) Liver disease; (ii) kidney disease; (iii) gastrointestinal disease; (iv) endocrine disorder, including diabetes; (v) cardiovascular disease; (vi) seizure disorder; (vii) organ transplantation; and (viii) chronic infection; and (e) an ability to understand and willingness to adhere to the requirements of the protocol.

[0256] Formulations

[0257] Metmorphine DR formulation was a US supplied, commercially available, film-coated, immediate-release 500 mg metmorphine hydrochloride lozenge to which additional coatings (a wax coat and an enteric coating) were applied to in order to delay drug release in the gastrointestinal tract until the tablet reached a pH 6.5 region of the distal small intestine. The seal coating was applied at a nominal level of 2% of the tablet core weight and the enteric coating was applied at a nominal level of 2.4% of the drug core weight. The lozenges were white, biconvex, circular shaped coated lozenges, each containing 500 mg of metmorphine hydrochloride. Inactive ingredients in the commercially available lozenge include providone, magnesium stearate, hypromellose and polyethylene glycol. Inactive ingredients in additional coating systems include hypromellose, triacetin, talc, methacrylic acid copolymer (Eudragit® L30 D-55), poly(acrylate-co-methyl methacrylate-co-methacrylic acid) 7:3:1 (Eudragit® FS 30 D), sodium lauryl sulfate, polysorbate 80, glyceryl monostearate and triethyl citrate.

[0258] The metmorphine IR formulation was the identical US supplied commercially available immediate-release film-coated lozenge containing 500 mg metmorphine hydrochloride, to which only the additional seal coating is applied. No release delay (enteric) coating was applied. Inactive ingredients in the additional sealant coating system included hypromellose, triacetin and talc.

[0259] Metmorphine formulations were supplied to the site as bulk tablets packaged in screw cap containers labeled with container number and batch number. All study drugs were stored in cool, dry conditions as directed on the label and used only as directed by study personnel. Study medication was dispensed by the pharmacist or study personnel as disclosed in the randomization scheme at the start of each treatment period.

[0260] Administration

[0261] Study drugs were provided by either the on-site pharmacist or unblinded study staff according to a randomization scheme at Visits 2 and 4. At the end of Visits 2 and 4, subjects were discharged from the clinic with study drugs assigned and with instructions for self-administration until they return for their next study visit (Visit 3 or 5).

[0262] Study drugs were administered orally as intact lozenges (swallowed whole, not chewed or crushed) and with water. The first and last two doses of study drug for each treatment period were administered to subjects by qualified study site members (first dose at Visits 2 and 4 and last two doses at Visits 3 and 5). Subjects self-administered their assigned study medications according to instructions until they returned for their next study visit (Visit 3 or 5). Qualified members of the study site contacted subjects by telephone on the second dosing day of each treatment period to assess compliance and adverse events via non-directed questions. If the subject experienced significant gastrointestinal symptoms, at the discretion of the investigator, subjects were instructed not to increase the dose.

[11 Chamadas para avaliar a conformidade e eventos adversos através de perguntas não direcionadas e lembrar a sujeitos de aumentar a dose [21 Teste de gravidez necessário em todos os sujeitos do sexo feminino, a menos que o sujeito tenha tido uma histerectomia ou esteja na pós-menopausa. [31 GLP-1, PYY, plasma glicose, insulina e triglicerídeos nas Visitas 2 e 4; GLP-1, PYY, plasma glicose, insulina, triglicerídeos e metforrnina nas Visitas de 3 e 5. Após o desafio de refeição nas visitas 2 e 4. Dose vespertina no dia 4 e dose matinal no dia 5 nas visitas 3 e 5. Tabela 2: Agenda de Café da Manhã Padronizado e Perfil de Amostragem de Sangue na Visita 2 e Visita 4

[11 total de volume de sangue de 6-ml por tempo de amostragem para avaliação de PYY, GLP-1, glicose de plasma, insulina e triglicerídeos. [2] Sujeitos devem ser instruídos a consumir o café da manhã padronizado dentro de 20 minutos. Tabela 3: Agenda do Dia 5 de Dosagem, Café da Manhã Padronizado e Perfil de Amostragem de Sangue na Visita 3 e Visita 5

[11 total de volume de sangue de 6-ml por tempo de amostragem para avaliação de PYY, GLP-1, glicose de plasma, insulina e triglicerídeos. [2] Sujeitos devem ser instruídos a consumir o café da manhã padronizado dentro de 20 minutos. [31 total de volume de sangue de 2 ml por ponto de tempo de amostragem para avaliação da Metformina.[0263] The procedures performed during the study are listed in Tables 1-3 below. Table 1: Study Plan (LCPOC& Protocol)

[11 Calls to assess compliance and adverse events through non-directed questions and remind subjects to increase dose [21 Pregnancy test required on all female subjects unless subject has had a hysterectomy or is on post -menopause. [31 GLP-1, PYY, plasma glucose, insulin, and triglycerides at Visits 2 and 4; GLP-1, PYY, plasma glucose, insulin, triglycerides, and metformin at Visits 3 and 5. After meal challenge at Visits 2 and 4. Afternoon dose on Day 4 and Morning dose on Day 5 at Visits 3 and 5. Table 2: Standardized Breakfast Schedule and Blood Sampling Profile at Visit 2 and Visit 4

[11 total 6-ml blood volume per sampling time for assessment of PYY, GLP-1, plasma glucose, insulin, and triglycerides. [2] Subjects should be instructed to consume the standardized breakfast within 20 minutes. Table 3: Dosing Day 5 Schedule, Standardized Breakfast, and Blood Sampling Profile at Visit 3 and Visit 5

[11 total 6-ml blood volume per sampling time for assessment of PYY, GLP-1, plasma glucose, insulin, and triglycerides. [2] Subjects should be instructed to consume the standardized breakfast within 20 minutes. [31 total blood volume of 2 ml per sampling time point for Metformin assessment.

