# Patent

## Title
Substituted benzimidazole-, benztriazole-, and benzimidazolone-O-glucosides

## Abstract

This invention relates to substituted benzimidazole-O-glucosides, benztriazole-O-glucosides, and benzimidazolone-O-glucosides, compositions containing them, and methods of using them, for example, for the treatment or prophylaxis of diabetes and Syndrome X.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a continuation application of U.S. Ser. No. 12/337,949, filed Dec. 18, 2008, now abandoned, which is a continuation application of U.S. Ser. No. 11/453,727, filed Jun. 15, 2006, now U.S. Pat. No. 7,511,020, which is a divisional of U.S. Ser. No. 10/903,234, filed Jul. 30, 2004, now U.S. Pat. No. 7,094,764, which claims benefit of provisional applications Ser. No. 60/579,792, filed Jun. 15, 2004; Ser. No. 60/519,209, filed Nov. 12, 2003; Ser. No. 60/491,523, filed Aug. 1, 2003; and Ser. No. 60/491,534, filed Aug. 1, 2003, each of which is incorporated by reference herein.


FIELD OF THE INVENTION
This invention relates to substituted benzimidazole-O-glucosides, benztriazole-O-glucosides, and benzimidazolone-O-glucosides, compositions containing them, and methods of using them, for example, for the treatment or prophylaxis of diabetes and Syndrome X.
BACKGROUND OF THE INVENTION
Diabetes is a chronic disorder affecting carbohydrate, fat and protein metabolism in animals.
Type I diabetes mellitus, which comprises approximately 10% of all diabetes cases, was previously referred to as insulin-dependent diabetes mellitus (“IDDM”) or juvenile-onset diabetes. This disease is characterized by a progressive loss of insulin secretory function by beta cells of the pancreas. This characteristic is also shared by non-idiopathic, or “secondary”, diabetes having its origins in pancreatic disease. Type I diabetes mellitus is associated with the following clinical signs or symptoms: persistently elevated plasma glucose concentration or hyperglycemia; polyuria; polydipsia and/or hyperphagia; chronic microvascular complications such as retinopathy, nephropathy and neuropathy; and macrovascular complications such as hyperlipidemia and hypertension which can lead to blindness, end-stage renal disease, limb amputation and myocardial infarction.
Type II diabetes mellitus (non-insulin-dependent diabetes mellitus or NIDDM) is a metabolic disorder involving the dysregulation of glucose metabolism and impaired insulin sensitivity. Type II diabetes mellitus usually develops in adulthood and is associated with the body's inability to utilize or make sufficient insulin. In addition to the insulin resistance observed in the target tissues, patients suffering from type II diabetes mellitus have a relative insulin deficiency—that is, patients have lower than predicted insulin levels for a given plasma glucose concentration. Type II diabetes mellitus is characterized by the following clinical signs or symptoms: persistently elevated plasma glucose concentration or hyperglycemia; polyuria; polydipsia and/or hyperphagia; chronic microvascular complications such as retinopathy, nephropathy and neuropathy; and macrovascular complications such as hyperlipidemia and hypertension which can lead to blindness, end-stage renal disease, limb amputation and myocardial infarction.
Syndrome X, also termed Insulin Resistance Syndrome (IRS), Metabolic Syndrome, or Metabolic Syndrome X, is recognized in some 2% of diagnostic coronary catheterizations. Often disabling, it presents symptoms or risk factors for the development of Type II diabetes mellitus and cardiovascular disease, including impaired glucose tolerance (IGT), impaired fasting glucose (IFG), hyperinsulinemia, insulin resistance, dyslipidemia (e.g., high triglycerides, low HDL), hypertension and obesity.
Therapy for IDDM patients has consistently focused on administration of exogenous insulin, which may be derived from various sources (e.g., human, bovine, porcine insulin). The use of heterologous species material gives rise to formation of anti-insulin antibodies which have activity-limiting effects and result in progressive requirements for larger doses in order to achieve desired hypoglycemic effects.
Typical treatment of Type II diabetes mellitus focuses on maintaining the blood glucose level as near to normal as possible with lifestyle modification relating to diet and exercise, and when necessary, the treatment with anti-diabetic agents, insulin or a combination thereof. NIDDM that cannot be controlled by dietary management is treated with oral antidiabetic agents.
Although insulin resistance is not always treated in all Syndrome X patients, those who exhibit a prediabetic state (e.g., IGT, IFG), where fasting glucose levels may be higher than normal but not at the diabetes diagnostic criterion, is treated in some countries (e.g., Germany) with metformin to prevent diabetes. The anti-diabetic agents may be combined with pharmacological agents for the treatment of the concomitant co-morbidities (e.g., antihypertensives for hypertension, hypolipidemic agents for lipidemia).
First-line therapies typically include metformin and sulfonylureas as well as thiazolidinediones. Metformin monotherapy is a first line choice, particularly for treating type II diabetic patients who are also obese and/or dyslipidemic. Lack of an appropriate response to metformin is often followed by treatment with metformin in combination with sulfonylureas, thiazolidinediones, or insulin. Sulfonylurea monotherapy (including all generations of drugs) is also a common first line treatment option. Another first line therapy choice may be thiazolidinediones. Alpha glucosidase inhibitors are also used as first and second line therapies. Patients who do not respond appropriately to oral anti-diabetic monotherapy, are given combinations of the above-mentioned agents. When glycemic control cannot be maintained with oral antidiabetics alone, insulin therapy is used either as a monotherapy, or in combination with oral antidiabetic agents.
One recent development in treating hyperglycemia is focused on excretion of excessive glucose directly into urine. Specific inhibitors of SGLTs have been shown to increase the excretion of glucose in urine and lower blood glucose levels in rodent models of IDDM and NIDDM.
SUMMARY OF THE INVENTION
One aspect of the present invention is directed to methods and compositions for the treatment or prophylaxis of diabetes, Syndrome X, or associated symptoms or complications. More specifically, this invention is directed to a novel method of treating diabetes or Syndrome X, or associated symptoms or complications thereof, in a subject afflicted with such a condition, said method comprising administering one or more glucose reabsorption inhibitors and administering one or more antidiabetic agent(s) for the treatment of diabetes or Syndrome X, or associated symptoms or complications thereof.
Another aspect of the invention features compounds of formula (III):




wherein:

    X is CH, N, or C═O;
    R1 is H or absent;
    R2 is H, F, Cl, OCH3, OCH2CH3, C1-4 alkyl, CF3, SCH3, substituted or unsubstituted phenyl and NR3R4;
    R3 and R4 are H, C1-6 alkyl, or taken together with the nitrogen atom to which they are both attached form a 5-6 membered heterocyclic ring with optionally 1-2 additional heteroatoms independently selected from O, S, and N;
    Q is —(CH2)n— where n is 1 or 2;
    P is H, C2-7 acyl, or (C1-6 alkoxy)carbonyl; and


Z is substituted or unsubstituted, and is selected from C3-7 cycloalkyl, phenyl, 5- or 6-membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, a biaryl, and a 9- or 10-membered fused bicyclyl or fused heterobicyclyl, wherein said fused heterobicyclyl has between 1 and 4 heteroatoms independently selected from N, O, and S; or a pharmaceutically acceptable salt, thereof.
One aspect of the invention features a pharmaceutical composition comprising a glucose reabsorption inhibitor, at least one additional antidiabetic agent, and a pharmaceutically acceptable carrier. The invention also provides a process for formulating a pharmaceutical composition, comprising formulating together a glucose reabsorption inhibitor, one or more antidiabetic agent(s), and a pharmaceutically acceptable carrier.
An embodiment of the invention is a method for treating diabetes or Syndrome X, or associated symptoms or complications thereof in a subject, said method comprising administering to said subject a jointly effective amount of a glucose reabsorption inhibitor and administering to said subject a jointly effective amount of at least one antidiabetic agent, said combined administration providing the desired therapeutic effect.
Another embodiment of the invention is a method for inhibiting the onset of diabetes or Syndrome X, or associated symptoms or complications thereof in a subject, said method comprising administering to said subject a jointly effective dose of a glucose reabsorption inhibitor and administering to said subject a jointly effective amount of one or more anti-diabetic agent(s), said combined administration providing the desired prophylactic effect.
In the disclosed methods, the diabetes or Syndrome X, or associated symptoms or complications thereof, is selected from IDDM, NIDDM, IGT, IFG, obesity, nephropathy, neuropathy, retinopathy, atherosclerosis, polycystic ovarian syndrome, hypertension, ischemia, stroke, heart disease, irritable bowel disorder, inflammation, and cataracts.
Also included in the invention is the use of one or more glucose reabsorption inhibitors in combination with one or more antidiabetic agents for the preparation of a medicament for treating a condition selected from IDDM, NIDDM, IGT, IFG, obesity, nephropathy, neuropathy, retinopathy, atherosclerosis, polycystic ovarian syndrome, hypertension, ischemia, stroke, heart disease, irritable bowel disorder, inflammation, and cataracts.


DETAILED DESCRIPTION OF THE INVENTION
All diabetics, regardless of their genetic and environmental backgrounds, have in common an apparent lack of insulin or inadequate insulin function. Because transfer of glucose from the blood into muscle and fatty tissue is insulin dependent, diabetics lack the ability to utilize glucose adequately, which leads to undesired accumulation of glucose in the blood (hyperglycemia). Chronic hyperglycemia leads to decrease in insulin secretion and contributes to increased insulin resistance, and as a result, the blood glucose concentration is increased so that diabetes is self-exacerbated (Diabetologia, 1985, “Hyperglycaemia as an inducer as well as a consequence of impaired isle cell function and insulin resistance: implications for the management of diabetes”, Vol. 28, p. 119); Diabetes Cares, 1990, Vol. 13, No. 6, “Glucose Toxicity”, pp. 610-630). Therefore, by treating hyperglycemia, the aforementioned self-exacerbating cycle is interrupted so that the prophylaxis or treatment of diabetes is made possible.
U.S. Pat. No. 6,153,632 to R. Rieveley discloses a method and composition stated to be for the treatment of diabetes mellitus (Type I, Impaired Glucose Tolerance [“IGT”] and Type II), which incorporates a therapeutic amount of one or more insulin sensitizers along with one or more of an orally ingested insulin, an injected insulin, a sulfonylurea, a biguanide or an alpha-glucosidase inhibitor for the treatment of diabetes mellitus.
According to one aspect, the invention features the combination of a PPAR modulator, preferably a PPAR δ agonist, and an SGLT inhibitor, preferably an SGLT 2 inhibitor or a selective SGLT 2 inhibitor.
A. Terms
Some terms are defined below and by their usage throughout this disclosure.
Unless otherwise noted, “alkyl” and “alkoxy” as used herein, whether used alone or as part of a substituent group, include straight, cyclic, and branched-chain alkyl having 1 to 8 carbon atoms, or any number within this range. For example, alkyl radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, 2-butenyl, 2-butynyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl and 2-methylpentyl. Alkoxy radicals are oxygen ethers formed from the previously described straight or branched chain alkyl groups. The alkyl and alkoxy group may be independently substituted with one to five, preferably one to three groups selected from halogen (F, Cl, Br, I), oxo, OH, amino, carboxyl, and alkoxy. The alkyl and alkoxy group may also be independently linked to one or more PEG radicals (polyethylene glycol).
The term “acyl” as used herein, whether used alone or as part of a substituent group, means an organic radical having a carbonyl group linked to hydrocarbyl group having 1 to 7 carbon atoms (branched or straight chain or cyclic) derived from an organic acid by removal of the hydroxyl group. For example C4 acyl can include (CO)CH2CH2CH2CH3 and (CO)(CH2 (CH)(CH3)2; similarly, C6 acyl includes both (CO)(C6H13) and (CO)(C6H5). The term “Ac” as used herein, whether used alone or as part of a substituent group, means acetyl.
“Aryl” is a carbocyclic aromatic radical including, but not limited to, phenyl, 1- or 2-naphthyl and the like. The carbocyclic aromatic radical may be substituted by independent replacement of 1 to 3 of the hydrogen atoms thereon with halogen, OH, CN, mercapto, nitro, amino, cyano, optionally substituted C1-C8-alkyl, optionally substituted alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkyl-amino, di(C1-C8-alkyl)amino, formyl, carboxyl, alkoxycarbonyl, alkoxycarbonyloxy, alkanoyloxy, phenyl, carbamoyl, carboxamide, di-lower alkylcarbamoyloxy, phenoxycarbonyloxy group, lower alkylenedioxy, benzoyloxy, alkyl-CO—O—, alkyl-O—CO—, —CONH2, alkyl-O—CO—O—, or alkyl-CO—NH—. Illustrative aryl radicals include, for example, phenyl, naphthyl, biphenyl, indene




indane





fluorophenyl, difluorophenyl, benzyl, benzoyloxyphenyl, carboethoxyphenyl, acetylphenyl, ethoxyphenyl, phenoxyphenyl, hydroxyphenyl, carboxyphenyl, trifluoromethylphenyl, methoxyethylphenyl, acetamidophenyl, tolyl, xylyl, dimethylcarbamylphenyl and the like. “Ph” or “PH” denotes phenyl.

