RU2800510C1 - C-aryl glucose sglt2 inhibitors and method of their use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a pharmaceutical composition having an inhibitory effect on SGLT2. This composition includes a compound of formula

and a pharmaceutically acceptable vehicle.

EFFECT: obtaining a pharmaceutical composition capable to inhibit sodium-dependent glucose transporters found in the intestines and kidneys of mammals and being of value for the treatment of diabetes mellitus and its micro- and macrovascular complications.

1 cl, 3 tbl, 1 ex

Description

Technical field

The present invention relates to C-aryl glucosides which are inhibitors of sodium-dependent glucose transporters found in the intestine and kidneys (SGLT2), as well as to a method for the treatment of diabetes mellitus, predominantly type 2 diabetes, hyperglycemia, hyperinsulinemia, obesity, hypertriglyceridemia , Syndrome X, complications of diabetes mellitus, atherosclerosis and related conditions, consisting in the use of such C-aryl glucosides both as monotherapy and in combination with one, two or more antidiabetic drugs and in combination with one, two or more other drugs such as antihyperlipidemic drugs.

Prerequisites for the creation of the invention

Approximately 100 million people worldwide suffer from type 2 diabetes mellitus (NIDDM), which is characterized by hyperglycemia resulting from excessive hepatic glucose production and peripheral tissue insulin resistance, the cause of which is still unknown. Hyperglycemia is considered a major risk factor for the development of complications of diabetes mellitus, and probably contributes directly to the deterioration in insulin secretion seen in advanced type 2 diabetes. Normalization of plasma glucose levels in NIDDM patients will improve insulin secretion and delay the development of complications of diabetes mellitus.

Presumably, inhibition of sodium-dependent glucose transporters SGLT2 in the kidneys will help normalize plasma glucose levels, and possibly also normalize body weight by increasing glucose excretion.

The development of new safe and oral antidiabetic agents is also desirable to complement existing therapies based on the use of drugs such as sulfonylurea derivatives, metformin and insulin, as well as to reduce the possible side effects that occur with the use of other indicated hypoglycemic drugs.

Hyperglycemia is a marker of type 2 diabetes mellitus (NIDDM); stable control of plasma glucose levels can slow down the complications and destruction of beta-cells, which is observed in advanced type 2 diabetes mellitus. Normally, plasma glucose is filtered in the renal glomeruli and actively reabsorbed in the proximal renal tubules. It is believed that the main transporters responsible for glucose reabsorption in the proximal renal tubules are SGLT2. The use of a specific SGDT inhibitor, phlorizin or its analogs, in rodents and dogs - models of diabetes - led to the normalization of blood plasma glucose levels by increasing urinary glucose excretion without the development of side effects associated with hypoglycemia. Long-term (for 6 months) treatment of Zucker rats - models of diabetes - with an SGLT2 inhibitor led to an increase in insulin response to glycemia, improved tissue sensitivity to insulin, and prevented the development of nephropathy and neuropathy in these animals, while the absence of any pathological changes in kidneys and electrolyte imbalance in blood plasma. It is assumed that selective inhibition of SGrT2 in patients with diabetes mellitus will normalize the level of glucose in blood plasma by increasing its excretion in the urine, as well as improve tissue sensitivity to insulin and delay the development of complications.

Ninety percent of all glucose reabsorbed in the kidney is reabsorbed in the epithelial cells lining the initial S1 segment of the renal cortical proximal tubules, and it is likely that SGLT2 is the primary transporter responsible for this reabsorption. SGDT2 is a 672 amino acid protein with 14 overlapping membrane segments that is predominantly expressed in the initial S1 segment of the proximal renal tubule. The substrate specificity, sodium ion dependence, and localization of SGLT2 are combined with their high capacity and low affinity properties. In addition, hybrid depletion studies have shown that SGLT2s are the predominant sodium-dependent glucose transporters in the S1 segment of the proximal tubule, since virtually all of the sodium-dependent glucose transport activity determined by mRNA isolated from rat kidney cortex is inhibited by the antisense oligonucleotide specific for rat SGLT2. It is possible that the SGLT2 gene is responsible for some forms of familial glucosuria, a genetically determined disease in which the reabsorption of glucose in the kidneys is impaired in various ways. None of these syndromes found to date are associated with the 36bT2 locus on chromosome 16. However, studies of highly homologous rodent SGLT2s clearly suggest that SGLT2s are the major sodium-dependent glucose transporters in the kidneys and suggest that mapped the glucosuria locus encodes the SGLT2 regulator. It can be assumed that inhibition of SGLT2 will reduce the level of glucose in blood plasma by increasing its excretion in patients with diabetes mellitus.