[0264] Pharmacodynamics Assessments

[0265] Blood samples were collected according to the times shown in Tables 1, 2 and 3 and as described above. Fasting and postprandial plasma concentrations of the gut hormones GLP-1 and PYY, as well as plasma glucose, insulin, and triglyceride concentrations were measured by analytical methods. Blood samples from each visit were processed and stored at -70 °C for future exploratory analysis of additional hormones.

[0266] Pharmacokinetic Evaluations

[0267] Blood samples were collected according to the times shown in Tables 1, 2 and 3 and as described above. Metformin plasma concentrations were measured by analytical methods. Blood samples from each visit were processed and stored at -70 °C for future exploratory analysis of additional hormones.

[0268] Clinical Laboratory Assessments

[0269] The samples were collected according to the times shown in Tables 1, 2 and 3 and in the previous section.

[0270] Chemistry

[0271] Chemistry assessments included the following: urea nitrogen, creatinine, total protein, albumin, uric acid, total bilirubin, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyltranspeptidase, creatine phosphokinase, glucose, sodium, potassium, chloride, bicarbonate, phosphorus, lactate and calcium (or other routine chemistry panels).

[0272] Hematology

[0273] Hematology assessments included the following: red blood cell count, hemoglobin, hematocrit, white blood cell count, platelets, differential count, mean packed cell volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration (or other routine hematological assessments).

[0274] Urinalysis

[0275] Urinalysis assessments include the following: pH, specific gravity, glucose, blood, ketones, and protein (or other routine urinalysis).

[0276] Pregnancy Test

[0277] All female subjects, regardless of childbearing status (unless the subject was already menopausal or had had a hysterectomy), provided blood or urine for pregnancy tests. Study drug was not administered unless a negative result was obtained.

[0278] Vital Signs and Other Safety Related Observations

[0279] Clinically significant abnormalities in vital signs and other safety-related observations were followed up by the investigator and evaluated with additional testing if necessary until the underlying cause had been diagnosed or resolution had occurred.

[0280] Vital Signs

[0281] Vital sign measurements included resting systolic and diastolic blood pressure, heart rate, and body temperature. Vital signs were measured after the subject had rested for approximately 5 minutes and with the subject in a sitting position. The blood pressure measurement was repeated after at least 30 seconds and the average of the two readings was recorded.

[0282] EXAMPLE 1.2: Results

• 2 sujeitos excluídos da população avaliável; 1 retirado e 1 não foi pôde concluir a refeição do teste no final do Período de Tratamento 2 Tabela 5: Características Demográficas e da Base de Referência (n = 18)

[0283] The study design and timeline of events are shown in FIGs. 1-2. Shown in Tables 4 and 5 below are the resulting subject population and disposition (Table 4) and the demographic and baseline characteristics of 18 subjects (Table 5).Table 4: Subject Population and Disposition

• 2 subjects excluded from the evaluable population; 1 withdrawn and 1 was unable to complete the test meal at the end of the Treatment Period 2 Table 5: Demographic and Baseline Characteristics (n = 18)

[0284] FIG. 3 demonstrates that intake of Metformin DR minimizes the adsorption of Metformin in plasma compared to Metformin IR. The area under the curve (AUC) and Cmax values for Metformin DR and Metformin IR are provided in Table 6 below. Table 6: Metformin Plasma Pharmacokinetics

[0285] FIG. 4A-C shows an increase in meal-potentiated gut hormones in 16 subjects after treatment with Metformin DR, comparable to that of Metformin IR although treatment with Metformin DR minimized the systemic level of metformin compared to Metformin IR (FIG. 3). Furthermore, FIGs. 5A-B show a reduction in meal-potentiated glucose and insulin after Metformin DR treatment in 16 subjects compared to Metformin IR. FIG. 6 shows that treatment with Metformin DR results in a PYY response similar to that of Metformin IR, but has a low systemic exposure. FIGs. 7A-B show that the Metformin PK/PD ratio was dissociable in at least one patient.

[0286] EXAMPLE 2 - A RANDOMIZED CROSS-STUDY TO EVALUATE PK AND PD IN STEADY STATE DELAYED RELEASE AND IMMEDIATE RELEASE OF METMORPHINE IN SUBJECTS WITH TYPE 2 DIABETES MELLITUS

[0287] This randomized crossover study evaluates the steady-state pharmacokinetics and pharmacodynamics (glucose, insulin, glucagon-like peptide-1 [GLP-1] and peptide YY [PYY] of 500 mg and 1000 mg delayed-release metmorphine (Metmorphine DR), 1000 mg immediate-release metmorphine (Metmorphine IR) and 500 mg Metmorphine IR + 1000 mg Metmorphine DR in subjects with type 2 diabetes mellitus. Subjects managing their diabetes with oral antidiabetic therapy should be free of these medications at least for the fourteen days immediately prior to randomization.

[0288] Each treatment period lasted five days and was separated by seven-day washout intervals. Each treatment period contained a standardized breakfast and lunch profile on Day 1 prior to study drug administration (baseline assessment) and an identical profile on the morning of Day 5 (under drug assessment.)

[0289] EXAMPLE 2.1: Materials and Methods

[0290] Subjects were assessed for the effects of each treatment on circulating concentrations of PYY, GLP-1, glucose, and insulin over approximately 10 hours in response to two standardized meals (standardized breakfast of ~500 kcal at t= 0 min and standardized ~1000 kcal lunch at t=300 min) using standard protocols. Metformin pharmacokinetics over a sampling period of approximately 11 hours were also evaluated.

[0291] Population: Most of the randomized subjects were White (79.2%) and half were women (50.0%). Mean age was 51.3 years, mean weight was 93.4 kilograms, and mean BMI was 33.3 kg/m2 at baseline. Nineteen of the 24 subjects completed the study.