The term “heteroaryl” as used herein represents a stable five or six-membered monocyclic or bicyclic aromatic ring system which consists of carbon atoms and from one to three heteroatoms selected from N, O and S. The heteroaryl group may be attached at any heteroatom or carbon atom, which results in the creation of a stable structure. Examples of heteroaryl groups include, but are not limited to benzofuranyl, benzothiophenyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, thiophenyl, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzisoxazolyl, benzoxazolyl, benzopyrazolyl, indolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl or quinolinyl. Preferred heteroaryl groups include pyridinyl, thiophenyl, furanyl, and quinolinyl. When the heteroaryl group is substituted, the heteroaryl group may have one to three substituents which are independently selected from halogen, OH, CN, mercapto, nitro, amino, cyano, optionally substituted C1-C8-alkyl, optionally substituted alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkyl-amino, di(C1-C8-alkyl)amino, formyl, carboxyl, alkoxycarbonyl, alkoxycarbonyloxy, alkanoyloxy, phenyl, carbamoyl, carboxamide, di-lower alkylcarbamoyloxy, phenoxycarbonyloxy group, lower alkylenedioxy, benzoyloxy, alkyl-CO—O—, alkyl-O—CO—, —CONH2, alkyl-O—CO—O—, or alkyl-CO—NH—.
The terms “heterocycle,” “heterocyclic,” and “heterocyclyl” refer to an optionally substituted, fully or partially saturated, aromatic or nonaromatic, cyclic group which is, for example, a 4- to 7-membered monocyclic, 7- to 11-membered (or 9- to 10-membered) bicyclic (or heterobicyclyl), or 10- to 15-membered tricyclic ring system, which has at least one heteroatom in at least one carbon atom containing ring. Each ring of the heterocyclic group containing a heteroatom may have 1, 2, or 3 heteroatoms selected from nitrogen atoms, oxygen atoms, and sulfur atoms, where the nitrogen and sulfur heteroatoms may also optionally be oxidized. The nitrogen atoms may optionally be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom.
Exemplary monocyclic heterocyclic groups include pyrrolidinyl; oxetanyl; pyrazolinyl; imidazolinyl; imidazolidinyl; oxazolyl; oxazolidinyl; isoxazolinyl; thiazolidinyl; isothiazolidinyl; tetrahydrofuryl; piperidinyl; piperazinyl; 2-oxopiperazinyl; 2-oxopiperidinyl; 2-oxopyrrolidinyl; 4-piperidonyl; tetrahydropyranyl; tetrahydrothiopyranyl; tetrahydrothiopyranyl sulfone; morpholinyl; thiomorpholinyl; thiomorpholinyl sulfoxide; thiomorpholinyl sulfone; 1,3-dioxolane; dioxanyl; thietanyl; thiiranyl; and the like. Exemplary bicyclic heterocyclic groups (or heterobicyclyls) include quinuclidinyl; tetrahydroisoquinolinyl; dihydroisoindolyl; dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl); dihydrobenzofuryl; dihydrobenzothienyl; dihydrobenzothiopyranyl; dihydrobenzothiopyranyl sulfone; dihydrobenzopyranyl; indolinyl; isochromanyl; isoindolinyl; benzimidazolyl, benzthiazolyl; piperonyl; tetrahydroquinolinyl; and the like. When the heteroaryl group is substituted, the heterocyclyl may be independently substituted with one to five, preferably one to three groups selected from halogen, OH, CN, mercapto, nitro, amino, cyano, optionally substituted C1-C8-alkyl, optionally substituted alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkyl-amino, di(C1-C8-alkyl)amino, formyl, carboxyl, alkoxycarbonyl, alkoxycarbonyloxy, alkanoyloxy, phenyl, carbamoyl, carboxamide, di-lower alkylcarbamoyloxy, phenoxycarbonyloxy group, lower alkylenedioxy, benzoyloxy, alkyl-CO—O—, alkyl-O—CO—, —CONH2, alkyl-O—CO—O—, or alkyl-CO—NH—.
The term “biaryl” includes a heteroaryl linked to a phenyl, a phenyl linked to a heteroaryl (such as thiophene, pyridine, and pyrazole), and a phenyl linked to a phenyl. Examples of phenyl-phenyl, heteroaryl-phenyl, heteroaryl-phenyl, and phenyl-heteroaryl, respectively, include:




The term “composition” is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combinations of the specified ingredients in the specified amounts.
The term “combined administration” includes co-administration wherein: 1) the two or more agents are administered to a subject at substantially similar times; and 2) the two or more agents are administered to a subject at different times, at independent intervals which may or may not overlap or coincide.
The term “subject” as used herein, refers to an animal, preferably a mammal, most preferably a human, who is the object of treatment, observation or experiment.
The term “RXR modulator” as used herein, refers to Retinoid-X receptor agonists, partial agonists, or antagonists. Preferably the modulator increases insulin sensitivity. According to one aspect, the modulator is an RXR agonist.
Diabetes, Syndrome X, and associated symptoms or complications include such conditions as IDDM, NIDDM, IGT, IFG, obesity, nephropathy, neuropathy, retinopathy, atherosclerosis, polycystic ovarian syndrome, hypertension, ischemia, stroke, heart disease, irritable bowel disorder, inflammation, and cataracts. Examples of a prediabetic state includes IGT and IFG.
Methods are known in the art for determining effective doses for therapeutic and prophylactic purposes for the disclosed pharmaceutical compositions or the disclosed drug combinations, whether or not formulated in the same composition. For therapeutic purposes, the term “jointly effective amount” as used herein, means that amount of each active compound or pharmaceutical agent, alone or in combination, that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated. For prophylactic purposes (i.e., inhibiting the onset or progression of a disorder), the term “jointly effective amount” refers to that amount of each active compound or pharmaceutical agent, alone or in combination, that inhibits in a subject the onset or progression of a disorder as being sought by a researcher, veterinarian, medical doctor or other clinician, the delaying of which disorder is mediated by the modulation of glucose reabsorption activity or other antidiabetic agent activity or both. Thus, the present invention provides combinations of two or more drugs wherein, for example, (a) each drug is administered in an independently therapeutically or prophylactically effective amount; (b) at least one drug in the combination is administered in an amount that is sub-therapeutic or sub-prophylactic if administered alone, but is therapeutic or prophylactic when administered in combination with the second or additional drugs according to the invention; or (c) both drugs are administered in an amount that is sub-therapeutic or sub-prophylactic if administered alone, but are therapeutic or prophylactic when administered together.
The term “protecting groups” refer to those moieties known in the art that are used to mask functional groups; protecting groups may be removed during subsequent synthetic transformations or by metabolic or other in vivo administration conditions. During any of the processes for preparation of the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J. F. W. McOmie, Plenum Press, 1973; and T. W. Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, Third Edition, John Wiley & Sons, 1999. The protecting groups may be removed at a convenient subsequent stage using methods known in the art. Examples of hydroxyl and diol protecting groups are provided below.
Protection for the hydroxyl group includes methyl ethers, substituted methyl ethers, substituted ethyl ethers, substitute benzyl ethers, and silyl ethers.
Substituted Methyl Ethers
Examples of substituted methyl ethers include methyoxymethyl, methylthiomethyl, t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyloxymethyl, p-methoxybenzyloxymethyl, (4-methoxyphenoxy)methyl, t-butoxymethyl, 4-pentenyloxymethyl, siloxymethyl, 2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl, tetrahydropyranyl, 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxido, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl and 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl.
Substituted Ethyl Ethers
Examples of substituted ethyl ethers include 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, and polyethyleneglycol ethers.
Substituted Benzyl Ethers
Examples of substituted benzyl ethers include p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxy)phenyldiphenylmethyl, 4,4′,4′′-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4′,4′′-tris(levulinoyloxyphenyl)methyl, 4,4′,4′′-tris(benzoyloxyphenyl)methyl, 3-(Imidazol-1-ylmethyl)bis(4′,4′′-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1′-pyrenyl methyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, and benzisothiazolyl S,S-dioxido.
Silyl Ethers
Examples of silyl ethers include trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl, and t-butylmethoxyphenylsilyl.
Esters
In addition to ethers, a hydroxyl group may be protected as an ester. Examples of esters include formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, p-P-phenylacetate, 3-phenylpropionate, 4-oxopentanoate(levulinate), 4,4-(ethylenedithio)pentanoate, pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate(mesitoate), and polyethyleneglycol esters.
Carbonates
Examples of carbonates include methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, 2-(triphenylphosphonio)ethyl, isobutyl, vinyl, allyl, p-nitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl, methyl dithiocarbonate, and polyethyleneglycol carbonates.
Assisted Cleavage
Examples of assisted cleavage include 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl carbonate, 4-(methylthiomethoxy)butyrate, and 2-(methylthiomethoxymethyl)benzoate.
Miscellaneous Esters
Examples of miscellaneous esters include 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate(tigloate), o-(methoxycarbonyl)benzoate, p-P-benzoate, α-naphthoate, nitrate, alkyl N,N,N′,N′-tetramethylphosphorodiamidate, N-phenylcarbamate, borate, dimethylphosphinothioyl, and 2,4-dinitrophenylsulfenate
Sulfonates
Examples of sulfonates include sulfate, methanesulfonate(mesylate), benzylsulfonate, and tosylate.
Protection for 1,2- and 1,3-Diols
Cyclic Acetals and Ketals
Examples of cyclic acetals and ketals include methylene, ethylidene, 1-t-butylethylidene, 1-phenylethylidene, (4-methoxyphenyl)ethylidene, 2,2,2-trichloroethylidene, acetonide (isopropylidene), cyclopentylidene, cyclohexylidene, cycloheptylidene, benzylidene, p-methoxybenzylidene, 2,4-dimethoxybenzylidene, 3,4-dimethoxybenzylidene, and 2-nitrobenzylidene.
Cyclic Ortho Esters
Examples of cyclic ortho esters include methoxymethylene, ethoxymethylene, dimethoxymethylene, 1-methoxyethylidene, 1-ethoxyethylidine, 1,2-dimethoxyethylidene, α-methoxybenzylidene, 1-(N,N-dimethylamino)ethylidene derivative, α-(N,N-dimethylamino)benzylidene derivative, and 2-oxacyclopentylidene.
Silyl Derivatives
Examples of silyl derivatives include di-t-butylsilylene group, and 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivative.
Glucose Reabsorption Inhibitors
One method of treating hyperglycemia is to excrete excessive glucose directly into urine so that the blood glucose concentration is normalized. For example, sodium-glucose cotransporters (SGLTs), primarily found in chorionic membrane of the intestine and the kidney, are a family of proteins actively involved in the normal process of glucose absorption. Among them, SGLT1 is present in intestinal and renal epithelial cells (Lee et al., 1994), whereas SGLT2 is found in the epithelium of the kidney (You et al., 1995, MacKenzie et al., 1994). Glucose absorption in the intestine is primarily mediated by SGLT1, a high-affinity low-capacity transporter with a Na+:glucose transport ratio of 2:1. SGLT2, also known as SAAT1, transports Na+ and glucose at a ratio of 1:1 and functions as a low-affinity high-capacity transporter. These SGLTs are characterized in Table 1:













TABLE 1








Preferred
Km*
TmG**
Km*


ISOFORM
TISSUE
Stoichiometry
Substrate
in vitro
in vitro
In vivo





















SGLT1
Sm. Intestine
2:1
D-glucose
0.1
nd
Nd





D-galactose



Kidney (S1, S3)
2:1
D-glucose
0.39
7.9
0.3





D-galactose


SGLT2
Kidney (S3)
1:1
D-glucose
1.64
83
6


(SAAT1)





