MXPA00005025A - Combination of an aldose reductase inhibitor and a glycogen phosphorylase inhibitor - Google Patents

Abstract

Pharmaceutical combination compositions and methods including aldose reductase inhibitors and glycogen phosphorylase inhibitors. The compositions and methods are useful for the treatment insulin resistant conditions such as diabetes and in reducing tissue damage due to ischemia.

Description

COMBINATION OF AN INHIBITOR OF ALDOSE REDUCTASE AND AN INHIBITOR OF GLUCOGEN PHOSPHORYLASE BACKGROUND OF THE INVENTION This invention relates to a pharmaceutical combination of an aldose reductase inhibitor and an inhibitor of glycogen phosphorylase, to kits containing such combinations and to the use of such combinations to treat diabetes, hyperglycemia, hypercholesterolemia, hypertension, hyperinsulinemia, hyperlipidemia, Atherosclerosis and tissue ischemia in mammals. Despite the early discovery of insulin and its extensive subsequent use in the treatment of diabetes, and the discovery and subsequent use of sulfonylureas (for example, Chlorpropamide ™ (Pfizer), Tolbutamide ™ (Upjohn), Acetohexamide ™ (El Lilly) ), Tolazamida ™ (Upjohn)), biguanides (e.g., Phenformin ™ (Ciba Geigy), Metformin ™ (GD Searle)), alpha glucosidase inhibitors (e.g., Precose ™ (Bayer)) and insulin sensitizers ( for example, Rezulin ™ (Parke Davis)) as oral hypoglycemic agents, there is a continuing need for treatments for diabetes. The use of insulin, necessary in about 10% of diabetic patients in which synthetic hypoglycemic agents are the not effective (Type I diabetes, insulin dependent diabetes mellitus), requires multiple daily doses, usually by self injection. The determination of the appropriate dose of insulin requires frequent estimates of sugar in urine or blood. The administration of an excess dose of insulin causes hypoglycemia, with effects ranging from mild abnormalities in blood glucose and coma or even death. The treatment of diabetes mellitus non-insulin dependent (Type II diabetes, NIDDM) usually consists of a combination of diet, exercise, oral agents, eg sulfonylureas, and in more severe cases, insulin. However, clinically available hypoglycemic agents may have other side effects that limit their use. In any case, when one of these agents fails in an individual case, another can succeed. Clearly, a continuing need for hypoglycaemic agents that may have fewer side effects or that may be satisfactory where others fail is evident. Aldose reductase inhibitors constitute a class of compounds that have become widely known for their usefulness in the prevention and treatment of conditions arising from complications of diabetes, such as neuropathy and diabetic nephropathy. Such compounds are well known to those skilled in the art and are easily identified by conventional biological assays. For example, the compound zopolrestat, 1-phthalazinoacetic acid, 3,4-dihydro-4-oxo-3 [[5-trifluoromethyl) -2-benzothiazolyl] methio] -, is known, for example, from the patent of USA 4,939,140 to Larson et al., Assigned jointly, (the description of which is incorporated herein by reference), together with various compounds related thereto, which have utility as inhibitors of aldose reductase. The zopolrestat has the structure and, as an aldose reductase inhibitor, it is useful in the treatment of the complications mentioned above that are caused by diabetes mellitus. Certain inhibitors of aldose reductase have been indicated for use in the reduction of lipid levels in mammals. See, for example, the US patent. 4,492,706 (the disclosure of which is incorporated herein by reference) of Kallai-Sanfacon and EP 0 310 931 A2 (Ethyl Corporation). The patent of E.U.A. 5,064,830 co-assigned (the disclosure of which is incorporated herein by reference) by Going, discloses the use of certain oxaphthalazinyl acetic acids, including zopolrestat, to reduce levels of uric acid in blood. The patent application of E.U.A. No. 08 / 059,688, jointly assigned, describes the use of certain inhibitors of aldose reductase, including zopolrestat, to reduce blood lipid levels in humans. The description indicates that the therapeutic utilities derive from the treatment of diseases caused by a higher level of triglycerides in the blood, including such diseases cardiovascular disorders such as thrombosis, arteriosclerosis, myocardial infarction and angina pectoris. Atherosclerosis, a disease of the arteries, is recognized as the leading cause of death in the United States and Western Europe. The pathological sequence leading to atherosclerosis and occlusive heart diseases is well known. The first stage of this sequence is the formation of "fatty streaks" in the carotid, coronary and cerebral arteries, and in the aorta. These lesions are yellow due to the presence of lipid deposits that are found mainly within the smooth muscle cells and in the macrophages of the intima of the arteries and the aorta. In addition, it is postulated that most of the cholesterol found within fatty streaks, in turn, leads to the development of "fibrous plaque", which consists of accumulated smooth muscle cells of the intimate lipid-laden and surrounded by lipids extracellular, collagen, elastin and proteoglycans. The cells plus the matrix form a fibrous covering that covers a deeper deposit of cellular debris and more extracellular lipids. Lipids consist mainly of free and esterified cholesterol. The fibrous plaque forms slowly and is likely to calcify and necrose over time, progressing to a "complicated lesion" that contributes to the occlusion of the arteries and the tendency to mural thrombosis and spasms of the arterial muscle that characterize advanced atherosclerosis . Epidemiological evidence has firmly established hyperlipidemia as a primary risk factor in the onset of cardiovascular diseases (CVD) due to atherosclerosis. In recent years, the leaders of the medical profession have placed a renewed emphasis on the reduction of plasma cholesterol levels and, in particular, of low density lipoprotein cholesterol, as an essential step in the prevention of CVD. It is now known that the upper limits of the "normal" range are significantly lower than those that had been assessed so far. As a result, it is now considered that large segments of Western populations have a particularly high risk. Such independent risk factors include glucose intolerance, hypertrophy of the left ventricle, hypertension and being male. Cardiovascular disease is especially frequent among diabetic subjects, at least in part, due to the existence of multiple independent risk factors in this population. Therefore, the successful treatment of hyperlipidemia in the general population, and in diabetic subjects in particular, is of exceptional medical importance. Hypertension (or high blood pressure) is a condition that occurs in the human population as a secondary symptom of several different disorders, such as stenosis of the renal arteries, pheochromocytoma or endocrine disorders. However, hypertension also appears in many patients in whom the causative agent of the disorder is unknown. Although such "essential" hypertension is often associated with disorders such as obesity, diabetes and hypertriglyceridemia, the relationship between these disorders has not been clarified. In addition, many patients exhibit the symptoms of high blood pressure in the complete absence of other signs of disease or disorder. It is known that hypertension can directly cause heart failure, kidney failure and stroke (cerebral hemorrhage). These conditions can cause short-term death in a patient. Hypertension can also contribute to the development of atherosclerosis and coronary heart disease. These conditions gradually weaken a patient and, in the long term, can cause death. The exact cause of essential hypertension is unknown, although several factors are thought to contribute to the onset of the disease. Among such factors are stress, uncontrolled emotions, unregulated release of hormones (the renin system, angiotensin and aldosterone), an excess of salts and water due to poor kidney function, thickening of walls and hypertrophy of the vasculature. they produce narrowing of blood vessels, and genetic factors. It is considered that the treatment of essential hypertension takes into account the above factors. Thus, a wide range of beta-blockers, vasoconstrictors, angiotensin-converting enzyme inhibitors and the like have been created and marketed as anti-hypertensives. The treatment of hypertension using these compounds has been shown to be beneficial in the prevention of short-term deaths, such as in heart failure, renal failure and cerebral hemorrhage. However, the development of atherosclerosis or heart disease due to hypertension for a long period of time remains a problem. This implies that although blood pressure is being reduced, the underlying cause of essential hypertension is not responding to this treatment. Hypertension has been associated with high levels of insulin in the blood, a condition known as hyperinsulinemia. Insulin, a peptide hormone whose main actions are to promote the use of glucose, the synthesis of proteins and the formation and storage of neutral lipids, also acts to promote the growth of vascular cells and increases the retention of sodium by the kidney, among others. things. These latter functions can be performed without affecting glucose levels and are known causes of hypertension. The growth of peripheral vasculature, for example, can cause narrowing of peripheral capillaries; while the retention of sodium increases the volume of blood. Thus, the reduction of insulin levels in hyperinsulinemic patients can prevent abnormal vascular growth and retention of sodium by the kidney caused by high insulin levels and, therefore, relieve hypertension. Cardiac hypertrophy is a significant risk factor in the onset of sudden death, myocardial infarction and congestive heart failure. These cardiac events are due, at least in part, to increased susceptibility to myocardial injury after ischemia and reperfusion, which may appear in outpatients as well as after surgery. There is a medical need, not satisfied, to prevent or minimize adverse myocardial perioperative cases, particularly, perioperative myocardial infarction. Both cardiac and non-cardiac surgery are associated with substantial risks of myocardial infarction or death. It is considered that some 7 million patients who undergo non-cardiac surgery are at risk, with incidences of perioperative death and serious cardiac complications as high as 20 to 25% in some series. In addition, of the 400,000 patients who undergo annual coronary bypass surgery, it is estimated that perioperative myocardial infarction occurs in 5% and death in 1-2%. Currently, no drug therapy is commercialized in this area that reduces lesions in the cardiac tissues of perioperative myocardial ischemia or increases cardiac resistance to ischemic episodes. It is hoped that such therapy will save lives and reduce hospitalizations, improve the quality of life and reduce the overall health costs of high-risk patients.

Hepatic glucose production is an important goal for NIDDM therapy. The liver is the main regulator of plasma glucose levels after absorption (fasting), and the rate of hepatic glucose production in patients with NIDDM is significantly higher than in normal individuals. Similarly, in the postprandial state (after eating), when the liver has a proportionally smaller role in the total plasma glucose supply, the production of hepatic glucose is abnormally high in patients with NIDDM. Glycogenolysis is an important goal for the interruption of hepatic glucose production. The liver produces glucose through glycogenolysis (breakage of the glucose polymer, glycogen) and gluconeogenesis (glucose synthesis from precursors with 2 and 3 carbons). Several lines of evidence indicate that glycogenolysis can make an important contribution to the production of hepatic glucose in the NIDDM. First, in normal human beings, after absorption, it is estimated that up to 75% of the hepatic glucose production comes from glycogenolysis. Second, patients who have liver diseases that affect the storage of glycogen, including Hers disease (glycogen phosphorylase deficiency), present episodic hypoglycaemia. These observations suggest that glycogenolysis may be a significant process for the production of hepatic glucose.

Glycogenolysis is catalyzed in the liver, muscle and brain by tissue-specific isoforms of the enzyme glycogen phosphorylase. This enzyme cleaves the macromolecule of glycogen by releasing glucose 1 -phosphate and a new macromolecule of shorter glycogen. To date, two types of glycogen phosphorylase inhibitors have been presented: glucose and glucose analogues [Martin, J.L. et al. Biochemistry 1991, 30, 10101] and caffeine and other purine analogues [Kasvinsky, P.J. et al. J. Biol. Chem. 1978, 253, 3343-3351 and 9102-9106]. It has been postulated that these compounds, and inhibitors of glycogen phosphorylase in general, are of potential use for the treatment of NIDDM, by reducing the production of glucose by the liver and reducing blood glucose. [Blundell, T.B. et al. Diabetology 1992, 35, Suppl. 2, 569-576 and Martin et al. Biochemistry 1991, 30, 10101]. The mechanism (s) responsible for the myocardial lesions observed after ischemia and reperfusion are not fully understood. It has been reported (MF Allard, et al., Am. J. Physiol. 267, H66-H74, 1994) that "the reduction of glycogen before ischemia ... is associated with a better functional recovery of the left ventricle after the ischemia in hearts of hypertrophied rats ". Thus, although there are a variety of therapies for hyperglycemia, hypercholesterolemia, hypertension, hyperinsulinemia, hyperlipidemia, atherosclerosis and ischemia, there is a continued need and a continuing search in this field for the technique of alternative therapies.

BRIEF DESCRIPTION OF THE INVENTION This invention is directed to pharmaceutical compositions comprising aldose reductase inhibitors and glycogen phosphorylase inhibitors, and to the use of such compositions for the treatment of insulin-resistant conditions, including diabetes, in mammals (e.g., humans, men or women) or to the use of such compositions to reduce tissue damage (e.g., substantially preventing tissue damage or inducing tissue protection) resulting from ischemia. The combinations comprise therapeutically effective amounts of an α-aldose reductase inhibitor and an inhibitor of glycogen phosphorylase. A preferred amount of aldose reductase inhibitor is from about 0.1 mg / kg to about 20 mg / kg and a preferred amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. An especially preferred inhibitor of aldose reductase is 1-phthalazinoacetic acid, 3,4-dihydro-4-oxo-3 - [[5-trifluoromethyl) -2-benzothiazolyl] methyl] -. Preferred glycogen phosphorylase inhibitors include compounds having the formula I.