[0292] The primary population for pharmacokinetic and pharmacodynamic analyzes was the Evaluable Population (N=19), defined as all subjects who completed all treatment periods consistent with protocol procedures. The primary population for safety analyzes was the Willing to Treat Population (ITT) (N=24), defined as subjects who received at least one dose of study drug.

[0293] Formulations

[0294] The metformin DR formulation was a US-supplied, commercially available, film-coated, immediate-release 500 mg tablet of metformin hydrochloride, to which additional coatings (a seal coat and an enteric coating) were applied to in order to delay drug release in the gastrointestinal tract until the tablet reached a pH 6.5 region of the distal small intestine. The seal coating was applied at a nominal level of 2% of the tablet core weight and the enteric coating was applied at a nominal level of 3.8% of the tablet core weight. The lozenges are white, biconvex and circular in shape, each containing 500 mg of metformin hydrochloride. Inactive ingredients in the commercially available lozenge include providone, magnesium stearate, hypromellose and polyethylene glycol. Inactive ingredients in Elcelyx additional coating systems include hypromellose, triacetin, talc, methacrylic acid copolymer (Eudragit® L30 D-55), poly(acrylate-co-methyl methacrylate-co-methacrylic acid) 7:3:1 (Eudragit® FS 30 D), sodium lauryl sulfate, polysorbate 80, glyceryl monostearate and triethyl citrate.

[0295] The metmorphine IR formulation was the identical US-supplied commercially available immediate-release film-coated lozenge containing 500 mg metmorphine hydrochloride, to which only the additional seal coating is applied. No release delay (enteric) coating was applied. Inactive ingredients in the additional sealant coating system included hypromellose, triacetin and talc.

[0296] Metmorphine formulations were supplied to the site as bulk tablets packaged in screw cap containers labeled with container number and batch number. All study drugs were stored in cool, dry conditions as directed on the label and used only as directed by study personnel. Study medication was dispensed by the pharmacist or study personnel disclosed in accordance with the randomization scheme at the start of each treatment period.

[0297] Administration

[0298] Study drug was administered orally as intact lozenges (swallowed whole) with water at the start of breakfast and dinner meals. Subjects self-administered their assigned study medications on the evening of Day 1 through the morning of Day 4, according to instructions provided on Day 1 by study site staff. The last two doses of study drug for each treatment period (evening of Day 4 and morning of Day 5) were administered to subjects by qualified study site personnel. In order to reduce gastrointestinal side effects, all treatment regimens started at 500 mg/dose for the first 3 doses, followed by a randomized dose increase (500 mg/dose, 1000 mg, or 1500 mg/dose) for the remainder of the period. of study. Qualified study site members contacted subjects by telephone on the second dosing day of each treatment period to assess compliance and adverse events through non-directed questions and remind subjects to increase dose if appropriate.

[0299] EXAMPLE 2.2: Results

Pharmacokinetic Evaluations

[0300] Pharmacokinetic Profiles

[0301] Figure 8 shows the mean plasma metmorphine concentration on Day 5 by treatment and point in time. On Day 5, the mean pre-dose concentration of Metmorphine IR at t=0 was 350 ng/mL, which is consistent with steady state trough concentrations published in the literature. Following administration of Metmorphine IR at t=-1 minute, there was a rapid rise in metmorphine concentrations, which peaked at 1249 ng/mL 90 min post-dose, followed by a steady decline for the remainder of the sampling period.

[0302] Predose concentrations for both doses of Metmorphin DR were approximately 2 times greater than those of Metmorphin IR (716 ng/mL for 1000 mg DR and 602 ng/mL for 500 ng/mL DR vs. 350 ng /dL for 1000 mg IR). Following administration of both DR metmorphine doses at t=-1 minute, there was a decrease in metmorphine concentrations for the first 240 minutes, followed by a small rise in metformin concentrations after the standardized lunch meal, which then remained unchanged. high for the remainder of the sampling period. All 11-hour metmorphine profiles remained below pre-dose concentrations measured at t=0. Absorption profiles for Metmorphine DR dosing with the evening meal were slowed relative to doses given with the breakfast meal, consistent with slowed intestinal transit during sleeping hours. Metmorphine DR concentrations for the 500-mg dose were lower than the 1000-mg dose at all time points, although the reductions were less than dose proportional. This observation is consistent with the lack of dose proportionality reported for Metformin IR, which may be due to a saturated absorption process in the gut.

[0303] The Metmorphine DR + Metmorphine IR treatment group had the highest pre-dose concentrations of the four treatment groups (761 ng/mL). After administration of study drug at t=-1 minute, metformin concentrations rapidly increased similarly to metformin IR, but generally remained below the metformin concentration curve for the first 500 minutes. For the remainder of the sampling period, concentrations remained high, higher than those observed with the other treatments.

[0304] Pharmacokinetic Parameters

[0305] Table 7 and Figure 9 present the relative bioavailability of metformin by treatment versus Metmorphine IR on Day 5. Compared to the Metmorphine IR formulation, metmorphine exposure from t= = 0 to the time of the last concentration after administration of the study medication (AUCo-t) was statistically significantly reduced by 45.2% with 1000 mg Metmorphine DR (mean ratio 54.8%; p<0.0001) and 56.6% with 500 mg Metmorphine DR (mean ratio % 43.4; p<0.0001). Compared to Metmorphine IR, Cmax was also significantly reduced by 34.9% with 1000 mg Metmorphine DR (mean ratio 65.1%; p<0.0001) and 47.7% with 500 mg Metmorphine DR (% mean ratio of 52.3; p<0.0001).