SGLT1, another sodium-dependent glucose transporter that is 60% identical in amino acid sequence to SGLT2, is expressed in the small intestine and the more distal S3 segment of the proximal renal tubule. Despite the similarity in amino acid sequences, SGLT1 and SGLT2 differ in their biochemical properties. For SGLT1, the molar ratio between Na ⁺ ions and transported glucose is 2:1, while for SGLT2 it is 1:1. Km for Na ⁺ ions is 32 and 250-300 mM for SGLT1 and SGLT2, respectively. The Km values for glucose uptake and the non-metabolizable glucose analogue α-methyl-D-glucopyranoside (AMG) are similar for SGLT1 and SGLT2, i.e. 0.8 and 1.6 mM (glucose) and 0.4 and 1.6 mM (AMG) for SGLT1 and SGLT2 transporters, respectively . However, the two transporters differ significantly in their substrate specificity for sugars such as galactose, which is a substrate only for SGLT1.

The use of phlorizin, a specific inhibitor of SGLT activity, allowed in vivo validation of this concept, as it resulted in increased glucose excretion, decreased fasting and postprandial plasma glucose levels, and increased glucose utilization without potential hypoglycemic side effects in several rodents and one dog - models of diabetes. Such possible consequences of the use of Florizin for two weeks, such as ion imbalance in blood plasma, impaired renal function and changes in the normal structure of the renal tissue, were not noted. In addition, hypoglycemia and other side effects were not observed even when using phlorizin in healthy animals, despite the presence of glucosuria. The use of a renal SGLT inhibitor for 6 months resulted in the normalization of fasting and postprandial plasma glucose levels, improved insulin secretion and increased glucose utilization, and also prevented the development of nephropathy and neuropathy in rat models of diabetes mellitus and obesity, while the absence of hypoglycemia and pathological kidney changes.

Florizin is not suitable for oral use because it is a non-specific SGLT1/SGLT2 inhibitor that undergoes hydrolysis in the gut to form the aglycone phloretin, which is a potent inhibitor of facilitated glucose transporters. Competitive inhibition of facilitated glucose transporters (GLUTs) is undesirable because it can lead to increased insulin resistance in peripheral tissues, as well as promote hyperglycemia in the central nervous system. Inhibition of SGLT1 can also have serious consequences, which can be traced to the hereditary syndrome of glucose-galactose malabsorption (GGM), in which mutation of the SGLT1 transporter leads to a decrease in glucose uptake in the intestine and the development of life-threatening diarrhea and dehydration. The different biochemical properties of SGLT1 and SGLT2, as well as the percentage of mismatches in their amino acid sequences, allow the identification of selective inhibitors of SGLT2.

Familial glucosuria syndromes are conditions in which the intestinal transport of glucose as well as the renal transport of other ions and amino acids is normal. Patients suffering from familial glucosuria develop without abnormalities, have normal plasma glucose levels and do not suffer from any serious health problems, despite sometimes quite high values of excreted glucose (110-140 g / day). The main symptoms observed in such patients are polyphagia, polyuria and polydipsia; while their kidneys function normally and have no morphological changes. Thus, from the available data, it can be concluded that impaired renal glucose reabsorption by the kidneys appears to have minimal long-term adverse effects in otherwise normal individuals.

The following references refer to C-aryl glucoside SGLT2 inhibitors for the treatment of diabetes mellitus.

WO 01/27128 refers to compounds having the following structure:

where A represents O, S, NH or (CH ₂ ) _n where n is 0-3; R ¹ , R ² , and R ^2a are independently hydrogen, OH, OR ⁵ , alkyl, CF ₃ , OCHF ₂ , OCF ₃ , SR ⁵ⁱ or halogen, and so on;

R ³ and R ⁴ are independently hydrogen, OH, OR ^5a , OAryl, OCH ₂ Aryl, alkyl, cycloalkyl, CF ₃ , -OCHF ₂ , OCF ₃ , halogen, etc. It is believed that these compounds are inhibitors of SGLT2 transporters and, therefore, may be agents for the treatment of diabetes mellitus and its complications.

WO 98/31697 refers to compounds having the following structure:

where Ar is, among other things, phenyl, biphenyl, diphenylmethane, diphenylethane and diphenylether, R ¹ is a glycoside, R ² is H, OH, amino, halogen, carboxy, alkyl, cycloalkyl or carboxamido and R ³ is halogen , alkyl, or acyl, and k, type are independently 1-4. A subset of the compounds described in WO 98/31697 includes compounds having the following structure:

where A is O or (CH ₂ ) _x where x=0-3

R ₃ is hydrogen, an alkyl or acyl group, where n=1-4

R ² is hydrogen, alkyl, OH, NH, halogen, CO ₂ H or carboximide, where k=1-4

These compounds are considered as agents for the treatment and prevention of inflammatory diseases, autoimmune diseases, infections, cancer and cancer metastases, reperfusion diseases, thrombosis, ulcers, wounds, osteoporosis, diabetes mellitus and atherosclerosis, etc.