Formula I and the pharmaceutically acceptable salts and prodrugs thereof in which the broken line (-) is an optional bond; A is -C (H) =, -C (alkyl (C? -C) = or -C (halo) = when the broken line (-) is a bond, or A is methylene or -CH (alkyl (C)) -? - C4) - when the broken line (-) is not a link, each of Ri, R-io or Rn is, independently, H, halo, 4-, 6-or 7-nitro, cyano, alkyl (C -? - C4), alkoxy (d-C4), fluoromethyl, difluoromethyl or trifluoromethyl; R2 is H; R3 is H or (C1-C5) alkyl; R4 is H, methyl, ethyl, n-propyl, hydroxy; alkyl (C? -C3), alkoxy (Cr C3) -alkio (C -? - C3), phenyl-alkyl (C? -C4), phenyl-hydroxy-aIcyl (C? -C), phenylthien-2 - or -3-α-alkyl (C -? - C4) or fur-2 or -3-yl-alkyl (C? -C4), wherein said R4 rings are mono-, di- or tri- substituted independently on carbon with H, halo, (C -? - C) alkyl, (C? - C4) alkoxy, trifluoromethyl, hydroxy, amino or cyano; or R is pyrid-2-, -3- or -4 -l- (C-C4) alkyl, thiazole-2-, -4- or -5-yl (C1-C) alkyl, midazole-1-, -2-, -4- or -5-yl -alkyl (CrC), pyrrol-2- or -3-yl-alkyl (C? -C), oxazole-2-, -4- or -5-yl-alkyl (Ct-C4), pi Razo I-3-, -4- or -5-α-alkyl (C?-C4), isoxazole-3-, -4- or -5-yl-alkyl (C?-C4), isothiazoI-3-, -4- or -5-yl-alkyl (C? -C4), pyridazin-3- or -4-yl- (C1-C4) alkyl, pyrimidin-2-, -4-, -5- or -6 -alkyl (C? -C4), pyrazin-2- or -3-yl-alkyl (C? -C4) or 1, 3,5-triazin-2-yl-alkyl (C? -C), in which said above R heterocycles are optionally mono- or di-substituted independently with halo, trifluoromethyl, (C? -C) alkyl, (C? -C4) alkoxy, amino or hydroxy, and said mono- or di-substituents are linked carbon R5 is H, hydroxy, fluoro, alkyl (C? -C5), alkoxy (C? -C5), alkanoyl (C? -C6), amino-alkoxy (C? -C4), mono-N- or di-N , N-alkyl (C? -C) amino-alkoxy (C? -C4), carboxy-alkoxy (CrC4), alkoxy (C? -C5) -carbonyl-alkoxy (C? -C), benzyloxycarbonyl-alkoxy (C C), or carbonyloxy, wherein said carbonyloxy is bonded via carbon-carbon with phenyl, thiazolyl, imidazolyl, 1 H-indolyl, furyl, pyrrolyl, oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl. or 1, 3,5-triazinyl, and wherein said R5 rings above are optionally mono-substituted with halo, alkyl (C? -C), alkoxy (C-? -C), hydroxy, amino or trifluoromethyl and said mono-substituents are attached to carbon; R7 is H, fluoro or alkyl (C? -C5); or Rs and R7 can be taken together to be oxo; R6 is carboxy, alkoxycarbonyl (C? -C8), C (O) NR8Rg or C (O) R12, wherein Rs is H, (C? -C3) alkyl, hydroxy or (C1-C3) alkoxy; and R9 is H, (C? -C8) alkyl, hydroxy, (C-? - C8) alkoxy, perfluorinated methylene (CrC8) alkyl, phenyl, pyridyl, thienyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl or 1, 3,5-triazinyl, wherein said Rg rings above are carbon-nitrogen bonds; or Rg is mono-, di- or tri-substituted (C? -C5) alkyl, wherein said substituents are independently of H, hydroxy, amino, mono-N- or di-N, N-alkyl (C ? -C5) amino; or R9 is mono- or di-substituted alkyl (C? -C5), wherein said substituents are independently phenyl, pyridyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, pyridinyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl or 1, 3,5-triazinyl, in which the non-aromatic rings Rg containing nitrogen are optionally mono-substituted on nitrogen with (C -? - C6) alkyl, benzyl, benzoyl or alkoxycarbonyl (C? -C6), and in which the Rg rings are optionally mono-substituted on a carbon with halo, alkyl (C? -C4), alkoxy (C?? C4), hydroxy, amino or mono-N- and d-N, N-alkyl (C -? - C5) amino, with the proviso that no quaternized nitrogen is included and there are no nitrogen-oxygen, nitrogen-nitrogen or nitrogen-nitrogen bonds halo; R-? 2 is piperazin-1-yl, 4-alkyl (C? -C4) piperazin-1-yl, 4-formylpiperazin-1-yl, morpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxo-thiomorpholino, thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl, 1, 1-dioxo-thiazolidin-3-yl, 2-alkoxy (C6-6) carbonylpyrrolidin-1-yl, oxazolidin-3-yl or (R) -hydroxymethylpyrrolidin-1-yl; or R 2 is oxazetidin-2-yl 3- and / or 4-mono- or di-substituted, oxazolidin-3-yl 2-, 4- and / or 5-mono- or di-substituted, thiazolidin-3- 2-, 4- and / or 5-mono- or di-substituted ilo, 2-, 4- and / or 5-mono- or di-substituted 1-oxothiazolidin-3-yl, 1,1-dioxothiazolidin-3-yl ilo 2-, 4- and / or 5-mono- or di-substituted, pyrrolidin-1-yl 3- and / or 4-mono- or di-substituted, piperidin-1-yl 3-, 4- and / or 5-mono-, di- or tri-substituted, piperazin-1-yl 3-, 4- and / or 5-mono-, di- or tri-substituted, azetidin-1-yl 3-substituted, 1, 2- oxazinan-2-yl 4- and / or 5-mono- or di-substituted, pyrazolidin-1-yl 3- and / or 4-mono- or di-substituted, isoxazolidin-2-yl 4- and / or 5- mono- or di-substituted, isothiazolidin-2-yl 4- and / or 5-mono- and / or di-substituted, wherein said substituents R12 are, independently, H, halo, alkyl- (C? -C5) , hydroxy, amino, mono-N- or di-NN-alkylaminoCCrCs), formyl, oxo, hydroxyimino, (C? -C5) alkoxy, carboxy, carbamoyl, mono-N- or di-N, N-alkyl (C? -C4) carbamoyl, (C? -C4) alkoxy imino, (C? -C4) alkoxy methoxy, (C? -C6) alkoxycarbonyl, carboxy-alkyl (C? -C5) or hydroxy-alkyl (C? -C5); with the proviso that if R is H, methyl, ethyl or n-propyl, R5 is OH; with the proviso that if R5 and R7 are H, then R4 is not H, methyl, ethyl, n-propyl, hydroxy-alkyl (C? -C3) or alkoxy (C? -C3) -alkyl (CrC3) and R &; is C (O) NR8R9, C (0) R? 2 or alkoxy (CrC4) carbonyl. A first group of preferred compounds of formula I consists of the compounds in which Ri is 5-H, 5-halo, 5-methyl or 5-cyano; Rio and R11 are each, independently, H or halo; A is -C (H) =; R2 and R3 are H; R is phenyl-alkyl (C? -C2), wherein said phenyl groups are mono-, di- or tri-substituted independently with H, halo or mono- or disubstituted independently with H, halo, alkyl (C? -C4) ), alkoxy (C? -C), trifluoromethyl, hydroxy, amino or cyano; or R is thien-2- or 3-yl-alkyl (C? -C2), pyrid-2, -3- or -4-yl-a-alkyl (C? -C2), thiazole-2-, -4 - or -5-yl-aIcyl (C? -C), imidazol-1-, -2-, -4- or -5-yl-alkyl (C2), fur-2- or -3-yl-alkyl (C C2), pyrroi-2- or -3-yl-alkyl (C? -C2), oxazol-2-, -4- or -5-yl-alkyl (C -? - C2), pyrazole-3- , -4- or -5-yl (C1-C2) alkyl, isoxazol-3-, -4- or -5-yl- (C-? - C2) alkyl in which said R heterocycles are optionally mono- or di-substituted independently with halo, trifluoromethyl, alkyl (CrC4), (C1-C4) alkoxy, amino or hydroxy and said mono- or di-substituents are bonded with carbon; R5 is hydroxy; R6 is C (O) NR8R9 or C (O) 12; and R7 is H. Within the first preferred group of preferred compounds of formula I, there is a first group of especially preferred compounds, wherein the carbon atom a has a stereochemistry (S); the carbon atom b has a stereochemistry (R); R4 is phenyl-alkyl (C? -C2), thien-2-yl (C1-C2) alkyl, thien-3-yl-alkyl (C -? - C2), fur ^ -yl-alkyl ^ - C ^ o fur-3-yl-alkyl (C? -C2), wherein said rings are mono- or di-substituted independently with H or fluoro; R6 is C (O) NR8R9; R8 is (C1-C3) alkyl, hydroxy or (C? -C3) alkoxy; and Rg is H, alky (CrC8), hydroxy, hydroxy (C-? - C6) alkyl, (CtC8) alkoxy, pyridyl, morpholinyl, piperazinyl, pyrrolidinyl, piperidinyl, imidazolyl or thiazolyl, or C ^ alkylcycloalkyl) mono- substituted with pyridyl, morpholinyl, piperazinyl, pyrrolidinyl, piperidinyl, imidazolyl or thiazolyl. Within the first group of especially preferred compounds, the [(1 S) - ((R) -Hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-ethyl] -amide compounds of 5-chloro-1 H-indole are particularly preferred. -2-carboxylic,. { (1S) - [(R) -hydroxy- (methoxy-methyl-carbamoyl) -methyl] -2-phenyl-ethyl} 5,6-dichloro-1 H-indole-2-carboxylic acid amide. { 5-Chloro-1 H-indole-2-carboxylic acid (1 SH (R) -hydroxy- (methoxy-methyl-carbamoyl) -methyl] -2-phenyl-ethyl} -amide; (1S) - { (R) -hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl] -2-phenyl-ethyl) -amide of 5-chloro-1 H- indole-2-carboxylic acid,. {(1 S) - [(R) -hydroxy- (methyl-pyridin-2-yl-carbamoyl) -methyl] -2-phenyl-ethyl} -amide of 5- chloro-1 H-indole-2-carboxylic acid or ((1 SH (R) -hydroxy- [methyl- (2-pyridin-2-yl-ethyl) -carbamoyl] -methyl] -2-phenyl-ethyl) 5-chloro-1 H-indole-2-carboxylic acid amide Within the first group of especially preferred compounds, there are those compounds in which a.Ri is 5-chloro, is benzyl, R8 is methyl, and Rg is methyl, Ri is 5-chloro Rn is H, R10 is 6-chloro, R is benzyl, R8 is methyl, and Rg is methoxy, Ri is 5-chloro, R4 is benzyl, Rs is methyl; and Rg is methoxy, Ri is 5-chloro, R4 is benzyl, R8 is methyl, and Rg is 2- (hydroxy) ethyl, Ri is 5-chloro, Rio and R.1 are H, R4 is benzyl,; R8 is methyl; and Rg is pyridin-2-yl; and f. Ri is 5-chloro; Rio and Rn are H; R is benzyl; Rs is methyl; and Rg is 2- (pyridin-2-yl) ethyl. Within the first group of especially preferred compounds above, there is a second group of especially preferred compounds in which the carbon atom a has a stereochemistry (S); the carbon atom b has a stereochemistry (R); R is phenyl-aikyl (C? -C2), thien-2-yl-alkyl (C-C2), thien-3-yl-alkyl (C? -C2), fur-2-yl-alkyl (C? C2) or fur-3-yl-alkyl (C? -C2), wherein said rings are mono- or di-substituted independently with H or fluorine; R6 is C (0) R? 2; and R-12 is morpholino, 4-alkyl (C -? - C) piperazin-1-yl, 3-substituted azetidin-1-yl, pyrrolidin-1-yl 3- and / or 4-mono- or di-substituted. , isoxazolidin-2-yl 4 and / or 5 mono- or di-substituted, 1,2-oxazinan-2-yl 4- and / or 5- mono- or di-substituted, in which each of said substituents is , independently, H, halo, hydroxy, amino, mono-N- or di- N, N-alkyl (C-pCe) amino, oxo, hydroxyimino or alkoxy. Within the second preferred group of especially preferred compounds are particularly preferred compounds [(1 S) -benzyl- (2R) -hydroxy-3- (4-methyl-piperazin-1-yl) -3-oxo-propyl hydrochloride. ] 5-Chloro-1 H-indole-2-carboxylic acid amide, [(1 S) -benzyl- (2R) -hydroxy-3- (3-hydroxy-azetidin-1-yl) -3-oxo -propyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid, ((1 S) -benzyl- (2R) -hydroxy-3-isoxazolidin-2-yl-3-oxo-propyl) -amide of 5-chloro-1 H-indole-2-carboxylic acid, ((1 S) -benzyl- (2R) -hydroxy-3- [1,2] oxazinan-2-yl-3-oxo-propyl) -amide of 5-chloro-1 H-indole-2-carboxylic acid, [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxy-pyrrolidin-1-yl) -3-oxo- 5-Chloro-1 H-indole-2-carboxylic acid propyrapide, [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxy-pyrrolidin-1 -ii) - (2R) -hydroxy- 5-Chloro-1 H-indole-2-carboxylic acid 3-oxo-propyl] -amide, [(1 S) -benzyl-3 - ((3R, 4R) -dihydroxy-pyrrolidin-1-yl) - ( 2R) -hydroxy-3-oxo-propyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid 5-Chloro-1H-indole-2-carboxylic acid [(1 S) -benzyl- (2R) -hydroxy-3-morpholin-4-yl-3-oxo-propyl) -amide. Within the second above group of especially preferred compounds are the compounds in which a. Ri is 5-chloro; R4 is benzyl; and R-? 2 is 4-methylpiperazin-1-yl; b. R1 is 5- chloro; R4 is benzyl; and R 2 is 3-hydroxyacetidin-1-yl; c. R1 is 5-chloro; Rio and R11 are H; R is benzyl; and R-? 2 is isoxazolidin-1-yl; d. R1 is 5-chloro; R10 and R11 are H; R4 is benzyl; and R12 is (1, 2) -oxazinan-2-yl; and. Ri is 5-chloro; Rio and R11 are H; R4 is benzyl; and R-? 2 is 3 (S) -hydroxypyrrolidin-1-yl; F. R-i is 5-chloro; R4 is benzyl; and R 2 is (3S, 4S) -dihydroxypyrrolidin-1 -lo; g. R-i is 5-chloro; R4 is benzyl; and R-? 2 is (3R, 4S) -dihydroxypyrrolidin-1-yl; h. R1 is 5-chloro; R4 is benzyl; and R 2 is morpholino. A second of preferred compounds of formula I, consists of those compounds wherein R 1 is H, halo, methyl or cyano; each of R-io and Rn is, independently, H or halo; A is -C (H) =; R2 and R3 are H; R4 is phenyl-alkyl (C? -C2), wherein said phenyl groups are mono-, di- or tri-substituted independently with H, halo or mono- or disubstituted independently with H, halo, alkyl (C? -C4) ), alkoxy (C -? - C); trifluoromethyl, hydroxy, amino or cyano; or R .; is tien-2- or -3-yl-alkyl (C? -C2), pyrid-2-, -3- or -4-yl-alkyl (Cr C2), thiazole-2-, -4- or -5-yl-alkyl (C? -C2), imidazol-1-, -2-, -4- or -5-yl-alkyl (Ci-C2), fur-2- or -3-yl-alkyl (C? -C2), pyrrol-2- or -3-yl-alkyl (C2), oxazole-2-, -4- or -5-yl-alkyl ( CrC2), pyrazoI-3-, -4- or -5-yl-alkyl (C? -C2), isoxazole-3-, -4- or -5-yl-alkyl (CrC), wherein said R4 heterocycles above are optionally mono- or di-substituted independently with halo, trifluoromethyl, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, amino or hydroxy and said mono- or di-substituents are attached to carbon; R5 is hydroxy; R6 is carboxy or (C? -C8) alkoxycarbonyl; and R7 is H, fluoro or (C1-C6) alkyl. Within the second group of preferred compounds of formula I is a group of especially preferred compounds in which the carbon atom a has a stereochemistry (S); the carbon atom b has a stereochemistry (R); R4 is phenyl-alkyl (C? -C2), thien-2-yl-alkyl (C? -C2), thien-3-yl-alkyl (C? -C2), fur-2-yl-alkyl (C -? - C2) or fur-3-yl-alkyl (C? -C2), wherein said rings are mono- or di-substituted independently with H or fluorine; R-io and R ?? they are H; Rβ is carboxy; and R7 is H. Within the independently above group, there is a compound in which Ri is 5-chloro; R-io and R11 are H; and R4 is benzyl. A third group of preferred compounds of formula I consists of those compounds wherein R-i is H, halo, methyl or cyano; each of R10 and Rn are, independently, H or halo; A is -C (H) =; R2 and R3 are H; R is phenyl-alkyl (CrC2), wherein said phenyl groups are mono-, di- or tri-substituted independently with H or halo, or are mono- or di-substituted independently with H, halo, (C1-C4) alkyl ), (C 1 -C 4) alkoxy, trifluoromethyl, hydroxy, amino or cyano; or R4 is thien- 2- or -3-yl-alkyl (CrC2), pyrid-2-, -3- or -4-yl-alkyl (Cr C2), thiazole-2-, -4- or -5- il-alkyl (C? -C2), midazole-1, -2-, -4- or -5-iI-alkyl (Cr C2), fur-2-or -3-yl-a-alkyl (C1-C2) ), pyrrole-2- or -3-alkyl (C1-C2), oxazole-2-, -4- or -5-yl-alkyl (C? -C2), pyrazole-3-, -4- or -5 -alkyl (C? -C2), or -soxazol-3-, -4- or -5-yl-alkyl (C? -C2), wherein said R4 heterocycles are optionally mono- or di-substituted , independently, with halo, trifluoromethyl, alkyl (CtC), alkoxy (CrC4), amio or hydroxy, and said mono- or di-substituents are attached to carbon; R5 is fluoro, alkyl (C? -C), alkoxy (C-1-C5), amino-alkoxy (C -? - C4), mono-N- or di-N, N-alkyl (C? -C4) amino-alkoxy (C? -C4), carboxy (C1-C4) alkoxy, alkoxy (CrC5) -carbonyl-(C1-C4) alkoxy, benzyloxycarbonyl-(C1-C4) alkoxy; R6 is carboxy or (C -? - C8) alkoxycarbonyl; and R7 is H, fluorine or alkyl (C-i-Cß). A fourth group of preferred compounds of formula I consists of those compounds wherein R 1 is halo, methyl or cyano; each of R-to and Rn is, independently, H or halo; A is -C (H) =; R2 and R3 are H; R is phenyl-alkyl (C? -C2), wherein said phenyl groups are mono-, di- or tri-substituted independently with H or halo, or are mono- or di-substituted independently with H, halo, alkyl ( C? -C4), (C? -C) alkoxy, trifluoromethyl, hydroxy, amino or cyano; or R4 is thien-2- or -3-yl-alkyl (CrC2), pyrid-2-, -3- or -4-yl-alkyl (Cr C2), thiazole-2-, -4- or -5- il-C1-C2 alkyl, midazole-1, -2-, -4- or -5-yl-alkyl (Cr C2), fur-2- or -3-yl-alkyl (C1-C2) , pyrrole-2- or -3-yl-alkyl (C? -C2), oxazole-2-, -4- or -5-yl-alkyl (C -? - C2), pyrazole-3-, -4- or -5-yl-alkyl (C? -C2) or isoxazole-3-, -4- or -5-yl-(C1-C2) alkyl, wherein said R4 heterocycles above are optionally mono- or di-substituted independently with halo, thifluoromethyl, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, amino or hydroxy, and said mono- or di-substituents are attached to carbon; R5 is fluoro, alkyl (C -? - C4), alkoxy (C -? - C5), amino-alkoxy (C1-C4), mono-N- or di-N, N-alkyl (C? -4) amino -alcoxy (C? -C), carboxy (C1-C4) alkoxy, alkoxy- (C? -C5) -carboni-(C1-C4) alkoxy, benzyloxycarbonyl-(C1-C4) alkoxy; and R6 is C (O) NR8Rg or C (O) R12; and R is H, fluorine or alkyl (Ci-Cß). Preferred glycogen phosphorylase inhibitors include compounds having the formula IA Formula IA and the pharmaceutically acceptable salts and prodrugs thereof in which the broken line (-) is an optional bond; A is -C (H) =, C (alkyl (C? -C4)) =, -C (Halo) = or -N =, when the broken line (-) is a bond, or A is methylene or - CH (alkyl (C? -C4)) -, when the dashed line (-) is not a bond; each of Ri, Rio or Rn is, independently, H, halo, cyano, 4-, 6- or 7-nitro, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, fluoromethyl, difluoromethyl or trifluoromethyl; R2 is H; R3 is H or alkyl (CrC5); R 4 is H, methyl, styrene, n-propyl, hydroxy (C 1 -C 3) alkyl, (C 1 -C 3) alkoxy-(C 1 -C 3) alkyl, phenyl-C 1 -C 4 alkyl, phenylhydroxy-alkyl (CrC), (phenyl) ((C? -C4) alkoxy) - (C4) alkyl, thien-2- or -3-yl-alkyl (C? -C4) or fur-2- or -3-yl -alkyl (C -? - C4), wherein said R4 rings are mono-, di- or tri-substituted independently on carbon with H, halo, (C1-C4) alkyl, (C1-C4) alkoxy, trifluoromethyl, hydroxy, amino, cyano or 4,5-dihydro-1 H-imidazol-2-yl; or R4 is pyrid-2-, -3- or -4-yl-alkyl (C- | -C4), thiazole-2-, -4- or -5-yl-alkyl (C? -C), imidazole -2-, -4- or -5-yl-alkyl (C? -C4), pyrrole-2- or -3-yl (C1-C4) alkyl. oxazole-2-, -4- or -5-yl-alkyl (C? -C4), pyrazole-3-, -4- or -5-yl- (C1-C4) alkyl, isoxazole-3-, - 4- or -5-yl-alkyl (C -? - C4), isothiazol-3-, -4- or -5-yl (C1-C4) alkyl, pyridazin-3- or -4-yl-alkyI ( C1-C4), pyrimidin-2-, -4-, -5- or -6-yl (C1-C4) alkyl, pyrazin-2- or -3-yl (C1-C4) alkyl, 1, 3,5-triazin-2-yl-alkyl (CrC4) or indole-2-alkyl (CtC), wherein said above R4 heterocycles are optionally mono- or di-substituted independently with halo, trifluoromethyl, alkyl ( C? -C4), (C-? - C4) alkoxy, amino, hydroxy or cyano and said substituents are attached to carbon; or R 4 is R-15-carbonyloxymethyl, wherein said R 15 is phenyl, thiazolyl, imidazolyl, 1H-indoline, furyl, pyrrolyl, oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or 1,3, 5-triazinyl and wherein said above R15 rings are optionally mono- or di-substituted independently with halo, amino, hydroxy, (C4) alkyl, (C1-C4) alkoxy or trifluoromethyl and said mono- or di-substituents are linked carbon R5 is H; R6 is carboxy, (C? -C8) alkoxycarbonyl, benzyloxycarbonyl, C (O) NR8R9 or C (O) R12; wherein R8 is H, (C? -C6) alkyl, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl (C1-C5) alkyl, hydroxy or (C? -C8) alkoxy; and Rg is H, cyclo (C3-C8) alkyl, cyclo (C3-C8) alkyl- (C1-C5) alkyl, (C4-C7) cycloalkenyl, (C3-C) cycloalkyl-alkoxy; (C1-C5), (C3-C7) cycloalkyloxy, hydroxy, perfluorinated methylene-(C-? - C8) alkyl, phenyl or a heterocycle, said pyridyl heterocycle, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, pyridinyl, piperidinyl, morpholinyl, pyridazinyl, primidinyl, pyrazinyl, piperazinyl, 1, 3,5-triazinyl, benzothiazolyl, benzoxazolyl, benzoimidazolyl, thiochromanyl or tetrahydrobenzothiazolyl, wherein said heterocyclic rings they are linked by carbon-nitrogen bond; or Rg is (Ci-Cβ) alkyl or (C? -C8) alkoxy, wherein said (C Cß) alkyl or (C? -C8) alkoxy are optionally monosubstituted with (C4-C7) cyclo-alkenyl-1- ilo, phenyl, thienyl, pyridyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, midazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl, piperidinyl, morpholinyl, thiomorpholinyl, 1-oxothiomorpholinyl, 1,1-dioxothiomorpholinyl, pyridazinyl, pyrimidinyl , pyrazinyl, piperazinyl, 1, 3,5-triazinyl or indolyl, and wherein said alkyl (Ct-Cß) or (C-? - C8) alkoxy are optionally, additionally and independently mono- or di-substituted with halo, hydroxy, (C1-C5) alkoxy, amino, mono-N- or di-N, N-alkyl (C? -C5) amino, cyano, carboxy or (C -? - C) alkoxycarbonyl; and wherein the Rg rings are optionally mono- or di-substituted independently, on carbon, with halo, (C? -C4) alkyl, (C1-C4) alkoxy, hydroxy, hydroxy (C1-C4) alkyl, amino -alkyl (C1-C4), mono-No di-N, N-alkyl (C? -C4) amino-alkyl (C? -C4), alkoxy (C? -C) -alkyl (C? -C4), amino, mono-N- or di-N, N-alkyl (C-C4) amino, cyano, carboxy, (C1-C5) alkoxycarbonyl, carbamoyl, formyl or trifluoromethyl, and optionally said Rg rings may additionally be mono- or independently substituted with alkyl (C? -C5) or halo; with the proviso that no quaternized nitrogen is included in any Rg heterocycle; R 2 is morpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, thiazolidin-1-yl, 1-oxothiazolidin-3-yl, 1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin- 1-yl, piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl, pyrazolidin-1-yl, isoxazolidin-2-yl, isothiazolidin-2-yl, 1,2-oxazetidin-2-yl, oxazolidin-3-yl, 3,4-dihydroisoquinolin-2-yl, 1,3-dihydroisoindol-2-yl, 3,4-dihydro-2H-quinol-1 - ilo, 2,3-dihydro-benzo [1,4] oxazin-4-ylo, 2,3-dihydro-benzo [1,4] -thiazine-4-yl, 3,4-dihydro- 2H-quinoxalin-1-yl, 3,4-dihydro-benzo [c] [1,2] oxazin-1-yl, 1,4-dihydro-benzo [d] [1, 2] oxazin-3-yl, 3,4-dihydro-benzo [e] [1,2] -oxazin-2-yl, 3H-benzo [d] isoxazol-2-yl, 3H-benzo [c] isoxazol-1-yl or azepane-1- ilo, wherein said R-? 2 rings are optionally mono-, di- or tri-substituted, independently, with halo, (C1-C5) alkyl, (C1-C5) alkoxy, hydroxy, amino, mono-N- or di-N, N-alkyl (Ci-CsJamino, formyl, carboxy, carbamoyl, mono-N- or di-N, N-alkyl) l (C? -C5) carbamoyl, (C-? - C6) alkoxy (C? -C3) alkoxy, (C? -C5) alkoxycarbonyl, benzyloxycarbonyl, alkoxy (CrC5) carbonyl-alkyl (C-1-C5) ), (C 1 -C 4) alkoxy carbonylamino, carboxy (C 1 -C 5) alkyl, carbamoyl (C 1 -C 5) alkyl, mono-N- or di-N, N-C 1 -C 5 alkylcarbamoyl-alkyl ( C ^ Cs), hydroxy (C1-C5) alkyl, (C? -C4) alkoxy- (C? -C4) alkyl, amino- (C-C4) alkyl, mono-N- or di-N, N- alkyl (C? -C4) amino-alkyl (C? -C4), oxo, hydroxyimino or alkoxyimino (C? -C6), and in which no more than two substituents are selected from oxo, hydroxyimino or alkoxy (C-? -C6) imino, and oxo, hydroxyimino or alkoxy (Ci-Cß) imino are not found on non-aromatic carbons; and wherein said R-? 2 rings are optionally mono- or disubstituted independently with alkyl (C Cs) or halo; with the proviso that when R6 is (C1-C5) alkoxycarbonyl or benzyloxycarbonyl, then Ri, is 5-halo, 5-aikyl (C? -C4) or 5-cyano and R4 is (phenyl) (hydroxy) -alkyl (C? -C), (phenyl) ((C -? - C4) alkoxy) (C 1 -C 4) alkyl, hydroxymethyl or Ar (C 1 -C 2) alkyl, where Ar is thien-2 or -3- ilo, fur-2- or -3-yl or phenyl, wherein said Ar is optionally mono- or di-substituted independently with halo; with the proviso that when R4 is benzyl and R5 is methyl, R-? 2 is not 4-hydroxy-piperidin-1-yl, or when R4 is benzyl and R5 is methyl, R6 is not C (0) N (CH3 )2; with the proviso that when R-t, R10 and Rn are H, R4 is not imidazol-4-ylmethyl, 2-phenylethyl or 2-hydroxy-2-phenylethyl; with the proviso that when R8 and Rg are n-pentyl, R1 is 5-chloro, 5-bromo, 5-cyano, 5-alkyl (C? -C5), 5-alkoxy (CrC) or trifluoromethyl; with the proviso that when R 2 is 3,4-dihydroisoquinol-2-yl, said 3,4-dihydroisoquinol-2-yl is not substituted with carboxy-alkyl (C 1 -C); with the proviso that when R8 is H and Rg is alkyl (C -? - C6), Rg is not substituted with carboxy or (C -? - C4) alkoxycarbonyl on the carbon that is attached to the nitrogen atom N of NHR9; and with the proviso that when R6 is carboxy and R-i, R10, Rn and R5 are H, then R4 is not benzyl, H, (phenyl) (hydroxy) methyl, methyl, ethyl or n-propyl. A first group of preferred compounds of formula IA coast of those compounds wherein R 1 is 5-H, 5-halo, 5-methyl, 5-cyano or 5-trifluoromethyl; each of R10 and Rn is, independently, H or halo; A is -C (H) =; R2 and R3 are H; R 4 is H, methyl, phenyl-C 1 -C 2 alkyl, wherein said phenyl groups are mono- or di-substituted independently with H, halo, alkyl (C1-C4), (C1-C4) alkoxy, trifluoromethyl, hydroxy, amino or cyano and wherein said R4 groups are optionally additionally mono-substituted with halo; or R.1 is thien-2- or -3-yl-alkyl (C? -C2), pyrid-2-, -3- or -4-yl-alkyl (d-C2), thiazole-2-, - 4- or -5-yl-alkyl (C? -C2), imidazole-2-, -4- or -5-yl-alkyl (C? -C2), fur-2- or -3-yl-alkyl ( C? -C2), pyrrole-2- or -3-yl-(C1-C2) alkyl, oxazole-2-, -4- or -5-yl-alkyl (C? -C2), pyrazole-3 , -4- or -5-yl-alkyl (C? -C2), isoxazole-3-, -4- or -5-yl-alkyl (C? -C2), isothiazole-3-, -4- or - 5-yl-alkyl (C? -C2), pyridazin-3-, or -4-yl-alkyl (C? -C2), pyrimidin-2-, -4- or -5- or 6-yl-alkyl ( C? -C2), pyrazin-2-, or 3-yl-alkyl (d-C2) or 1, 3,5-triazin-2-yl-alkyl (C? -C2), wherein said R4 heterocycles above optionally mono- or di-substituted optionally and independently with halo, trifluoromethyl, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, amino or hydroxy and said mono- or di-substituents are attached to carbon; R5 is H; and R6 is C (O) NR8R9 or C (O) R12. Within the first preferred group of preferred compounds of formula IA there is a first group of especially preferred compounds wherein R4 is H, phenyl-alkyl (d-C2), thien-2- or -3-yl-alkyl (d-C2) ), fur-2-or -3-yl-alkyl (d-C2), wherein said R rings are mono- or di-substituted independently with H or fluorine; R6 is C (O) R12; and R-? 2 is morpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, thiazoiidin-3-yl, 1-oxothiazolidin-3-yl, 1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl, isoxazolidin-2-yl, isothiazolidin-2-yl, 1, 2- oxazetidin-2-yl, oxazolidin-3-yl, 1,3-dihydroisondol-2-ylo or azepan-1-yl, wherein said R 2-rings are optionally mono- or disubstituted independently with halo, alkyl (d-C5), (d-C5) alkoxy, hydroxy, amino, mono-N- or di-N, N-alkyl (C? -C5) amino, formyl, carboxy, carbamoyl, mono-N- or di- N, N-alkyl (C? -C5) carbamoyl, (C1-C5) alkoxycarbonyl, hydroxy (C1-C5) alkyl, amino-(C1-C4) alkyl, mono-N- or di-N, N- alkyl (C? -C4) amino-alkyl (C? -C4), oxo, hydroxyimino or alkoxyimino (d-C6), with the proviso that only the heterocycles R? 2 thiazolidin-3-yl, pyrrolidin-1-yl , piperidin-1-yl, piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl, isoxazolidi n-2-yl or oxazolidin-3-yl are optionally mono- or di-substituted with oxo, hydroxyimino or (C? -C6) imino alkoxy; and wherein said R-? 2 rings are optionally and additionally mono- or di-substituted independently with (C1-C5) alkyl. Within the above group of especially preferred compounds are the compounds [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H -indole-2-carboxylic acid, [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid 5-Chloro-1 H-indole-2-carboxylic acid [2 - (4S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide, [(1 S) -benzyl] -2- (5-Chloro-1 H -indole-2-carboxylic acid cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide, [2- (1, 5-Chloro-1 H-indole-2-carboxylic acid 1-dioxo-thiazolidin-3-yl) -2-oxo-ethyl] -amide, [2-oxo-2-thiazolidin-3-yl-ethyl) - 5-Chloro-1 H-indole-2-carboxylic acid amide, [(1 S) - (4-fluoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo- ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid, [(1 S) -benzyl-2 - ((3RS) -hydroxy-p -peridin-1-yl) -2-oxo -ethyl] -amino-5-chloro-1 H-indole-2-carboxylic acid, [2-oxo-2 - ((1-RS) -oxo-1-thiazolidin-3-yl) -ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid, [(1 S) - (2-fIuoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo-ethyl ] -5-chloro-1 H-indole-2-carboxylic acid amide, [(1 S) -benzyl-2 - ((3S, 4S) -dihydroxy-pyrroiidin-1-yl) -2-oxo-ethyl] 5-Chloro-1 H-indole-2-carboxylic acid amide, [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-ethyl] -amide of 5 -chloro-1 H-indole-2-carboxylic acid, [(1 S) -benzyl-2- (3-hydroxyimino-azetidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H -indole-2-carboxylic acid or [(1 S) -benzyl-2- (4-hydroxyimino-piperidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-acid carboxylic, Within the above group of especially preferred compounds, there is found a first group of particularly preferred compounds wherein R4 is H; and R-I2 is thiazolidin-3-yl, 1-oxo-thiazoiidin-3-yl, 1,1-dioxo-thiazolidin-3-yl or oxazolidin-3-yl, or said substituents R12 are optionally mono- or di- independently substituted with carboxy, (C Cs) alkoxycarbonyl, hydroxy-alkyl (dd), amino-alkyl (d-C3), mono-N- or di-N, N-alkyl (d-C3) amino-alkyl (dd) ) or R 2 is mono- or di-substituted pyrrolidin-1-yl, wherein said substituents are independently carboxy, alkoxy (dd) carbonyl, alkoxy (C1-C5), hydroxy, hydroxy-alkyl (dd), amino, amino-alkyl (dd), mono-No di-N, N-alkyl (CrC3) amino-alkyl (CrC3) or mono-N- or di -N, N-alkyl (CrC4) amino; and R12 rings are optional, additionally and independently disubstituted with alkyl (d-d). Preferred compounds within the immediately preceding group of particularly preferred compounds are the compounds in which a. Ri is 5-chloro; R 12 is cis-3,4-dihydroxy-pyrrolidin-1-yl; b. Ri is 5-chloro; R10 and Rn are H; and R12 is (3S, 4S) -dihydroxy-pyrrolidin-1-yl; c. Ri is 5-chloro; Rio and R11 are H; and R12 is 1,1-dioxo-thiazolidin-3-yl; d. Ri is 5-chloro; Rio and 11 are H; and R-I2 is thiazolidin-3-yl; and e. Ri is 5-chloro; Rio and Rn are H; and R 2 is 1-oxo-thiazolidin-3-yl. Within the above group of especially preferred compounds, there is a second group of particularly preferred compounds wherein R4 is phenylmethyl, thien-2- or -3-ylmethyl, wherein said R4 rings are optionally mono- or di-substituted with fluorine; and R-? 2 is thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl, 1,1-dioxo-thiazolidin-3-yl or oxazolidin-3-yl, or said R? 2 substituents are optionally mono- or di-substituted independently with carboxy, alkoxy (dd) carbonyl, hydroxy-alkyl (dd), amino-alkyl (dd), mono-N- or di-N, N-alkyl (Ci-d) amino-alkyl (dd) ) or R 2 is mono- or di-substituted azetidin-1-yl, mono- or di-substituted pyrrolidin-1-yl or mono- or di-substituted piperidin-1-yl, wherein said substituents are independently carboxy, alkoxy (dd) carbonyl, hydroxy-alkyl (dd), amino-alkyl (dd), mono-N-od-N, N-alkyl (Crd) amino-alkyl (dd), hydroxy, alkoxy (CrCs), amino, mono-N- or di-N, N-alkyl (CrC5) amino, oxo, hydroxyimino or alkoxy (dd) imino; and the R 2 rings are optionally and additionally mono- or di-substituted, independently, with alkyl (d-d). Within the immediately preceding group of particularly preferred compounds are the compounds in which a. Ri is 5-chloro; R 4 is 4-fluorobenzyl; R12 is 4-hydroxipiperdin-1 -lo; and the stereochemistry of carbon (a) is (S); b. R1 is 5-chloro; R4 is benzyl; R 12 is 3-hydroxypiperidin-1-yl; and the stereochemistry of carbon (a) is (S); c. R1 is 5-chloro; R is benzyl; R-I2 is cis-3,4-dihydroxy-pyrrolidin-1-yl; and the stereochemistry of carbon (a) is (S); Ri is 5-chloro; Rio and Rn are H; and R 4 is benzyl R 12 is 3-hydroxyimino-pyrrolidin-1-yl; and the stereochemistry of carbon (a) is (S); R1 is 5-chloro; R-io and R11 are H; R 4 is 2-fluorobenzyl; R 12 is 4-hydroxypiperidin-1-yl; and the stereochemistry of carbon (a) is (S); R1 is 5-chloro; R10 and R11 are H; and R 4 is benzyl; R 12 is (3S, 4S) -dihydroxy-pyrrolidin-1-yl; and the stereochemistry of carbon (a) is (S); R1 is 5-chloro; R10 and R11 are H; R4 is benzyl; R 2 is 3-hydroxy-azetidin-1-yl; and the stereochemistry of carbon (a) is (S); h. Ri is 5-chloro; R4 is benzyl; R 12 is 3-hydroxyimino-azetidin-1-yl; and the stereochemistry of carbon (a) is (S); and i. R1 is 5-chloro; R4 is benzyl; R-? 2 is 4-hydroxyimino-piperidin-1-yl; and the stereochemistry of carbon (a) is (S). A second group of especially preferred compounds within the first group of preferred compounds are those compounds wherein R4 is H, phenyl-alkyl (dd), thien-2- or -3-yl-alkyl (CrC2), fur-2 -o-3-yl-alkyl (dd) wherein said R4 rings are independently mono- or di-substituted with H or fluorine; R8 is H, alkyl (d-d), hydroxy or alkoxy (d-d); and Rg is H, cycloalkyl (dd), cycloalkyl (d-C6) -alkyl (dd), methylene-alkyl (dd), perfluorinated, pyridyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, piperidinyl, benzothiazolyl or thiochromanyl; or Rg is alkyl (dd) wherein said alkyl (dd) is optionally substituted by cycloalkenyl (C4-d), phenyl, thienyl, pyridyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, piperidinyl, morpholinyl, thiomorpholinyl, 1-oxothiomorpholinyl or 1,1-dioxothiomorpholinyl and wherein said alkyl (dd) or alkoxy (dd) is optional, additionally and independently mono- or di-substituted with halo, hydroxy, alkoxy (dd), amino, mono-N - od-N, N-alkyl (dd) amino, cyano, carboxy or alkoxy (dd) carbonyl; and wherein the Rg rings are optionally mono- or di-substituted independently on carbon, with halo, alkyl (dd), alkoxy (rC4), hydroxy, amino, mono-N- or di- N, N-alkyl (dd) amino , carbamoyl, alkoxy (dd) carbonyl or carbamoyl. Within the second immediately preceding group of especially preferred compounds are those compounds in which Ri is 5-chloro; R4 is benzyl; R8 is methyl; and R9 is 3- (dimethylamino) propyl; the stereochemistry of carbon (a) is (S); R1 is 5-chloro; R.o and R11 are H; R is benzyl; R8 is methyl; and Rg is 3-pyridyl; c. the stereochemistry of carbon (a) is (S); Ri is 5-chloro; R4 is benzyl; R8 is methyl; and Rg is 2-hydroxyethyl; and d. the stereochemistry of carbon (a) is (S); R1 is 5-chloro; R10 and R11 are H; R 4 is 4-fluorophenylmethyl; R8 is methyl; and Rg is 2-morpholinoethyl A third group of especially preferred compounds within the first group of preferred compounds are the compounds wherein R4 is H, phenyl-alkyl (dd), thien-2- or -3-yl-alkyl ( dd), fur-2-or -3-yl-alkyl (dd), wherein said R4 rings are mono- or disubstituted independently with H or fluorine; R6 is C (O) NR8R9; R8 is H, alkyl (d-d), hydroxy or alkoxy (d-d); and Rg is alkoxy (dd), wherein said alkoxy (dd) is optionally substituted with cycloalkenyl (dd), phenyl, thienyl, pyridyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, piperidinyl, morpholinyl, thiomorpholinyl, -oxothiomorpholinyl or 1,1-dioxothiomorpholinyl and wherein said alkyl (dd) or alkoxy (dd) is optional, additionally and independently mono- or di-substituted with halo, hydroxy, alkoxy (dd), amino, mono-N- or di-N, N-alkyl (dd) amino, cyano, carboxy or alkoxy (dd) carbonyl; and wherein the Rg rings are optionally mono- or disubstituted independently on carbon with halo, alkyl (dd), alkoxy (dd), hydroxy, amino, mono-N- or di- N, N-alkyl (dd) amino, carbamoyl, (C1-C5) alkoxycarbonyl or carbamoyl. Within the third group of especially preferred compounds immediately above are the compounds in which a. R1 is 5-chloro; R-io and R11 are H; R4 is benzyl; Rs is methyl; and Rg is 2-hydroxyethoxy; b. the stereochemistry of carbon (a) is (S); R1 is 5-chloro; R 4 is 4-fluorophenylmethyl; R8 is methyl; and Rg is methoxy; c. the stereochemistry of carbon (a) is (S); Ri is 5-chloro; R4 is benzyl; R8 is methyl; and R9 is methoxy; A second group of preferred compounds of formula IA are those compounds wherein R 1 is 5-halo, 5-methyl, 5-cyano or trifluoromethyl; each of R10 and Rn is, independently, H or halo; A is -C (H) =; R2 and R3 are H; R4 is H, phenyl-alkyl (CrC2), thien-2- or -3-yl alkyl (d-C2), fur-2- or -3-yl-alkyl (d-C2), wherein said rings are mono- or di-substituted independently with H or fluorine; R5 is H; and R6 is alkoxy (Crd) carbonyl. A third group of preferred compounds of formula IA are those compounds wherein R 1 is 5-halo, 5-methyl, 5-cyano or trifluoromethyl; Each of R-1 and R-11 is, independently, H or halo; A is -C (H) =; R2 and R3 are H; R 4 is H, methyl or phenyl-alkyl (CrC 2), wherein said phenyl groups are mono- or di-substituted independently with H, halo, alkyloid-d), alkoxy (dd), trifluoromethyl, hydroxy, amino or cyano, and wherein said phenyl groups are additionally mono- or di-substituted independently with H or halo; or R is thien-2- or -3-yl-alkyl (CrC2), pyrid-2-, -3- or -4-yl-alkyl (d-C2), thiazole-2-, -4- or -5 -alkyl (d-C2), imidazole-2-, -4- or -5-yl-alkyl (d-C2), fur-2- or -3-yl-a-alkyl (C C2), pyrrole -2- or -3-yl- (d-C2) alkyl, oxazoI-2-, -4- or -5-yl alkyl (CrC2), pyrazole-3-, -4- or -5-yl-alkyl ( CrC2), isoxazol-3-, -4- or -5-yl-alkyl (CrC2), isothiazol-3-, -4- or -5-yl-alkyla (d-C2), pyridazin-3- or -4-alkyl (CrC2), pyrimidin-2-, -4-, -5- or -6-yl-alkyl (CrC2), pyrazin-2- or -3-yl-alkyl (CrC2) or 1.3, 5-triazin-2-yl-alkyl (d-C2), wherein said above R4 heterocycles are optionally mono- or di-substituted independently with halo, trifluoromethyl, alkyl (CrC4), alkoxy ^ dd), amino or hydroxy, and said mono- or di-substituents are attached to carbon; R5 is H; and Re is carboxy. Within the third group of preferred compounds there is a first group of especially preferred compounds, wherein R10 and R11 are H; and R4 is H. Within the especially preferred group immediately above, there is a compound in which Ri is 5-chloro. Another aspect of this invention is a method of treating mammals that have insulin resistant conditions, which comprises administering to a mammal having an insulin resistant condition therapeutically effective amounts of a. a first compound, said first compound being an inhibitor of aldose reductase; and b. a second compound, said second compound being an inhibitor of glycogen phosphorylase. Preferred insulin-resistant conditions, taken individually or as a group, include diabetes, hyperinsulinemia, glucose intolerance, hypergiukaemia and / or hyperlipidemia after meals, type II diabetes, altered body composition, reduced lean mass bodily, obesity (especially visceral abdominal obesity), hypertension, dyslipidemia (for example, increased levels of free fatty acids, triglycerides, cholesterol VLDL and LDL cholesterol, and reduction of HDL cholesterol levels), atherosclerosis, ischemia of tissues and diseases cardiovascular, obesity, syndrome X (also called "metabolic syndrome"), pregnancy, infection disorders, uremia, hyperandrogenism, hypercortisolemia or other conditions of excess adrenocortical hormones, acromegaly, excess growth hormone or polycystic ovarian disease.