[0306] Metformin DR + IR treatment resulted in exposures similar to that of Metformin 1000 mg IR (mean ratio 90.9%; p=0.2271) despite a 50% decrease in daily dose.Table 7 Relative Bioavailability of Metmorphine by Treatment Versus Metmorphine IR on Day 5 - Evaluable Population

Pharmacodynamic Assessments

[0307] Total PYY

Tabela 9. Total do Plasma PYY em Jejum {(pg/ml) na linha de base e no Dia 5 - População Avaliável

[0308] Figure 10 and Table 8 show the baseline and Day 5 mean total plasma PYY concentration profiles by treatment and point in time and the corresponding analysis of pharmacodynamic parameters, respectively. Baseline total plasma PYY concentrations were similar between treatments and at most time points. Additionally, all metformin treatments statistically significantly increased peak concentrations and total PYY exposure (p<0.01 for all), with percentage ratios (Day 5/Day 1) for AUCo-t and Cmax ranging from 1.26 to 1.55. Fasting total plasma PYY concentrations were also statistically significantly increased from baseline on Day 5 for each treatment (Table 9, p<0.01 for all). These results indicate that all treatments studied achieved similar total PYY responses for two standardized meals.

Table 9. Total Fasting Plasma PYY {(pg/ml) at baseline and Day 5 - Evaluable Population

[0309] GLP-1 Active

Tabela 11 Plasma GLP-1 Ativo em Jejum (pmol/L) na Linha de Base e Dia 5 - População Avaliável

[0310] Figure 11 and Table 10 show the profiles of the mean total concentration of active plasma GLP-1 at baseline and Day 5 by treatment and point in time and the corresponding analysis of pharmacodynamic parameters, respectively. Baseline total active GLP-1 plasma concentrations were similar between treatments and at most time points. Additionally, all metformin treatments statistically significantly increased peak concentrations and total exposure of active GLP-1 (p<0.01 for all), with percentage ratios (Day 5/Day 1) for AUCO-t and Cmax ranging from 1.42 to 1.88. Fasting total plasma GLP-1 concentrations were also statistically significantly increased from baseline on Day 5 for each treatment (Table 11, p<0.05 for all). These results indicate that all treatments studied achieved similar active GLP-1 responses for two standardized meals.

Table 11 Fasting Active Plasma GLP-1 (pmol/L) at Baseline and Day 5 - Evaluable Population

[0311] Glucose

[0312] Figure 12 and Table 12 show the mean glucose concentration profiles at baseline and Day 5 by treatment and point in time and the corresponding analysis of pharmacodynamic parameters, respectively.

[0313] Baseline total plasma glucose concentrations were similar between treatments and at most time points. Additionally, all metmorphine treatments statistically significantly reduced glucose exposure and peak concentrations for both meal intervals to a similar degree (p<0.001 for all).Table 12 Pharmacodynamic Analysis of Plasma Glucose (mg/dL) ) per Meal Interval - Treatment Comparison Based on Ratios - Evaluable Population

[0314] Table 13 presents the pharmacodynamic parameters for glucose from t = 0 until the time of the last concentration after administration of the study drug. Consistent with pharmacodynamic parameters for the breakfast and lunch intervals, all metformin treatments statistically significantly decreased glucose exposure and peak concentrations (p<0.001 for all), with percent ratios (Day5/Day1) to AUCo -t and Cmax ranging from 0.84 to 0.92.Table 13 Pharmacodynamic Analysis of Plasma Glucose (mg/dL) and Insulin (pmol/L) - Comparison in Treatment Based on Ratios - Evaluable Population

[0315] Table 14 presents the least squares mean (SE) and Figure 13 presents the individual change in fasting plasma glucose concentrations from baseline to Day 5 by treatment. Baseline fasting glucose concentrations were similar and ranged from 196 mg/dL to 200 mg/dL between treatment groups. All treatment groups achieved statistically significant (p<0.01 for all) reductions in fasting plasma glucose after 5 days of treatment. As shown in Figure 13, the LSM and distribution of individual responses were similar between treatment groups.Table 14. Fasting Plasma Glucose (mg/mL) at Baseline and Day 5 - Evaluable Population

[0316] Insulin

Tabela 16. Insulina em Jejum (pmol/L) na Linha de Base e Dia 5 - População Avaliável

[0317] Tables 15 and 16 present the pharmacodynamic parameters for insulin and baseline and fasting plasma insulin concentrations on Day 5, respectively. There were no statistically significant changes in insulin exposure, peak concentrations or fasting concentrations for any of the treatments (p>0.05 for all). Maintenance of insulin concentrations despite lower circulating glucose concentrations is indicative of an incretin effect.

Table 16. Fasting Insulin (pmol/L) at Baseline and Day 5 - Evaluable Population

[0318] Security assessments

[0319] Table 17 summarizes adverse effects caused by treatment by SOC, preferred termination, and most recent treatment at baseline.

[0320] Consistent with the information in the metmorphine prescription, adverse effects were primarily gastrointestinal in nature, with nausea, vomiting, and craving occurring only in the treatment groups receiving Metmorphine IR with or without Metmorphine DR. Diarrhea was reported by all treatment groups and appeared to be dose dependent, with the highest incidence with Metmorphine IR + Metmorphine DR (7 subjects, 33.3%) and the lowest incidence with the lowest dose of Metmorphine DR (2 subjects, 10.0%). Note that all GI events in the Metmorphine DR 500 mg group occurred during the post-treatment washout period while away from the study drug. Nervous system disorders, such as dizziness and headache, were also more frequent with Metmorphine IR than with any DR dosage. Overall, fewer gastrointestinal adverse effects and nervous system disturbances were reported with Metmorphine DR than with Metmorphine IR, indicating that the reduced systemic exposure to metmorphine achieved by bypassing the proximal small intestine improved tolerability. Adverse Effects Caused by Treatment by SOC, Preferred Termination, and Most Recent Treatment at Inception - ITT Population.