Description of the invention

The present invention relates to C-aryl glucosidic compounds having the formula I:

as well as their pharmaceutically acceptable salts, all their possible stereoisomers and prodrug esters of these compounds.

The compound of formula I has the ability to inhibit sodium-dependent glucose transporters found in the intestines and kidneys of mammals and is of value in the treatment of diabetes mellitus and its micro- and macrovascular complications such as retinopathy, neuropathy, nephropathy, as well as in the treatment of wounds.

The present invention relates to a compound of formula I, pharmaceutical compositions comprising this compound, and methods of using the same.

In addition, the present invention relates to a method for treating and slowing the progression or onset of diabetes, preferably type 1 and type 2 diabetes, including complications of diabetes such as retinopathy, neuropathy, nephropathy; to a method for treating wounds, as well as to a method for treating concomitant conditions such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, increased blood levels of fatty acids and glycerol, obesity, hyperlipidemia, including hypertriglyceridemia, Syndrome X, atherosclerosis and hypertension, and also to a method for increasing the level of high-density lipoproteins in the blood, which consists in applying to a person who needs treatment, a compound of formula I.

The present invention also relates to a method for the treatment of diabetes and associated conditions as described above, comprising administering to a person in need of treatment a therapeutically effective amount of a mixture of a compound of formula I and another type of antidiabetic agent and/or another drug, such as a lipid-lowering agent. means.

The conditions, diseases, and disorders collectively referred to as "Syndrome X" (also known as metabolic syndrome) are described in detail in Johannsson J. Clin. Endocrinol. Metab., 82, 121-34 (1991).

The term "other drugs" as used herein refers to one or more antidiabetic agents (not an SGLT2 inhibitor of formula I), one or more antiobesity agents, an antihypertensive agent, an antiplatelet agent, an antiatherosclerotic agent, and/or one or more lipid-lowering agents ( including anti-atherosclerotic agents).

According to the method described above, the compound of formula I, declared in accordance with the present invention, will be administered together with one or two or more antidiabetic agents and / or together with one, two or more other drugs (depending on the method of administration) , in ratios from 0.01:1 to 300:1, preferably from 0.1:1 to 10:1.

Detailed description of the invention

The compound of formula I, declared in accordance with the present invention, can be obtained according to the scheme below, and its description, while the temperature is expressed in centigrade values.

A compound of formula I can be prepared according to Scheme 1 by treating a compound of formula II

base, such as LiOH or NaOH, in the presence of a solvent, such as a mixture of H ₂ O/THF/MeOH in a ratio of 1:2:3 or aq. MeOH, or aq. EtOH.

The compound of formula II (which is a readily crystallizable new intermediate) provides the necessary conditions for the purification of the compound of formula Ia, obtained as a mixture of α- and β-anomers.

A compound of formula II can be prepared by treating compound Ia with Ac ₂ O in a solvent such as CH ₂ Cl ₂ containing pyridine and a catalyst such as dimethylaminopyridine (DMAP).

Compounds of formula Ia can be prepared by reducing a compound of formula III with a reducing agent such as Et ₃ SiH in a solvent such as CH ₂ Cl ₂ /MeCN 1:1 at -10° in the presence of a Lewis acid catalyst such as as BF ₃ ⋅Et ₂ O.

A compound of formula II may alternatively be prepared from a compound of formula III by acetylation of a compound of formula III using Ac ₂ O in a solvent such as toluene or CH ₂ Cl ₂ containing a base such as Hunig's base or Et ₃ N and a catalyst such as DMAP, to obtain a compound of formula IV.

Subsequent conversion of compound IV to compound II can be achieved by treating compound IV at 20° with a reducing agent such as Et ₃ SiH in a solvent such as MeOH containing 1 eq. H ₂ O and a Lewis acid catalyst such as BF ₃ ⋅ Et ₂ O.

Scheme 1

A compound of formula III can be prepared according to Scheme 2 by 1) adding a cold THF solution of an aryl lithium of formula V to persylated gluconolactone of formula VI in a solvent such as toluene at -75°. Then, after 30 minutes, a solution of a protic acid such as methanesulfonic acid (MSA) in methanol is added and the solution is stirred at 20° until complete transformation of the lactol intermediate into compound III.

A compound of formula VI can be prepared by treating commercially available D-gluconolactone with a silating agent such as trimethylsilyl chloride in a solvent such as THF containing a base such as N-methylmorpholine.