Especially preferred insulin-resistant conditions, taken individually or in groups, include dyslipidemia, tissue ischemia, obesity, polycystic ovarian disease, syndrome X, and hypertension. In particular, diabetes is especially preferred. A preferred aldose reductase inhibitor is 1-phthalazineacetic acid, 3,4-dihydro-4-oxo-3 - [[5-trifluoromethyl] -2-benzothiazolyl] methyl- or a pharmaceutically acceptable salt thereof. A preferred inhibitor of glycogen phosphorylase is 5-chloro-1 H- [(IS) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyrrolidin-1-yl) -3-oxopropyl] -amide. indole-2-carboxylic; [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1-yl) - 5-chloro-1 H-indole-2-carboxylic acid (2R) -hydroxy-3-oxopropy-amide; [(1 S) - ((R) -hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy-methoxy-methyl-carbamoyl) -methyl) -2-phenyl-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl] -2-phenyl-ethyl] -amide of 5-chloro-1 H- acid indole-2-carboxylic; [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) -benzyl-3 - ((cis) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Doro-1 H-indole-2-carboxylic acid [2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2- (1, 1-dioxo-thiazolidin-3-yl) -2-oxo-ethyl] -amide; [(1 S) - (4-fluoro-benzyl) -2- (4-hydroxy-p -peridin-1-l) -2-oxo-ethyl] -amide of 5-chloro-1 H- indole-2-carboxylic; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-2 - ((3RS) -hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2-oxo-2 - ((1-RS) -oxo-thiazolidin-3-yl) -ethyl] -amide; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-ethyl] -amide of the acid -chloro-1 H-indole-2-carboxylic acid; A particularly preferred mammal is a man or a woman. Another preferred aspect of this method is a process in which the first compound and the second compound are administered in a substantially simultaneous manner.

Another aspect of this invention is a synergistic pharmaceutical composition for achieving an insulin sensitizing effect in a mammal, comprising a. an amount of a first compound, said first compound being an aldose reductase inhibitor; and b. an amount of a second compound, said second compound being an inhibitor of glycogen phosphorylase. wherein the amount of the first compound alone and the amount of the second compound alone is insufficient to achieve the effect of insulin sensitization if administered individually and in which the combined effect of the amounts of the first and second compounds is greater than the sum of the effects of insulin sensitization that can be achieved with the individual amounts of the first and second compounds, and a pharmaceutically acceptable diluent or carrier. Another aspect of this invention is a team comprising: a. a therapeutically effective amount of an aldose reductase inhibitor and a pharmaceutically acceptable carrier in a first unit dosage form; b. a therapeutically effective amount of a glycogen phosphorylase inhibitor and a pharmaceutically acceptable carrier in a second unit dosage form, and c. a container means for containing said first and second dosage forms.

Yet another aspect of this invention is a synergistic process for achieving an insulin sensitizing effect in a mammal having insulin resistant conditions, which comprises administering to said mammal a. an amount of a first compound, said first compound being an inhibitor of aldose reductase, and b. an amount of second compound, said second compound being an inhibitor of glycogen phosphorylase in which the amount of the first compound alone and the amount of the second compound alone is insufficient to achieve said insulin sensitizing effect and in which the combined effect of the amounts of the first and second compounds is greater than the sum of the insulin sensitization effects that can be achieved with the individual amounts of the first and second compounds. Another aspect of this invention is directed to a method of reducing tissue damage resulting from or resulting from ischemia, which comprises administering to a mammal in need of such treatment a therapeutically effective amount of a. an inhibitor of reductose aldose, and b. an inhibitor of glycogen phosphorylase. Preferred ischemic tissues taken individually or in groups are those in which the ischemic tissues are cardiac, cerebral, hepatic, renal, pulmonary, intestinal, skeletal muscle, splenic, pancreatic, nervous, spinal cord, retinane tissues, the vasculature or the intestinal tissue. A particularly preferred ischemic tissue is cardiac tissue. Preferably, the combination of this invention is administered prophylactically. The ischemic lesion that can be treated in accordance with this invention can occur during an organ transplant. Preferably, the combination of this invention is administered before cardiac surgery. The term "insulin resistance conditions" refers to conditions (a syndrome or insulin resistance state) in which the sensitivity and / or responsiveness to insulin in organs, tissues or cells of a mammalian body is reduced in comparison with the normal state (or insulin sensitive). This resistance results in multiple abnormalities in the metabolism of glucose, proteins and lipids, imbalances of electrolytes and ions, and growth, for example, of organs, tissues and cells, which may manifest in one or more of the following disorders (but without limitation: hyperinsulinemia, glucose intolerance (IGT), hypergiucemia and / or hyperlipidemia after meals, type II diabetes, altered body composition, reduction of lean body mass, obesity (especially visceral abdominal obesity), hypertension, dyslipidemia (for example, increased levels of free fatty acids, triglycerides, VLDL cholesterol and LDL cholesterol, and reduction of HDL cholesterol levels), atherosclerosis, tissue ischemia and cardiovascular diseases (Kopelman and Albon, 1977; DeFronzo and Ferrannini, 1991; Reaven, 1991; Malmstrom, et al., 1997). As a result of the state of insulin resistance, a greater amount of insulin is required to achieve or achieve the same biological action of insulin in organs, tissues and cells, compared to the normal (or insulin-sensitive) state, which leads to to an increased amount in the pancreas to secrete more insulin (impaired hyperinsulinemia) and, in the most extreme circumstance, pancreatic failure and insulin insufficiencies that lead to a type I diabetes disorder. Insulin-resistant states, for example, include obesity, syndrome X (also called "metabolic syndrome"), pregnancy or infection conditions, uremia, hyperandrogenism, hypercortisolemia or other adrenocortical hormone excess disorders, acromegaly or excess growth hormone, polycystic ovaries, or may be associated with advanced age or specific ethnic groups (Kopelman and Albon, 1997). The term "insulin sensitization effect" refers to a state in which the tissues of patients are made to give a normal or better than normal biological response to a given amount of insulin. The term "reduction" is intended to include partial prevention or prevention, which, although greater than what could be obtained without taking drugs or taking a placebo, is less than 100%, in addition to substantially total prevention. The term "lesions resulting [...] from ischemia" as used herein, refers to conditions directly associated with reduced blood flow in the tissues, for example, due to a clot or blockage of the blood vessels. they deliver blood to the target tissue and which result, inter alia, in a reduction of oxygen transport to such tissue, defective tissue behavior, tissue dysfunction and neocrosis. Alternatively, when the blood flow or organ perfusion may be quantitatively adequate, the oxygen transport capacity of the blood or organ perfusion medium may be reduced, for example, in a hypoxic medium, such that the Oxygen supply to the tissue is reduced, and tissue malfunction, tissue dysfunction and tissue necrosis occur. The term "aldose reductase inhibitor" refers to compo that inhibit the bioconversion of glucose to sorbitol, catalyzed by the enzyme aldose reductase. The term glycogen phosphorylase inhibitor refers to any substance, agent or any combination of substances and / or agents that reduces, retards or eliminates the enzymatic action of glycogen phosphorylase. The currently known enzymatic action of glycogen phosphorylase is the degradation of glycogen by catalysis of the reversible reaction of a macromolecule of glycogen and inorganic phosphate to give glucose-1-phosphate and a glycogen macromolecule which is a glucosyl residue shorter than that of glycogen. original glycogen macromolecule (direction of advance of glycogenolysis). The term "treatment" or "treating", as used herein, includes preventive (for example prophylactic) and palliative treatment. By "pharmaceutically acceptable" is meant the carrier, diluent, excipients and / or salts which have to be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. The term "prodrug" refers to compo that are drug precursors which, after administration, release the drug in vivo by some chemical or physiological processes (e.g., a prodrug, after being brought to physiological pH or through an action enzyme becomes the desired drug). Exemplary pro-drugs, after cleavage, release the corresponding free acid. By "alkylene" is meant a saturated hydrocarbon (straight or branched chain) in which a hydrogen atom is removed from each of the terminal carbons. Examples of such groups are (assuming that the designated length encompasses the particular example), methylene, ethylene, propylene, butylene, pentylene, hexylene and heptylene. By "halo" is meant chlorine, bromine, iodine or fluorine. By "alkyl" is meant a saturated straight-chain hydrocarbon or a branched-chain saturated hydrocarbon. Examples of such alkyl groups (assuming that the designated length encompasses the particular example) methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tere-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, 1-methylbutyl, -methylbutyl, 3-methylbutyl, hexyl, isohexyl, heptyl and octyl. By "alkoxy" is meant a straight chain saturated alkyl or a branched chain saturated alkyl linked through an oxy. Examples of such alkoxy groups (assuming that the designated length encompasses the particular example) methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentoxy, isopentoxy, neopentoxy, tert-pentoxy, hexoxy, isohexoxy, heptoxy and octoxy . As used herein, the term mono-N- or di-N, N-alkyl (dd) ... refers to the alkyl radical (d-Cx) taken independently when it is di-N, N-alkyl (d) -Cx) ... (x refers to integers). It should be understood that if a carbocyclic or heterocyclic radical can bind or otherwise form bonds with a designated substrate, through different ring atoms without indicating a specific point of attachment, then all possible points are included, either through a carbon atom or, for example, a trivalent nitrogen atom. For example, the term "pyridyl" means, for example, 2-, 3- or 4-pyridyl, the term "thienyl" means, for example, 2- or 3-thienyl, and so on.

The term "pharmaceutically acceptable salt" refers to non-toxic anionic salts containing anions such as (but not limited to) chloride, bromide, iodide, sulfate, bisulfate, phosphate, acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate , gluconate, methanesulfonate and 4-toluene sulfonate. The term also refers to non-toxic cationic salts such as (but not limited to) sodium, potassium, calcium, magnesium, ammonium or protonated benzathine (NN-dibenzylethylenediamine), choline, ethanolamine, diethanolamine, ethylenediamine, meglamine (N-methyl-glucamine ), benetamine (N-benzylphenethylamine), piperazine or tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol). As used herein, the terms "reaction inert solvent" and "inert solvent" refer to a solvent or a mixture of solvents that do not interact with the starting materials, reagents, intermediates or products in a manner that affects adversely to the performance of the desired product. The negative or positive sign in parentheses used in this document in the nomenclature indicates the direction of the plane in which light polarized by the particular stereoisomer rotates. The chemist with ordinary skill will recognize that certain compounds of this invention will contain one or more atoms that may be in a particular stereochemical or geometric configuration, giving rise to stereoisomers and configurational isomers. All these isomers and mixtures thereof are included in this invention. Also included are hydrates and solvents of the compounds of this invention. DTT means dithiothreitol, DMSO means dimethyl sulfoxide, EDTA means ethylenediamine tetraacetic acid. From the specification and claims describing the invention, other features and advantages will become apparent.

DETAILED DESCRIPTION OF THE INVENTION In general, the compounds of this invention can be obtained by methods including methods known in the chemical arts, particularly in light of the description contained herein. Certain processes for the manufacture of the compounds of this invention are provided as further features of the invention and are illustrated by the following reaction schemes. In the experimental section, other procedures can be described. Any inhibitor of aldose reductase can be used as the compound (active agent) of this invention. The term "aldose reductase inhibitor" refers to compounds that inhibit the bioconversion of glucose to sorbitol, catalyzed by the enzyme aldose reductase. Such inhibition is easily determined by those skilled in the art in accordance with conventional assays (J. Malone, Diabetes, 29: 861-864, 1980. "Red Cell Sorbitol, an Indicator of Diabetic Control"). A variety of aldose reductase inhibitors are described and mentioned below, however, other aldose reductase inhibitors will be known to those skilled in the art. The disclosures of the United States patents presented below are incorporated herein by reference. In addition, the common chemical names USAN or other names are in parentheses when they are applicable, together with the reference to the appropriate patent literature describing the compound. The activity of an aldose reductase inhibitor in a tissue can be determined by assaying the amount of aldose reductase inhibitor that is necessary to reduce sorbitol in tissue (ie, by inhibiting the additional production of sorbitol after blockage of the aldose reductase) or to reduce fructose in the tissue (inhibiting the production of sorbitol after the blockade of aldose reductase and, consequently, the production of fructose). Although no limitation is desired by any particular theory or mechanism, it is believed that an inhibitor of aldose reductase, by inhibiting aldose reductase, prevents or reduces ischemic lesions as described below. Accordingly, examples of the aldose reductase inhibitors useful in the compositions and methods of this invention include: 1. 3- (4-Bromo-2-fluorobenzyl) -3,4-dihydro-4-oxo- 1-phthalazinoacetic acid (ponalrestat, US Pat. No. 4,251,528); 2. N [[(5-trifluoromethyl) -6-methoxy-1-naphthalenyl] thoxomethyl} -N-methylglycine (tolrestat, document E.U.A. 4,600,724); 3. 5 - [(Z, E) - ^ methyl cinnamidene] -4-oxo-2-thioxo-3-thiazolideneacetic acid (epalrestat, documents E.U.A. 4,464,382, E.U.A. 4,791,126, E.U.A. 4,831,045); 4.- 3- (4-Bromo-2-fluorobenzyl) -7-chloro-3,4-dihydro-2,4-dioxo-1 (2H) -quinazolinoacetic acid (zenarestat, US documents 4,734,419 and 4,883,800) ); 5.- 2R, 4R-6,7-dichloro-4-hydroxy-2-methylchroman-4-acetic acid (document E.U.A. 4,883,410); 6.- 2R, 4R-6,7-dichloro-6-fluoro-4-hydroxy-2-methyl-4-acetic acid (document U.S.A. 4,883,410); 7.- 3,4-Dihydro-2,8-diisopropyl-3-oxo-2H-1,4-benzoxacin-4-acetic acid (document E.U.A. 4,771,050); 8.- 3,4-Dihydro-3-oxo-4 - [(4,5,7-trifluoro-2-benzothiazolyl) methyl] -2H-1,4-benzothiazine-2-acetic acid (SPR-210, document US 5,252,572); 9. N- [3,5-dimethyl-4 - [(nitromethyl) sulfonyl] phenyl] -2-methyl-benzeneacetamide (ZD5522, documents U.S.A. 5,270,342 and U.S.A. 5,430,060); 10.- (S) -6-fluorospiro [chroman-4,4'-amidazolidin] -2,5'-dione (sorbinil, documents E.U.A. 4,130,714); 1 1 .- d-2-methyl-6-fluoro-spiro (chroman-4 ', 4'-imidazolidine) -2', 5'-dione (document E.U.A. 4,540,704); 12- 2-fluoro-spiro (9H-fluorene-9,4'-amidazoiidine) -2 ', 5'-dione (document U.S.A. 4,438,272); 13.- 2,7-di-fluoro-spiro (9H-fluorene-9,4'-imidazolidine) -2 ', 5'-dione (documents U.S. 4,436,745, U.S. 4,438,272); 14.- 2,7-di-fluoro-5-methoxy-spiro (9H-fluorene-9,4'-imidazolidine) -2 ', 5'-dione (US documents 4,436,745, US 4,438,272); 15.- 7-fluoro-spiro (5H-indenol [1,2-b] pyridine-5,3'-pyrrolidine) -2,5'-dione (documents E.U.A.4,436,745 and E.U.A. 4,438,272); 16.- d-cis-6'-chloro-2 ', 3'-dihydro-2'-methyl-spiro- (imidazolidin-4,4'-4'-H-pyran (2,3-b) pyridine) -2,5-dione (document US 4,980,357); 17. Spiro [imidazolidine-4,5 '(6H) -quinoline] 2,5-dione-3'-chloro-7', 8'-dihydro-7'-methylene- (5'-cis) ( U.S. Patent 5,066,659); 18. (2S, 4S) -6-fluoro-2 ', 5'-dioxo-spiro (chroman-4,4'-imidazolidine) -2-carboxamide (E.U.A. 5,447,946); and 19.- 2 - [(4-bromo-2-fluorophenyl) methyl] -6-fluorospiro [isoquinoline-4 (1 H), 3'-pyrrolidine] -1, 2 ', 3,5' (2H) - Tetrona (ARI-509, document US 5,037,831). Other aldose reductase inhibitors include compounds having the formula I or pharmaceutically acceptable salts thereof, wherein Z is O or S; R1 is hydroxy or a group capable of being removed in vivo to produce a compound of formula I wherein R1 is OH; and X and Y are the same or different and are selected from hydrogen, trifluoromethyl, fluorine and chlorine. A preferred subgroup within the above group of aldose reductase inhibitors include the compounds with the numbers 1, 2, 3, 4, 5, 6, 9, 10 and 17 and the following compounds of formula I: acid 3.4 -dihydro-3- (5-fluorobenzothiazol-2-ylmethyl) -4-oxophthalazin-1-yl-acetic acid [R1 = hydroxy; X = F; Y = H]; 21. 3- (5,7-difluorobenzothiazol-2-ylmethyl) -3,4-dihydro-4-oxophthalazine-1-acetic acid [R1 = hydroxy; X = Y = F]; 22. 3- (5-Chlorobenzothiazol-2-ylmethyl) -3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1 = hydroxy; X = Cl; Y = H]; 23. 3- (5,7-Dichlorobenzothiazol-2-ylmethyl) -3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1 = hydroxy; X = Y = CI]; 24. 3,4-Dihydro-4-oxo-3- (5-trifluoromethyl-benzoxazol-2-ylmethyl) -phthalazin-1-ylacetic acid [R1 = hydroxy; X = CF3; Y = H]; . 3,4-dihydro-3- (5-fluorobenzoxazol-2-ylmethyl) -4-oxophthalazin-1-ylacetic acid [R1 = hydroxy; X = F; Y = H]; 26. 3- (5,7-difluorobenzoxazol-2-ylmethyl) -3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1 = hydroxy; X = Y = F]; 27. 3- (5-Chlorobenzoxazol-2-ylmethyl) -3,4-dihydro-4-oxophthalazin-1-ylacetic acid [R1 = hydroxy; X = CI; Y = H]; 28.- 3- (5,7-dichlorobenzoxazol-2-ylmethyl) -3,4-dihydro-4-oxophthalazin-1-acetic acid [R1 = hydroxy; X = Y = CI]; and 29. zopolrestat, 1-phthalazinoacetic acid, 3,4-dihydro-4-oxo-3 - [[5- (trifluoromethyl) -2-benzothiazole] methyl] - [R1 = hydroxy; X = trifluoromethyl; Y = H]. In compounds 20-23 and 29, Z is S. In compounds 24-28, Z is O. Of the above subgroup, compounds 20-29 are more preferred, 29 being especially preferred. The aldose reductase inhibitor compounds of this invention are readily available or can be easily synthesized by those skilled in the art using standard methods. conventional organic synthesis, particularly in view of the descriptions of the specifications of the relevant patents. Any inhibitor of glycogen phosphorylase can be used as the second compound of this invention. The term glycogen phosphorylase inhibitor refers to any substance or agent, or to any combination of substances and / or agents, that reduces, retards or eliminates the enzymatic action of glycogen phosphorylase. The currently known enzymatic action of glycogen phosphorylase is the degradation of glycogen by the catalysis of the reversible reaction of a macromolecule of glycogen and inorganic phosphate to give glucose-1-phosphate and a glycogen molecule having a glucosyl residue less than the macromolecule of the original glycogen (direction of advance of glycogenolysis). Such actions are readily determined by those skilled in the art in accordance with conventional tests (for example, as described below). A variety of compounds are included in the following published PCT patent applications: publications of PCT patent applications WO 96/39384 and WO 96/39385. However, other inhibitors of glycogen phosphorylase will be known to those skilled in the art. In general, the compounds of formula I and IA can be obtained by methods including the methods known in the chemical arts, particularly in light of the description contained herein. In the following reaction schemes, certain processes for the manufacture of the compounds of formula I and IA are illustrated as additional features of the invention.