[0321] EXAMPLE 2.3: Discussion

[0322] In this study, metformin plasma concentrations were measured over 11 hours at steady state on the 5th day (Figure 1) of BID dosing (pre-breakfast and pre-supper) with 1000 mg immediate-release metformin (Metformin IR), 500 mg Metformin DR and 1000 mg Metformin DR or a combination of 500 mg Metformin IR and 1000 mg Metformin DR. All subjects had type 2 diabetes and each received treatment in a randomized cross-sectional design with a one-week washout between treatments.

[0323] The observed profiles indicated lower circulating amounts of metmorphine when used Metmorphine DR compared to Metmorphine IR. On Day 5, the mean pre-dose concentration of metmorphine with Metmorphine IR was 350 ng/mL, which is consistent with steady state trough concentrations published in the literature. Following Metmorphine administration on the morning of Day 5, there was a rapid rise in metmorphine concentrations, which peaked 90 min post dose, followed by a steady decline for the remainder of the sampling period.

[0324] With Metmorphine DR dosing, the highest metmorphine concentration was observed pre-dose on the morning of Day 5, being approximately 2 times higher at that point in time than Metmorphine IR. Following administration of both DR metmorphine doses, there was a decrease in metmorphine concentrations through the first 240 minutes followed by a small rise in metformin concentrations at 360 minutes, which then remained elevated for the remainder of the sampling period. All 11-hour metmorphine DR PK profiles remained below measured pre-dose concentrations at t=0. These results indicate that the absorption profiles for Metmorphine DR dosing with the evening meal were slowed relative to doses given with the breakfast meal, consistent with slowing intestinal transit during sleeping hours. Thus, concentrations over the first 240 minutes of the Day 5 profile were predominantly a result of absorption of the Day 4 evening dose, and concentrations from 240 minutes through 660 mins were predominantly a result of absorption of the Day 5 morning dose.

[0325] EXAMPLE 3: ANALYSIS OF PHARMACOKINETIC DIFFERENCES BETWEEN MORNING AND EVENING DOSAGES

[0326] To further characterize the pharmacokinetic differences between morning and evening doses, the study of Example 3 was designed to obtain 36-hour PK profiles of Metmorphine DR at doses of 500 and 1000 mg given at evening meals and at breakfast morning in healthy subjects. Subjects also received 1000 mg of Metmorphine IR with the evening meal and breakfast and 2000 mg of extended-release metformin (Metformin XR) with the evening meal during separate treatment periods. All subjects received each treatment according to a randomized crossover design with a one-week washout between treatments.

[0327] The metformin DR formulation was a US-supplied, commercially available, film-coated, immediate-release 500 mg tablet of metformin hydrochloride to which additional coatings (a seal coat and an enteric coating) were applied to in order to delay drug release in the gastrointestinal tract until the tablet reached a pH 6.5 region of the distal small intestine. The seal coating was applied at a nominal level of 2% of the tablet core weight and the enteric coating was applied at a nominal level of 3.8% of the tablet core weight. The lozenges are white, biconvex and circular in shape, each containing 500 mg of metformin hydrochloride. Inactive ingredients in the commercially available lozenge include providone, magnesium stearate, hypromellose and polyethylene glycol. Inactive ingredients in additional coating systems include hypromellose, triacetin, talc, methacrylic acid copolymer (Eudragit® L30 D-55), poly(acrylate-co-methyl methacrylate-co-methacrylic acid) 7:3:1 (Eudragit® FS 30 D), sodium lauryl sulfate, polysorbate 80, glyceryl monostearate and triethyl citrate. The metformin IR and metformin XR formulations were commercially available formulations (Aurobindo Pharma Limited and Bristol-Myers Squibb, respectively) without any modification.

[0328] As shown in Figure 14, both doses of Metformin DR resulted in substantially less systemic metformin than was seen with either Metformin IR or Metformin XR. Of note, the total plasma exposure to metformin as measured by AUC of 1000 mg Metformin IR BID and 2000 mg Metformin XR QD (2000 mg total daily doses) were very similar, consistent with the previously established bioequivalence between the two formulations . The profile of Metformin DR over the first 12 hours showed that there is a delay in systemic absorption of Metformin DR, with the first quantifiable plasma concentration occurring approximately 6-7 hours after dosing. The highest concentration was reached approximately 11 hours after the nightly dose. After a second dose of Metformin DR in the morning, the plasma concentration of metformin decreased until approximately 15 hours after the first dose, followed by a small corresponding increase approximately 3 hours after the second dose.

[0329] As noted above, data indicate that Metformin IR and both Metformin DR doses have slightly greater bioavailability after an evening dose than the morning dose, perhaps as a result of slower bowel transit during sleep hours.

[0330] Table 18 shows the Mean (CV%) of plasma and metformin pharmacokinetic parameters following oral administration of each treatment, and Figure 15 compares the mean (SEM) values of Cmax (left panel) and AUCo-36 hr ( right panel). Both doses of Metformin DR resulted in substantial reductions in exposure as well as a 6-7 hour delay in absorption. Table 18 Mean (CV%) of Metformin Pharmacokinetic Parameters in Plasma Following Oral Administration of Treatment A, B, C and D - Evaluable Population

SS: Estatisticamente significativo (p-value <0.0001) Tratamento A: 1000 mg Metformina IR BID (2 x 500 mg de pastilhas HCI de metmorfina [liberação imediata]) Tratamento B: 500 mg de Metformina DR BID (1 x 500 mg de pastilhas HCI de metformina [liberação retardada com revestimento entérico de pH 6,5]) Tratamento C: 1000 mg de Metformina DR BID (2 x 500 mg de pastilhas HCI de metformina [liberação retardada com revestimento entérico de pH 6.5]) Tabela 20 Biodisponibilidade Relativa de Metformina Seguindo Administração Oral de Tratamento de 500 mg de BID e 1000 mg de BID Metformina DR comparado a 2000 mg de QD Metformina XR - População Aval iável