*(mM) for D-glucose


**Maximal transport rate pmol/min/mm






Renal reabsorption of glucose is mediated by SGLT1 and SGLT2 (Silverman et al., 1992; Deetjen et al., 1995). Plasma glucose is filtered in the glomerulus and is transepithelially reabsorbed in the proximal tubules. SGLT1 and SGLT2 are located in the apical plasma membranes of the epithelium and derive their energy from the inward sodium gradient created by the Na+/K+ ATPase pumps located on the basolateral membrane. Once reabsorbed, the elevated cytosolic glucose is then transported to the interstitial space by facilitated glucose transports (GLUT1 and GLUT2). Therefore, inhibition of SGLTs reduces plasma glucose through suppression of glucose reabsorption in the kidney. A therapeutically or prophylactically effective amount of an SGLT inhibitor, such as that sufficient to increase urine glucose excretion, or to decrease plasma glucose, in a subject by a desired amount per day, can be readily determined using methods established in the art. Recently, it has been found that phlorizin, a natural glycoside present in barks and stems of Rosaceae (e.g., apple, pear, etc.), inhibits Na+-glucose co-transporters located in chorionic membrane of the intestine and the kidney. By inhibiting Na+-glucose co-transporter activity, phlorizin inhibits the renal tubular glucose reabsorption and promotes the excretion of glucose so that the glucose level in a plasma is controlled at a normal level for a long time via subcutaneous daily administration (Journal of Clinical Investigation, 1987, Vol. 79, p. 1510).
Other SGLT inhibitors include alkyl- and phenyl-glucosides, 1-5-isoquinolinesulfonyl)-2-methylpiperazine-HCl (indirectly via protein kinase C), p-chloromercuribenzoate (PCMB), N,N′-dicyclohexylcarbodiimide (DCCD), copper and cadmium ions, and trivalent lanthanides.
B. Compounds
The invention features compounds of Formula (III):




wherein:

    X is CH, N, or C═O;
    R1 is H or absent;
    R2 is H, F, Cl, OCH3, OCH2CH3, C1-4 alkyl, CF3, SCH3, substituted or unsubstituted phenyl and NR3R4;
    R3 and R4 are H, C1-6 alkyl, or taken together with the nitrogen atom to which they are both attached form a 5-6 membered heterocyclic ring with optionally 1-2 additional heteroatoms independently selected from O, S, and N;
    Q is —(CH2)n— where n is 1 or 2;
    P is H, C2-7 acyl, or (C1-6 alkoxy)carbonyl; and


Z is substituted or unsubstituted, and is selected from C3-7 cycloalkyl, phenyl, 5- or 6-membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, a biaryl, and a 9- or 10-membered fused bicyclyl or fused heterobicyclyl, wherein said fused heterobicyclyl has between 1 and 4 heteroatoms independently selected from N, O, and S; or a pharmaceutically acceptable salt, thereof.
Examples of compounds of Formula (III) include those wherein: (a) R1 is H or absent; (b) R2 is H, methyl, or ethyl; (c) Q is —(CH2)n— and n is 1; (d) Z is independently substituted with between 1 and 3 substituents independently selected from C1-4 alkoxy, phenoxy, C1-4 alkyl, C3-6 cycloalkyl, halo, hydroxy, cyano, amino, C1-4 alkylthio, C1-4 alkylsulfonyl, C1-4 alkylsulfinyl, C1-4 aminoalkyl, mono- and di(C1-4 alkyl)amino, phenyl, C1-4 alkylaminosulfonyl (SO2NHR), amino-(alkylsulfonyl) (—NHSO2R—), C1-4 dialkylaminosulfinyl (SONHRR), C1-4 alkylamido (NHCOR), C1-4 alkylcarbamido (CONHR), 5-6 membered heterocyclyl containing between 1 and 3 heteroatoms independently selected from N, S, and O; and wherein the substituent(s) on Z can be further independently substituted with between 1 and 3 substituents independently selected from C1-4 alkoxy, C1-4 alkyl, halo, hydroxy, cyano, amino, mono or di C1-4 alkyl amino and C1-4 alkylthio; (e) Z is 4-substituted phenyl, 3,4-disubstituted phenyl, benzhydryl, substituted or unsubstituted thiophene, biaryl, benzofuranyl, dihydrobenzofuranyl, 4-substituted pyridyl, benzo[b]thienyl, chromanyl, benzothiophenyl, indenyl, indanyl, naphthyl, or 2,3-dihydro-benzo[1,4]dioxanyl; (f) Z is unsubstituted or substituted with between 1 and 2 substituents independently selected from methoxy, ethoxy, fluoro, chloro, methyl, ethyl, propyl, butyl and isopropyl; (g) Z is biphenyl, 4-(3-pyridyl)phenyl, 4-(2-thienyl)phenyl), 4-(1H-imidazole-1-yl)-phenyl, 4-(1H-pyrazol-1-yl)-phenyl, (4-propyl)phenyl, (4-ethyl)phenyl, (4-methoxyphenyl), dihydrobenzofuran-5-yl, or dihydrobenzofuran-6-yl; (h) R1 is absent, X is CH; and R2 is H, methyl, ethyl, or methoxy; (i) Q is —(CH2)n—; n is 1 or 2; and R2 is H, methyl, or ethyl; (j) limitations of (i) and R1 is absent; (k) R1 is absent; Q is —(CH2)n—; n is 1 or 2; and R2 is H, methyl, or ethyl; and Z is 4-substituted phenyl, 3,4-disubstituted phenyl, benzhydryl, substituted or unsubstituted thiophene, biaryl, benzofuranyl, dihydrobenzofuranyl, 4-substituted pyridyl, benzo[b]thienyl, chromanyl, benzothiophenyl, indenyl, indanyl, naphthyl, or 2,3-dihydro-benzo[1,4]dioxanyl; (l) Z is biphenyl, 4-(3-pyridyl)phenyl, 4-(2-thienyl)phenyl), 4-(1H-imidazole-1-yl)-phenyl, 4-(1H-pyrazol-1-yl)-phenyl, (4-ethyl)phenyl, (4-methoxyphenyl), dihydrobenzofuran-5-yl, or dihydrobenzofuran-6-yl; and Z is unsubstituted or substituted with between 1 and 2 substituents independently selected from methoxy, ethoxy, fluoro, chloro, methyl, ethyl, propyl, butyl and isopropyl; (m) and combinations of the above.
Examples of most preferred compounds include those selected from 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3H-benzoimidazol-4-yloxy}-β-D glucopyranoside; 2-[3-(4-Ethyl-benzyl)-3H-benzotriazol-4-yloxy]-D-glucopyranoside and 2-[3-(4-Ethyl-benzyl)-3H-benzoimidazol-4-yloxy]-D-glucopyranoside.
Additional examples of preferred compounds include those selected from 2-[3-(4-Ethyl-benzyl)-6-methyl-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-{3-[2-(4-Methoxyphenyl)-ethyl]-3H-benzoimidazol-4-yloxy}-β-D-glucopyranoside; 2-{3-[2-(4-Methoxyphenyl)-ethyl]-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside; 2-[3-(2-Naphthalen-2-yl-ethyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside and 2[3-(4-Ethyl-benzyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D-glucopyranoside.
Further examples of compounds of the invention include those selected from 2-[3-(2-Naphthalen-2-yl-ethyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D-glucopyranoside and 2-[3-(2-(4-Methoxyphenyl)-ethyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D glucopyranoside.
Additional preferred compounds include those selected from 2-[3-(5-Ethyl-thiophen-2-ylmethyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-6-methyl-3H-benzoimidazol-4-yloxy}-β-D-glucopyranoside; 2-[3-(4-Thiophen-3-yl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-[3-(4-Pyrazol-1-yl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-[3-(4-Pyridin-3-yl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-[3-(4-Pyrrol-1-yl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-[3-(4-Imidazol-1-yl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-(3-Biphenyl-4-ylmethyl-3H-benzoimidazol-4-yloxy)-β-D-glucopyranoside; 2-[3-(4-Ethyl-benzyl)-6-methoxy-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-[3-(4-Ethyl-benzyl)-6-trifluoromethyl-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-6-methyl-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside and 2-[3-(4-Ethyl-benzyl)-6-methyl-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside.







The more preferred compounds of the invention include those selected from 2-[3-(4-Ethyl-benzyl)-3H-benzoimidazol-4-yloxy]-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3H-benzoimidazol-4-yloxy}-β-D glucopyranoside; 2-[3-(4-Ethyl-benzyl)-3H-benzotriazol-4-yloxy]-D-glucopyranoside; 2-[3-(4-Ethyl-benzyl)-6-methyl-3H-benzoimidazol-4-yloxy]-D-glucopyranoside; 2-{3-[2-(4-methoxy-phenyl)-ethyl]-3H-benzotriazol-4-yloxy}-D-glucopyranoside; 2-[3-(5-Ethyl-thiophen-2-ylmethyl)-3H-benzoimidazol-4-yloxy]-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-6-methyl-3H-benzoimidazol-4-yloxy}-D-glucopyranoside; 2-[3-(4-Thiophen-3-yl-benzyl)-3H-benzoimidazol-4-yloxy]-D-glucopyranoside; 2-[3-(4-Pyrrol-1-yl-benzyl)-3H-benzoimidazol-4-yloxy]-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-6-methyl-3H-benzoimidazol-4-yloxy}-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-6-methyl-3H-benzotriazol-4-yloxy}-D-glucopyranoside; 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3H-benzotriazol-4-yloxy}-D-glucopyranoside and 2-[3-(4-Ethyl-benzyl)-6-methyl-3H-benzotriazol-4-yloxy}-D-glucopyranoside.
C. Synthetic Methods
One aspect of the invention features compounds of formula (III). These compounds can be made according to traditional synthetic organic methods or combinatorial or matrix synthesis methods. The following schemes and chemical Examples 1-10 provide general guidance.




Compounds of this invention where n is 1 or 2 and R1, R2, X and Z are defined as in the Formula (III) can be prepared as outlined in Scheme 1. Compounds of Formula 1 wherein R2 is H can be prepared through a Curtius rearrangement of commercially available 3-methoxy-2-nitrobenzoic acid followed by treatment with t-butanol at refluxing temperatures as described by Orjales et. al. [J. Med. Chem. 1997, 40, 586] and palladium catalyzed hydrogenation to reduce the nitro group. Subsequent acylation of the resulting diamine intermediate with substituted acyl chlorides in the presence of a base such as triethylamine in a chlorinated solvent or treatment with substituted carboxylic acid, EDCI and HOBt in an appropriate solvent such as DMF followed by reduction of the resulting amide with borane tetrahydrofuran complex or lithium aluminum hydride in a solvent such as tetrahydrofuran (THF) at temperatures from 20° C. to reflux can afford compounds of Formula 2 wherein n is 1 or 2. Compounds of Formula 2 can be formed directly by treatment of the diamine intermediate with substituted benzaldehydes in the presence of acetic acid and sodium cyanoborohydride. Removal of the t-butoxycarbonyl (BOC) protecting group in the presence of an acid such as hydrochloric or trifluoroacetic acid provides compounds of Formula 4 wherein R2 is H, and P1 is methyl.