SCHEME I SCHEME II SCHEME SCHEME IV XX XXI XXII XXIV XXVI SCHEME V SCHEME VI 3. H2, Pd / C Exhaustive XLIV IIIA SCHEME Vil SCHEME VIII LXVI SCHEME IX LXI LXVII LXIX LXVIII SCHEME X According to reaction scheme I, the compounds of formula I wherein R., R.0 > R-n, A, R2, R3, R, R5, R6 and R7 are as defined above, they can be prepared with any of two general procedures. In the first method, the desired compound of formula I can be prepared by coupling the indolyl-2-carboxylic acid or the appropriate indoline-2-carboxylic acid of formula I with the appropriate amine of formula III (ie, acylation of the amine ). In the second method, the desired compound of formula I can be prepared by coupling the appropriate compound of formula IV (ie, a compound of formula I in which R6 is carboxy) with the appropriate alcohol or the amine or alcohol of formula R8RgNH or R 2 H, wherein R 8, R 9 and R 2 are as defined above (ie, acylation of the amine or alcohol). Typically, the compound of formula II is combined with the compound of formula III (or the compound of formula IV is combined with the appropriate amine (eg, R.2H or R2R9NH)) or with alcohol in the presence of a suitable coupling agent. . A suitable coupling agent is an agent that transforms a carboxylic acid into a reactive species that forms an amide or ester bond brings the reaction with an amine or an alcohol, respectively. The coupling agent can be a reagent which effects this condensation in a single container process when mixed together with the carboxylic acid and the amine or the alcohol. If the acid is to be condensed with an alcohol, it is preferable to use a large excess of the alcohol as the reaction solvent, adding or not adding 1.0 to 1.5 equivalents of dimethylaminopyridine. Exemplary coupling reagents are 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide-hydroxybenzotriazole hydrochloride (DEC / HBT), carbonyldiimidazole, dicyclohexylcarbodiimide / hydroxybenzotriazole (HBT), 2-ethoxy-1-ethoxycarbonyl-2, 2- dihydroquinoline (EEDQ), carbonyldiimidazole / HBT and diethylphosphorylnide. The coupling is carried out in an inert solvent, preferably in an aprotic solvent at a temperature from about -20 ° C to about 50 ° C, for about 1 to about 48 hours. Examples of solvents include acetonitrile, dichloromethane, dimethylformamide and chloroform. The coupling agent can also be an agent that converts the carboxylic acid into an activated intermediate, which is isolated and / or formed in a first step and allowed to react with the amine or alcohol in a second step. Examples of such coupling agents and activated intermediates are thionyl chloride or oxalyl chlorine to form the acid chloride, cyanuric fluoride to form an acid fluoride or an alkyl chloroformate, such as isobutyl chloroformate or isopropenyl (with a tertiary amine base) to form a mixed anhydride of the carboxylic acid. If the coupling agent is oxalyl chloride, it is advantageous to employ a small amount of dimethylformamide as a co-solvent with another solvent (such as dichloromethane) to catalyze the formation of the acid chloride. The use of these coupling agents and the proper selection of solvents and temperatures are known to those skilled in the art or can be readily determined in the literature. In Houben-Weyl, Vol XV, part II, E. Wunsch, Ed., G, Theime Verlag, 1974, Stuttgart, and M.

Bodansky, Principies of Peptide Synthesis, Springer-Verlag Berlin, 1984 and The Peptides. Analysis, Synthesis and Biology (ed. E. Gross and J. Meienhofer), vols 1-5 (Academic Press NY 1979-1983), describe these and other exemplary conditions useful for the coupling of carboxylic acids. The compounds of formula IV wherein R., R.0, R-n, A, R2, R3, R4.R5 and R7 are as defined above, can be prepared from the corresponding ester of formula V (ie, the compounds of formula I wherein R6 is (C .. C5) alkoxycarbonyl or benzyloxycarbonyl) by hydrolysis with an alkali! aqueous at a temperature from about -20 ° C to about 100 ° C, typically at about 20 ° C, for about 30 minutes to about 24 hours. Alternatively, compounds of formula IV are prepared by activating an indole carboxylic acid of formula II with a coupling agent (as described above), which gives an activated intermediate (such as an acid chloride, acid fluoride or mixed anhydride), which is then allowed to react with a compound of formula III in which R3, R4, R5 and R7 are as described above and R6 is carboxy, in a suitable solvent, in the presence of a suitable base. Suitable solvents include water or methanol or a mixture thereof, together with a cosolvent such as dichloromethane, tetrahydrofuran or dioxane. Suitable bases include sodium, potassium or lithium hydroxides, sodium or potassium bicarbonate, sodium or potassium carbonate or potassium carbonate together with tetrabutyl ammonium bromide (1 equivalent), in an amount sufficient to consume the acid liberated in the reaction (generally, enough to maintain the pH of the reaction above 8). The base may be added in an increasing manner together with the activated intermediate to effect appropriate control of the pH of the reaction. The reaction is generally carried out between -20 ° C and 50 ° C. The isolation procedures are those created by those skilled in the art to remove impurities, but typically consist of the elimination of cosolvents miscible with water by evaporation, the extraction of impurities at high pH with an organic solvent, acidification at a low pH ( 1-2) and filtration or extraction of the desired product with a suitable solvent such as ethyl acetate or dichloromethane. The compound of formula V can be prepared by coupling the appropriate compound of formula III, wherein Rβ is alkoxycarbonyl, and the appropriate compound of formula II, in a procedure analogous to that described above (e.g., procedure A). Alternatively, compounds of formula I containing sulfur atoms in the oxidation state of sulfoxide or sulfone can be prepared from the corresponding compounds of formula I having the sulfur atom in the non-oxidized form, by treatment with an agent suitable oxidant, such as with m-chloroperoxybenzoic acid, in dichloromethane, at a temperature from about 0 ° C to about 25 ° C, for about 1 to about 48 hours, using from about 1 to about 1.3 equivalents for the conversion into the of oxidation of the sulfoxide, and more about 2 equivalents for the conversion in the oxidation state of the sulfone. Alternatively, the compounds of formula I that are mono- or di-alkylated on aminoalkoxy R5, can be prepared from the corresponding compound of formula I wherein R5 is aminoalkoxy, by monoalkylation or dialkylation at the amine R5 to prepare the compound of desired formula I. Such mono- or di-alkylation can be carried out by treating the aminoalkoxy compound R5 with 1 equivalent of the appropriate carbonyl compound (for monoalkylation) or with more than 2 equivalent of the appropriate carbonyl compound (for dialkylation) and a suitable reducing agent in a solvent suitable. Suitable reducing conditions include sodium cyanoborohydride sodium borohydride in methanol or ethanol, or hydrogen / hydrogenation catalyst (such as palladium on carbon), in a polar solvent such as water, methanol or ethanol at about 0 ° C-60 ° C, for 1 to 48 hours. Alternatively, the compounds of formula I, wherein R5 is alkanoyloxy (RCOO-), are prepared by O-acylation of the appropriate compound of formula I with an appropriate acid chloride or other derivative of activated acid in the presence, if necessary , of a suitable base (eg, a tertiary amine base such as trialkylamine or pyridine), preferably in an aprotic solvent such as tetrahydrofuran or dichloromethane, at a temperature of from about 0 ° C to about 50 ° C, for about 0.5 to approximately 48 hours.

Alternatively, compounds of formula I in which R5 and R7 are taken together to be oxo, are prepared by oxidation of the corresponding compound of formula I, for example, wherein R5 is hydroxy and R is H, with an oxidizing agent suitable. Examples of the oxidizing agents include the "Dess-Martin reagent in dichloromethane, a carbodiimide and dimethyl sulfoxide and an acid catalyst (Pfitzner-Moffatt conditions or modifications thereof, such as by using a water-soluble carbodiimide) or reactions of the type from Swern (eg oxalyl chloride / DMSO / triethylamine) Compounds of formula I having other functionality sensitive to oxidation can take advantage of the appropriate protection and deprotection of such functionality, for example, in reaction scheme I, certain compounds of formula I contain a primary amine, a secondary amine or a carboxylic acid functionality in the part of the molecule defined by R5 or Rβ, which may interfere with the desired coupling reaction of reaction scheme I if the intermediate of formula III or the amine R.sub.2 H or RsRgNH is left unprotected.Therefore, the primary or secondary amine functionality can be protected, It is present in the radicals R5 or Rβ of the intermediate of formula III or of the amine (R8RgNH or R-? 2H), with a suitable protecting group, during the coupling reaction of reaction scheme I. The product of such coupling is a compound of formula I containing the protecting group. This protecting group is removed at a later stage to provide the compound of formula I. Suitable protecting groups for the protection of amine or carboxylic acid include the protecting groups commonly used in the synthesis of peptides (such as Nt-butoxycarbonyl, N- carbobenzyloxy and 9-fluorenylmethyleneoxycarbonyl for amines and lower alkyl or benzyl esters for carboxylic acids) that are not chemically reactive under the coupling conditions described above (and immediately preceding the examples herein as process A) and can be removed without the chemical alteration of another functionality of the compound of formula I. The indole-2-carboxylic acids and the starting indole-2-carboxylic acids used in the reaction scheme I, when they can not be purchased on the market or are not known in the prior art (such technique is widely published), can be acquired by procedures conventional synthetics. For example, according to reaction scheme II, the indole ester of formula VII can be prepared from the compound of formula VI (wherein Q is selected to achieve the desired A, as defined above), by a Fischer indole synthesis (see The Fischer Nature Svnthesis Robinson, B. (Willwy, New York, 1982)) followed by saponification of the resulting indole ester of formula VII to produce the corresponding acid of formula VIII. The starting aryl hydrazone can be prepared by condensation of a readily available hydrazine with the appropriate carbonyl derivative or by the Japp-Klingeman reaction (see Orqanic Reactions, Phillips, R. R., 1959, , 143). Alternatively, the indole-2-carboxylic acid of formula VI HA can be prepared by condensation of an ortho-methyl-nitro compound of formula IX with an oxalate ester to produce the indole ester of formula X, followed by reduction of the nitro group and subsequent hydrolysis. This three-step procedure is known as the indole synthesis of Reissert (Reissert, Chemische Berichte 1897, 30, 1030. The conditions for carrying out this sequence and references thereto are described in the literature (Kermack et al., J. Chem. Soc. 1921, 1 19, 1602; Cannon et al., J. Med. Chem. 1981, 24, 238; Julián, et al in Heterocyclic Compounds, Vol 3 (Willey, New York, NY, 1962, R.C. Elderfield, ed.) P 18). An example of the specific embodiment of this sequence is shown in examples 10A-10B of this document. Also, 3-halo-5-chloro-1 H-indole-2-carboxylic acids can be prepared by halogenation of 5-chloro-1 H-indole-2-carboxylic acids. As an alternative, (to reaction scheme II) the substituted indolines of formula XIV can be prepared by reduction of the corresponding indoles of formula XV with a reducing agent, such as magnesium in methanol, at a temperature of from about 25 ° C to about 65 ° C, for about 1 to about 48 hours (reaction scheme III).

The indolincarboxylic acids of formula XVI are prepared by saponification of the corresponding ester of formula XVII (reaction scheme III). The compound of formula XVII are prepared by reduction of the corresponding indole ester of formula VII with a reducing agent, such as magnesium in methanol, as described for the conversion of the compound of formula XV to the compound of formula XIV above. The following paragraphs describe how to prepare the various amines that are used in the previous reaction schemes. According to the reaction scheme IV, the compounds of formula XXII (the amines of formula III of reaction scheme I in which R5 is OH, R7 is H and R6 is an ester) or the compounds of formula XXVI (R6 is C (0) NR8R9 or C (0) R.2) are prepared starting from an N-protected aldehyde (designated PT) of formula XX. The aldehyde of formula XX or the sodium bisulfite adduct of an aldehyde of formula XX is treated with potassium or sodium cyanide in an aqueous solution, with a cosolvent such as dioxane or ethyl acetate, at a temperature of about 0 ° C to about 50 ° C, to provide a cyanohydrin of formula XXI. The cyanohydrin of formula XXI is treated with an alcohol (for example, alkanol (dd), such as methanol) and a strong acid catalyst, such as hydrogen chloride, at a temperature of from about 0 ° C to about 50 ° C, followed by by the addition of water, if necessary. The protecting group (PT) is then removed, if still present, by a suitable deprotection procedure, yielding a compound of formula XXII. For example, if the PT N-protecting group of formula XX is tert-butoxycarbonyl (t-Boc), the compound of formula XXIII is formed directly from the compound of formula XXI and the addition of water is not necessary. The compound of formula XXII can be protected in nitrogen with a suitable protecting group to form a compound of formula XXIII, followed by hydrolysis of the ester with an alkali accusative at a temperature from about 0 ° C to about 50 ° C in an inert solvent the reaction, resulting in the corresponding hydroxy acid of formula XXIV. The compound of formula XXIV is coupled (in a procedure analogous to the coupling process described in reaction scheme I) with an appropriate amine R8RgNH or HR.2 to form a compound of formula XXV, which is then deprotected resulting in the compound of formula XXVI (ie, the compound of formula III in which R5 is OH, R7 is H and R6 is C (O) R.2 or C (0) NR8Rg). An example of the conversion of a cyanohydrin of formula XXI to the corresponding methyl ester of formula XXII with removal of the t-Boc protecting group is provided in PCT publication WO / 9325574, example 1a. Other examples can be found in which the cyanohydrin is converted to the lower alkyl esters of formula XXIII, in the patent of E.U.A. No. 4,814,342 and in EPO publication 0438233. Certain compounds of formula I are stereoisomeric by virtue of the stereochemical configuration in the carbons marked as a and b. A person skilled in the art can prepare the intermediates of formula XXII and XXVI with the desired stereochemistry, according to the reaction scheme IV. For example, the aldehyde of formula XX can be purchased in any enantiomeric form (stereochemistry in a) by literature procedures indicated below (see reaction scheme V): The cyanohydrin of formula XXI can be prepared from the compound of formula XX by treatment with sodium or potassium cyanide, as described above, while maintaining stereochemistry at carbon a, resulting in a mixture of stereoisomers on carbon b. The specialist chemist can employ crystallization in this phase to separate isomers or purify an isomer. For example, the preparation of the compound of formula XXI wherein PT is Boc, R3 is H and R4 is benzyl, and the stereochemistry of carbons a and b is (S) and (R) respectively, employing this route together with purification by recrystallization, is described in Biochemistry 1992, 31, 8125-8141. Alternatively, the separation of isomers can be performed by chromatography or recrystallization techniques, after conversion of a compound of formula XXI (mixture of isomers) into a compound of formula XXII, XXIII, XXIV, XXV, XXVI, V, IV, I, by the procedures and / or sequences described in this document. The intermediates of formula XXI of a specific stereochemistry in the carbons a and b convert the intermediates of formula XXII with the retention of this stereochemistry, by treatment with an alcohol and a strong acid catalyst, followed by the addition of water, if necessary, as It has been described above.

Alternatively, the desired isomer of the compound of formula XXI can also be obtained by derivatization of the intermediate of formula XXI and chromatographic separation of the diastereomeric derivatives (for example, with trimethylsilyl chloride (TMS) or t-butyldimethylsilyl chloride (TBDMS) to give O-TMS or O-TBDMS derivatives). A silyl derivative of an intermediate of formula XXI having a single stereoisomeric form in carbons a and b is converted, with retention of stereochemistry, into an intermediate of formula XXII (if the silyl group has not been removed in this step, it is subsequently removed by an appropriate procedure, such as treatment with tetrabutylamine fluoride in tetrahydrofuran), by the procedure described above for the conversion of the compound of formula XXI into the compound of Formula XXII. According to reaction scheme V, aldehydes of formula XX (starting materials for reaction scheme IV) are prepared from the corresponding amino acids of formula XXX. The amino acid of formula XXX is protected at the nitrogen with a protecting group (PT) (such as Boc). The protected compound is esterified with an alcohol and converted to an ester, preferably the methyl or ethyl ester of the compound of formula XXXI. This can be done by treating the compound of formula XXX with methyl or ethyl iodide in the presence of an appropriate base (for example, K2CO3) in a polar solvent such as dimethylformamide. The compound of formula XXXI is reduced, for example, with diisobutylaluminum hydride in hexane, toluene or a mixture thereof, at a temperature from about -78 ° C to about -50 ° C, followed by inactivation with methanol at -78. ° C, as described in J. Med, Chem., 1985, 28, 1779-1790, to form the aldehyde of formula XX. Alternatively, (not shown in reaction scheme V), analogous N-methoxymethylamides are formed corresponding to the compound of formula XXXI, wherein the ester alcohol substituent is replaced by N (OMe) Me, from a compound of formula XXX, NO-dimethylhydroxylamine and a suitable coupling agent (for example, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (DEC) .The resulting compound is reduced, for example, with a lithium aluminum hydride in a solvent inert to the reaction, such as ether or tetrahydrofuran, at a temperature from about 0 ° C to about 25 ° C, to form the aldehyde of formula XX.This two step process is general for the conversion of α-amino acids N- protected in aldehydes of formula XX (Fehrentz and Castro, Synthesis 1983, 676-678) Alternatively, the aldehydes of formula XX can be prepared by oxidation of protected aminoalcohols of formula XXXII, for example, with pyridine-S03 , at a temperature of about -10 ° C to about 40 ° C, in a solvent inert to the reaction, preferably dimethylsulfoxide. Protected aminoalcohols of formula XXXII, if they can not be purchased on the market, can be prepared by protecting aminoalcohols of formula XXXII. The aminoalcohols of formula XXXII are prepared by reduction of amino acids of formula XXX. This reduction is carried out by treating compounds of formula XXX with lithium aluminum hydride according to the method described by Dickman et al., Organic Synthesis; Wiley; New York, 1990; Collect. Vol Vil, p 530, or with sulfuric acid-sodium borohydride by the procedure of Abiko and Masamune, Tetrahedron Lett. 1992 333, 5517-5518, or with sodium borohydride-iodine according to the method of McKennon and Meyers, J. Org. Chem. 1993, 58, 3568-3571, who also reviewed other suitable methods for converting the amino acids of formula XXX into aminoalcohols of formula XXXII. According to reaction scheme VI, the compounds of formula XXX used in the reaction scheme V can be prepared as indicated below. The amino acids of formula XLl can be prepared by N-alkylation of the protected amino acids (PT) of formula XL by treatment with an appropriate base and alkylating agent. The specific procedures for this alkylation are described by Benoiton, Can. J. Chem. 1977, 55, 906-910, and Hansen, J. Org. Chem. 1985, 50 945-950. For example, when R3 is methyl, sodium hydride and methyl iodide are used in tetrahydrofuran. Deprotection of the compounds of formula XLl produces the desired compound of formula XXX. Alternatively, an amino acid of formula XLl I can be N-alkylated by a three-step sequence involving reductive benzylation (such as with benzaldehyde, Pd / C catalyzed hydrogenation) to give the mono-N-benzyl derivative, and reductive amination with the appropriate acyl compound (for example, with formaldehyde and sodium cyanoborohydride, to introduce R 3 as methyl), to give the amino acid N-benzyl, N-R 3 -substituted. The N-benzyl protecting group is conveniently removed (for example, by hydrogenation with an appropriate catalyst) to produce the compound of formula XXX. The specific conditions for this three-step alkylation process are described by Reinhold et al., J. Med. Chem., 1968, 11, 258-260. The immediately preceding preparation can also be used to introduce a radical R3 into the intermediate of formula XLIV, to form the intermediate of formula XLV (which is an intermediate of formula III in which R7 is OH). The immediately preceding preparation can also be used to introduce a radical R3 into an intermediate of formula Illa (which is an intermediate of formula III) in which R3 is H). The amino acids used in the schemes of this document (for example, XL, XLII) if they can not be purchased commercially or are presented in the literature, can be prepared by a variety of methods known to those skilled in the art. For example, the Strecker synthesis or variations thereof can be used. Accordingly, an aldehyde (R-tCHO), sodium or potassium cyanide and ammonium chloride react to form the corresponding aminonitrile. The aminonitrile is hydrolyzed with a mineral acid to form the desired amino acid of formula XLI I R4C (NH2) COOH. Alternatively, the Bucherer-Berg process in which a hydantoin is formed can be used by heating an aldehyde (R4CHO) with ammonium carbonate and potassium cyanide, followed by hydrolysis (for example, with barium hydroxide in reflux dioxane) with an acid or a base to form the desired amino acid of formula XLII R4C (NH2) COOH. Other procedures for the synthesis of alpha-amino acids which will enable a person skilled in the art to prepare the desired intermediate of formula XLI I R4C (NH2) COOH necessary for the synthesis of compounds of formula I are also presented in the literature. from Duthaler (Tetrahedron 1994, 50, 1539-1650) or from Williams (RM Williams, Synthesis of optically active amino acids, Pergamon: Oxford, United Kingdom 1989), suitable methods are found for the synthesis or resolution of compounds of formula XLl I A specific procedure for the synthesis of an intermediate of formula XLII in any enantiomeric form of the corresponding intermediate RX (X = CI, Br, or I), is the procedure of Pirrung and Krishnamurthy (J. Org. Chem. 1993, 58, 957-958), or the procedure of O'Donnell, et al. (J. Am. Chem. Soc. 1989, 11, 2353-2355). The required R4X intermediates are readily prepared by many procedures familiar to chemists skilled in the art. For example, compounds in which R4X is ArCH2X can be prepared by radical hologenation of the ArCH3 compound or by Ar-H arene formulation and conversion of the alcohol to the bromide. Another specific procedure for the synthesis of the intermediates of formula XLll in any enantiomeric form is that of Corey and Link (J.

Am. Chem. Soc. 1992, 114, 1906-1908). Thus, an intermediate of formula R4COCCI3 is reduced enantiospecifically to the intermediate R4CH (OH) CCI3, which is converted by treatment with azide and base to an intermediate R4CH (NH3) COOH, which is reduced by catalytic hydrogenation to the desired compound of formula XLll. The required trichloromethyl ketone R COCCI3 is obtained by reaction of the aldehyde R4CHO with the trichloromethane anion, followed by oxidation (Gallina and Giordano, Synthesis 1989, 466-468). The intermediate amines of formula III (used in reaction scheme I) in which R5 and R7 are H, can be prepared according to reaction scheme VII. An amino acid of formula L (suitably protected (PT)) is activated by the conversion to the acid chloride, fluoride or mixed anhydride (for example, with isobutyl chloroformate and triethylamine in an inert solvent, such as tetrahydrofuran or dioxane, to a temperature from about -0 ° C to about -40 ° C) and the activated intermediate is treated with diazamethane to give the diazoketone of formula Ll. The diazoketone of formula Ll is treated with an alcohol (ROH) (for example, alkanol (d-d), such as methanol) and a suitable catalyst such as heat, silver oxide, to prepare the ester of formula LN. The ester of formula LN is deprotected to form the compound of formula IIIA (by rearrangement of Wolff). Alternatively, the ester of formula Lll is hydrolysed, for example, with alkali and coupled with the appropriate amine R- | 2H or HNR8Rg, to prepare the compound of formula IIIB as previously described.