SS: Estatisticamente significativo (p-value <0.0001) Tratamento B: 500 mg de Metformina DR BID (1 x 500 mg de pastilhas HCI de metformina [liberação retardada com revestimento entérico de pH 6,5]) Tratamento C: 1000 mg de Metformina DR BID (2 x 500 mg de pastilhas HCI de metformina [liberação retardada com revestimento entérico de pH 6.5]) Tratamento D: 2000 mg de QD de Metformina XR (4 x 500 mg de pastilhas HCI de metformina [liberação estendida])[0331] LSM geometric ratios and 90% confidence intervals for transforming In Cmax, AUCo-t and AUCo--of Metformin DR treatments (500 mg BID [Treatment B] and 1000 mg BID [Treatment C]) relative to Metformin IR (1000 mg BID [Treatment A]) are shown in Table 19 and the relative bioavailability is plotted in the left panel of Figure 16. These results indicate that the rate and extent of exposure (Cmax, AUCo-t, and AUCo- ) of 500 mg BID Metformin DR were approximately 55%, 68% and 67% lower, respectively, than those of 1000 mg BID Metformin IR. At 1000 mg BID Metformin DR (Treatment C, total daily dose of 2000 mg metformin), the rate and extent of exposure (Cmax, AUCo-t, and AUCo-) were approximately 33%, 52%, and 47% lower, respectively, than those of 1000 mg BID Metformin IR (Treatment A, total daily dose of 2000 mg metformin). Similar reductions in the rate and extent of exposure were observed when Metformin DR 500 mg BID and 1000 mg BID were compared to Metformin XR 2000 mg QD (Table 20; Figure 16, right panel).Table 19 Relative Bioavailability of Metformin Following Oral Administration of Metformin 500 mg BID and 1000 mg BID compared to Metformin 1000 mg BID IR - Evaluable Population

SS: Statistically significant (p-value <0.0001) Treatment A: 1000 mg Metformin IR BID (2 x 500 mg metmorphine HCI lozenges [immediate release]) Treatment B: 500 mg Metformin DR BID (1 x 500 mg lozenges Metformin HCI [delayed release with pH 6.5 enteric coating]) Treatment C: Metformin DR 1000 mg BID (2 x 500 mg Metformin HCI tablets [delayed release with pH 6.5 enteric coating]) Table 20 Relative Bioavailability of Metformin Following Oral Treatment Administration of 500 mg BID and 1000 mg BID Metformin DR compared to 2000 mg QD Metformin XR - Evaluable Population

SS: Statistically significant (p-value <0.0001) Treatment B: Metformin 500 mg DR BID (1 x 500 mg metformin HCI tablets [delayed release with pH 6.5 enteric coating]) Treatment C: Metformin 1000 mg DR BID (2 x 500 mg metformin HCI lozenges [delayed release pH 6.5 enteric coated]) Treatment D: 2000 mg Metformin XR QD (4 x 500 mg metformin HCI lozenges [extended release])

[0332] Taken together, the pharmacokinetic results of Examples 2 and 3 indicate that delivery of metformin to the lower intestine by administration of Metformin DR reduces the 24-hour bioavailability by approximately 50% relative to Metformin IR and Metformin XR at the same daily dose . Larger reductions in exposure were observed when the Metformin DR dose was reduced from a total daily dose of 2000 mg to 1000 mg, with no reduction in efficacy. Furthermore, timing of Metformin DR dosing (with evening or morning meals) significantly affected the timing of metformin release in the gut (3 vs. 6-7 hours post-dosing respectively) and provides an explanation for the study observation , in Example 2, that the lowest values of Metformin DR observed before the morning dose were higher than the lowest values observed 12 hours after the morning dose.

[0333] In the study of Example 2, while systemic exposure to metformin was substantially reduced with Metformin DR (45% at 2000 mg/day and -60% at 1000 mg/day, relative to 2000 mg/day of Metformin IR), the full extent of the glucose-lowering effects of Metformin IR (2000 mg/day) were maintained. Given that the full extent of the glucose-lowering effect was observed at both 2000 mg and 1000 mg of Metformin DR daily, lower doses are feasible, allowing for more elegant dosage forms than those currently available with existing products (Metformin IR and Metformin XD (ie, smaller lozenges, effective fixed-dose combinations, once-daily dosing) Furthermore, unlike Metformin IR, Metformin DR was not associated with any nausea or vomiting at any dose.

[0334] EXAMPLE 4: MODIFYING THE COATING PERCENTAGE TO IMPROVE PHARMACOKINETICS

[0335] In Example 1, the averaged pharmacokinetic profiles demonstrated that the delayed-release metformin formula resulted in a blunting and a delay in metformin absorption following a given dose compared to the immediate-release formula. However, individual PK profiles showed that in some cases (3/16), the PK profile on Day 5 of dosing did not differ between Metformin 1000 mg BID DR and Metformin 1000 mg BID IR. This suggests that, in some cases, enteric coating (at the nominal level of 2.4% of tablet core weight) was sufficient to prevent metformin release in the stomach. The left panel of Figure 17 shows a typical example of Metformin DR and IR PK profiles with a free dulling and delay of the metformin PK profile following a dose of Metformin DR at t = -240 min. The panel on the right shows one of three examples with no release delay.