Alternatively, commercially available 2-amino-3-nitrophenol can be acylated with substituted acyl chlorides or carboxylic acids as described above then treated with TBDMSCl or MOMBr to give compounds of Formula 3 wherein m is 0 or 1. The nitro group can then be reduced using palladium catalyzed hydrogenation conditions followed by borane reduction of the amide to provide compounds of Formula 4 wherein R2 is H, and P1 is TBDMS or MOM.
Compounds of Formula 4 wherein R2 are groups other than H defined in the Claims above, can be prepared by acylation of commercially available 5-substituted 2-aminophenols with substituted acyl chlorides or carboxylic acids as described above followed by protection of the phenolic hydroxyl group with a methyl or a pivaloyl group using procedures known in the art. The starting substituted phenols that are not readily obtained from commercial sources, can be prepared by starting with the appropriately substituted phenols, protecting the hydroxyl group, then applying standard techniques known in the art (Ogawa, M. et. al. EP 579204; Widdowson, et. al. U.S. Pat. No. 5,780,483) to inter-change or expand functionality at the R2 position to give compounds of Formula 5 wherein R2 is as defined in Formula (III).
The compounds of Formula 5 can be converted to compounds of Formula 6 by direct nitration or by an initial bromination step using bromine or N-bromosuccinamide (NBS) in solvents such as carbon tetrachloride or chloroform followed by nitration. Suitable reagents for nitration include nitronium tetrafluoroborate which can be added to a solution of the intermediate in a solvent such as acetonitrile at temperatures between 0° C. and 20° C. or ammonium nitrite in the presence of trifluoroacetic anhydride in a chlorinated solvent. Palladium-catalyzed hydrogenation in a polar solvent like methanol, ethanol or 2-propanol can simultaneously reduce the nitro group and remove the bromine of compounds of Formula 6. Amide reduction using borane tetrahydrofuran complex as previously described can provide the diamine compounds of Formula 4 where R2 encompasses groups defined in the Formula (III).
Compounds of Formula 2 can be dissolved in DMF and heated in the presence of a base such as potassium carbonate in a 130° C. oil bath for one to two hours to directly provide compounds of Formula 7 where P1 is methyl. Alternatively compounds of Formula 4 can be converted to compounds of Formula 7, where P1 is TBDMS, MOM or Pivaloyl, by treatment with triphosgene, urea or carbonyldiimidazole using known procedures.
Furthermore compounds of Formula 4 can be heated with triethylorthoformate in the presence of a catalytic amount of p-toluenesulfonic acid at 120° C. for three to four hours to give compounds of Formula 8 or treated with sodium nitrite under acidic conditions at 0° C. to provide compounds of Formula 9.
Removal of the protecting groups P1 to provide compounds of Formula 10 can be accomplished using known methods described by Greene and Wuts (Protecting Groups in Organic Chemistry, 3rd. Ed; Wiley Interscience).
Compounds of Formula 11 can be obtained from compounds of Formula 10 by glycosidation of the phenol group with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide in an appropriate solvent, such as acetone, acetonitrile or DMF under basic conditions, such as potassium carbonate, lithium carbonate or lithium hydroxide, followed by deprotection of acetyl groups in an alcoholic solvent such as methanol using mild basic conditions such as potassium carbonate or sodium methoxide at room temperature.
Compounds of Formula 12 may be prepared by the addition of mixed imidazolyl carbonates to a mixture of compounds of Formula 11 and a base such as NaH. The mixed imidazolyl can be prepared by reacting equimolar amounts of an alcohol with carbonyldiimidazole at 0° C. and may be preferred over nonsterically demanding reagents such as alkyl chloroformate to regioselectively acylate the 6-OH group of the glucose (Bertolini, et. al. JOC 1998, 63, 6031).
D. Additional Antidiabetic Agents
Antidiabetic agents that can be used according to the invention, as a second, third, or subsequent antidiabetic agent, in a composition, formulation, or combination method of treatment (dosing regimen) include, but are not limited to the classes and compounds exemplified in Table 2.







TABLE 2







Combination Therapies with SGLT Inhibitors










Mechanism or Class
Drug/Compound







Biguanide (class)
metformin




Fortamet (metformin XT)




metformin GR




metformin XL




NN-414




fenofibrate/metformin combo



Insulin Secretagogue
glimeparide



(mech), Sulfonylureas



(class)




glyburide/glibenclamide combo




glyburide/metformin combo




glipizide




glipizide/metformin combo




gliclazide




chlorpropamide




tolbutamide




tolazamide



Insulin Secretagogue
repaglinide



(mech), Meglitinides



(class)




nateglinide




mitiglinide



Alpha-glucosidase
acarbose



inhibitors (mech)




miglitol




voglibose




emiglitate



Insulin and Insulin
insulin lispro



analogues (class)




insulin glargine




insulin detemir




insulin glulisine




insulin aspart




human insulin (Humulin R)




human insulin (Novolin R




human insulin (Novolin BR)




insulin, zinc suspension




(Humulin L)




insulin NHP (Humulin N)




insulin, zinc suspension




(Novolin L)




insulin NHP (Novolin N)




insulin, zinc suspension




(Humulin U)




human insulin, regular and NHP




mix (Humulin 50/50)




human insulin, regular and NHP




mix (Humulin 70/30)




human insulin, regular and NHP




mix (Novolin 70/30)



Inhaled insulin (class)
Exubera




AERx Insulin Diabetes




Management System




AIR inhaled insulin



Oral insulin (class)
Oralin



PPARgamma (mech)
rosiglitazone




rosiglitazone/metformin combo




pioglitazone




isaglitazone (netoglitazone,




MCC-555)




rosiglitazone/sulfonylurea




ragaglitazar




balaglitazone (NN-2344)




R-483




rivoglitazone (CS-011)




FK-614




SCD-DKY




tesaglitazar




T131




CLX0921




LY-293111 (VML-295)




MBX 102




AA10090




CDDO (TP-155C)




DRF-2189




PHT-46




farglitazar




GW-7845




L-764406




NC-2100




PN 2022 (PN 2034)



PPARalpha/gamma dual
MK767/MK0767 (KRP 297)



agonists (mech)




muraglitazar (BMS-298585)




tesaglitazar




LY-818




oxeglitazar (EML-4156)




LY-929




BVT-142




DRF-2655




DRF-4832




DRF-4158




LY-465608




KT6-207




LSN-862



PPARalpha Agonist (mech)
Fenofibrate




Gemfibrozil




Clofibrate




Ciprofibrate




Benzafibrate




K-111




LY518674 (LY674)




KRP-101




NS-220




GW-9578




GW-7647




GW-9820




LF-200337




ST-1929




Wy-14643



PPARdelta Agonist (mech)
GW501516




GW-1514




L-165041




GW 8547



PPARalpha/delta Dual
GW-2433



Agonist (mech)



PPARgamma/delta Dual
none in the last PPAR CEA



Agonist (mech)



PPARalpha/gamma/delta
CLX-0940



Modulator (mech)



RXR Agonist (mech)



Insulin Seretagogue
Exanatide injectable



(mech), GLP-1 analogue



(class)




Exanatide LAR injectable




Exanantide oral




Liraglutide



GLP-1 agonist (mech)
exenatide (AC2993)




liraglutide (NN2211)




LY-307161




CJC-113




ZP10




GLP-1




BIM-51077



DPPIV Inhibitor (mech)
LAF-237




P32/98




P93/01




NVP-728



Lipase Inhibitor (mech)
Orlistat




ATL962



Glucokinase Activator
Ro 28-1675



(mech)




Ro 27-4375



beta-3 Agonist (mech)
LY-337604




L-796568




CP-331684




CP-331679




CP-114271




Rafabegron (TAK-677)




YM-178




N5984




GW427353



IBAT Inhibitor (mech)
AZD-7806




SC-990




SC-017




GW-264



HM74a/HM74 Agonist
Acipimox



(mech)



Glucocorticoid
A348441



Antagonist (mech)




A362947




CP394531




CP409069




CP472555



Glycogen Phosphorylase a
NN4201



Inhibitor (mech)




Ingliforib (CP368296)



FXR Antagonist (mech)
GW-4064



LXR Agonist (mech)
GW-3965




T-0901317




T-0314407



FXR Antagonist (mech)



GLP-1 Analogue (class)
Albugon



GSK-3beta Inhibitor



(mech)



PTP-1b Inhibitor (mech)
ISIS-113715




KP102



Amylin Receptor Agonist
Pramlintide (symlin/amylin)



NO Scavenger (mech)
NOX-700



11beta-Hydroxysteroid
BVT-3498



Dehydrogenase Inhibitor



Peptide YY hormone
AC162325



Glucagon Antagonist
NN-2501



(mech)



PEPCK Inhibitor (mech)
R1438



Somatotropin Release-
SOM230



inhibiting Factor (mech)



CPT-1 Inhibitor (mech)
ST1326



Carboxypeptidase
MLN-4760



Inhibitor (mech)



Leptin analogue (class)
Metrileptin










E. Combinations
The invention features a combination therapy comprising administering a glucose reabsorption inhibitor, such as an SGLT inhibitor, and one or more antidiabetic agent(s) for the treatment of diabetes or Syndrome X, or associated symptoms or complications thereof. The demonstrated efficacy of SGLT inhibitors in numerous models of NIDDM validates the utility of this drug alone for the treatment of NIDDM in humans. Since glucose reabsorption inhibitors have a mechanism of action distinct from that of other antidiabetic agents, such as RXR modulators, the disclosed combination may have the advantage of reducing the amount of either drug necessary to achieve combined therapeutic or pharmaceutical efficacy, relative to the use of either drug alone, thereby reducing one or more adverse side-effects, which often include weight gain, edema, cardiac hypertrophy, hepatohypertrophy, hypoglycemia, or hepatotoxicity, or any combination thereof.
The invention provides a method for treating diabetes or Syndrome X, or complications thereof in a subject, said method comprising administering to said subject a jointly effective amount of a glucose reabsorption inhibitor in combination with a jointly effective amount of an antidiabetic agent, such as an RXR modulator. In one aspect of the invention, the antidiabetic agent is an RXR agonist or RXR antagonist that increases insulin sensitivity in the subject. For example, an insulin sensitizer can increase glucose tolerance in a subject in an oral glucose tolerance test.
Preferably, the diabetes or Syndrome X, or associated symptoms or complication thereof is selected from IDDM, NIDDM, IGT, and IFG.
This invention also provides a pharmaceutical composition comprising one or more glucose reabsorption inhibitors (alone or in combination with one or more antidiabetic agents), and a pharmaceutically acceptable carrier. In one aspect of the invention, the antidiabetic agent is an RXR agonist or RXR antagonist that increases insulin sensitivity in the subject.
In particular, the glucose reabsorption inhibitor is a SGLT1 and/or SGLT2 inhibitor.
For use in medicine, the salt or salts of the compounds of Formula (V) refer to non-toxic “pharmaceutically acceptable salt or salts.” Other salts may, however, be useful in the preparation of compounds according to this invention or of their pharmaceutically acceptable salts. Representative organic or inorganic acids include, but are not limited to, hydrochloric, hydrobromic, hydroiodic, perchloric, sulfuric, nitric, phosphoric, acetic, propionic, glycolic, lactic, succinic, maleic, fumaric, malic, tartaric, citric, benzoic, mandelic, methanesulfonic, hydroxyethanesulfonic, benezenesulfonic, oxalic, pamoic, 2-naphthalenesulfonic, p-toluenesulfonic, cyclohexanesulfamic, salicylic, saccharinic or trifluoroacetic acid. Representative basic/cationic salts include, but are not limited to, benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, or zinc. The compounds of Formula (V) or a pharmaceutically acceptable salt thereof, may include an intramolecular salt thereof, or a solvate or hydrate thereof.
F. Administration, Formulation, and Dosages
The utility of the disclosed compounds, compositions, and combinations to treat disorders in glucose and lipid metabolism can be determined according to the procedures well known in the art (see the references listed below), as well as all the procedures described in U.S. Pat. Nos. 5,424,406, 5,731,292, 5,767,094, 5,830,873, 6,048,842, WO01/16122 and WO01/16123 which are incorporated herein by reference. The compound may be administered to a patient by any conventional route of administration, including, but not limited to, intravenous, oral, subcutaneous, intramuscular, intradermal and parenteral administration. Preferably, formulations are for oral administration.
The present invention also provides pharmaceutical compositions comprising one or more glucose reabsorption inhibitors and one or more RXR modulators in association with a pharmaceutically acceptable carrier.
The daily dosage of the products may be varied over a wide range from 1 to 1000 mg per adult human per day. For oral administration, the compositions are preferably provided in the form of tablets containing, 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 150, 200, 250 or 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of 1 to 2 times per day. The dosages, however, may be varied depending upon the requirement of the patients, the severity of the condition being treated and the compound being employed. The use of either daily administration or post-periodic dosing may be employed. Preferably these compositions are in unit dosage forms such as tablets, pills, capsules, powders, granules, sterile parenteral solutions or suspensions, metered aerosol or liquid sprays, drops, ampoules, auto-injector devices or suppositories; for oral, parenteral, intranasal, sublingual or rectal administration, or for administration by inhalation or insufflation. Alternatively, the composition may be presented in a form suitable for once-weekly or once-monthly administration; for example, an insoluble salt of the active compound, such as the decanoate salt, may be adapted to provide a depot preparation for intramuscular injection. For preparing solid compositions such as tablets, the principal active ingredient or ingredients are mixed with a pharmaceutical carrier, e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and other pharmaceutical diluents, e.g. water, to form a solid preformulation composition containing a homogeneous mixture of one or more glucose reabsorption inhibitors and one or more antidiabetic agents, or a pharmaceutically acceptable salt thereof. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient or ingredients are dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective dosage forms such as tablets, pills and capsules. This solid preformulation composition is then subdivided into unit dosage forms of the type described above containing from 0.1 to about 500 mg of the active ingredient or ingredients of the present invention. The tablets or pills of the novel composition can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of material can be used for such enteric layers or coatings, such materials including a number of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.
The liquid forms in which the novel compositions of the present invention may be incorporated for administration orally or by injection include, aqueous solutions, suitably flavoured syrups, aqueous or oil suspensions, and flavoured emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles. Suitable dispersing or suspending agents for aqueous suspensions, include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinyl-pyrrolidone or gelatin. The liquid forms in suitably flavored suspending or dispersing agents may also include the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like. For parenteral administration, sterile suspensions and solutions are desired. Isotonic preparations which generally contain suitable preservatives are employed when intravenous administration is desired.
Advantageously, the combinations of one or more glucose reabsorption inhibitors of the present invention, alone or in combination with one or more additional antidiabetic agents, may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, one or more glucose reabsorption inhibitors and/or one or more antidiabetic agents according to the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders; lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
Wherein the present invention is directed to the administration of a combination, the compounds may be co-administered simultaneously, sequentially, or in a single pharmaceutical composition. Where the compounds are administered separately, the number of dosages of each compound given per day, may not necessarily be the same, e.g. where one compound may have a greater duration of activity, and will therefore, be administered less frequently.
Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular compound used, the strength of the preparation, the mode of administration, and the advancement of the disease condition. In addition, factors associated with the particular patient being treated, including patient age, weight, diet and time of administration, will result in the need to adjust dosages.
The novel compositions of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of lipids, including but not limited to amphipathic lipids such as phosphatidylcholines, sphingomyelins, phosphatidylethanolamines, phophatidylcholines, cardiolipins, phosphatidylserines, phosphatidylglycerols, phosphatidic acids, phosphatidylinositols, diacyl trimethylammonium propanes, diacyl dimethylammonium propanes, and stearylamine, neutral lipids such as triglycerides, and combinations thereof. They may either contain cholesterol or may be cholesterol-free.
From Formula (III) and other disclosed formulae it is evident that some compounds in the compositions of the invention may have one or more asymmetric carbon atoms in their structure. It is intended that the present invention include within its scope the stereochemically pure isomeric forms of the compounds as well as their racemates. Stereochemically pure isomeric forms may be obtained by the application of art known principles. Diastereoisomers may be separated by physical separation methods such as fractional crystallization and chromatographic techniques, and enantiomers may be separated from each other by the selective crystallization of the diastereomeric salts with optically active acids or bases or by chiral chromatography. Pure stereoisomers may also be prepared synthetically from appropriate stereochemically pure starting materials, or by using stereospecific reactions.
Some compounds in the compositions of the present invention may have various individual isomers, such as trans and cis, and various alpha and beta attachments (below and above the plane of the drawing). In addition, where the processes for the preparation of the compounds according to the invention give rise to mixture of stereoisomers, these isomers may be separated by conventional techniques such as preparative chromatography. The compounds may be prepared as a single stereoisomer or in racemic form as a mixture of some possible stereoisomers. The non-racemic forms may be obtained by either synthesis or resolution. The compounds may, for example, be resolved into their components enantiomers by standard techniques, such as the formation of diastereomeric pairs by salt formation. The compounds may also be resolved by covalent linkage to a chiral auxiliary, followed by chromatographic separation and/or crystallographic separation, and removal of the chiral auxiliary. Alternatively, the compounds may be resolved using chiral chromatography. Unless otherwise noted, the scope of the present invention is intended to cover all such isomers or stereoisomers per se, as well as mixtures of cis and trans isomers, mixtures of diastereomers and racemic mixtures of enantiomers (optical isomers) as well.
The therapeutic effect of the glucose reabsorption inhibitor administered in combination with one or more antidiabetic agent(s) in treating diabetes, Syndrome X, or associated symptoms or complications can be shown by methods known in the art. The following examples of combination treatment with SGLT inhibitors and other antidiabetic agents such as RXR modulators are intended to illustrate the invention but not to limit it.
G. Synthetic Chemical Examples
One aspect of the invention features compounds of formula (III) as described above in the Summary section, the description, and the appended claims. These disclosed compounds may be made according to traditional synthetic organic chemistry methods or according to matrix or combinatorial chemistry methods. The Schemes and Examples 1-9 below provide general guidance and detailed examples of how the disclosed compounds may be prepared.
1HNMR spectra were measured on a Brucker AC-300 (300 MHz) spectrometer using tetramethylsilane (TMS) as an internal standard.
Example 1
2-[3-(2-Naphthalen-2-yl-ethyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D-glucopyranoside