According to reaction scheme VIII, intermediate amines of formula III, wherein R5 is an oxygen-bonded substituent (eg, alkoxy) (used in reaction scheme I) can be prepared as indicated below. The compound of formula XLl is alkylated by oxygen, by treatment with an appropriate alkylating agent (for example, alkyl iodide, alkyl bromide, alkyl chloride or alkyl tosylate) and sufficient base to form the alkoxide (sodium hydride or potassium), in a suitable polar aprotic solvent (eg, dimethylformamide or tetrahydrofuran), at a temperature from about 0 ° C to about 150 ° C, resulting in a compound of formula LXII. The compound of formula LXII is deprotected to produce the desired amine intermediate. The intermediate amines of formula III in which R5 is alkoxycarbonylalkoxy (d-d) (used in reaction scheme I), can be prepared as indicated below. The compound of formula LXl is alkylated with a haloalkanoate ester to form a compound of formula LXIII, which is then deprotected to form the desired amine. The corresponding acid can be prepared by hydrolysis of the ester using an aqueous alkali in an appropriate solvent. Amines of formula III in which Rβ contains an ester and R 5 contain a carboxy can be prepared from the amines of formula LX 11 (as prepared above in this paragraph), wherein R 5 contains the carboxylic acid functionality protected as t-butyl ester, by treatment with an anhydrous acid to provide the corresponding acid in R5 without hydrolyzing the ester in the R6 position. Compounds of formula LXVI (intermediate amines of formula III in which R5 is protected aminoalkoxy) can be prepared from the compound of formula LXI. The compound of formula LXl is alkylated with a haloalkane-nitrile to form the compound of formula XLIV. The compound of formula XLIV is reduced to the primary amine by treatment with hydrogen and an appropriate catalyst (for example, rhodium on carbon) in the presence of ammonia in a protic, preferably polar solvent, such as water, methanol or ethanol, to give the primary amine of formula LXV. The compound of formula LXV is protected in nitrogen with a protecting group (Pt?), Which is orthogonal with respect to the other protective group (Pt), followed by deprotection of the protective group (Pt) to produce the desired compound of formula III. The protected compound of formula III is coupled with the appropriate compound of formula II and the resulting protected compound of formula I is deprotected. Compounds of formula LXIII and LXIV wherein n is two, preferably are prepared by treatment of the compound of formula LXl with an excess of acrylate or acrylonitrile ester, respectively, in the presence of a suitable base, such as potassium or sodium hydroxide, in a suitable solvent, preferably a polar protic solvent. According to reaction scheme IX, the compounds of formula XLXVII and of formula LXIX (compounds of formula III wherein R5 is F or R5 and R7 are both F), can be prepared from the compound of formula LXI. The compound of formula LX1 is treated with a suitable fluorinating agent, such as diethylaminosulfur trifluoride in a reaction-inert solvent, such as an aprotic solvent, preferably dichloromethane, to form the compound of formula LXVII. The compound of formula LXVII is conveniently deprotected. The compound of formula LX1 is oxidized to give the compound of formula LXVIII, using the conditions described above for the preparation of compounds of formula I in which R5 and R7 together form oxo. The compounds of formula LXVIII are defluorinated under suitable conditions (for example, diethylaminosulfur trifluoride in dichloromethane). According to reaction scheme X, the compound of formula LXXIII or the compound of formula LXIV, wherein R7 is alkyl (ie, the compound of formula III wherein R7 is alkyl), are prepared from the compound of formula LXX (see also reaction scheme V for the analogous preparation of amine). The compound of formula LXX is treated with an organometallic reagent R7M and the resulting secondary alcohol is oxidized as in the immediately preceding paragraph to form the compound of formula LXXI. The compound of formula LXXI is converted, by the cyanohydrin of formula LXXII, to the compound of formula LXXIII, using the same conditions as those used to convert the compound of formula XXI to the compound of formula XXII in reaction scheme IV .

Alternatively, the compound of formula LXXN is converted to the compound of formula LXIV, as described for the conversion of the cyano intermediate to the amide, in the reaction scheme V. A compound of the formula R8NH2 or R9NH2 is monoalkylated with a carbonyl compound corresponding to R8 or Rg, respectively, under appropriate reductive amination conditions, to give an amine of formula R8RgNH. To avoid dialkylation, it may be preferable to protect the amines (R8NH2 or R9NH2) with a suitable protecting group PT, to give Rβ (Pt) NH or Rg (Pt) NH, for example, by reaction with benzaldehyde and a reducing agent. The protected amines are monoalkylated with a carbonyl compound corresponding to R9 or R8, respectively, under suitable reductive amination conditions, to give R8RgN (Pt). The protecting group (Pt) is removed (for example, by exhaustive catalytic hydrogenation when Pt is benzyl) to give a compound of the formula R8RgNH. A person skilled in the art can obtain the appropriate reductive amination conditions in the literature. These conditions include those presented by Borch et al. (J. Am. Chem. Soc. 1971, 2897-2904) and those indicated by Emerson (Organic Reactions, Willey: New York, 1948 (14), 174), Hutchins et al. (Org. Prep. Int.Procedures 1979 (11), 20, and Lane et al. (Synthesis, 1975, 135) The conditions of reductive amination that favor N-monoalkylation, include those presented by Morales et al. (Synthetic Communications 1984, 1213-1220) and Verardo et al. (Synthesis 1992, 121-125) The amines R8NH2 or RgNH2 can also be monoalkylated with RgX or R8X, respectively, in which X is chloride, bromide, tosylate or mesylate. Alternatively, an intermediate of the formula R8 (PT) NH or Rg (Pt) NH can be alkylated with RgX or R8X and the protecting group can be removed to give a compound of the formula R8RgNH. Additional processes can be used to prepare amines of the formula R8R9NH, wherein R8-NH or R9-NH are linked by an oxygen-nitrogen bond Thus, a readily available compound of the formula alkoxy (dd) carbonyl-NHOH or NH2CONHOH is dialkylated on nitrogen and oxygen by treatment with a base and a excess of suitable alkylating agent (RX), to give the c or alkoxy (d-d) carboniI-N (R) OR which is then hydrolyzed to give a compound of the formula R8RgNH (where R8 = Rg = R). Suitable conditions, the base and the alkylating agent include those described by Goel and Krolls (Org. Prep. Proced. Int. 1987, 19, 75-78) and Major and Fleck (J. Am. Chem. Soc. 1928, 50 , 1479). Alternatively, an amine of formula NH2CONH (OH) can be alkylated sequentially, first in oxygen to give NH2CONH (OR ') and then in nitrogen to give NH2CON (R ") (OR'), by successive treatment with the alkylating agents R ' X and R "X, respectively, in the presence of a suitable base. The base and suitable alkylating agents include those described by Kreutzkamp and Messinger (Chem. Ber. 100, 3463-3465 (1967) and Danen et al (J. Am. Chem. Soc. 1973, 95, 5716-5724). Hydrolysis of these alkylated hydroxyurea derivatives yields the amines RONH2 and R'ONHR ", which correspond to certain amines of the formula R8RgNH, The chemists skilled in the art can adapt the procedures described in this paragraph to other alkylating agents R, R 'and R "-X, to prepare other amines of the formula R8RgNH, in which R8-N or Rg-N are linked by oxygen-nitrogen bonds, One et al (SynLett 1991, 559-560) describes the addition catalyzed by BF3 of a reagent organometallic R-Li to an O-alkyl oxime of the formula R'CH = N-OR ", to give compounds of formula R'RCH-NH (OR") This route can also be used to give compounds of the formula R8R9NH, in the that one of R 8 -NH or Rg-NH are linked by oxygen-nitrogen bonds The prodrugs of this invention in those which a carboxyl group in a carboxylic acid of formula I is replaced by an ester, can be prepared by combining the carboxylic acid with the appropriate alkyl halide in the presence of a base, such as potassium carbonate, in an inert solvent, such as dimethylformamide, at a temperature of about 0 ° C to 100 ° C, for about 1 to about 24 hours. Alternatively, the acid is combined with the appropriate alcohol as solvent in the presence of a catalytic amount of acid, such as concentrated sulfuric acid, at a temperature of about 20 ° C to 120 ° C, preferably at reflux, for about 1 hour at approximately 24 hours. Another method is the reaction of the acid with a stoichiometric amount of the alcohol in the presence of a catalytic amount of acid in an inert solvent, such as tetrahydrofuran, with the co-elimination of the water that is produced by physical means (eg, Dean-purification). Stark) or chemicals (for example, molecular sieves).

The prodrugs of this invention in which an alcohol function has been transformed into an ether, can be prepared by combining the alcohol with the appropriate alkyl bromide or iodide, in the presence of a base such as potassium carbonate, in an inert solvent such as dimethylformamide, at a temperature of about 0 ° C to 100 ° C, for about 1 to about 24 hours. The alkanoylaminomethyl ethers can be obtained by reaction of the alcohol with a bis- (alkanoylamino) methane, in the presence of a catalytic amount of acid, in an inert solvent such as tetrahydrofuran, according to a procedure described in US 4,997,984. Alternatively, these compounds can be prepared by the methods described by Hoffman et al. in J. Org. Chem, 1994, 59, 3530. The dialkyl phosphate esters can be prepared by reacting the alcohol with a dialkyl chlorophosphate, in the presence of a base, in an inert solvent such as tetrahydrofuran. The dihydrogen phosphates can be prepared by reaction of the alcohol with a diaryl chlorophosphate or dibenzyl, as described above, followed by hydrolysis or hydrogenation in the presence of a noble metal catalyst, respectively. The glycosides are prepared by reaction of the alcohol and a carbohydrate in an inert solvent, such as toluene, in the presence of acid. Typically, the water formed in the reaction is removed as it is formed, as described above. An alternative procedure is the reaction of the alcohol with a suitably protected glycosyl halide in the presence of a base followed by deprotection. The N- (1-hydroxyalkyl) amides, the N- (1-hydroxy-1- (alkoxycarbonyl) methyl) amides or the compounds in which R2 has been replaced by C (OH) C (O) OY, can be prepared by the reaction of the parent amide or indole with the appropriate aldehyde, under neutral or basic conditions (for example, sodium ethoxide in ethanol) at temperatures between 25 and 70 ° C. The N-alkoxymethyl or N-1- (alkoxy) alkyl groups can be obtained by reacting the N-unsolyl indole with the necessary alkyl halide, in the presence of a base, in an inert solvent. 1- (N, N-dialkylaminomethyl) indole, 1- (1 - (N, N-dialkylamino) ethyl) nol and N, N-dialkylaminomethyl amides (e.g. R3 = CH2N (CH3) 2) can be prepared by the reaction of the parent NH compound with the appropriate aldehyde and amine, in an alcohol solvent at 25-70 ° C. The clinical prodrugs (e.g., the prodrugs of this invention wherein R2 and R3 are a common carbon) can be prepared by reaction of the parent compound (drug) with an aldehyde or ketone or its dimethylacetal, in an inert solvent, in the presence of a catalytic amount of acid, with the joint elimination of water and methanol. Alternatively, these compounds can be prepared by reaction of the aminoalcohol or the hydroxyamide with a gem-dibromo alkane, in the presence of a base (eg, potassium carbonate) in an inert solvent (eg, dimethylformamide).

The compounds of formula IA can be prepared as described below. The numbers of the schemes and the numbers of the formulas mentioned after this point of the text refer to the numbers of the schemes and to the numbers of the formulas that appear after this point of the text (that is, they should not be confused with the previous discussion).

SCHEME XI SCHEME XII SCHEME XIII SCHEME XIV SCHEME XV est SCHEME XVI appropriate 3. H2, Pd / C Exhaustive Illa According to reaction scheme XI, the compounds of formula IA, wherein R ^ R10, Rn, A, R2, R3, R4, R5 and R6 are as defined above, can be prepared by either of two general procedures . In the first method, the desired compound of formula IA can be prepared by coupling indole-2-carboxylic acid, indoline-2-carboxylic acid or benzimidazole-2-carboxylic acid, of formula II, with the appropriate amine of formula III (ie, acylation of the amine). In the second method, the desired compound of formula IA can be prepared by coupling the appropriate compound of formula IV (ie, a compound of formula IA in which R6 is carboxy), with the appropriate alcohol or the amine of formula R8RgNH or R 2H, wherein R8, R9 and R12 are as defined above (ie, acylation of the amine or alcohol). The first method (coupling of compounds of formula II with compounds of formula III) is typically preferred when R4 is not H and R5 is H. Typically, the compound of formula II is combined with the compound of formula III (or the compound of formula IV is combined with the amine (e.g., R12H or R8RgNH)) or the appropriate alcohol, in the presence of a suitable coupling agent. A suitable coupling agent is an agent that transforms a carboxylic acid into a reactive species, which forms an amide or ester bond upon reaction with an amine or alcohol, respectively. The coupling agent can be a reagent that performs this condensation in a single container process when mixed together with the carboxylic acid and the amine or alcohol. If the acid is to be condensed with an alcohol, it is preferable to use a large excess of the alcohol as the reaction solvent, adding or not adding 1.0 or 1.5 equivalents of dimethylaminopyridine. Exemplary coupling reagents are 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide-hydroxybenzotriazole hydrochloride (DEC / HBT), carbonyldiimidazole, dicyclohexylcarbodiimide / hydroxybenzotriazole (HBT), 2-ethoxy-1-ethoxycarbonyl-1, 2-dihydroquinoline (EEDQ), carbonyldiimidazole / HBT, propanophosphonic anhydride (propanophosphonic acid anhydride, PPA) and diethylphosphorylnide. The coupling is carried out in an inert solvent, preferably in an aprotic solvent, at a temperature from about -20 ° C to about 50 ° C, for about 1 to about 48 hours, in the optional presence of a tertiary amine base, such as triethylamine. Examples of solvents include acetonitrile, dichloromethane, ethyl acetate, dimethylformamide, chloroform or mixtures thereof. The coupling agent can also be an agent that converts the carboxylic acid into an activated intermediate, which is isolated and / or formed in a first step and allowed to react with an amine or alcohol in a second step. Examples of such coupling agents and activated intermediates are thionyl chloride or oxalyl chloride to form the acid chloride, cyanuric fluoride to form an acid fluoride or an alkyl chloroformate, such as isobutyl chloroformate or isopropenyl (with a tertiary amine base) to form a mixed carboxylic acid anhydride. If the coupling agent is oxalyl chloride, it is advantageous to employ a small amount of dimethylformamide as a co-solvent with another solvent (such as dichloromethane) to catalyze the formation of the acid chloride. This acid chloride can be coupled by mixing with the intermediate of formula III in an appropriate solvent, together with an appropriate base. Suitable solvent / base combinations are, for example, dichloromethane, dimethylformamide or acetonitrile, or mixtures thereof, in the presence of a tertiary amine base, for example, triethylamine. Other suitable solvent / base combinations include water, an alcohol (dd) or a mixture thereof, together with a cosolvent such as dichloromethane, tetrahydrofuran or dioxane, and a base such as sodium or potassium carbonate, sodium hydroxide, potassium or lithium, or sodium bicarbonate in an amount sufficient to consume the acid released in the reaction. The use of phase transfer catalyst (typically from 1 to 10% by mole), such as quaternary ammonium halide (for example, tetrabutylammonium bromide or methyl trioctylamino chloride) when using a mixture of cosolvents is advantageous only partially miscible (for example, dichloromethane-water or dichloromethane-methanol). The use of these coupling agents and the proper selection of solvents and temperatures are known to those skilled in the art or can be readily determined in the literature. In Houben-Weyl, Vol XV, part II, E. Wunsch, Ed., G, Theime Verlag, 1974, Stuttgart, and M. Bodansky, Principies of Peptide Synthesis, Springer-Verlag Berlin, 1984 and The Peptides. Analysis, Shynthesis and Biology (ed. E. Gross and J. Meienhofer), vols 1-5 (Academic Press NY 1979-1983), describe these and other exemplary conditions useful for the coupling of carboxylic acids. Compounds of formula IV in which Ri, R 10f R n, A, R 2, R 3, R and R 5 are as defined above, can be prepared from the corresponding esters of formula V (ie, compounds of formula IA in that Rβ is alkoxy (dd) carbonyl or benzyloxycarbonyl) by hydrolysis with an aqueous alkali at a temperature from about -20 ° C to about 100 ° C, typically at about 20 ° C, for about 30 minutes to about 24 hours. Alternatively, compounds of formula IV are prepared by activation of an indole carboxylic acid of formula II with a coupling agent (as described above) which gives activated intermediate (such as an acid chloride, acid fluoride or mixed anhydride) ) which is then allowed to react with a compound of formula III in which R3? R and R5 are as described above and R6 is carboxy, in a suitable solvent, in the presence of a suitable base. Suitable solvents include water, methanol or a mixture thereof, together with a cosolvent such as dichloromethane, tetrahydrofuran or dioxane. Suitable bases include sodium, potassium or lithium hydroxides, sodium or potassium bicarbonate, sodium or potassium carbonate, or potassium carbonate together with tetrabutyl ammonium bromide (1 equivalent) in an amount sufficient to consume the acid liberated in the reaction (usually enough to maintain the pH of the reaction above 8). The base can be added incrementally together with the activated intermediate to perform the proper pH control of the reaction. The reaction is generally carried out between -20 ° C and 50 ° C. The isolation procedures are those created by those skilled in the art to remove impurities, but typically consist of the elimination of cosolvents miscible with water by evaporation, the extraction of impurities at high pH with an organic solvent, acidification at a low pH ( 1-2) and filtration or extraction of the desired product with a suitable solvent such as ethyl acetate or dichloromethane. The compound of formula V can be prepared by coupling the appropriate compound of formula III, wherein R6 is alkoxycarbonyl, and the appropriate compound of formula II, in a procedure analogous to that described above. Alternatively, compounds of formula IA containing sulfur atoms in the oxidation state of sulfoxide or sulfone, can be prepared from the corresponding compounds of formula IA having the sulfur atom in the non-oxidized form, by treatment with a suitable oxidizing agent, such as m-chloroperoxybenzoic acid in dichloromethane, at a temperature from about 0 ° C to about 25 ° C, for about 1 to about 48 hours, using from about 1 to about 1.3 equivalents for the conversion into the of sulfoxide oxidation, and more than about 2 equivalents for the conversion to the sulfone oxidation state. For example, in reaction scheme XI, certain compounds of formula IA contain a primary amine, a secondary amine or a carboxylic acid functionality in part of the molecule defined by RQ, which may interfere with the desired coupling reaction of the reaction XI, if the intermediate of formula III or the amine R? 2H or R8RgNH is left unprotected. Therefore, the primary amine, the secondary amine or the carboxylic acid functionality can be protected, when present in the Rβ radicals of the intermediate R8R9NH or R-? 2H amine of the formula III, by an appropriate protecting group, during the coupling reaction of reaction scheme XI. The product of such coupling reaction in such a case is a compound of formula IA which contains the protecting group. This protecting group is removed at a later stage to provide the compound of formula IA. Protective groups suitable for the protection of amine and carboxylic acid include the protecting groups commonly used in the synthesis of peptides (such as Nt-butoxycarbonyl, N-carbobenzyloxy and 9-fluorenylmethyleneoxycarbonyl for amines, and lower alkyl or benzyl esters for acids carboxylic) which are not chemically reactive under the coupling conditions described above and which can be removed without chemically altering another functionality of the compound of formula IA. The indole-2-carboxylic acids and the starting indolin-2-carboxylic acids used in the reaction scheme XI, when they can not be purchased on the market or are not known in the prior art (such a technique is widely published), are can be purchased by conventional synthetic procedures. For example, according to reaction scheme XII, the indole ester of formula VII (wherein A is not nitrogen), can be prepared from the compound of formula VI (wherein Q is selected to achieve the desired A , as defined above, except for N) by a Fischer indole synthesis (see The Fischer Nature Robinson Syndrome, B. (Wiley, New York, 1982)) followed by saponification of the resulting indole ester of formula VII to produce the corresponding acid of formula VIII. The starting aryl hydrazone can be prepared by condensation of a readily available hydrazine with the appropriate carbonyl derivative, or by the Japp-Klingeman reaction (see Organic Reactions, Phillips, R.R., 1959, 10, 143). Alternatively, the indole-2-carboxylic acid of formula VINA can be prepared by condensation of an ortho-methyl-nitro compound of formula IX with an oxalate ester, to produce the indole ester of formula X, followed by reduction of the nitro group and the subsequent hydrolysis. This three-step procedure is known as the Reissert indole synthesis (Reissert, Chemische Berichte 1897, 30, 1030). The conditions for carrying out this sequence and references thereto are described in the literature (Kermack, et al., J. Chem Soc. 1921, 1 19, 1602; Cannon et al., J. Med Chem 1981, 24, 238; Julian, et al in Heterocyclic Compounds, vol. 3 (Wiley, New York, NY, 1962, R. C. Elderfield, ed.) 18). Also, 3-halo-5-chloro-1 H-indole-2-carboxylic acids can be prepared by halogenation of 5- chloro-1 H-indole-2-carboxylic acids. According to reaction scheme XIII, the intermediates of the benzimidazole-2-carboxylic acid of formula XI can be prepared by condensation of an ortho-diamine compound of formula XIII with glycolic acid, followed by oxidation of the resulting benzimidazole-2-methane of formula XII ( Bistrzycki, A. and Przeworski, G. Ber. 1912, 45, 3483). Alternatively, (to reaction scheme XII) the substituted indolines of formula XIV can be prepared by reducing the corresponding ones of formula XV with a reducing agent, such as magnesium in methanol, at a temperature of about 25 ° C to about 65 °. C, for about 1 to about 48 hours (reaction scheme III). The indolecarboxylic acids of formula XVI are prepared by saponification of the corresponding ester of formula XVII (reaction scheme XIII). The ester of formula XVII is prepared by reduction of the corresponding idol ester of formula VII, with a reducing agent such as magnesium, in methanol, as described above for the conversion of the compound of formula XV to the compound of formula XIV. The following paragraphs describe ways to prepare the various amines that are used in the above reaction schemes. According to reaction scheme XIV, an alpha-amino acid of formula XXIII can be protected in nitrogen with an appropriate protecting group (PT) (eg, t-Boc) to form a compound of formula XXIV. One skilled in the art can easily select an appropriate protecting group and a method for its introduction. For example, two common protecting groups are t-Boc (introduced by treatment of the amino acid with di-t-butyl dicarbonate in a suitable solvent or solvent mixture, preferably protic, at high pH) and CBZ (introduced by amino acid treatment with benzyl chloroformate in a suitable solvent or solvent mixture, preferably protic, and a base). The compound of formula XXIV is coupled (in a procedure analogous to the coupling process described in reaction scheme XI) with an appropriate R8RgNH or R- | 2H amine, to form a compound of formula XXV, which is then deprotected resulting in the compound of formula IIIb (ie, the compound of formula III in which R6 is C (0) R? 2 or C (0) NR8Rg). If the protecting group is t-Boc, by treating the compound of formula XXV with an acid of a suitable solvent, preferably aprotic. Acids for that deprotection include HCl, MeS03H or trifluoroacetic acid. According to the reaction scheme XV, a compound of formula XXXI (N-protected amine of formula III wherein R6 is (C? -C8) alkoxycarbonyl or benzyloxycarbonyl) can be prepared from the corresponding unprotected amino acid of formula XXX, by N-protection (producing a protected amino acid of formula XXXII) followed by esterification. For example, the compound of formula XXXIII can be esterified with the appropriate alcohol and an acid catalyst, such as hydrogen chloride or thionyl chloride or, in the case of tert-butanol, by treatment of the amino acid with isobutylene and an acid catalyst such as concentrated sulfuric acid or by treatment with an alkyl halide (e.g., methyl iodide) and a base (e.g., potassium carbonate). Alternatively, the esterification may precede the protection stage.