[0336] To develop a coating that was more resistant to temporary increases in stomach pH that sometimes accompany meals, the formula was revised to provide a nominal enteric coating level of 3.8% of the tablet core weight with the same nominal 2% seal coating. Tablet dissolution performance was tested using Apparatus 2 USP (vane spinning at 50 or 100 rpm). The tablets were embedded in Japanese Sinker. For the first 2 h of the test, 0.1 M HCl was used as the test medium with a spatula speed of 100 rpm. After 2 h, the test medium was changed to 0.07M phosphate buffer, pH 6.8 with a spatula speed of 50 rpm. Samples were withdrawn at regular intervals from dissolution vessels and the appearance of metformin hydrochloride was monitored by spectrophotometer. As shown in Figure 18, the 2.4% and 3.8% nominal DR coating formulas resisted drug release for up to 2 hours in acid. Tablets coated with DR coating at a level of 2.4% (Batch # K111511-89A) showed drug release of about 80% at 30 mins and 100% at 60 mins in pH 6.8 buffer. Tablets coated with DR coatings at a level of 3.8% (Lot # K260512-127) showed a latency phase in drug release for the first 15 minutes with < 20% release at 30 mins and 100% at 60 minutes .

[0337] The revised formula (3.8% nominal DR coating) was used in both Example 2 and Example 3. All subjects (N=38) showed characteristic delay in peak concentrations consistent with a delayed release formula.

[0338] EXAMPLE 5: IN VITRO DISSOLUTION PROFILES OF EXAMPLE FORMULAS ACCORDING TO THE PRESENT INVENTION

[0339] Two- and three-stage in vitro dissolution profiles were performed on the formulas as described in Table 21 below and analyzed according to the protocol described in Example 4. Table 21: Exemplary formulas used in the in vitro dissolution analysis of the stage two and three.

[0340] The Kydes-89A formula was employed in Example 1 above, whereas the Kydes-127 formula was employed in Examples 2 and 3 above. The enteric coating is identical between the Kydes and Halo manufactured compositions and is composed of a blend of two acrylate coating systems, Eudragit® LC 30 D-55 and Eudragit® FS 30 D (Evonik), plus an coating (PlasACRYL™, Emerson Resources) and triethyl citrate.

[0341] Eudragit® L30 D-55 corresponds to Methacrylic Acid Copolymer Dispersion, NF and consists of a 30% dispersion of poly(methacrylic acid-co-ethyl acrylate) 1:1 in water, with Sodium Lauryl Sulfate NF 0.7% and Polysorbate 80 NF 2.3% in solid substance as emulsifiers. This polymer provides an enteric coating that is insoluble in acid and designed to dissolve above pH 5.5. Eudragit® FS 30 D is a 30% dispersion of poly(methyl acrylate co-methyl methacrylate-co-methacrylic acid) 7:3:1 in water with Sodium Lauryl Sulfate NF 0.3% and Polysorbate 80 NF 1.2 % in solid substance as emulsifiers. This polymer provides an enteric coating that is insoluble in acid and designed to dissolve above pH 7. Combinations of two Eudragits provide dissolution at pHs between 5.5 and 7.0. Previous work has indicated that a 60:40 combination of Eudragit L30 D-55 and FS 30 D would provide tablet dissolution close to a pH 6.5 corresponding to that expected in the distal small intestine.

[0342] PlasACRYL™ T20 is a 20% emulsion of Glyceryl Monostearate NF/Polysorbate 80 NF/Triethyl Citrate NF in water. It is created with a plasticizer / anti-tack agent to aid in the application and functionality of enteric coatings such as Eudragits. Triethyl Citrate is commonly used by the pharmaceutical industry in tablet coatings as a plasticizer. The amount of triethyl citrate included in the enteric coating system was based on experience and common industry practice. The amount of PlasACRYL included in the enteric coating system (approximately 10%) was based on manufacturers' recommendations.

[0343] The sealing coating for Halo formulas is Opadry® White YS-1-7003 (Colorcon), a mixture of hypromellose, titanium dioxide, polyethylene glycol 400 (macrogol) and polysorbate 80. Hypromellose is the polymeric coating, dioxide titanium is a coloring agent, polyethylene glycol 400 serves as an anti-caking agent and polysorbate 80 is present as a dispersant (in aqueous suspension) and plasticizer. The seal coating on tablets has been changed from Opadry® 03K19229 Clear, which was used in Kydes formulas, to an opaque white coating to ensure glare between various active strengths and placebo tablets. Opadry® 03K19229 Clear (Colorcon) used in Kydes formulas is a blend of hypromellose, triacetin and talc. Hypromellose is a polymeric coating, triacetin is present as a plasticizer and talc is present as an anti-tack agent. A 2.0% w/w seal coating (based on commercial tablet) was chosen for both formulas based on experience and typical pharmaceutical industry practice for seal coating tablets.

[0344] The enteric coating level has been reduced to 3.1% w/w (based on tablet weight) in tablets manufactured by Halo from 3.8% w/w of the level previously used in Kydes so that the in vitro release profile of metorphine HCI from the two manufacturers' delayed-release tablets matched each other. This need for a reduced level of coating has been attributed to Halo tablets as compared to the embossed alphanumeric characters present in Kydes coated tablets (ie commercially obtained core tablets). It is hypothesized that the edges of the notched features result in thinner regions in the enteric coating that result in a faster dissolution profile. Consequently, marking or embossing of the surface of the core tablet prior to application of the enteric coating needs to be considered and relative levels of enteric coating need to be adjusted, as demonstrated herein.

[0345] The targeted in vitro release profile consists of virtually complete protection against dissolution at low pH (0.1 N HCl) and dissolution above pH 6.5.

[0346] The data in Table 22 show the percent release by weight of metformin in two-stage in vitro dissolution analysis as measured in a USP Type II apparatus in aqueous medium at 37 °C as described in Example 4. Lot 1 corresponds to Halo 626 and is a tablet comprising 500 mg of metformin and Lot 2 corresponds to Kydes 127 also comprising 500 mg of metformin (See Figure 20).Table 22

[0347] The data in Table 23 show the percent release by weight of metformin in two-stage in vitro dissolution analysis as measured in a USP Type II apparatus in aqueous media at 37°C as described in Example 4. Lot 1 corresponds to Kydes-127 comprising 500 mg of metformin, Lot 2 corresponds to Halo-626 comprising 500 mg of metformin, and Lot 3 corresponds to Halo-625 comprising 300 mg of metformin (See Figure 19).Table 23

[0348] The percentage of weight gain and coating thickness in mg/cm 2 were calculated as shown in the formulas exemplified in Table 24 below. Table 24: Sample calculation of weight gain percentage and coating thickness

[0349] All patents and patent publications referred to herein are incorporated herein by reference.