    A. (2-Amino-3-methoxy-phenyl)-carbamic acid tert-butyl ester 1a: Thionyl chloride (2.3 mL, 32 mmol) was added to a suspension of commercially available 3-methoxy-2-nitrobenzoic acid (4.2 g, 21.3 mmol) in toluene (20 mL). The mixture was stirred a reflux for 30 minutes, cooled to room temperature (RT) and concentrated to dryness. The acid chloride was then dissolved in acetone (HPLC grade, 20 mL) and added dropwise through an addition funnel to a cold (0° C.) solution of sodium azide (2.2 g, 33.8 mmol) in water (30 mL) with stirring. The resulting suspension was stirred at 0° C. for 10 minutes then at RT for 50 min. Water was added (150 mL) and the white solid product was collected by filtration. The solid product was diluted with t-butanol (25 mL) and heated to reflux in an oil bath. After 20 minutes at reflux the excess t-butanol was removed under reduced pressure and the residue purified by chromatography on silica gel (EtOAc:hexane; 5:100) to provide the desired product as a yellow solid (4.59 g, 80%). This product (2.48 g, 9.2 mmol) was then dissolved in ethanol (40 mL) and hydrogenated over 10% Pd/C (250 mg) under 40 psi of H2 pressure for 3 hours. The catalyst was removed by filtration and the filtrated concentrated in vacuo to give the title compound as a solid (2.2 g, 100%).
    B. [3-Methoxy-2-(2-naphthalen-2-yl-acetylamino)-phenyl]-carbamic acid tert-butyl ester 1b: A mixture of 1a (0.413 g, 1.74 mmol), prepared in Part A, 2-naphthylacetic acid (0.32 g, 1.74 mmol), 1-hydroxybenzotriazole hydrate (0.53 g, 3.5 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimde hydrochloride (0.83 g, 4.3 mmol) in dry DMF (4 mL) was stirred at RT overnight, poured into H2O (40 mL) and extracted with EtOAc (2×60 mL). The combined EtOAc extract was washed with brine, dried over MgSO4, filtered and concentrated to give a crude oil. Column chromatography provided the title compound as a white solid (0.5 g, 70%).
    C. [3-Methoxy-2-(2-naphthalen-2-yl-ethylamino)-phenyl]-carbamic acid tert-butyl ester 1c and 3-Methoxy-(2-naphthalen-2-yl-ethyl)-benzene-1,2-diamine 1d: To a cold (0° C.) solution of 1b (0.423 g, 1.04 mmol), prepared in Part B, in dry tetrahydrofuran (30 mL) was slowly added borane tetrahydrofuran complex (1M solution, 2 mL). The reaction mixture was stirred at reflux for 2 hours, cooled to 0° C. and 1N NaOH solution (3 mL) was slowly added. Water (5 mL) was added and the mixture was stirred at RT overnight. The mixture was diluted with water (50 mL) and the aqueous mixture extracted with ethyl acetate (2×60 mL). The combined organic extracts were washed with brine, dried over MgSO4, concentrated and chromatographed (silica gel, EtOAc/Hexane; 1:4 ratio) to afford the title compounds, [3-Methoxy-2-(2-naphthalen-2-yl-ethylamino)-phenyl]-carbamic acid tert-butyl ester 1c (0.26 g, 64%) as a clear oil and 3-Methoxy-N2-(2-naphthalen-2-yl-ethyl)-benzene-1,2-diamine 1d (80 mg, 20%) as a white solid.
    D. 7-Methoxy-1-(2-naphthalen-2-yl-ethyl)-1,3-dihydro-benzoimidazol-2-one 2e: A mixture of the carbamate 1c (0.48 g, 1.2 mmol), prepared in Part C, and K2CO3 (0.24 g, 1.5 mmol) in DMF (10 mL) was heated in a 150° C. oil bath for 1.5 hours, cooled to RT, poured into H2O (30 mL), stirred at RT for 20 minutes and filtered to give crude solid product. The solids were washed with Et2O and dried to give the title compound as an off-white solid (0.28 g, 72%).
    E. 7-Hydroxy-1-(2-naphthalen-2-yl-ethyl)-1,3-dihydro-benzoimidazol-2-one 1f: A mixture of 1e (0.28 g, 0.88 mmol), prepared in Part D, in dichloromethane (50 mL) was cooled to −78° C. and boron tribromide (1M solution in CH2Cl2; 4.4 mL) was slowly added. The reaction mixture was stirred at −78° C. for 30 minutes then slowly warmed up to RT and stirred at RT for 24 hours. A 1M HCl solution (5 mL) was added dropwise followed by ice H2O (50 mL). The methylene chloride was removed under reduced pressure and the aqueous mixture extracted with EtOAc (2×60 mL). The combined EtOAc extract was washed with brine, dried over MgSO4 and concentrated to give the title compound as a yellow solid (0.267 g, 100%).
    F. 2-[3-(2-Naphthalen-2-yl-ethyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D-glucopyranoside 1: To a solution of 1f (0.26 g, 0.87 mmol), prepared in Part E, in DMF (5 mL) was added potassium carbonate (0.30 g, 2.2 mmol), followed by addition of 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide (0.54 g, 1.3 mmol). The reaction mixture was stirred at room temperature for 24 h. Then solids were filtered and washed with ethyl acetate. The ethyl acetate solution was washed with water and brine, dried over MgSO4 and concentrated in vacuo. The residue was chromatographed (silica gel, EtOAc/Hexane, 3:2) to give pure product (24%—based on recovered starting material) as a solid. The solid product was diluted with methanol (3 mL) and sodium methoxide (1M solution in MeOH, 0.2 mL) was added. The resulting solution was stirred at RT for 1 hour, diluted with water (20 mL) and evaporated to remove MeOH. The aqueous mixture was extracted with EtOAc (2×30 mL). The combined EtOAc extracts was washed with brine, dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel: eluting with 5% methanol in chloroform to give the title compound (0.023 g, 57%) as a white solid. 1HNMR (300 MHz, CD3OD) δ 7.82-7.78 (m, 3H), 7.72 (s, 1H), 7.47-7.42 (m, 3H), 7.04 (m, 2H), 6.82-6.79 (dd, J=6.25 HZ, J=2.31 Hz, 1H), 5.18 (d, J=7.73 Hz, 1H), 4.47-4.41 (m, 2), 3.92 (d, J=10.9 Hz, 1H), 3.74-3.63 (m, 2H), 3.54-3.46 (m, 3H), 3.28-3.23 (m, 2H). MS: m/z (MH+) 467.


Example 2
2-[3-(2-naphthalen-2-yl-ethyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside




    A 7-Methoxy-1-(2-naphthalen-2-yl-ethyl)-1H-benzoimidazole 2a: To 1d (0.66 g, 2.26 mmol), prepared in Part C of Example 1, and triethylorthoformate (0.45 ml, 2.71 mmol) was added catalytic amount of p-toluenesulfonic acid. The mixture was heated in a 120° C. oil bath for 5 hours then cooled to RT. The residue was purified by chromatography (silica gel, 10% EtOAC in Hexane) to afford the title compound (0.513 g, 75%).
    B 2-[3-(2-naphthalen-2-yl-ethyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside 2: The compound 2a prepared in the previous step was demethylated then glycosylated in the same manner as described in Parts E through F to provide the title compound. 1HNMR (400 MHz, CD3OD) δ 7.79-7.68 (m, 3H), 7.58 (s, 1H), 7.47 (s, 1H), 7.42-7.39 (m, 2H), 7.13-7.15 (m, 4H), 5.24 (d, J=7.91 Hz, 1H), 4.93 (m, 1H), 4.74 (m, 1H), 3.96-3.92 (dd, J=11.9 Hz, J=1.8 Hz, 1H), 3.77-3.65 (m, 2H), 3.57-3.33 (m, 5H). MS: m/z (MH+) 451.