According to reaction scheme XVI, the compounds of formula XXX in R3 is not H, used in the reaction scheme V, can be prepared as indicated below. The amino acids of formula XLl can be prepared by N-alkylation of the protected amino acids (Pt) of formula XL by treatment with an appropriate base and an alkylating agent. The specific procedures for this alkylation are described by Benoiton, Can. J. Chem. 1977, 55, 906-910 and Hansen, J. Org. Chem. 1985, 50 945-950. For example, when R3 is methyl and Pt is Boc, sodium hydride and methyl iodide are used in tetrahydrofuran. Deprotection of the compounds of formula XLl yields the desired compound of formula XXX. Alternatively, an amino acid of formula XLll can be N-alkylated by a three-step sequence involving a reductive benzylation (such as with belzaldehyde or Pd / C catalyzed hydrogenation) to give the mono-N-benzyl derivative, and a reductive amination with the appropriate carbonyl compound (for example with formaldehyde and sodium cyanoborohydride, to introduce R 3 as methyl) to give the amino acid N-benzyl, N-R 3 substituted. The N-benzyl protecting group is conveniently removed (for example by hydrogenation with an appropriate catalyst) to yield the compound of formula XXX. Specific conditions for this three-step alkylation are described by Reinhold et al., J. Med. Chem., 1968,, 258-260. The immediately preceding preparation can also be used to introduce a radical R3 into an intermediate of formula Illa (which is an intermediate of formula III in which R3 is H). The amino acids used in the schemes of this document (eg, XL, XLll), if they can not be purchased commercially or are not presented in the literature, can be prepared by a variety of methods known to those skilled in the art. For example, the Strecker synthesis or variations thereof may be used. Accordingly, an aldehyde (R4CHO), sodium or potassium cyanide and ammonium chloride react to form the corresponding aminonitrile. The aminonitrile is hydrolyzed with a mineral acid to form the desired amino acid of the formula XLll R4C (NH2) COOH. As an alternative, the procedure of Bucherer-Berg, in which a hydantoin is formed by heating an aldehyde (R4CHO) with ammonium carbonate and potassium cyanide, followed by hydrolysis (for example, with barium hydroxide in reflux dioxane), with an acid or a base for form the desired amino acid of formula XLIII RC (NH2) COOH. The literature also presents other methods for the synthesis of α-amino acids that would allow one skilled in the art to prepare the desired intermediate of formula XLIII R C (NH2) COOH, necessary for the synthesis of the compounds of formula IA. Suitable methods for the synthesis and / or resolution of the compounds of formula XLll are found in reports by Duthaler (Tetrahedron 1994, 50, 1539-1650) or Williams (RM Williams, Synthesis of optically active amino acids.) Pergamon: Oxford, UK 1989). A specific procedure for the synthesis of the intermediate of formula XLll in any enantiomeric form from the corresponding intermediate R4X (X = Cl, Br or I) is the procedure of Pirrung and Krishnamurthy (J. Org. Chem. 1993, 58, 957- 958), or by the procedure of O'Donnell, et al., (J. Am. Chem. Soc. 1989, 1 1 1, 2353-2355). The necessary P ^ X intermediates are readily prepared by many procedures familiar to chemists skilled in the art. For example, compounds in which R4X is ArCH2X can be prepared by radical halogenation of the ArCH3 compound or by Ar-H arene formulation and conversion of the alcohol to the bromide. Another specific procedure for the synthesis of the intermediates of Formula XLll in any enantiomeric form is that of Corey and Link (J. Am. Chem. Soc. 1992, 14, 1906-1908). Thus, an intermediate of formula R4COCCI3 is enantiospecifically reduced to the intermediate R4CH (OH) CCI3, which is converted after treatment with an azide and a base, into an intermediate R CH (N3) COOH, which is reduced by catalytic hydrogenation to the compound of formula XLll desired. The required trichloromethyl ketone R4COCCI3 is obtained by reaction of the aldehyde R4CHO with the trichloromethanide anion, followed by oxidation (Gallina and Giordanio, Synthesis 1989, 466-468).

A compound of the formula R8NH2 or RgNH2 is monoalkylated with a carbonyl compound corresponding to R8 or Rg, respectively, under suitable reductive amination conditions, to give an amine of the formula R8R9NH. To avoid dialkylation, it may be preferable to protect the amines (R8NH2 or R9NH2) with a suitable protecting group PT to give R8 (PT) NH or Rg (Pt) NH, for example, by reaction with belzaldehyde and a reducing agent. The protected amines are monoalkylated with a carbonyl compound corresponding to R8 or Rg, respectively, under suitable reductive amination conditions, to give R8RgN (Pt). The protecting group (Pt) is removed (for example, by exhaustive catalytic hydrogenation when Pt is benzyl) to give a compound of the formula R8R9NH. A person skilled in the art can achieve the appropriate reductive amination conditions in the literature. These conditions include those presented by Borch et al. (J. Am. Chem. Soc. 1971, 2897-2904) and those indicated by Emerson (Organic Reactions, Wiley: New York, 1948 (14), 174), Hutchins et al. (Org Prep, Int.Process 1979 (11), 20, and Lane et al. (Synthesis 1975, 135) Reductive amination conditions that favor N-monoalkylation include those presented by Morales et al. (Synthetic Communications 1984, 1213-1220) and Verardo et al (Synthesis 1992 121-125) The amines R8NH2 or R9NH2 can also be monoalkylated with R9X or RaX, respectively, in which X is chloride, bromine, tosylate or mesylate. , an intermediate of the formula R8 (Pt) NH or R9 (PT) NH can be alkylated with R9X or R8X, and the protecting group can be removed to give a compound of the formula R8R9NH. Additional processes can be used to prepare amines of the formula R8RgNH, in the Thus, a readily available compound of the formula alkoxy (dd) carbonyl-NH 3 or NH 2 CONOHH is dialkylated to nitrogen and oxygen by treatment with a base and an excess of suitable alkylating agent (RX), to give the corr alkoxy (d-d) carbonyI-N (R) OR which is then hydrolyzed to give a compound of the formula R8R9NH (where R8 = Rg = R). Suitable conditions, the base and the alkylating agent include those described by Goel and Krolls (Org. Prep. Proced. Int. 1987, 19, 75-78) and Major and Fleck (J. Am. Chem. Soc. 1928, 50 , 1479). Alternatively, an N-hydroxyurea (NH2CONH (OH) can be alkylated sequentially, first in oxygen to give NH2CONH (OR ') and then in nitrogen to give NH2CON (R ") (OR'), by successive treatment with the alkylating agents R 'X and R' X, respectively, in the presence of an appropriate base The base and suitable alkylating agents include those described by Kreutzkamp and Messinger (Chem. Ber. 100, 3463-3465 (1967) and Danen et al (J. Am. Chem. Soc. 1973, 95, 5716-5724) The hydrolysis of these alkylated hydroxyurea derivatives produces the amines R'ONH2 and R'ONHR ", which correspond to certain amines of the formula R8R9NH. can adapt the procedures described in this paragraph to other alkylating agents R, R 'and R "-X, to prepare other amines of the formula R8RgNH, in which R8-N or R9-N are linked by oxygen-nitrogen bonding. al (SynLett 1991, 559-560) describes the addition catalyzed by BF3 of an organomet reagent lic R-Li or an O-alkyl oxime of formula R'CH = N-OR ", to give compounds of formula R'RCH-NH (OR"). This route can also be used to give compounds of formula R8R9NH, wherein one of R8-NH or Rg-NH is linked by oxygen-nitrogen bond. Prodrugs of this invention in which a carboxyl group in a carboxylic acid of formula IA is replaced with an ester, can be prepared by combining the carboxylic acid with the appropriate alkyl halide in the presence of a base, such as potassium carbonate, in a solvent inert, such as dimethylformamide, at a temperature of about 0 ° C to 100 ° C, for about 1 to about 24 hours. As an alternative, the acid is combined with the appropriate alcohol as a solvent in the presence of a catalytic amount of acid such as concentrated sulfuric acid, at a temperature of about 20 ° C to 120 ° C, preferably at reflux, for about 1 hour to about 24 hours . Another method is the reaction of the acid with a stoichiometric amount of the alcohol, in the presence of a catalytic amount of acid in an inert solvent, such as tetrahydrofuran, with the co-elimination of the water that is produced by physical means (eg, Dean purification). -Stark) or chemicals (for example, molecular sieves).The prodrugs of this invention in which an alcohol function has been transformed into an ether, can be prepared by combining the alcohol with the appropriate alkyl bromide or iodide, in the presence of a base such as potassium carbonate, in an inert solvent such as dimethylformamide, at a temperature of about 0 ° C to 100 ° C, for about 1 to about 24 hours. Alkanoylaminomethyl ethers can be obtained by reaction of the alcohol with a bis- (alkanoylamino) methane, in the presence of a catalytic amount of acid, in an inert solvent such as tetrahydrofuran, according to a procedure described in US 4,997,984. Alternatively, these compounds can be prepared by the methods described by Hoffman et al. in J. Org. Chem, 1994, 59, 3530. The dialkyl phosphate esters can be prepared by reacting the alcohol with a dialkyl chlorophosphate, in the presence of a base, in an inert solvent such as tetrahydrofuran. The dihydrogen phosphates can be prepared by reaction of the alcohol with a diaryl chlorophosphate or dibenzyl, as described above, followed by hydrolysis or hydrogenation in the presence of a noble metal catalyst, respectively. The glycosides are prepared by reaction of the alcohol and a carbohydrate in an inert solvent, such as toluene, in the presence of acid. Typically, the water formed in the reaction is removed as it is formed, as described above. An alternative procedure is the reaction of the alcohol with a suitably protected glycosyl halide, in the presence of a base, followed by deprotection. The N- (1-hydroxyalkyl) amides, the N- (1-hydroxy-1- (alkoxycarbonyl) methyl) amides or the compounds in which R2 has been replaced by C (OH) C (0) OY, can be prepared by the reaction of the parent amide or indole with the appropriate aldehyde, under neutral or basic conditions (for example, sodium ethoxide in ethanol) at temperatures between 25 and 70 ° C. The N-alkoxymethyl indoles or N-1- (alkoxy) alkyls can be obtained by reaction of the N-unsolyl indole with the necessary alkyl halide, in the presence of a base, in an inert solvent. 1- (N, N-dialkylaminomethyl) indole, 1- (1-N, N-dialkylamino) ethyl) indole and N, N-dialkylamino-ethyl-amides (for example, R 3 = CH 2 N (CH 3) 2) can be prepared, the reaction of the parent NH compound as the appropriate aldehyde and amine, in an alcohol solvent at 25-70 ° C. The prodrugs of this invention in which R2 and R3 are a common carbon, can be prepared by reaction of the parent compound (drug) with benzaldehyde or ketone or its dimethylacetal, in an inert solvent, in the presence of a catalytic amount of acid, with the joint removal of water or methanol. Although the preparation for most of the starting materials and reagents for the reaction schemes described above (for example, amines, substituted indolecarboxylic acids, substituted indolinecarboxylic acids and amino acids) has been described, they can also be easily acquired or synthesized by the skilled in the art using conventional organic synthesis methods. For example, many of the intermediates used here to prepare compounds of formula I and IA are, are related to or are derived from amino acids found in nature, in which there is great scientific interest and a great commercial need and, consequently, many of such intermediates can be purchased on the market, presented in the literature or easily prepared from other substances commonly available by methods indicated in the literature. Some of the preparation methods useful for the preparation of the compounds described herein (for example, the compounds of formula I and the compounds of formula IA) may require the protection of a remote functionality. The need for such protection will vary depending on the nature of the remote functionality and the conditions of the preparation procedures. The need for such protection is easily determined by one skilled in the art. The use of such protection / deprotection procedures is also within the skill in the art. For a general description of protective groups and their use, see T.W. Greene, Protective Groups in Organic Svnthesis, John Wiley & amp; amp;; Sons, New York, 1991. Some of the compounds of this invention have asymmetric carbon atoms and, therefore, are enantiomers or diastereomers. Diastereomeric mixtures can be separated into their individual diastereomers based on their physical-chemical differences by methods known per se. For example, by chromatography and fractional crystallization. The enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (eg, an alcohol), separating the diastereomers and converting (eg, hydrolyzing) the individual diastereomers into the corresponding pure enantiomers. All such isomers, including the diastereomers, enantiomers and mixtures thereof are considered part of this invention. In addition, some of the compounds of this invention are atropisomers (e.g., substituted biaryls) and are considered part of this invention. Many of the compounds of this invention are acidic and form a salt with a pharmaceutically acceptable cation. Some of the compounds of this invention are basic and form a salt with a pharmaceutically acceptable anion. All such salts are within the scope of this invention and can be prepared by conventional methods. For example, they can be prepared simply by contacting the acidic and basic entities, usually in a stoichiometric ratio, in an aqueous, non-aqueous or partially aqueous medium, as appropriate. The salts are recovered by filtration, by precipitation with a non-solvent followed by filtration, by evaporation of the solvent or, in the case of aqueous solutions, by lyophilization, as appropriate.

In addition, when the compounds of this invention form hydrates or solvates, they are also within the scope of the invention. The utility of the combinations of the present invention as medical agents in the treatment of diseases as detailed herein in mammals (e.g., humans) is demonstrated by the activity of the compounds of this invention in conventional assays and in in vitro assays. vitro and in vivo described below. Such assays also provide a means by which the activities of the compounds of this invention can be compared with the activities of other known compounds. The results of these comparisons are useful for determining dosage levels in mammals, including humans, for the treatment of such diseases.

Assays of aldose reductase inhibitors The activity of an aldose reductase inhibitor can be determined by the amount of aldose reductase inhibitor that is required to reduce sorbitol in tissues and, therefore, to reduce fructose in the tissues of according to the following essay. Male Sprague-Dawley rats are diabetic by injection of streptozocin at 55 mg / kg, i.v., in citrate buffer pH 4.5. The rats are fed without limitation under controlled housing conditions, lighting temperature. After 5 weeks of diabetes, the rats are anesthetized with an overdose of pentobarbital, tissue is rapidly extracted and analyzed for the content of sorbitol and fructose. The sorbitol levels are analyzed according to the procedure of Donald M. Eades et al., "Rapid Analysis of Sorbitol, Galactitol, Mannitol and Myoinositol Mixtures From Bilogical Sources", Journal of Chromatography, 490, 1-8 (1989). The fructose from the tissues of the rats is measured enzymatically using a modification of the Ameyama method (Methods in Enzymology, 89: 20-29, 1982), in which ferricyanide was replaced by resazurin, a dye that is reduced to resorufin, which has a high fluorescence. The amount of resorufin fluorescence is stoichiometric with the amount of fructose oxidized by the fructose dehydrogenase. The assay contains 0.1 ml of nerve extract in neutralized 6% perchloric acid, in a final volume of 1.5 ml. After incubation for 60 minutes at room temperature in a closed box, the fluorescence of the sample is determined at excitation = 560 nm, emission = 580 nm with 5 mm slots each, in a Perkin-Elmer model 650 fluorescence spectrometer -40. Fructose concentrations are calculated by comparison with a series of known fructose standards.

Glycogen phosphorylase inhibition assays The three different enzymes of purified glycogen phosphorylase (GP), in which glycogen phosphorylase is in the activated state "a" (called glycogen phosphorylase a, or abbreviated as GPa), and named here hepatic to human glycogen phosphorylase (HLGPa), human muscle glycogen phosphorylase a (HMGPa) and human brain glycogen phosphorylase (HBGPa), can be obtained by the following procedures.

Expression and fermentation The HLGP and HMGP cDNAs are expressed in plasmid pKK233-2 (Pharmacia Biotech, Inc., Piscataway, New Jersey) in E. coli strain XL-1 Blue (Stratagene Cloning Systems, LaJolla, CA). The strain is inoculated in LB medium (consisting of 10 g of tryptone, 5 g of yeast extract, 5 g of NaCl and 1 ml of 1 N NaOH per liter) plus 100 mg / l of ampicillin, 100 mg / l of pyridoxine and 600 mg / l of MnCl2, and allowed to grow at 37 ° C to a cell density of OD550 = 1.0. At this time, cells are induced with 1 mM isopropyl-1-thio-β-D-galactoside (IPTG). Three hours after induction, the cells are harvested by centrifugation and the cell pellets are frozen at -70 ° C until they are needed for purification. The HBGP cDNA can be expressed by several methodologies, for example, by the procedure described by Crerar et al., (J. Biol. Chem. 270: 13748-13756). The procedure described by Crerar et al., (J. Biol. Chem. 270: 13748-13756) for the expression of HBGP is as follows: the HBGP cDNA can be expressed in plasmid pTACTAC in strain 25A6 of E. coli. The strain is inoculated in LB medium (consisting of 10 g of tryptone, 5 g of yeast extract, 5 g of NaCl and 1 ml of NaOH per liter) plus 50 mg / l of ampicillin, it is developed overnight, then resuspend in a fresh LB medium plus 50 mg / l of ampicillin, re-inoculate in a volume 40X of medium LB / amp containing isopropyl-1-thio- ^ D-galactoside (IPTG) 250 μM, 0.5 mM pyridoxine and 3 mM MgCl 2 and cultivated at 22 ° C for 48-50 hours. The cells can then be harvested by centrifugation and the cell pellets are frozen at -70 ° C until they are needed for purification. The HLGP cDNA is expressed in the plasmid pBlueBac III (Invitrogen Corp. San Diego, CA) which is used to cotransfect with the BaculoGold Viral Linear DNA (Pharmingen, San Diego, CA) Sf9 cells. Subsequently, the recombinant viruses are purified on plates. For the purification of proteins, Sf9 cells developed are infected in a medium without serum with a multiplicity of infection (moi) of 0.5 and a cell density of 2 x 106 cells / ml. After being grown for 72 hours at 27 ° C, the cells are centrifuged and the cell pellets are frozen at -70 ° C until they are needed for purification.

Purification of Glycogen Phosphorylase Expressed in E. coli E. coli cells in the sediments described above are resuspended in 25 mM? -glycerophosphate (pH 7.0 with 0.2 mM DTT, 1 mM MgCl2, plus the following protease inhibitors: 0.7 μg / ml Pepstatin A 0.5 μg / ml Leupeptin 0.2 mM Phenylmethylsulfonyl fluoride (PMSF), 0. 5 mM EDTA are lysed by pretreatment with 200 μg / ml lysozyme and 3 μg / ml DNase, followed by sonication in 250 ml batches for 5 x 1.5 minutes on ice, using a Branson Model 450 ultrasonic cell breaker (Branson Sonic Danbury Power Co. CT). The lysates of E. coli cells are then rinsed by centrifugation at 35,000 X g for 1 hour, followed by filtration through 0.45 μm filters. The GP in the soluble fraction of the lysates (estimated to be less than 1% of the total protein) is purified by controg the enzymatic activity (as described in the GPa Activity Assay section, shown below) in a series of chromatographic steps detailed below.

Immobilized metal affinity chromatography (IMAC) This step is based on the procedure of Luong et al (Luong et al, Journal of Chromatography (1992) 584, 77-84). 500 ml of the filtered soluble fraction from the cell lysates (prepared from approximately 160-250 g of original cell pellet) are introduced into a 130 ml column of IMAC Chelating-Sepharose (Pharmacia LKB Biotechnology, Piscataway, New Jersey) which was loaded with 50 mM CuCI2 and 25 mM? -glycerophosphate, 250 mM NaCl and 1 mM midazole in equilibrium buffer at pH 7. The column is washed with equilibration buffer until the A280 returns to the baseline. Then, the sample is eluted from the column with the same buffer containing 100 mM imidazole to remove the bound GP and other bound proteins. Fractions containing GP activity are pooled (approximately 600 ml) and ethylenediaminetetraacetic acid (EDTA), DL-dithiothreitol (DTT), phenylmethylsulfonyl fluoride (PMSF), leupeptin and pepstatin A are added to obtain concentrations of 0.3 mM, 0.2, 0.2 mM, mM, 0.5 μg / ml and 0.7 μg / ml, respectively. The combined GP is desalted on a Sephadex G-25 column (Sigma Chemical Co., St. Louis, Missouri) equilibrated with 25 mM Tris-HCl (pH 7.3) and 3 mM DTT buffer (Buffer A) to remove imidazole and stores on ice until the second chromatographic stage.

Chromatography at 5'-AMP-Sepharose The pooled and desalted GP sample (approximately 600 ml) is then mixed with 70 ml of 5'-AMP Sepharose (Pharmacia LKB Biotechnology, Piscataway, New Jersey) which has been equilibrated with Buffer A ( see above). The mixture is gently stirred for 1 hour at 22 ° C and then introduced into a column and washed with Buffer A until the A280 returns to the baseline. The GP and other proteins are eluted from the column with 25 mM Tris-HCl, 0.2 mM DTT and 10 mM adenosine-5'-monophosphate (APM) at pH 7.3 (Buffer B). Fractions containing GP are pooled after identification, determining enzymatic activity (described below) and visualizing the GP protein band of approximately 97 Kdaltons Mr by polyacrylamide and sodium dodecyl sulfate gel electrophoresis (SDS-PAGE) followed by staining with silver (2D-silver Stain II "Daiichi Kit", Daiichi Pure Chemicals Co., LTD., Tokyo, Japan) and then meet. The pooled GP is dialyzed into 25 mM glycerophosphate, 0.2 mM DTT, 0.3 mM EDTA, 200 mM NaCl, pH 7.0 buffer (Buffer C) and stored on ice until used. Prior to the use of the GP enzyme, the enzyme is converted from the inactive form which is expressed in strain XL-1 Blue of £. coli (called GPb) (Stratagene Cloning Systems, La Jolla, California), in the active form (named GPa) by the procedure described in section (A) Activation of GP, shown below.

Purification of glycogen phosphorylase expressed in Sf9 cells The Sf9 cells of the sediments described above are resuspended in 25 mM β-glycerol phosphate (pH 7.0) with 0.2 mM DTT and 1 mM MgCl 2., plus the following protease inhibitors: 0.7 μg / ml Pepstatin A 0.5 μg / ml Leupeptin 0.2 mM Phenylmethylsulfonyl fluoride (PMSF), and 0.5 mM EDTA are lysed by pretreatment with 3 μg / ml DNase followed by sonication in batches , for 3 x 1 minutes on ice, using a Branson Model 450 ultrasonic cell breaker (Branson Sonic Power Co. Danbury CT). The Sf9 cell lysates are then rinsed by centrifugation at 35,000 x g for 1 hour, followed by filtration through 0.45 μm filters. The GP in the soluble fraction of the lysates (estimated to be 1.5% of the total protein) is purified by controlling the enzymatic activity (as described in the Gpa Activity Assay section, shown below) in a series of chromatographic steps detailed below.

Immobilized metal affinity chromatography (IMAC) Immobilized metal affinity chromatography is performed as described in the previous section. The collected and desalted GP is then stored on ice until further processed.

Activation of GP Before performing another chromatography, the fraction of the inactive enzyme, as expressed in the Sf9 cells (called GPb), is converted into the active form (called GPa) by the following procedure described in section (A) Activation of GP shown below.

Anion exchange chromatography After activation of the GPb purified by IMAC to GPa by reaction with the immobilized phosphorylase kinase, the pooled GPa fractions are dialyzed against 25 mM Tris-HCl, pH 7.5 containing 0.5 mM DTT, 0.2 mM EDTA , phenylmethylsulfonyl fluoride (PMSF) 1.0 mM, 1.0 μg / ml leupeptin and 1.0 μg / ml pepstatin. The sample is then introduced into a MonoQ Anion Exchange Chromatography column (Pharmacia Biotech, Inc., Piscataway, New Jersey). The column is washed with equilibration buffer until the A280 returns to the baseline. Then, the sample is eluted from the column with a linear gradient of 0-0.25 M NaCl to remove the bound GP and other bound proteins. Fractions containing GP elute within the NaCl range of 0.1-0.2 M, as detected by controlling the eluent with respect to the absorbance of the peaks of the A28o proteins. The GP protein is then identified by visualizing the GP Mr protein band of approximately 97 kdaltons by polyacrylamide and sodium dodecyl sulfate gel electrophoresis (SDS-PAGE), followed by silver staining (2D-silver Stain II "Daiichi Kit", Daiichi Puré Chemicals Co., LTD., Tokyo, Japan) and then meets. The pooled GP is dialyzed into 25 mM N, N-bis [2-hydroxyethyl-2-aminoethanesulfonic acid, 1.0 mM DTT, 0.5 mM EDTA, 5 mM NaCl, buffer pH 6.8 and stored on ice until use.

Determination of the enzymatic activity of GP A) Activation of GP: Conversion of GPb into GPa Before the determination of the enzymatic activity of GP, the enzyme is converted from the inactive form, as expressed in strain XL-1 Blue of E Coli (called GPb) (Stratagene Cloning Systems, La Jolla, California), in the active form (designated GPa) by phosphorylation of Gp using the phosphorylase kinase as indicated below. The fraction of the inactive enzyme as expressed in Sf9 cells (referred to as GPb) is also converted to the active form (termed GPa) by the following procedure.

Gp reaction with immobilized phosphorylase kinase Phosphorylase kinase (Sigma Chemical Company, St. Louis, MO) is immobilized in Affi-Gel 10 (BioRad Corp., Melville, NY) following the manufacturer's instructions. Briefly, the enzyme phosphorylase kinase (10 mg) is incubated with washed Affi-Gel beads (1 ml) in 2.5 ml of 100 mM HEPES and 80 mM CaC.2 at pH 7.4, for 4 hours at 4 ° C. The Affi-Gel beads are then washed once with the same buffer before blocking with 50 mM HEPES and 1 M glycine methyl methyl ester glycine at pH 8.0, for one hour at room temperature. The blocking buffer is removed and replaced with 50 mM HEPES (pH 7.4), 1 mM mercaptoethanol and 0.2% NaN3 for storage. Before use to convert GPb to GPa, Affi-Gel beads with the immobilized phosphorylase kinase are equilibrated by washing them in the buffer used to perform the kinase reaction, which consists of 25 mM glycerophosphate, 0.3 mM DTT and 0.3 mM EDTA. at pH 7.8 (kinase assay buffer). The inactive and partially purified GP obtained from the chromatography on 5'-AMP-Sepharose above (from E. coli) or the mixture of GPa and GPb obtained from the previous IMAC (from Sf9 cells) is diluted 1: 10 with the kinase assay buffer and then mixed with the aforementioned phosphorylase kinase enzyme immobilized on the Affi-Gel beads. NaATP is added at a concentration 5 mM and MgCl 2 at a concentration of 6 mM. The resulting mixture is mixed gently at 25 ° C for 30-60 minutes. The sample is removed from the beads and the percentage of activation of GPb is estimated by conversion to GPa determining the enzymatic activity of GP in the presence and absence of 3.3 mM AMP. The percentage of total GP enzymatic activity due to GPa enzyme activity (independent of AMP) is then calculated as follows:% Total HLGPa = HLGP activity - AMP HLGP activity + AMP As an alternative, the conversion of GPb into Gpa can be controlled by isoelectric focusing, based on the displacement of the electrophoretic mobility that is detected after the conversion of Gpb into GPa. The GP samples are analyzed by isoelectric focusing (IEF) using the Pharmacia pfastGel system (Pharmacia Biotech, Inc., Piscataway, New Jersey), using pre-molded gels (range pl 4-6.5) and the procedure recommended by the manufacturer. The resolved bands of GPa and GPb are then visualized on the gels by silver staining (2D-silver Stain II "Daiichi Kit", Daiichi Pure Chemicals Co., LTD., Tokyo, Japan). The identification of GPa and GPb is carried out by comparison with the GPa and GPb standards derived from E. coli that are tested in parallel on the same gels as the experimental samples.