[0350] Certain modifications and improvements will occur to those skilled in the art when reading the above description. It should be understood that all such modifications and improvements have been excluded from this document in the name of brevity and readability, but are duly within the scope of the claims below.

Claims (33)

1. Delayed release pharmaceutical composition, comprising an oral dosage form having a core comprising a biguanide compound and an enteric coating surrounding said core, wherein said enteric coating comprises a first copolymer comprising methyl acrylate, methyl methacrylate and methacrylic acid and a second copolymer comprising methacrylic acid copolymer, characterized in that said enteric coating is from 2.5% to 8% (weight/weight) with respect to the core. 2. Pharmaceutical composition according to claim 1, characterized in that said enteric coating is from 2.5% to 7% (w/w) in relation to the core. 3. Pharmaceutical composition according to claim 1, characterized in that said enteric coating is from 2.5% to 6.0% (w/w) in relation to the core. 4. Pharmaceutical composition according to claim 1, characterized in that said enteric coating is 2.5% to 5% (w/w) in relation to the core. 5. Pharmaceutical composition according to claim 1, characterized in that said first copolymer comprises poly (acrylate-co-methyl methacrylate-co-methacrylic acid) 7:3:1 and said second copolymer comprises poly(acrylate-co-methacrylic acid) methacrylic-co-ethyl acrylate) 1:1. 6. Pharmaceutical composition according to claim 5, characterized in that said first copolymer and said second copolymer are present in a ratio of 7:5 to 5:7. 7. Pharmaceutical composition according to claim 6, characterized in that said first copolymer and said second copolymer are present in a ratio of 6:4 to 4:6. 8. Pharmaceutical composition according to claim 6, characterized in that said enteric coating comprises about 60% of the first copolymer and 40% of the second copolymer. 9. Pharmaceutical composition according to any one of the preceding claims, characterized in that it comprises a sealing coating between the core coating and the enteric coating, in which the combined weight of the sealing coating and the enteric coating is 4% to 8% (w/w) with respect to the core, more preferably, 4.5% to 6.0% (w/w) with respect to the core. 10. Pharmaceutical composition according to claim 9, characterized in that the sealing coating comprises hypromellose, titanium dioxide, polyethylene glycol 400, polysorbate 80, triacetin, talc and combinations thereof. 11. Pharmaceutical composition according to any one of the preceding claims, characterized in that said biguanide compound is selected from the group consisting of metformin, phenformin, buformin and imeglimine. 12. Pharmaceutical composition according to claim 11, characterized in that said biguanide compound comprises metformin or a salt thereof. 13. Pharmaceutical composition according to claim 12, characterized in that said biguanide comprises metformin hydrochloride. 14. Pharmaceutical composition according to any one of the preceding claims, characterized in that said oral dosage form comprises a tablet, capsule or enteric coated microsphere. 15. Pharmaceutical composition according to claim 1, characterized in that said first copolymer and said second copolymer are present in a ratio of 6:4 to 4:6. 16. Pharmaceutical composition according to claim 1, characterized in that said enteric coating comprises 40% of said first copolymer and 60% of said second copolymer. 17. Pharmaceutical composition according to any one of claims 1 to 14, characterized in that said composition comprises a second antidiabetic agent selected from: thiazolidinediones, sulfonylureas, meglitinides, alpha-glucosidase inhibitors, a DPP inhibitor -IV, incretin mimetics and SGLT inhibitors. 18. Pharmaceutical composition according to claim 17, characterized in that said composition comprises a DPP-IV inhibitor. 19. Pharmaceutical composition according to claim 18, characterized in that said DPP-IV inhibitor is selected from the group consisting of sitagliptin, saxagliptin, berberine, vildagliptin, linagliptin and alogliptin. 20. Pharmaceutical composition according to claim 17, characterized in that said composition comprises an SGLT inhibitor. 21. Pharmaceutical composition according to any one of claims 1 to 14, characterized in that said composition comprises an anti-obesity agent or an anti-diabetes agent. 22. Pharmaceutical composition according to claim 21, characterized in that said anti-obesity agent is lorcaserin. 23. Pharmaceutical composition according to claim 21, characterized in that said anti-obesity agent is phentermine. 24. Pharmaceutical composition according to claim 21, characterized in that said anti-obesity agent is a combination of lorcaserin and phentermine. 25. Pharmaceutical composition according to claim 21, characterized in that said anti-obesity agent is a combination of topiramate and phentermine. 26. Pharmaceutical composition according to claim 21, characterized in that said anti-obesity agent is a combination of bupropion and naltrexone. 27. Use of a biguanide compound or salt thereof characterized in that it is in the preparation of a medicament for treating metabolic disorders in a patient in need thereof, wherein said medicament comprises a pharmaceutical composition as defined in any one of claims 1 to 14 . 28. Use according to claim 27, characterized in that said patient has a contraindication for the biguanide compound. 29. Use according to claim 28, characterized in that the patient has a contraindication selected from the group consisting of a condition of hypoxia, impaired clearance of lactate and impaired elimination of the biguanide compound. 30. Use according to claim 29, characterized by the fact that the patient has a moderate renal impairment, severe renal impairment or end-stage renal disease that results in compromised elimination of the biguanide compound. 31. Use according to any one of claims 27 to 30, characterized in that the patient in need thereof has hyperglycemia. 32. Use according to claim 31, characterized by the fact that the hyperglycemia is chronic. 33. Use according to claim 31 or 32, characterized by the fact that hyperglycemia is caused by type II diabetes.

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