Example 3
2[3-(4-Ethyl-benzyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D-glucopyranoside




    A [2-(4-Ethyl-benzoylamino)-3-methoxy-phenyl]-carbamic acid tert-butyl ester 3a: To a solution of 1a (1.5 g, 6.3 mmol), prepared in Part A of Example 1, and triethylamine (1.3 mL, 9.45 mmol) in methylene chloride (40 mL) was added 4-Ethyl benzoyl chloride (1.06 mL, 6.3 mmol) dropwise. The resulting mixture was stirred at RT for 6.25 hours, poured into water (50 mL) and layers were separated. The organic layer was dried over MgSO4, filtered and concentrated to give a crude mixture of the title compound in quantitative yields.
    B [2-(4-Ethyl-benzylamino)-3-methoxy-phenyl]-carbamic acid tert-butyl ester (3b) and N2-(4-Ethyl-benzyl)-3-methoxy-benzene-1,2-diamine 3c: Treatment of the compound 3a prepared in the previous step with borane tetrahyrofuran complex using the same procedure described in Part C of Example 1 provided the title compounds in quantitative yields.
    C 2[3-(4-Ethyl-benzyl)-1,3-dihydro-benzoimidazol-2-one-4-yloxy]-β-D-glucopyranoside (3): The title compound was prepared in the same manner as described in Part D-F of Example 1 substituting 1c with 3b prepared in the previous step. 1HNMR (300 MHz, CD3OD) δ 7.24 (d, J=7.90 Hz, 2H), 7.12 (d, J=8.14 Hz, 2H), 7.02-6.93 (m, 2H) 6.80 (d, J=7.69 Hz, 1H) 5.44 (d, 15.51 Hz, 1H) 5.27 (d, J=15.61 Hz, 1H) 5.00 (d, J=7.33 Hz, 1H) 3.90-3.86 (dd, J=1.93, J=12.05, 1H) 3.69-3.66 (dd, J=5.53, J=12.00, 1H) 3.51-3.36 (m, 4H) 2.59 (q, 2H) 1.19 (t, 3H). MS: m/z (MH+) 431.


Example 4
2-[3-(4-Ethyl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside




2-[3-(4-Ethyl-benzyl)-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside 4: The compound 3c prepared in Part B of Example 3 was treated as described in Example 2 replacing 1d with 3c to provide the title compound. 1HNMR (300 MHz, CD3OD) δ 8.09 (s, 1H) 7.34 (d, J=8.00, 1H), 7.20-7.13 (m, 5H), 7.09 (d, J=7.95, 1H), 5.86 (d, J=15.29, 1H), 5.61 (d, J=15.27, 1H), 5.09 (d, J=7.28, 1H), 3.89 (d, J=11.91, 1H), 3.71-3.67 (dd, J=5.48, J=11.86, 1H), 3.55-3.36 (m, 4H), 2.59 (q, J=7.7 Hz, 2H), 1.18 (t, J=7.4 Hz, 3H). MS: m/z (MH+) 415.
Example 5
2-{3-[2-(4-Methoxyphenyl)-ethyl]-3H-benzoimidazol-4-yloxy}-β-D-glucopyranoside




    A N-(2-Hydroxy-6-nitro-phenyl)-2-(4-methoxy-phenyl)-acetamide 5a: To a mixture of commercially available 2-Amino-3-nitrophenol (5 g, 32.4 mmol) and triethylamine (9 ml, 64.9 mmol) in methylene chloride (20 ml) was slowly added p-methoxyphenylacetyl chloride (5 ml, 32.4 mmol). The resulting mixture was stirred at RT overnight, poured into ice water (30 ml) and a 0.5M HCl solution (20 ml) was added. An additional 30 ml of methylene chloride was added and layers were separated. The methylene chloride layer was washed with water, brine and dried over MgSO4. The solution was filtered and solvent removed under reduced pressure to give the title compound as a dark solid (9.45 g, 96%).
    B N-[2-(tert-Butyl-dimethyl-silanyloxy)-6-nitro-phenyl]-2-(4-methoxy-phenyl)-acetamide 5b: To a cold (0° C.) solution of 5a (3.1 g, 10.3 mmol), prepared in Part A, and tert-Butyldimethylsilyl chloride (1.86 g, 12.3 mmol) in dry DMF (10 ml) was slowly added imidazole (1.7 g, 25.7 mmol). Stirring was continued at 0° C. for 30 minutes then at RT for 3 hours. The resulting mixture was poured into ice water (80 ml) and extracted with Et2O (2×80 ml). The combined ether extracts were dried over MgSO4 and concentrated to provide the product as an oil in quantitative yield.
    C N-[2-Amino-6-(tert-butyl-dimethyl-silanyloxy)-phenyl]-2-(4-methoxy-phenyl)-acetamide 5c: A solution of 5b (1.4 g, 3.36 mmol), prepared in Part B, in ethanol was hydrogenated over 10% Pd/C at 40 psi of hydrogen pressure for 4 hours. The catalyst was filtered off through Celite and the filtrate concentrated in vacuo. The title compound was isolated as an off-white solid (0.693 g, 53%) by recrystallization from Et2O.
    D 3-(tert-Butyl-dimethyl-silanyloxy)-N2-[2-(4-methoxy-phenyl)-ethyl]-benzene-1,2-diamine 5d and 3-amino-2-[2-(4-methoxy-phenyl)-ethylamino]-phenol 5e: To a cold (0° C.) solution of 5c (0.693 g, 1.8 mmol), prepared in Part C, in dry tetrahydrofuran (20 mL) was slowly added borane tetrahydrofuran complex (1M solution, 2.7 mL). The reaction mixture was stirred at reflux for 2 hours, cooled to 0° C. and 1N NaOH solution (1.5 mL) was slowly added. Water (5 mL) was added and the mixture was stirred at RT overnight. The mixture was diluted with water (25 mL) and the aqueous mixture extracted with ethyl acetate (2×30 mL). The combined organic extracts were washed with brine, dried over MgSO4, concentrated and chromatographed (silica gel, EtOAc/Hexane; 1:4 ratio) to afford the title compound 5d (0.4 g, 60%) as a white solid and the title compound 5e (0.7 g, 35%).
    E 7-(tert-Butyl-dimethyl-silanyloxy)-1-[2-(4-methoxy-phenyl)-ethyl]-1H-benzoimidazole 5f: To a mixture of 5d (0.38 g, 1.02 mmol), prepared in Part D, and triethylorthoformate (023 ml, 1.23 mmol) was added catalytic amount of p-toluenesulfonic acid. The mixture was heated in a 120° C. oil bath for 5 hours then cooled to RT. The residue was purified by chromatography (silica gel, EtOAC/Hexane, 1:4) to afford the title compound (0.33 g, 84%).
    F 3-[2-(4-Methoxy-phenyl)-ethyl]-3H-benzoimidazol-4-ol 5g: To a cold (0° C.) solution of 5f (0.30 g, 0.78 mmol), prepared in Part E, in dry tetrahydrofuran (10 ml) was added tetrabutlyammonium fluoride (2 ml, 1.96 mmol). The resulting mixture was warm to RT, stirred for 11⁄2 hours and water (15 ml) was added. The solid precipitates were collected by filtration and dried under reduced pressure to give the title compound (0.206 g, 99%) as a tan solid.
    G 2-{3-[2-(4-Methoxyphenyl)-ethyl]-3H-benzoimidazol-4-yloxy}-β-D-glucopyranoside 5: The title compound was prepared by treating 5g with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide as described in Part F of Example 1. 1HNMR (400 MHz, CD3OD) δ 7.59 (s, 1H), 7.31 (d, J=8.1 Hz, 1H), 7.20 (t, J=7.9 Hz, 1H), 7.13 (d, 7.8 Hz, 1H), 6.95 (d, J=8.4, 2H), 6.78 (d, J=8.5 Hz, 2H), 5.21 (d, H=7.8 Hz, 1H), 4.62-4.58 (m, 1H), 3.95-3.91 (m, 1H), 3.76-3.72 (m, 4H), 3.65-3.45 (m, 5H), 3.27-3.12 (m, 2H). MS: m/z (MH+) 431.


Example 6
2-[3-(2-(4-Methoxyphenyl)-ethyl)-3H-benzoimidazol-4-yloxy]-β-D glucopyranoside




    A 7-(tert-Butyl-dimethyl-silanyloxy)-1-[2-(4-methoxy-phenyl)-ethyl]-1,3-dihydro-benzoimidazol-2-one 6a: Triphosgene (0.40 g, 1.35 mmol) was added to a cold (0° C.) solution of 5d (0.50 g 1.35 mmol), prepared in Part D of Example 5, in dry THF (20 ml). Stirring was continued at 0° C. for 30 minutes then at RT for 2 hours. A saturated NaHCO3 solution (60 ml) was added slowly and the mixture extracted with EtOAc (2×60 ml). The combined EtOAc extracts were washed with brine and dried over MgSO4. The mixture was filtered and the filtrate concentrated under reduced pressure to provide the title compound as a solid (0.53 g, 98%).
    B 2-[3-(2-(4-Methoxyphenyl)-ethyl)-3H-benzoimidazol-4-yloxy]-β-D glucopyranoside 6: The title compound was prepared in the same manner as described in Part F in Example 5 substituting compound 5f with 6a followed by treatment with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide as described in Part F of Example 1. 1HNMR (400 MHz, CD3OD) δ 7.17 (d, J=8.38 Hz, 2H), 7.03-6.98 (m, 2H), 6.84-6.78 (m, 3H), 5.14 (d, J=7.7 Hz, 1H), 4.37-4.26 (m, 2H), 3.90 (d, J=12.1 Hz, 1H), 3.76 (s, 3H), 3.73-3.69 (m, 1H), 3.62-3.43 (m, 4H), 3.14-2.96 (m, 2H). MS: m/z (MH+) 447.


Example 7
2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3H-benzoimidazol-4-yloxy}β-D glucopyranoside




    A. 2-[2-(2,3-Dihydro-benzofuran-5-yl)-ethylamino]-3-nitro-phenol 7a: A mixture of the 2-Amino-3-nitrophenol (1.0 g, 6.6 mmol), (2,3-Dihydro-benzofuran-5-yl)-acetic acid (1.07 g, 6.5 mmol; Dunn, et. al. J. Med. Chem. 1986, 29, 2326), 1-Hydroxybenzotriazole hydrate (3.0 g, 19.5 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimde hydrochloride (3.73 g, 19.5 mmol) in dry DMF (10 mL) was stirred at RT overnight, poured into H2O (80 mL) and extracted with EtOAc (2×80 mL). The combined EtOAc extract was washed with brine, dried over MgSO4, filtered and concentrated to give a crude oil. Column chromatography provided the title compound as a white solid (0.3 g, 15%).
    B 2-{3-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3H-benzoimidazol-4-yloxy}β-D glucopyranoside 7: The title compound was prepared in the same manner as described in Parts B through G of Example 5 substituting compound 5a with 7a prepared in the previous step. 1HNMR (300 MHz, Aceton-d6) δ 7.71 (s, 1H), 7.32-7.29 (m, 1H), 7.09-7.04 (m, 3H), 6.87 (d, J=7.98 Hz, 1H), 6.58 (d, J=8.14 Hz, 1H), 5.24 (d, J=7.5 Hz, 1H), 4.79-4.75 (m, 2H), 4.59-4.45 (m, 5H), 3.89 (d, J=11.95 Hz, 1H), 3.77-3.54 (m, 5H), 3.16-3.08 (m, 4H) MS: m/z (MH+) 443.