B) GPa activity assay The disease / condition treatment / prevention activities described herein of the glycogen phosphorylase inhibitor compounds of this invention can be determined indirectly by evaluating the effect of the compounds of this invention on the activity of the activated form of glycogen phosphorylase (GPa) by one of two procedures; the activity of glycogen phosphorylase a is measured in the forward direction by controlling the production of glucose-1-phosphate from glycogen, or following the reverse reaction, measuring the synthesis of glycogen from glucose-1-phosphate by the release of inorganic phosphate. All reactions can be performed in triplicate in 96-well microtiter plates and the absorbance change due to the formation of the reaction product at the wavelength specified below is measured in an MCC / 340 MKII Elisa Reader (Lab Systems, Finland) connected to a Titertech Microplate Stacker (ICN Biomedical Co., Huntsville, Alabama). To measure the enzymatic activity of GPa in the forward direction, the production of glucose-1-phosphate from glycogen is controlled by the general multienzyme coupled procedure of Pesce et al. [Pesce, MA, Bodourian, SH, Harris, RC and Nicholson, JF (1977) Clinical Chemistry 23, 1711-1717] modified as follows: diluted from 1 to 100 μg of GPa, 10 units of phosphoglucomutase and 15 glucose-6-phosphate dehydrogenase units (Boehringer Mannheim Biochemicals, Indianapolis, IN) at 1 ml, in buffer A (described below). Buffer A is at pH 7.2 and contains 50 mM HEPES, 100 mM KCl, 2.5 mM ethylene glycolite-acetic acid (EGTA), 2.5 mM MgCl2, 3.5 mM KH2P04 and 0.5 mM dithiothreitol. 20 μl of this stock solution is added to 80 μl of buffer A containing 0.47 mg / ml glycogen, glucose 9.4 mM and 0.63 mM of the oxidized form of nicotinamide adenine dinucleotide phosphate (NADP +). The compounds to be tested are added in the form of 5 μl of solution in dimethyl sulfoxide (DMSO) at 14% before the addition of the enzymes. The basal proportion of enzymatic activity of GPa in the absence of inhibitors is determined by adding 5 μl of 14% DMSO and a completely inhibited proportion of the GPa enzyme activity is obtained by adding 20 μl of the positive control test substance, caffeine, to a concentration 50 mM. The reaction is followed at room temperature by measuring the conversion of oxidized NADP + to reduced NADPH at 340 nm. To measure the enzymatic activity of GPa in the reverse direction, the conversion of glucose-1-phosphate to glycogen + inorganic phosphate is measured by the general procedure described by Engers et al [Engers, HD, Shechosky, S. and Madsen, NB (1970) Can J. Biochem, 48, 746-754], modified as follows: diluted from 1 to 100 μg of GPa in 1 ml of buffer B (described below). Buffer B is at pH 7.2 and contains 50 mM HEPES, 100 mM KCl, 2.5 mM EGTA, 2.5 mM MgCl 2, and 0.5 mM dithiothreitol. 20 μl of this stock solution is added to 80 μl of buffer B with 1.25 mg / ml of glycogen, glucose 9.4 mM and glucose-1-phosphate 0.63 mM. The compounds to be tested are added in the form of 5 μl of solution in 14% DMSO before the addition of the enzyme. The basal proportion of the GPa enzyme activity in the absence of added inhibitors is determined by adding 5 μl of 14% DMSO, and the fully inhibited proportion of the GPa enzyme activity is obtained by adding 20 μl of 50 mM caffeine. This mixture is incubated at room temperature for 1 hour and the inorganic phosphate released from glucose-1-phosphate is measured by the general procedure of Lanzetta et al. [Lanzetta, P. A., Alvarez, L. J., Reinach, P. S. and Candia, O. A. (1979) Anal. Biochem. 100, 95-97] modified as follows: 150 ml of 10 mg / ml ammonium molybdate and 0.38 mg / ml malachite green in 1 N HCl are added to 100 μl of the enzyme mixture. After an incubation period of 20 minutes at room temperature, the absorbance at 620 nm is measured. The above tests carried out with a range of concentrations of test compounds allow the determination of an IC 50 value (concentration of the test compound necessary for a 50% inhibition) for the in vitro inhibition of the enzymatic activity of GPa by that compound of test.

Insulin resistance indication assays The combinations of this invention readily adapt to clinical use as hypoglycaemic agents. The hypoglycaemic activity of the combinations of this invention can be determined by the amount of test compound that reduces glucose levels relative to a vehicle without test compound, in male ob / ob mice. The assay also allows the determination of an approximate minimum effective dose (MED) value for the in vivo reduction of the plasma glucose concentration in such mice for such test compounds. As blood glucose concentration is closely related to the development of diabetic disorders, these combinations, by virtue of their hypoglycaemic action, prevent, stop and / or reverse diabetic disorders. Five male mice aged 5 to 8 weeks, C57BL / 6J-ob / ob (obtained from Jackson Laboratory, Bar Harbor, ME) are enclosed per cage, under conventional animal care practices. After a one-week acclimatization period, the animals are weighed and 25 μl of retro-orbital breast blood is removed before any treatment. The blood sample is immediately diluted 1: 5 with saline containing 0.025% sodium heparin and kept on ice for analysis of metabolites. Animals are assigned to treatment groups so that each group has a similar average for plasma glucose concentration. After the allocation of the groups, the animals receive orally, every day, for 4 days, the vehicle consisting of: 1) 0.25% w / v of hypromellose in water without pH adjustment; or 2) 0.1% Pluronic® P105 Block Copolymer Sufactant (BASF Corporation, Parsippany, NJ) in 0.17% saline without pH adjustment. On day 5, the animals are weighed again and then orally receive the test compound or the vehicle alone. All drugs are administered in a vehicle that consists of: 1) 0.25% w / v of hypromellose in water without pH adjustment; or 2) 10% DMSO / 0.1% Pluronic® P105 (BASF Corporation, Parsippany, NJ) in 0.1% saline without pH adjustment. Three hours later, blood is drawn from the animals through the retro-orbital sinus to determine the levels of the metabolites in the blood. The freshly collected samples are centrifuged for two minutes at 10,000 x g at room temperature. In the supernatant, glucose is analyzed, for example, by the Abbott VP ™ automatic analyzers (Abbott Laboratories, Diagnostics Division, Irving, TX) and VP Super System® (Abbott Laboratories, Irving, TX) or the Abbott Spectrum CCX ™ (Abbott Laboratories, Irving, TX) using the A-Gent ™ Glucose-UV reagent system (Abbott Laboratories, Irving, TX) (a modification of the Richterich and Dauwalder procedure, Schweizerische Medizinishe Wochenschrift, 101, 860 (1971)) (hexokinase procedure ), using a 100 mg / dl standard. The plasma glucose is then calculated by the equation: Plasma glucose (mg / dl) = sample value x 8.14 where 8.14 is the dilution factor, adjusted for the plasma hematocrit (assuming that the hematocrit is 44% ).

The animals that received the vehicle maintained substantially unchanged hyperglycemic glucose levels (eg, greater than or equal to 250 mg / dL), animals treated with test compounds at adequate doses have significantly reduced glucose levels. The hypoglycaemic activity of the test compounds is determined by statistical analysis (unpaired t test) of the mean plasma glucose concentration between the group treated with test compounds and the group treated with vehicles on day 5. The previous test carried out with a range of doses of test compounds, allows the determination of an approximate value of the minimum effective dose (MED) for in vivo reduction of plasma glucose concentration. The compounds of this invention readily adapt to clinical use as agents to reverse hyperinsulinemia, as agents to reduce triglycerides and as hypocholesterolemic agents. Such activity can be determined by the amount of test compound that reduces the levels of insulin, triglycerides or cholesterol with respect to a control vehicle without test compound in male ob / ob mice. Since the concentration of cholesterol in the blood is closely related to the development of cardiovascular, cerebrovascular or peripheral vascular disorders, the combinations of this invention, by virtue of their hypocholesterolemic action, prevent, arrest and / or reverse atherosclerosis.

As the concentration of insulin in blood is closely related to the promotion of vascular cell growth and the increase in sodium retention by the kidney, (in addition to other actions, for example, the promotion of glucose utilization) and these functions are known causes of hypertension, the combinations of this invention, by virtue of their hypoinsulinemic action, prevent, stop and / or reverse hypertension. Since the concentration of triglycerides and free fatty acids i in the blood contributes to the overall levels of blood lipids, the combinations of this invention, by virtue of their triglyceride and fatty acid reducing activity, prevent, stop and / or reverse hyperlipidemia. Free fatty acids contribute to the overall level of blood lipids and, independently, have been negatively correlated with insulin sensitivity in a variety of physiological and pathological states. Five male mice between 5 and 8 weeks of age are enclosed, C57BL 6J-ob / ob (obtained from Jackson Laboratoy, Bar Harbor, ME) per cage, under conventional animal care practices and fed without limitation with a conventional diet for rodents. After an acclimatization period of one week, the animals are weighed and extracted μl of retro-orbital sinus blood before any treatment. The blood sample is immediately diluted 1: 5 with saline containing 0.025% sodium heparin and kept on ice for anas of metabolites. Animals are assigned to treatment groups so that each group has a similar average for plasma glucose concentration. The compound to be tested is administered by an oral probe as a solution of approximately 0.02% to 2.0% (w / v (w / v)) in 1) 10% DMSO / 0.1% Pluronic® P105 Block Copolymer Surfactant (BASF Corporation, Parsippany, NJ) in 0.1% saline without pH adjustment, or 2) 0.25% w / v of hypromellose in water without pH adjustment. Dosage is maintained once a day (s.i.d.) or dosing twice a day (b.i.d.) for 1 a, for example, 15 days. Control mice receive only 10% DMSO / 0.1% Pluronic® P105 in 0.1% saline without pH adjustment or 0.25% w / v hypromellose in water without pH adjustment. Three hours after the last dose is administered, the animals are sacrificed by decapitation and blood is collected from the trunk in 0.5 ml serum separator tubes containing 3.6 mg of a 1: 1 weight / weight mixture of sodium fluoride: potassium oxalate. Freshly collected samples are centrifuged for 2 minutes at 10,000 x g at room temperature and the serum supernatant is transferred and diluted 1: 1 volume / volume with a 1TIU / ml solution of aprotinin in 0.1% saline without pH adjustment. The diluted serum samples are then stored at -80 ° C until anas. The diluted and thawed serum samples are analyzed with respect to insulin, triglycerides, free fatty acids and cholesterol levels. Serum insulin concentration is determined using Equate® RIA INSULIN equipment (double antibody procedure, as specified by the manufacturer) purchased from Binax, South Portland, ME. The coefficient of variation between tests is < 10% Serum triglycerides are determined using the Abbot VP ™ and VP Super System® automatic analyzers (Abbot Laboratories, Irving, TX) or the Abbot Spectrum CCX ™ (Abbot Laboratories, Irving, TX) using the A-Gent ™ Reagent Triglycerides Test system. (Abbot Laboratories, Diagnostics Division Irving, TX) (lipase-coupled enzyme method; a modification of the procedure of Sampson et al., Clinical Chemistry 21, 1983 (1975)). Total serum cholesterol levels are determined using the Abbot VP ™ and VP Super System® automatic analyzers (Abbot Laboratories, Irving, TX) or the Abbot Spectrum CCX ™ (Abbot Laboratories, Irving, TX) and the A-Gent reagent system ™ Cholesterol Test (enzyme procedure coupled to cholesterol esterase; a modification of the procedure of Allain, et al., Clinical Chemistry 20, 470 (1974)) using 100 and 300 mg / dl standards. The concentration of free acids in serum is determined using a team from Amano International Enzyme Co., Inc., adapted for use with the Abbot VP ™ and VP Super System® automatic analyzers (Abbot Laboratories, Irving, TX) or the Abbot Spectrum CCX ™ (Abbot Laboratories, Irving, TX). Then the levels of insulin, triglycerides, free fatty acids and total cholesterol in serum are calculated by the equations, Insulin in serum (μU / ml) = Sample value X 2 Triglycerides in serum (mg / dl) = Sample value X 2 Total serum cholesterol (mg / dl) = Sample value X 2 Free fatty acids in serum (μEq / l) = Sample value X 2 where 2 is the dilution factor. The animals that received the vehicle maintained substantially unchanged and high levels of serum insulin (e.g., 275 μU / ml), serum triglycerides (e.g., 235 mg / dl), free fatty acids in serum (e.g. , 1500 μEq / ml) and total serum cholesterol (for example, 190 mg / dl), while animals treated with the compounds of this invention generally have lower levels of insulin, triglycerides, free fatty acids and total serum cholesterol . The activity of reducing the insulin, triglycerides, free fatty acids and total cholesterol in serum of the test compounds is determined by statistical analyzes (unpaired t tests) of the mean concentration of insulin, triglycerides, free fatty acids or total cholesterol in serum, between the group treated with the test compound and the control group with vehicle. The activity to provide protection from ischemia (eg, cardiac tissue lesions) for the compounds of this invention can be demonstrated in vitro together with the lines presented in Butwell et al., Am, J. Physiol., 264, H1884-H1889, 1993 and Allard et al., Am. J. Physio., 1994, 267, H66-H74. The experiments are performed using an isolated isovolumic cardiac preparation of rat, essentially as described in the article mentioned above. Previously normal male Sprague-Dawley rats, male Sprague-Dawley rats treated by an aortic bandage operation to have cardiac hypertrophy, male BB / W rats with acute diabetes or non-diabetic male BB / W rats, of control, are treated. similar age, with heparin (1000 u, ip), followed by pentobarbital (65 mg / kg, ip). After achieving deep anesthesia, as determined by the absence of the leg reflex, the heart is rapidly excised and placed in ice-cold saline. The heart undergoes retrograde perfusion through the aorta for 2 minutes. The heart rate and ventricular pressure are determined using a latex balloon in the left ventricle with a high-pressure tube connected to a pressure transducer. The heart is perfused with an infusion solution consisting of (mM) NaCl 118, KCl 4.7, CaCl 2 1.2, MgCl 2 1.2, NaHCO 3 25, glucose 1 1. The temperature of the perfusion apparatus is controlled very precisely with heated baths used for the perfusate and for the water in the heating jacket surrounding the perfusion tube, to maintain the heart temperature at 37 ° C. Oxygenation of the perfusate is provided by a pediatric hollow fiber oxygenator (Capiax, Terumo Corp., Tokyo, Japan) located very close to the heart. The hearts are exposed to the perfusion solution ± test compound for approximately 10 minutes or more, followed by 20 minutes of global ischemia and 60 minutes of reperfusion in the absence of the test compound. The cardiac rhythms of the control and of the hearts treated with test compound are compared in the post-ischemia period. The left ventricular pressure of the control hearts and of the hearts treated with test compound are compared in the post-ischemia period. At the end of the experiment, the hearts are also perfused and stained to determine the relationship between the area of infarction with respect to the area at risk (% IA / AAR) as described below. The therapeutic effects of the compounds of this invention to prevent lesions of cardiac tissues that would otherwise be obtained in an ischemic case, can also be demonstrated in vivo by the guidelines presented in Liu et al., Circulation, Vol 84. No. 1, (July 1991), as specifically described in this document. The in vivo assay tests the cardioprotection of the test compound with respect to the control group receiving saline vehicle. As prior information, it should be noted that brief periods of myocardial ischemia followed by reperfusion of the coronary artery, protect the heart from a more severe later myocardial ischemia (Murry, et al., Circulatio? 74: 1 124-1 136, 1986) . Cardioprotection, indicated by a reduction in infarcted myocardium, can be induced pharmacologically using adenosine receptor agonists administered intravenously in intact anesthetized rabbits studied as an in situ model of preconditioning myocardial ischemia (Liu et al., Circulation 84: 350 -356, 1991). The in vivo test tests whether the compounds can pharmacologically induce cardioprotection, that is, reduce the size of the myocardial infarction, when administered parenterally to intact anesthetized rabbits. The effects of the compounds of this invention can be compared to ischemic preconditioning using the Al adenosine agonist, N6-1- (phenyl-2R-isopropyl) adenosine (PIA), which has been shown to pharmacologically induce cardioprotection in intact anesthetized rabbits studied in situ (Liu et al., Circulation 84: 350-356, 1991). The exact methodology is described below.

Surgery: New Zeland male white rabbits (3-4 kg) are anesthetized with sodium pentobarbital (30 mg / kg, i.v.). A tracheotomy is performed through a cervical incision in the ventral midline and the rabbits are ventilated with 100% oxygen using a positive pressure ventilator. Catheters are placed in the left jugular vein to administer the drug and in the left carotid artery to measure blood pressure. The hearts are then exposed through a left thoracotomy and a loop (silk 00) is placed around a prominent branch of the left coronary artery. Ischemia is induced by tightening the loop and fixing it in place. The release of the loop allows reperfusion in the affected area. Myocardial ischemia is evidenced by regional cyanosis; reperfusion is evidenced by reactive hyperemia.

Protocol: Once the blood pressure and heart rate have been stable for at least 30 minutes, the experiment is started. Ischemic preconditioning is induced by clogging the coronary artery twice for 5 minutes, followed by a 10-minute reperfusion. The pharmacological preconditioning is induced by infusing the test compound twice, for example, for 5 minutes and leaving 10 minutes before another intervention, or infusing the adenosine agonist, PIA (0.25 mg / kg). After ischemic preconditioning, a pharmacological preconditioning or non-conditioning (unconditioned, vehicle control), the artery is obstructed for 30 minutes and then reperfused for 2 hours to induce a myocardial infarction. The test compound and the PIA are dissolved in saline or another suitable vehicle and released at 1-5 ml / kg, respectively.

Staining: (Liu et al., Circulation 84: 350-356, 1991). At the end of the 2-hour reperfusion period, the hearts are rapidly removed, deposited in a Langendorff apparatus and washed for 1 minute with normal saline solution warmed to body temperature (38 ° C). Then, the silk suture used as a loop is tied tightly to re-obstruct the artery and a suspension is infused with 0.5% fluorescent particles (1-10 μm) Duke Scientific Corp. (Palo Alto, Ca) with the perfusion liquid, for stain all the myocardium except the area at risk (non-fluorescent ventricle). The hearts are then frozen rapidly and stored overnight at -20 ° C. The next day, 2 mm cuts of the hearts are made and stained with 1% triphenyl tetrazolium chloride (TTC). As TTC reacts with living tissue, this staining differentiates between living tissue (dyed red) and dead tissue (non-stained infarcted tissue). The infarcted area (unstained) and the area at risk (without fluorescent particles) are calculated for each left ventricle cut using a precalibrated image analyzer. To normalize the ischemic lesion and determine the differences of the area at risk between hearts, the data are expressed as the relationship between the infarcted area versus the area at risk (% IA / AAR). All data are expressed as mean ± SEM and are statistically compared using a single-factor ANOVA or an unpaired t-test. It is considered statistical significance when p < 0.05. The invention can be tested with respect to its utility to reduce or prevent ischemic lesions in non-cardiac tissues, for example, in brain or liver, using methods presented in the scientific literature. The compounds of this invention can be administered by the preferred route and administration vehicle and at the preferred time of administration, either before the ischemic episode, during the ischemic episode, after the ischemic episode (reperfusion period) if included, or during any of the experimental phases mentioned above. The beneficial effects of the invention for reducing ischemic lesions in the brain can be demonstrated, for example, in mammals, using the procedure of Park, et al. (Ann Neurol, 1988; 24: 543-551). In summary, in the procedure of Park, et al., Adult male Sprague-Dawley rats are anesthetized, initially with 2% halothane and subsequently by mechanical ventilation with a mixture of nitrous oxide-oxygen (70%: 30%). which contains 0.5-1% halothane. Then a tracheotomy is performed. The output volume of the ventilator is adjusted to maintain a carbon dioxide blood pressure of approximately 35 mm Hg and adequate arterial oxygenation (PaO 2> 90 mm Hg). The body temperature can be controlled by the use of a rectal thermometer and the animals can be kept normothermic, if necessary, by external heating. The animals are then subjected to a subtemporal craniectomy to expose the main trunk of the left middle cerebral artery (MCA) under an operating microscope and the exposed artery is obstructed with microbipolar coagulation to generate large ischemic lesions in the cerebral cortex and in the basal ganglia. After 3 hours of obstruction of the MCA, the rats are deeply anesthetized with 2% halothane and a thoracotomy is performed to infuse heparinized saline solution into the left ventricle. The effluent is collected by an incision in the right atrium. Washing with saline is followed by approximately 200 ml of a solution with 40% formaldehyde 40%, glacial acetic acid and absolute methanol (FAM, 1: 1: 8, v / v / v), then the animals They decapitate and the head is stored in fixative agent for 24 hours. The brain is then removed, dissected, processed, placed in paraffin wax and sectioned (approximately 100 sections per brain). The sections are then stained with hematoxylin-eosin or with a combination of cresyl violet and Luxol fast blue and examined by light microscopy to identify and quantify the ischemic lesion, using an image analyzer (eg, the Quantimet 720). Volumes and ischemic areas are expressed in absolute units (mm3 and mm2) and as a percentage of the total region examined. The effect of the compositions and methods of this invention for reducing ischemic brain injuries induced by MCA obstruction is detected based on a reduction in the area or volume of relative or absolute ischemic injury in the cerebral sections of the rats in the group of treatment compared to the cerebral sections of the rats of the control group treated with placebo. Other methods that could be used as an alternative to demonstrate the beneficial effect of the invention for reducing ischemic brain injuries include those described by Nakayama, et al., In Neurology 1988; 38: 1667-1673, Memezawa, et al. in Stroke 1992; 23: 552-559; Folbergrova, et al. in Proc. Nati Acad. Sci 1995; 92: 5057-5059, and Gotti, et al. in Brain Res. 1990; 522: 290-307. The beneficial effect of the compositions and methods of this invention for reducing ischemic liver injury can be demonstrated, for example, in mammals, using the method of Yokoyama et al. (Am. J. Physiol., 1990; 258: G564-G570). In summary, in the procedure of Yokoyama et al., Adult male Sprague-Dawley rats are anesthetized with sodium pentobarbital (40 mg / kg, i.p.), then the animals are tracheotomized and mechanically ventilated with ambient air. The liver is removed and placed in an environmental chamber maintained at a constant temperature (37 ° C), then perfused through the portal vein at a constant pressure of 15 cm H20 with a modified Krebs-Henseleit buffer, without hemoglobin, (in mM: NaCl 1 18, KCl 4.7, NaHCO3 27, CaCl2 2.5, MgSO4 1.2, KH2P04 1.2, EDTA 0.05 and glucose 1 1 mM, plus 300 U of heparin). The pH of the perfusate is maintained at 7.4 gassing the buffer with 95% 02-5% C02. Each liver is perfused at a flow rate of 20 ml / min in a one-step manner, during a wash and balance period of 30 minutes (pre-ischemic period), followed by a period of 2 hours of global ischemia and, then, for a period of 2 hours of reperfusion under conditions identical to those of the pre-ischemic period. Aliquots (20 ml) are collected from the perfusate during the preischemic period, immediately after the obstructive ischemic period and every 30 minutes of the reperfusion period of 2 hours. Perfusate samples are analyzed for the presence of hepatocellular enzymes, for example, aspartate amino-transferase (AST), alanine amino-transferase (ALT) and lactate dehydrogenase (LDH), which are taken to reflect quantitatively the degree of injury ischemic tissue of the liver during the procedure. The AST, ALT and LDH activities of the perfusate can be determined by various methods, for example, by the reflectometry method, using a Kodak Ektachem 500 automatic analyzer presented by Nakano, et al. (Hepatology 1995; 22: 539-545).

The effect of the compositions and methods of this invention for reducing ischemic liver injury induced by obstruction is detected based on the reduction of hepatocellular enzyme release immediately after the period of obstruction and / or during the reperfusion period after ischemia, in the livers subjected to perfusion of the rats of the treatment group compared to the livers perfused in the rats of the control group treated with placebo. Other methods and parameters that could be used as an alternative to demonstrate the beneficial effect of the compositions and methods of this invention for reducing ischemic liver injury, include those described by Nakano et al. (Hepatology 1995; 22: 539-545). Administration of the compounds of this invention can be performed by any method that releases a compound of this invention, preferably in the desired tissue (e.g., liver and heart tissues). These procedures include oral, parenteral, intraduodenal, etc. routes. Generally, the compounds of the present invention are administered in a single dose (for example once a day) or in multiple doses. The combinations of this invention are useful for reducing or minimizing lesions appearing directly in any tissue that may be susceptible to ischemia / reperfusion injury (eg, heart, brain, lung, kidney, liver, intestine, skeletal muscle or retina). ) as a result of ischemia (for example, myocardial infarction).