Example 8
2-[3-(4-Ethyl-benzyl)-6-methyl-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside




    A 4-Ethyl-N-(2-hydroxy-4-methyl-phenyl)-benzamide 8a: To a solution of commercially available 2-Amino-5-methylphenol (6 g, 48.7 mmol) and triethylamine (7 ml, 97.4 mmol) in methylene chloride (80 mL) was added 4-Ethyl benzoyl chloride (7.2 ml, 48.7 mmol) dropwise. The resulting mixture was stirred at RT for 2 hours, poured into water (100 mL) and layers were separated. The methylene chloride layer was washed with dilute HCl solution then again with H2O and dried over MgSO4. The mixture was filtered and concentrated to dryness to give a semi-solid. Recrystallization from hexane provided the title compound in quantitative yield as a solid.
    B 4-Ethyl-N-(2-methoxy-4-methyl-phenyl)-benzamide 8b: Methyliodide (1.2 ml, 19.2 mmol) was added dropwise to a mixture of 8a (4.0 g, 15.7 mmol), prepared in Part A, and potassium carbonate (5 g, 36.2 mmol) in DMF (20 mL). The resultant mixture was stirred at RT for 6 hours and water (60 mL) was added. The aqueous mixture extracted with Et2O (2×60 mL). The combined EtOAc extracts were dried over MgSO4, concentrated in vacuo and chromatographed (silica gel; 3% EtOAc in Hexane) to provide the title compound (3.58 g, 85%).
    C N-(5-Bromo-2-methoxy-4-methyl-phenyl)-4-ethyl-benzamide 8c: To a solution of 8b (3.58 g, 13.3 mmol), prepared in Part B, and iodine (3 crystals) in carbon tetrachloride (100 ml) was slowly added bromine (0.75 ml, 14.6 mmol). An addition 100 ml of carbon tetrachloride was added and the resulting suspension stirred at RT for 6 hours. The mixture was poured in a 10% sodium sulfite solution (400 ml), dichloromethane (200 ml) was added and layers were separated. The organic layer was washed with saturated NaHCO3 (1×), dried over MgSO4, filtered and concentrated under reduced pressure. The white solid product was precipitated with hexane and isolated by filtration to yield the title compound (3.1 g, 67%).
    D N-(3-Bromo-6-methoxy-4-methyl-2-nitro-phenyl)-4-ethyl-benzamide 8d: Nitronium tetrafluoroborate (0.434 g, 3.3 mmol) was added in one portion to a cold (0° C.) solution of 8c (1.13 g, 3.2 mM), prepared in Part C, in acetonitrile. The resulting mixture was stirred at 0° C. for 11 minutes then at RT for 15 minutes and H2O (30 ml) was added. The solid precipitates were collected by filtration and dried under reduced pressure to provide the title compound (0.93 g, 73%) as an off-white solid.
    E N-(2-Amino-6-methoxy-4-methyl-phenyl)-4-ethyl-benzamide 8e: A solution of 8d (1.2 g, 3.1 mmol), prepared in Part D, and K2CO3 (0.5 g, 3.6 mmol) in ethanol was hydrogenated over 10% Pd/C at 40 psi of hydrogen pressure for 6 hours. The catalyst was filtered off through Celite and the filtrate concentrated in vacuo. The residue was diluted with EtOAc (30 ml), washed with H2O (1×30 ml), and dried over MgSO4. The mixture was filtered and concentrated to provide the title compound (0.728 g, 83%) as a solid.
    F N2-(4-Ethyl-benzyl)-3-methoxy-5-methyl-benzene-1,2-diamine 8f: To a cold (0° C.) solution of 8e (0.69 g, 2.4 mmol), prepared in Part E, in dry tetrahydrofuran (25 mL) was slowly added borane tetrahydrofuran complex (1M solution, 4.8 mL). The reaction mixture was stirred at RT for 6 hours, cooled to 0° C. and 1N NaOH solution (1.5 mL) was slowly added. Water (5 mL) was added and the mixture was stirred at RT overnight. The mixture was diluted with water (25 mL) and the aqueous mixture extracted with ethyl acetate (2×30 mL). The combined organic extracts were washed with brine, dried over MgSO4, concentrated and chromatographed (silica gel, EtOAc/Hexane; 1:4 ratio) to afford the title compound (0.31 g, 46%) as a white solid.
    G 1-(4-Ethyl-benzyl)-7-methoxy-5-methyl-1H-benzoimidazole 8g: To 8f (0.275 g, 1.0 mmol), prepared in Part F, and triethylorthoformate (0.20 ml, 1.2 mmol) was added catalytic amount of p-toluenesulfonic acid. The mixture was heated in a 120° C. oil bath for 3 hours then cooled to RT. The residue was purified by chromatography (silica gel, EtOAC/Hexane, 1:4) to afford the title compound (0.253 g, 88%).
    H 2-[3-(4-Ethyl-benzyl)-6-methyl-3H-benzoimidazol-4-yloxy]-β-D-glucopyranoside 8: The title compound was prepared by treating 8 g as described in Parts E through F of Example 1 substituting compound 1e with 8 g 1HNMR (400 MHz, CD3OD) δ 8.03 (s, 1H), 7.21-7.14 (m, 5H), 6.95 (s, 1H), 5.81 (d, J=15.4 Hz, 1H), 5.58 (d, J=15.2 Hz, 1H), 5.07 (d, J=7.37 Hz, 1H), 3.88-3.92 (dd, J=12.1 Hz, J=2.2 Hz, 1H), 3.66 3.71 (dd, J=12.0 Hz, J=5.8 Hz, 1H), 3.52-3.37 (m, 4H), 2.63-2.58 (q, J=7.45 Hz, 2H), 2.43 (s, 3H), 1.19 (t, J=7.62 Hz, 3H). MS: m/z (MH+) 429.


Example 9
2-{3-[2-(4-methoxy-phenyl)-ethyl]-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside




    A 3-[2-(4-Methoxy-phenyl)-ethyl]-3H-benzotriazol-4-ol (9a): To a cold (0° C.) mixture of 5e (0.49 g, 1.9 mmol), prepared in Part D of Example 5, in a 3N HCl solution (20 ml) was rapidly added sodium nitrite (0.152 g, 2.0 mmol). The resulting mixture was stirred at 0° C. for 11⁄2 hour, neutralized to pH 6 with 3 N NaOH and diluted with H2O (40 ml). The aqueous mixture was extracted with EtOAc (2×60 ml). The combined EtOAc extracts were dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by chromatography (silica gel, 10% EtOAc/Hexane) to provide the title compound (0.25 g, 49%).
    B 2-{3-[2-(4-Methoxyphenyl)-ethyl]-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside (9): The title compound was prepared by treating 9a with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide as described in Part F of Example 1. 1HNMR (400 MHz, CD3OD) δ 7.59 (d, J=8.12 Hz, 1H), 7.35 (t, J=8.1 Hz, 1H), 7.27 (d, J=7.67 HZ, 1H), 7.08 (d, J=8.51 Hz, 2H), 6.79 (d, J=8.5 Hz, 2H), 5.24 (d, J=7.95 Hz, 1H), 5.17 (m, 1H), 5.06 (m, 1H), 3.93 (d, J=12.02 Hz, 1H), 3.74 (s, 3H), 3.73-3.64 (m, 2H), 3.57-3.46 (m, 3H), 3.28-3.26 (m, 2H). MS: m/z (MH+) 432.


Example 10
2-[3-(4-Ethyl-benzyl)-3H-benzotriazol-4-yloxy]-β-D-glucopyranoside




    A 1-(4-Ethyl-benzyl)-7-methoxy-1H-benzotriazole 10a: To a cold (0° C.) mixture of 3c, prepared in Part B of Example 3, (1.3 g, 5.1 mmol) in a 3 N HCl solution (40 ml) was rapidly added sodium nitrite (0.39 g, 5.6 mmol). The resulting mixture was stirred at 0° C. for 2 hours, neutralized to pH 6 with 3 N NaOH and diluted with H2O (100 ml). The aqueous mixture was extracted with EtOAc (3×60 ml). The combined EtOAc extracts were dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by chromatography (silica gel, 10% EtOAc/Hexane) to provide the title compound (0.69 g, 51%).
    B 3-(4-Ethyl-benzyl)-3H-benzotriazol-4-ol 10b: A mixture of 10a, prepared in Part A (0.69 g, 2.58 mmol) in dichloromethane (20 mL) was cooled to −78° C. and boron tribromide (1M solution in CH2Cl2; 10 mL) was slowly added. The reaction mixture was stirred at −78° C. for 30 minutes then slowly warmed up to RT and stirred at RT for 24 hours. A 1M HCl solution (20 mL) was added dropwise followed by ice H2O (20 mL). The dichloromethane was removed under reduced pressure and the aqueous mixture extracted with EtOAc (3×30 mL). The combined EtOAc extract was washed with brine, dried over MgSO4 and concentrated to give the title compound as a yellow solid (0.5 g, 77%).
    C 2-[3-(4-Ethyl-benzyl)-3H-benzotriazol-4-yloxy}-β-D-glucopyranoside 10c: To a solution of 10b (0.4 g, 1.58 mmol), prepared in Part B, in MeOH (10 mL) was added lithium hydroxide (0.42 g, 1.74 mmol) and the solution was stirred at RT. After 5 min, the solution was evaporated to dryness. 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide (3.25 g, 7.91 mmol) was added after diluting the residue in DMF (12 mL). After stirring the reaction mixture at RT overnight, potassium carbonate (2.2 g, 1.58 mmol) and MeOH (5 mL) was added and stirred at RT overnight. The resulting solution was poured into water (50 mL) and the product was extracted with EtoAc (3×50 mL), ether was added and the organic extract was washed with water (4×50 mL) and brine. The combined extract was dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by chromatography (silica gel, dichloromethane: methanol, 97:3) to give the title compound (0.1 g, 15%) as a white solid. 1HNMR (300 MHz, CD3OD) δ 7.60 (d, J=7.78, 1H) 7.29 (m, 4H) 7.14 (d, J=8.09, 2H) 6.17 (d, J=15.04, 1H) 6.01 (d, J=15.06, 1H) 5.16 (d, J=7.70, 1H) 3.90 (dd, J=1.95, J=12.13, 1H) 3.68 (m, 2H) 3.51 (m, 3H) 2.59 (q, J=7.64, J=15.19, 2H) 1.17 (t, J=7.59, 3H). MS: m/z (MH+) 416.


H. Biological Examples
Example 1
Materials and Methods
Cloning of the human SGLT1 and human SGLT2 cDNAs and construction of the mammalian expression vector: The human SGLT1 cDNA (Genbank M24847) was cloned from human small intestine. Human SGLT2 cDNA (Genbank M95549) was cloned from human kidney. Both full cDNAs were subcloned into pcDNA and sequenced to verify the integrity of the construct.
Generation of CHO-K1 cells stably expressing human SGLT1 or human SGLT2: Transfection of CHO-K1 cells was performed using DMRIE-C reagent (Life Technologies, Gaithersburg, Md.). Transfectants were then selected in the presence of the antibiotic G418 (Gibco-BRL, Grand Island, N.Y.) at 400 μg/ml. Individual clones were then characterized using the functional assay described below.
Cell-based assay for sodium-dependent glucose transport: Cell lines stably expressing human SGLT1 or SGLT2 were then used for functional analysis of Na+-dependent glucose uptake. Briefly, cells were plated at a density of 65,000 cells per well in a 96-well plate and allowed to grow for 48 hours. Cells were subsequently washed one time with Assay Buffer (50 mM HEPES pH 7.4, 20 mM Tris, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2 and 137 mM NaCl) and treated with compound in the absence or presence of NaCl for 15 minutes. Cells were then labeled with 14C-α-methylglucopyranoside (AMG, Sigma, St. Louis, Mo.), a non-metabolizable glucose analog specific for sodium-dependent glucose transporters as previously described (Peng, H. and Lever J. E. Post-transcriptional regulation of Na+/glucose cotransporter (SGLT1) gene expression in LLC-PK1 cells. J Biol Chem 1995; 270:20536-20542.). After 2 hours the labelled cells were washed three times with ice-cold PBS. After aspiration, cells were solubilized using Microscint 20 (Packard, Meriden, Conn.) and Na-dependent 14C-AMG uptake was quantified by measuring radioactivity. Plates were counted in a TopCount (Packard, Meriden, Conn.). Results are reported as the % inhibition or IC50 value from a representative experiment. Variability for the functional assay was typically within 20%.
Example 2
In Vivo Assay for Efficacy
Male Zucker Diabetic Fatty (ZDF) rats (7-8 weeks) were obtained from Charles River. Animals were maintained on a 12-hour light/dark cycle in a temperature-controlled room. Animals were given ad libitum access to food (standard rodent diet Purina 5008) and water. Animals were fasted for 12 hours prior to initiation of the experiment. On the morning of the experiment, animals were administered vehicle (0.5% methylcellulose) or compound by oral gavage (1 ml/kg). After one hour, animals received an oral glucose challenge (4 ml/kg of 50% solution) and were immediately placed in metabolism cages. Animals were given free access to water and urine was collected for 4 hours. Urinary glucose was quantified using the Trinder Reagent (Sigma).
Example 3
Effects on Plasma Glucose, Plasma Insulin, Plasma Triglycerides, Plasma Free Fatty Acids, Liver Weight, and Body Weight
To examine the effect of an SGLT inhibitor in combination with an RXR agonist, female db/db mice (6-7 weeks of age/Jackson Labs, ME) are treated daily for 11 days with vehicle (0.5% methylcellulose), an RXR agonist (0.1-10 mpk (mg/kg)), an SGLT inhibitor (100 mpk), or an RXR agonist plus SGLT inhibitor. Mice (n=8 animals/group) receive the test compounds or vehicle by oral gavage in a volume of 10 ml/kg of body weight. Body weight is recorded on day 1, prior to dosing, and days 4, 8 and 11. Eighteen hours after the final dose, mice are weighed and anesthetized with CO2/O2 (70:30). Mice are then bled by retro-orbital sinus puncture into 2 mL heparinized polypropylene tubes on ice. Plasma samples are then assayed for glucose, insulin, triglycerides, and free fatty acids. Livers are excised, weighed and frozen.
The SGLT inhibitors and RXR agonists have distinct mechanisms of action. Improved glycemic control, measured as a decrease in plasma glucose, plasma insulin, plasma free fatty acids, or plasma triglycerides, or a combination thereof, can be observed at lower concentrations of an RXR agonist when given in combination with an SGLT inhibitor. Therefore, a leftward shift in the dose-response curve for effect of an RXR agonist on the above parameters can become apparent. In addition, the weight gain observed following treatment with RXR agonists is less pronounced when given with the SGLT inhibitor, since SGLT inhibitors' promotion of the urinary excretion of glucose and loss of calories from the body is demonstrated by reduction in weight or weight gain. Also, since SGLT inhibitors promote a mild diuresis, the edema (and the edematous weight gain) commonly observed after treatment with RXR agonists can be less pronounced or absent. A reduction in the amount of an RXR agonist necessary to achieve efficacy in turn improves the side-effect profile. The decreased side effects can include such conditions as fatty liver, increased liver weight, body weight gain, heart weight gain, edema, cardiac hypertrophy, hepatohypertrophy, hypoglycemia, and hepatotoxicity, or any combination thereof.
Example 4
Effects on Plasma Glucose, HbA1c, Hematocrit, Plasma Insulin, Plasma Triglycerides, Plasma Free Fatty Acids, Total Cholesterol, HDL, Plasma Drug Levels, Liver Weight, Heart Weight, Fat Content and Body Weight
To examine the effect of an SGLT inhibitor in combination with an RXR agonist, male ZDF rats (6 weeks of age/GMI) are treated daily for 28 days with vehicle (0.5% methylcellulose), an RXR agonist (0.1 mpk-10 mpk), SGLT inhibitor (3-100 mpk), or an RXR agonist plus SGLT inhibitor. Rats (n=8 animals/group) receive the test compounds or vehicle by oral gavage in a volume of 2 ml/kg of body weight. Body weight is recorded on day 1, prior to dosing, and twice a week for the duration of the study. On the day prior to the final dose, animals are fasted overnight. One hour after the final dose, rats are weighed and anesthetized with CO2/O2 (70:30). Rats are then bled by retro-orbital sinus puncture into 2 mL heparinized polypropylene tubes on ice. Rats then receive a glucose challenge (2 g/kg p.o) and are placed in metabolism cages for the urine collection (4 hours). Animals are then sacrificed and epididymal fat pads, livers, and hearts are excised, weighed and frozen for histological examination. Plasma samples are then assayed for glucose, HbA1c, insulin, hematocrit, plasma drug levels, total cholesterol, HDL, free fatty acids, and triglycerides. Urine volume and urinary glucose, protein, osmolarity, electrolytes (Na, K, Cl), BUN and creatinine are measured.
The SGLT inhibitors and RXR agonists have distinct mechanisms of action. Improved glycemic control, measured as a decrease in plasma glucose, HbA1c, plasma insulin, or plasma triglycerides, or a combination thereof, can be observed at lower concentrations of RXR agonists when given in combination with an SGLT inhibitor. Therefore, a leftward shift in the dose-response curve for effect of RXR agonists on the above parameters can become apparent. In addition, the weight gain observed following treatment with RXR agonists is less pronounced when given with the SGLT inhibitor, since SGLT inhibitors' promotion of the urinary excretion of glucose and loss of calories from the body is demonstrated by reduction in weight or weight gain. Also, since SGLT inhibitors promote a mild diuresis, the edema (and the edematous weight gain) commonly observed after treatment with RXR agonists can be less pronounced or absent. This can be demonstrated by a reduction in the RXR agonist-induced increase in heart weight. A reduction in the amount of RXR agonists necessary to achieve efficacy in turn improves the side-effect profile. The decreased side effects can include such conditions as fatty liver, increased liver weight, body weight gain, heart weight gain, edema, cardiac hypertrophy, hepatohypertrophy, hypoglycemia, and hepatotoxicity, or any combination thereof.
The above examples can also show that the oral administration of an SGLT inhibitor in combination with an antidiabetic agents, such as an RXR modulator, improve the status of other markers of diabetes mellitus including glycosylated hemoglobin (Hgb A1C) levels. Particularly, the oral administration of an SGLT inhibitor in combination with one or more RXR modulators can reduce body weight or body weight gain as well as liver weight or liver weight gain, compared to administration of one or more RXR modulators alone.
Thus, for treating diabetes, particularly Type II diabetes mellitus, or Syndrome X, a compound of Formula (III) in combination with one or more antidiabetic agents, such as an RXR agonist that increases insulin sensitivity, may be employed comprising administering repeated oral doses of the compound of Formula (III) in the range of about 25 to 1000 mg once or twice daily and repeated doses of the anti-diabetic agent or agents at jointly effective dosages. The jointly effective dosage for antidiabetic agents disclosed herein may be readily determined by those skilled in the art based on standard dosage guidelines. In particular, such combined administration can be effective to accomplish reduction of body weight, body weight gain, liver weight, or liver weight gain in the subject.
Additionally, a method comprising (a) administering to a subject a jointly effective amount of a glucose reabsorption inhibitor; and (b) administering to the subject a jointly effective amount of an antidiabetic agent such as an RXR modulator can be used to reduce body weight, body weight gain, or liver weight of the subject in need thereof, wherein the combined administration can be in any order and the combined jointly effective amounts provide the desired therapeutic effect.
Also, a method comprising (a) administering to a subject a jointly effective amount of a glucose reabsorption inhibitor; and (b) administering to the subject a jointly effective amount of an antidiabetic agent can be used to control body weight, body weight gain, liver weight, or liver weight gain of the subject having diabetes, Syndrome X, or associated symptoms or complications, wherein the combined administration can be in any order and the combined jointly effective amounts providing the desired therapeutic effect.
Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular compound used, the mode of administration, the strength of the preparation and the advancement of the disease condition. In addition, factors associated with the particular patient being treated, including patient's sex, age, weight, diet, time of administration and concomitant diseases, will result in the need to adjust dosages.
While the foregoing specification teaches the principles of the present invention, with examples provided for the purpose of illustration, it will be understood that the practice of the invention encompasses all of the usual variations, adaptations and/or modifications as come within the scope of the following claims and their equivalents.









TABLE 3





EXAMPLE
CHOK-SGLT1
CHOK-SGLT2


#
IC50 (uM)
IC50 (uM)

















7
0.70
0.038


4
1.12
0.016


8
1.12
0.061


5
61% Inh. @10 μM
0.365


2
55% Inh. @10 μM
0.334


9
40% Inh. @10 μM
0.39


3
 1% Inh. @10 μM
1.02


1
0
31% Inh. @10 μM


6
0
18% Inh. @10 μM


10
0.59
0.008

## Claims

1. A method for treating diabetes or Syndrome X, or associated symptoms or complications thereof in a subject, comprising
(a) administering to said subject a jointly effective amount of a glucose reabsorption inhibitor of formula (III)





wherein:
X is CH, N, or C═O;
R1 is H or absent;
R2 is H, F, Cl, OCH3, OCH2CH3, or C1-4 alkyl, CF3, SCH3, substituted or unsubstituted phenyl or NR3R4;
R3 and R4 are H, C1-C6 alkyl, or taken together with the nitrogen atom to which they are both attached form a 5-6 membered heterocyclyl with optionally 1-2 additional heteroatoms independently selected from O, S, and N;
Q is —(CH2)n— where n is 1 or 2;
P is H, C1-7 acyl, or (C1-6 alkoxy)carbonyl; and


Z is substituted or unsubstituted, and is selected from C3-7 cycloalkyl, phenyl, 5- or 6-membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, a biaryl, and a 9- or 10-membered fused bicyclyl or fused heterobicyclyl, wherein said fused heterobicyclyl has between 1 and 4 heteroatoms independently selected from N, O, and S; or a pharmaceutically acceptable salt thereof;
(b) administering to said subject a jointly effective amount of a second antidiabetic agent, and
(c) administering to said subject a jointly effective amount of a third antidiabetic agent;

said co-administration being in any order and the combined jointly effective amounts providing the desired therapeutic effect.

2. The method of claim 1, wherein the third antidiabetic agent is selected from
(aa) insulins,
(bb) insulin analogues;
(cc) insulin secretion modulators, and
(dd) insulin secretagogues.

3. The method of claim 1, wherein the second antidiabetic agent is an RXR agonist.

4. The method of claim 1, wherein the diabetes or Syndrome X, or associated symptoms or complications thereof is selected from IDDM, NIDDM, IGT, IFG, obesity, nephropathy, neuropathy, retinopathy, atherosclerosis, polycystic ovarian syndrome, hypertension, ischemia, stroke, heart disease, irritable bowel disorder, inflammation, and cataracts.

5. The method of claim 1, wherein the diabetes or Syndrome X, or associated symptoms or complication thereof is IDDM.

6. The method of claim 1, wherein the diabetes or Syndrome X, or associated symptoms or complications thereof is NIDDM.

7. The method of claim 1, wherein the diabetes or Syndrome X, or associated symptoms or complications thereof is IGT or IFG.

8. The method of claim 1, wherein R1 is H or absent.

9. The method of claim 1, wherein R2 is H, methyl, or ethyl.

10. The method of claim 1, wherein Q is —(CH2)n— and n is 1.

11. The method of claim 1, wherein Z is independently substituted with between 1 and 3 substituents independently selected from C1-4 alkoxy, phenoxy, C1-4 alkyl, C3-6 cycloalkyl, halo, hydroxy, cyano, amino, C1-4 alkylthio, C1-4 alkylsulfonyl, C1-4 alkylsulfinyl, C1-4 aminoalkyl, mono- and di(C1-4 alkyl)amino, phenyl, C1-4 alkylaminosulfonyl (SO2NHR), amino-(alkylsulfonyl) (—NHSO2R—), C1-4 dialkylaminosulfinyl (SONHRR), C1-4 alkylamido (NHCOR), C1-4 alkylcarbamido (CONHR), 5-6 membered heterocyclyl containing between 1 and 3 heteroatoms independently selected from N, S, and O; and wherein the substituent(s) on Z can be further independently substituted with between 1 and 3 substituents independently selected from C1-4 alkoxy, C1-4 alkyl, halo, hydroxy, cyano, amino, mono or di C1-4 alkyl amino and C1-4 alkylthio.

12. The method of claim 1, wherein Z is 4-substituted phenyl, 3,4-disubstituted phenyl, benzhydryl, substituted or unsubstituted thiophene, biaryl, benzofuranyl, dihydrobenzofuranyl, 4-substituted pyridyl, benzo[b]thienyl, chromanyl, benzothiophenyl, indenyl, indanyl, naphthyl, or 2,3-dihydro-benzo[1,4]dioxanyl.

13. The method of claim 1, wherein Z is unsubstituted or substituted with between 1 and 2 substituents independently selected from methoxy, ethoxy, fluoro, chloro, methyl, ethyl, propyl, butyl and isopropyl.

14. The method of claim 1, wherein Z is biphenyl, 4-(3-pyridyl)phenyl, 4-(2-thienyl)phenyl, 4-(1H-imidazole-1-yl)-phenyl, 4-(1H-pyrazol-1-yl)-phenyl, (4-ethyl)phenyl, (4-propyl)phenyl, (4-methoxyphenyl), dihydrobenzofuran-5-yl, or dihydrobenzofuran-6-yl.

15. The method of claim 1, wherein R1 is absent; X is CH and R2 is H, methyl, ethyl, or methoxy.