Therefore, the active compounds are used prophylactically useful to prevent, ie (prospectively or prophylactically) to prevent or arrest, tissue damage (e.g., in myocardial tissue) in patients at risk of ischemia (e.g., ischemia) of myocardium). Generally, the compounds of this invention are administered orally, but parenteral administration (eg, intravenous, intramuscular, subcutaneous or intramedullary) can be used, for example, when oral administration is unsuitable for the present purpose or when the patient You can not ingest the drug. Topical administration may also be indicated, for example, when the patient is suffering from gastrointestinal disorders or when the medication is best applied to the surface of a tissue or organ, as determined by the corresponding physician. The two different compounds of this invention may be co-administered simultaneously or sequentially in any order, or a single pharmaceutical composition comprising an aldose reductase inhibitor, as described above, and a glycogen phosphorylase inhibitor, may be administered as described above. described above, in a pharmaceutically acceptable vehicle. In any case, the amount and duration of the administered compounds, of course, will be dependent on the subject to be treated, the severity of the affliction, the manner of administration and / or the criterion of the corresponding physician. Thus, due to the variability of one patient to another, the dosages given below are a guideline and the doctor can assess doses of the compounds to achieve the treatment (for example, an activity of reducing glucose levels or insulin levels) that is considered appropriate for the patient. Considering the degree of the desired treatment, the doctor has to balance a variety of factors such as the age of the patient, the presence of a pre-existing disease, as well as the presence of other diseases (for example, cardiovascular diseases). Thus, for example, in a mode of administration, the combination of this invention can be administered just before cardiac surgery (eg, in the 24 hours prior to surgery) when there is a risk of myocardial ischemia. In an exemplary alternative mode, the compounds can be administered after cardiac surgery (for example, within 24 hours after surgery), when there is a risk of myocardial ischemia. The compounds of this invention can also be administered in a chronic daily manner. In general, an amount of a combination of this invention is used which is sufficient to achieve an appropriate insulin sensitization effect. Alternatively, an amount of a combination of this invention is used which is sufficient to achieve the normal biological actions of insulin at "normal concentration" which would be evidenced by the maintenance of euglycemia, normoglycemia and normal lipidemia (e.g. triglycerides, cholesterol and free fatty acids) as well as normal and normotensive glucose tolerance. An amount of the combination that is effective for the protection of ischemia is also used. An amount of the aldose reductase inhibitor of this invention is used which is effective for the activities of this invention, for example, triglyceride and cholesterol reduction activities and the reversal activities of hyperinsulinemia. Typically, an effective dose for the aldose reductase inhibitors of this invention is in the range of about 0.1 mg / kg / day to 100 mg / kg / day, in a single dose or in divided doses, preferably 0.1 mg / kg / day at 20 mg / kg / day in a single dose or in divided doses. In general, an effective dosage for the activities of this invention, for example, the activities of reducing glucose, triglycerides, free fatty acids and blood cholesterol, and the reversal activities of hyperinsulinemia, glycogen inhibitor compounds Phosphorylase of this invention is in the range of 0.005 to 50 mg / kg / day, preferably 0.01 to 25 mg / kg / day and, even more preferably, 0.1 to 15 mg / kg / day. The compounds of the present invention are generally administered in the form of a pharmaceutical composition comprising at least one of the compounds of this invention together with a pharmaceutically acceptable diluent carrier. Thus, the compounds of this invention can be administered individually or in combination in any conventional oral, parenteral, rectal or transdermal dosage form. For oral administration, a pharmaceutical composition can take the form of solutions, suspensions, tablets, pills, capsules, powders and the like. Tablets containing various excipients such as sodium citrate, calcium carbonate and calcium phosphate are used together with various disintegrants such as starch and preferably potato or tapioca starch, and certain complex silicates, together with binding agents such as polyvinylpyrrolidone, sucrose, gelatin and gum arabic. In addition, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for forming tablets. Solid compositions of a similar type are also employed as fillers in soft and hard gelatin capsules; Preferred materials in this regard also include lactose or milk sugar, as well as high molecular weight polyethylene glycols. When aqueous suspensions and / or elixirs are desired for oral administration, the compounds of this invention can be combined with various sweetening agents, flavoring agents, coloring agents, emulsifying agents and / or suspending agents, as well as diluents such as water, ethanol, propylene glycol, glycerin and various combinations thereof. For the purposes of parenteral administration, solutions in sesame or peanut oil or in aqueous propylene glycol, as well as sterile aqueous solutions of the corresponding water-soluble salts may be employed. Such aqueous solutions may conveniently be buffered, if necessary, and the liquid diluent may first be made isotonic with sufficient saline or glucose. These aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal injection purposes. In this regard, all of the sterile aqueous media employed can be readily obtained by conventional techniques well known to those skilled in the art. For the purposes of transdermal administration (eg, topical), dilute, aqueous or partially aqueous solutions are prepared (normally at a concentration of about 0.1% to 5%), otherwise similar to the parenteral solutions above. Methods for preparing various pharmaceutical compositions with a certain amount of active ingredient are known or will become apparent in light of this description, for those skilled in the art, for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975), for examples of procedures for preparing pharmaceutical compositions. The pharmaceutical compositions according to this invention may contain from 0.1% to 95% of the compound (s) of this invention, preferably from 1% to 70%. In any case, the composition or formulation to be administered will contain an amount of compound (s) according to the invention effective to treat the condition / disease of the subject being treated.

As the present invention has an aspect that relates to the treatment of, for example, an insulin resistant condition, by treatment with a combination of active ingredients that can be administered separately, the invention also relates to a combination of the pharmaceutical compositions separated in the form of equipment. The kit includes two separate pharmaceutical compositions: an aldose reductase inhibitor and an inhibitor of glycogen phosphorylase, as described above. The equipment comprises a container for containing the separate compositions, such as a divided bottle or a divided container with a sheet. Typically, the equipment includes instructions for the administration of the separate components. The form of the kit is particularly advantageous when the separate components are preferably administered in different dosage forms (eg, oral and parenteral), when administered at different dosage intervals, or when the corresponding physician wishes to assess the individual components of the combination . An example of such equipment is the so-called blister. Blisters are well known in the packaging industry and are widely used for the packaging of pharmaceutical unit dosage forms (tablets, capsules and the like). Blisters generally consist of a sheet of relatively rigid material covered with a sheet of preferably transparent plastic material. During the packaging process, cavities are formed in the plastic sheet. The cavities have the size and shape of the tablets or capsules to be packaged. Then, the tablets or capsules are placed in the cavities and the sheet of relatively rigid material is sealed with the plastic sheet on the side of the sheet opposite the direction in which the cavities were formed. As a result, the tablets or capsules are sealed in the cavities between the plastic sheet and the sheet. Preferably, the strength of the sheet is such that the tablets or capsules can be removed from the blister by manually pressing on the cavity, whereby an opening is formed in the sheet at the cavity site. The tablet or capsule can then be removed through said opening. It may be desirable to provide a reminder on the equipment, for example, in the form of numbers near the tablets or capsules, the numbers corresponding to the days of the regimen in which the tablets or capsules so specified should be ingested. Another example of such a reminder is a calendar printed on the card, for example, as indicated below: "First week, Monday, Tuesday, ... etc Second week, Monday, Tuesday ..." etc. Other variations of reminders will be evident. A "daily dose" may be a single tablet or capsule, or several tablets or capsules to be taken on a given day. In addition, a daily dose of the first compound may consist of a tablet or capsule, while a daily dose of the second compound may consist of several tablets or capsules and vice versa. The reminder should reflect this.

In another specific embodiment of this invention, a dispenser designed to deliver the daily doses, one at a time, is provided in relation to the intended use. Preferably, the dispenser has a reminder, so that it further facilitates compliance with the regime. An example of such a reminder is a mechanical counter that indicates the number of daily doses that have been delivered. Another example of such a reminder is a battery-powered microchip memory, coupled to a liquid crystal display, or a reminder auditory signal that, for example, shows the data of the last daily dose taken and / or remembers when it should be taken. the next dose. The compounds of this invention either alone or in combination with each other or with other compounds, will generally be administered in a convenient formulation. The following formulation examples are illustrative only and are not intended to limit the scope of the present invention. In the formulations shown below, "active ingredient" refers to compounds (s) of this invention and, therefore, may refer to an aldose reductase inhibitor, an inhibitor of glycogen phosphorylase or a combination of the two.

Formulation 1: Gelatin capsules Hard gelatin capsules are prepared using the following: Ingredient Quantity (mg / capsule) Active ingredient 0.25-100 Starch, NF 0-650 Fluid starch powder 0-50 Silicone fluid 350 centistokes 0-15 A tablet formulation is prepared using the following ingredients: Ingredient Quantity (mg / tablet) Active ingredient 0.25-100 Cellulose, microcrystalline 200-650 Silicon dioxide, pyrolyzed 10-650 Stearic acid 5-15 The components are mixed and pressed to form tablets. As an alternative, tablets are made each containing 0.25-100 mg of active ingredients, as indicated below: Formulation 3: Tablets Ingredient Quantity (mg / tablet) Active ingredient 0.25-100 Starch 45 Cellulose, microcrystalline 35 Polyvinylpyrrolidone (in the form of a 4% solution in water) Carboxymethylcellulose sodium 4.5 Magnesium stearate 0.5 Talcum 1 The active ingredients, starch and cellulose are passed through an E.U.A. No. 45 and they mix thoroughly. The solution of polyvinylpyrrolidone is mixed with the resulting powders and the mixture is then passed through an E.U.A. No. 14. The granules thus produced are dried at 50 ° C-60 ° C and passed through an E.U.A. mesh screen. No. 18. Next, the sodium carboxymethyl starch, the magnesium stearate and the talcum, previously passed through an E.U.A. No. 60, are added to granules which, after mixing, are pressed into a tablet machine to produce tablets. Suspensions each containing 0.25-100 mg of active ingredient per 5 ml dose are manufactured as follows: Formulation 4: Suspensions Ingredient Quantity (mg / 5 ml) Active ingredient 0.25-100 mg Carboxymethromellose sodium 50 Syrup 1.25 mg Benzoic acid solution 0.10 ml Flavoring agent c.v. Colorant c.v. Purified water, up to 5 ml The active ingredient is passed through a No. 45 mesh screen and mixed with the sodium carboxymethylcellulose and the syrup to form a uniform paste. The benzoic acid, flavoring and coloring solution is diluted with some water and added, with stirring. After adding enough water to produce the necessary volume. An aerosol solution is prepared containing the following ingredients: Formulation 5: Aerosol Ingredient Quantity (% by weight) Active ingredient 0.25 Ethanol 25.75 Propellant 22 (chlorodifluoromethane) 70.00 The active ingredient is mixed with ethanol and the mixture is added to a portion of the propellant 22, cooled to 30 ° C and transferred to a refilling device. The necessary amount is then introduced into a stainless steel container and diluted with the rest of the propellant. Then the valve units in the container are adjusted. The suppositories are manufactured as follows: Formulation 6: Suppositories Ingredient Quantity (mg / suppository) Active ingredient 250 Saturated fatty acid glycerides 2,000 The active ingredient is passed through a mesh screen E.U.A. No. 60 and is suspended in the glycerides of saturated fatty acids, previously melted using the minimum necessary heat. The mixture is then poured into a suppository mold with a nominal capacity of 2 g and allowed to cool. An intravenous formulation is prepared as follows: Formulation 7: Intravenous solution Ingredient Quantity Active ingredient 20 mg Saline saline solution 1,000 ml The solution of the above ingredients is administered intravenously to a patient at a rate of about 1 ml per minute. The above active ingredient can also be a combination of agents.

Claims (3)

1. NOVELTY OF THE INVENTION CLAIMS 1. - A pharmaceutical composition comprising a therapeutically effective amount of: a) an aldose reductase inhibitor; b) an inhibitor of glycogen phosphorylase; and c) a pharmaceutical vehicle.
2. 2. A pharmaceutical composition as recited in claim 1, wherein the inhibitor of aldose reductase is 1-phthalazineacetic acid, 3,4-dihydro-4-oxo-3 [[5- (trifluoromethyl) -2 -benzothiazolyl] methyl] - or a pharmaceutically acceptable salt thereof.
3. 3. A pharmaceutical composition as recited in claim 2, wherein the glycogen phosphorylase inhibitor is [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyrrolidin-1-yl) 5-chloro-1 H-indole-2-carboxylic acid) -3-oxopropyl] -amide; [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1-yl) - 5-chloro-1 H-indole-2 (2R) -hydroxy-3-oxopropyl] -amide -carboxylic; [(1 S) - ((R) -hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy- (methoxy-methyl-carbamoyl) -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R-hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl] -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole 2-carboxylic; [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid (2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; [(1 S) -benzyl- 5 - Chloro-1 H-indole-2-carboxylic acid 3 - ((cis) -dihydroxypyrrolidin-1 -yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1 S) -benzyl) 2-Chloro-1 H-indole-2-carboxylic acid (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-etl] -amide; [2- (1, 5-Chloro-1 H-indole-2-carboxylic acid 1-dioxo-thiazolidin-3-yl) -2-oxo-ethyl] -amide; [(1S) - (4-phenyl-benzyl) -2- ( 5-Chloro-1 H-indole-2-carboxylic acid 4-hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide; [(1 S) -benzyl-2 - ((3RS) hydroxy) 5-Chloro-1 H-indole-2-carboxylic acid piperidin-1-yl) -2-oxo-ethyl] -amide; [2-oxo-2 - ((1-RS) -oxo-1-thiazolidin-3) 5-Chloro-1 H-indole-2-carboxylic acid amyl; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo- ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid 4.- A pharmaceutical composition as recited in claim 3, wherein the amount of the inhibitor of the aldose reductase is from about 0.1 mg / kg to about 20 mg / kg. 5. - A pharmaceutical composition as recited in claim 4, wherein the amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. 6. A method for treating a mammal having an insulin resistant condition, comprising administering to said mammal a therapeutically effective amount of a) an aldose reductase inhibitor; and b) an inhibitor of glycogen phosphorylase. 7. A method as claimed in claim 6, wherein the insulin-resistant condition is diabetes, hyperinsulinemia, glucose intolerance, hypergiukaemia and / or hyperlipidemia after meals, type II diabetes, alteration of body composition , reduction of lean body mass, obesity, hypertension, dyslipidemia, atherosclerosis, tissue ischemia, cardiovascular diseases, obesity, syndrome X, pregnancy, infection states, uremia, hyperandrogenism, hypercortisolemia or other adrenocortical hormone excess states, acromegaly, excess of growth hormone or polycystic ovarian disease. 8. A method as claimed in claim 6, wherein the insulin resistant condition is diabetes. 9. A method as recited in claim 8, wherein the aldose reductase inhibitor is 3,4-dihydro-4-oxo-3 - [[5- (trifluoromethyl) -2-benzothiazolyl] methyl ] -1-phthalazinoacetic acid, or a pharmaceutically acceptable salt thereof. 10. - A method as recited in claim 9, wherein the glycogen phosphorylase inhibitor is [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyridin-1-yl) -3 5-chloro-1 H-indole-2-carboxylic acid -oxopropyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1 -yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1 S) - ((R) -hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy- (methoxy-methyl-carbamoyl) -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indol-2 acid -carboxylic; [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; [(1 S) -benzyl-2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [2- (1, 1-dioxo-thiazolidin-3-yl) -2-oxo-etiI] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) -benzyl-3 - ((cis) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - (4-fluoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-acid carboxylic acid, [(1 S) -benzyl-2 - ((3RS) -hydroxy] -piperidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2- carboxylic; 5-Chloro-1 H-indole-2-carboxylic acid [2-oxo-2 - ((1-RS) -oxo-1-thiazolidin-3-yl) -ethyl] -amide; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid. 1 1.- A procedure as mentioned in the claim 10, wherein the amount of aldose reductase inhibitor is from about 0.1 mg / kg to about 20 mg / kg and the amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. 12. A procedure as mentioned in the claim 11, in which the mammal is a man or a woman. 13. A method as claimed in claim 6, wherein the insulin-resistant condition is obesity. 14. A method as recited in claim 6, wherein the insulin-resistant condition is polycystic ovary disease. 15. - A method as recited in claim 6, wherein the insulin resistant condition is syndrome X. 16. A method as recited in claim 6, wherein the insulin resistant condition is hypertension. 17.- A team that understands; a) a therapeutically effective amount of an aldose reductase inhibitor and a pharmaceutically acceptable carrier in a first unit dosage form; b) a therapeutically effective amount of a glycogen phosphorylase inhibitor and a pharmaceutically acceptable carrier in a second unit dosage form; and c) a container means for containing said first and second dosage form. 18. A kit as claimed in claim 17, wherein the inhibitor of aldose reductase is 1-phthalazinoacetic acid, 3,4-dihydro-4-oxo-3 - [[5- (trifluoromethyl) -2 -benzothiazole] methyl] - or a pharmaceutically acceptable salt thereof. 19. A kit as claimed in claim 18, wherein the inhibitor of glycogen phosphorylase is [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyrrolidin-1-yl) 5-chloro-1 H-indole-2-carboxylic acid-3-oxopropyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1 -yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1S) - ((R) -hydroxy-dimethylcarbamoylmethyl) -2-phenol-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy- (methoxy-methyl-carbamoyl) -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1S) - ((R) -hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2- carboxylic; [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2- (1, 1-dioxo-thiazolidin-3-yl) -2-oxo-ethyl] -amide; [(1 S) -benzyl-3 - ((cis) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - 5-chloro-1 H-indole-2-carboxylic acid (4-fluoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide , [(1 S) -benzyl-2 - ((3RS) -hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2-oxo-2 - ((1-RS) -oxo-1-thiazolidin-3-yl) -ethyl] -amide; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid. 20. A kit as claimed in claim 19, wherein the amount of aldose reductase inhibitor is from about 0.1 mg / kg to about 20 mg / kg. 21. A kit as claimed in claim 20, wherein the amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. 22. A pharmaceutical composition for achieving an insulin sensitization effect in a mammal, comprising, a) an amount of a first compound, said first compound being an inhibitor of aldose reductase; and b) an amount of a second compound, said second compound being an inhibitor of glycogen phosphorylase in which the amount of the first compound alone and the amount of the second compound alone is insufficient to achieve the effect of insulin sensitization and in which the The combined effect of the amounts of the first and second compounds is greater than the sum of the effects of insulin sensitization achievable with the individual amounts of the first and second compounds, and a pharmaceutically acceptable diluent or carrier. 23. A pharmaceutical composition as recited in claim 1, wherein the inhibitor of aldose reductase is 3,4-dihydro-4-oxo-3 - [[5- (trifluoromethyl) -2-benzothiazolyl] methyl] -1-phthalazinoacetic acid, or a pharmaceutically acceptable salt thereof. 24. - A pharmaceutical composition as recited in claim 23, wherein the glycogen phosphorylase inhibitor is [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyrrolidin-1-yl) - 5-Chloro-1 H-indole-2-carboxylic acid 3-oxopropyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1 -yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1 S) - ((R) -hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy- (methoxy-methyl-carbamoyl) -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H- indole-2-carboxylic; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; [(1 S) -benzyl-2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-etl] -amide of 5-chloro-1 acid H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-3 - ((cis) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2- (1, 1-dioxo-thiazoiidin-3-yl) -2-oxo-etl] -amide; [(1 S) - 5-chloro-1 H-indole-2-carboxylic acid (4-fluoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide , [(1 S) -benzyl-2 - ((3RS) -hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2-oxo-2 - ((1-RS) -oxo-1-thiazoidin-3-yl) -ethyl] -amide; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid. 25. A pharmaceutical composition as recited in claim 24, wherein the amount of aldose reductase inhibitor is from about 0.1 mg / kg to about 20 mg / kg. 26. A pharmaceutical composition as recited in claim 25, wherein the amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. 27. A method for achieving an insulin sensitization effect in a mammal having an insulin resistant condition, comprising administering to said mammal, a) an amount of a first compound, said first compound being an inhibitor of aldose reductase; and b) an amount of a second compound, said second compound being an inhibitor of glycogen phosphorylase, wherein the amount of the first compound alone and the amount of the second compound alone is insufficient to achieve the effect of insulin sensitization and in which the combined effect of the amounts of the first and second compounds is greater than the sum of the insulin sensitization effects that can be achieved with the individual amounts of the first and second compounds. 28.- A procedure as mentioned in the claim 27, wherein the inhibitor of aldose reductase is 3,4-dihydro-4-oxo-3 - [[5- (trifluoromethyl) -2-benzothiazolyl] methyl] -1-phthalazinoacetic acid, or a salt pharmaceutically acceptable thereof. 29. A procedure as mentioned in the claim 28, wherein the glycogen phosphorylase inhibitor is [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyrrolidin-1-yl) -3-oxopropyl] -amide 5- chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1 -yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1 S) - ((R) -hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1S) - ((R) -hydroxy-methoxy-methyl-carbamoyl) -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indol-2 acid -carboxylic; [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-etl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2- (1, 1-dioxo-thiazolidin-3-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indoi-2-carboxylic acid [(1 S) -benzyl-3 - ((cis) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1 S) - (4-Fluoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole -2-carboxylic; 5-Chloro-1 H-indole-2-carboxylic acid [(1 S) -benzyl-2 - ((3RS) -hydroxy-piperidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2-oxo-2 - ((1-RS) -oxo-1-thiazolidin-3-yl) -ethyl] -amide; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-ethyl] -amide of 5-cyclo-1 H-indole-2-carboxylic acid. 30. A process as recited in claim 29, wherein the amount of aldose reductase inhibitor is from about 0.1 mg / kg to about 20 mg / kg. 31. - A method as recited in claim 30, wherein the amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. 32. A method as claimed in claim 31, wherein the mammal is a man or a woman. 33.- A method for reducing tissue lesions resulting from ischemia, which comprises administering to a mammal in need of such treatment, a therapeutically effective amount of, a) an aldose reductase inhibitor; and b) an inhibitor of glycogen phosphorylase. 34.- A procedure as mentioned in the claim 33, in which the tissue is cardiac, cerebral, hepatic, renal, pulmonary, intestinal, skeletal muscle, splenic, pancreatic, nervous, spinal, retinal, vasculature or intestinal tissue. 35. A process as recited in claim 34, wherein the inhibitor of aldose reductase is 3,4-dihydro-4-oxo-3 - [[5- (trifluoromethyl) -2-benzothiazolyl] methyl ] -1-phthalazinoacetic acid, or a pharmaceutically acceptable salt thereof. 36.- A method as recited in claim 35, wherein the glycogen phosphorylase inhibitor is [(1 S) -benzyl- (2R) -hydroxy-3 - ((3S) -hydroxypyrrolidin-1-yl) 5-chloro-1 H-indole-2-carboxylic acid-3-oxopropyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid ((1 S) -benzyl-3 - ((3S, 4S) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide; [(1 S) - ((R) -hydroxy-dimethylcarbamoyl-methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy-methoxy-methyl-carbamoyl) -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - ((R) -hydroxy - [(2-hydroxy-ethyl) -methyl-carbamoyl] -methyl) -2-phenyl-ethyl] -amide of 5-chloro-1 H-indol-2 acid -carboxylic; [(1 S) -benzyl-2- (3-hydroxyimino-pyrrolidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2 - ((3S, 4S) -dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [(1S) -benzyl-2- (cis-3,4-dihydroxy-pyrrolidin-1-yl) -2-oxo-ethyl] -amide; 5-Chloro-1 H-indole-2-carboxylic acid [2- (1, 1-dioxo-thiazolidin-3-yl) -2-oxo-ethyl] -amide; [(1 S) -benzyl-3 - ((cis) -dihydroxypyrrolidin-1-yl) - (2R) -hydroxy-3-oxopropyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; [(1 S) - (4-fluoro-benzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxo-etl] -amide of 5-chloro-1 H-indole-2-acid -carboxylic; [(1 S) -benzyl-2 - ((3RS) -hydroxy-p -peridin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid; 5-Chloro-1 H-indole-2-carboxylic acid [2-oxo-2 - ((1-RS) -oxo-1-thiazolidin-3-yl) -ethyl] -amide; or [(1 S) -benzyl-2- (3-hydroxy-azetidin-1-yl) -2-oxo-ethyl] -amide of 5-chloro-1 H-indole-2-carboxylic acid. 37.- A procedure as mentioned in the claim 36, wherein the amount of aldose reductase inhibitor is from about 0.1 mg / kg to about 20 mg / kg and the amount of glycogen phosphorylase inhibitor is from about 0.1 mg / kg to about 15 mg / kg. 38.- A procedure as mentioned in the claim 37, in which the mammal is a man or a woman. 39.- A procedure as mentioned in the claim 38, wherein said tissue is cardiac tissue. 40.- A method as recited in claim 38, wherein said tissue is brain tissue. 41. A method as recited in claim 38, wherein said tissue is liver tissue. 42. A method as recited in claim 38, wherein said tissue is renal tissue. 43.- A procedure as mentioned in the claim 38, wherein said tissue is lung tissue. 44. A method as recited in claim 38, wherein said tissue is intestinal tissue. 45. - A method as recited in claim 38, wherein said tissue is skeletal muscle tissue. 46. A method as recited in claim 38, wherein said tissue is splenic tissue. 47.- A procedure as mentioned in the claim 38, wherein said tissue is pancreatic tissue. 48. A method as recited in claim 38, wherein said tissue is retinal tissue. 49.- A method as recited in claim 39, wherein said combination is administered prophylactically. 50.- A method as recited in claim 38, wherein said combination is administered before cardiac surgery. 51. A method as recited in claim 33, wherein the tissue injury resulting from ischemia is an ischemic injury that occurs during organ transplantation. 52.- A method for treating a mammal having an insulin resistant condition, comprising administering to said mammal the composition mentioned in claim 1. 53.- A method for reducing tissue lesions resulting from ischemia, comprising administering to a mammal in need of such treatment the composition mentioned in claim 1.