A compound of formula V can be prepared by treating a compound of formula VII with an alkyl lithium such as n-BuLi or t-Buli in a solvent such as THF at -75°.

A compound of formula VIII can be readily prepared by treating a compound of formula III with a reducing agent such as Et ₃ SiH in a solvent such as CH ₂ Cl ₃ /MeCN 1:1 at 0°-20° in the presence of a Lewis acid catalyst, such as BF ₃ ⋅Et ₂ O.

A compound of formula VIII can be prepared by a Friedel-Craft acylation reaction using commercially available ethoxybenzene (phenetol) and 2-chloro-5-bromobenzoyl chloride in a solvent such as CH ₂ Cl ₂ containing the equivalent of a Lewis acid catalyst such as AlCl ₃ or AlBr ₃ .

2-chloro-5-bromobenzoyl chloride can be easily prepared from commercially available 2-chloro-5-bromobenzoic acid by treating the latter with oxalyl chloride in a solvent such as CH ₂ Cl ₂ containing catalytic DMF.

Scheme 2

The following is a list of chemical compound abbreviations and terms used to characterize the present invention. These abbreviations correspond to the indicated compounds and terms throughout the text of the description (unless specifically stated otherwise) and can be used either alone or in combination with each other.

The following abbreviations are used to characterize the present invention:

Ph = phenyl

Bn = benzyl

t-Bu = tertiary butyl

Me = methyl

Et = ethyl

TMS = trimethylsilyl

TBS - tert-butyldimethylsilyl

THF = tetrahydrofuran

Et ₂ O = diethyl ether

EtOAc = ethyl acetate

DMF = dimethylformamide

MeOH = methanol

EtOH = ethanol

i-PrOH = isopropanol

HOAc or AcOH = acetic acid

TFA - trifluoroacetic acid

i-Pr ₂ NEt = diisopropylethylamine

Et ₃ N = triethylamine

DMAP = 4-dimethylaminopyridine

NaBH ₄ = sodium borohydrate

n-BuLi = n-butyllithium

Pd/C = palladium on carbon

KOH = potassium hydroxide

NaOH = sodium hydroxide

LiOH = lithium hydroxide

K ₂ CO ₃ \u003d potassium carbonate

NaHCO ₃ = sodium bicarbonate

Ar = argon

N ₂ = nitrogen

min = minute(s)

h = hour (hours)

l = liter

ml = milliliter

µl = microliter

g = gram

mg = milligram

they say = mole

mmol. = millimole

meq. = milliequivalent

RT = room temperature

us. = saturated

aq. = water

TLC = thin layer chromatography

HPLC = high performance liquid chromatography

LC/MS = high performance liquid chromatography/mass spectrometry

MS or Mass Spec = mass spectrometry

NMR = nuclear magnetic resonance

m.p. = melting point

The term "lower alkyl", as used here either alone or as part of other terms, unless otherwise specified, refers to straight and branched hydrocarbon chains containing from 1 to 8 carbon atoms, and the term "alkyl" or "alk", used here either alone or as part of other terms, refers to straight and branched hydrocarbon chains containing from 1 to 20 carbon atoms, preferably from 1 to 10 carbon atoms, even more preferably from 1 to 8 carbon atoms in the normal chain, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, a also to their branched chain isomers and the like, as well as to groups having from 1 to 4 substituents such as halogen, for example F, Br, Cl or J or CF ₃ , alkyl alkoxy, aryl, aryloxy, aryl(aryl ) or diaryl, arylalkyl, arylalkyloxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylalkyloxy, optionally substituted amine, hydroxy, hydroxyalkyl, acyl, alkanoyl, heteroaryl, heteroaryloxy, cycloheteroalkyl, arylheteroaryl, arylalkoxycarbonyl, heteroarylalkyl, heteroaryl coxy, aryloxyalkyl, aryloxyaryl, alkylamino, alkanoylamino, arylcarbonylamino, nitro, cyano, thiol, haloalkyl, trihaloalkyl and/or alkylthio.

The term "cycloalkyl", as used here either alone or as part of other terms, unless otherwise specified, refers to saturated or partially saturated (having 1 or 2 double bonds) cyclic hydrocarbon groups having from 1 to 3 rings, and including monocyclic alkyl, bicyclic alkyl and tricyclic alkyl, the rings of which consist of a total of 3-20 carbon atoms, more preferably 3-10 carbon atoms and can be combined with 1-2 aromatic rings, as described for aryl, which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, cyclohexenyl,

wherein any of these groups may optionally be substituted with 1-4 substituents such as halogen, alkyl, alkoxy, hydroxy, aryl, aryloxy, arylalkyl, cycloalkyl, alkylamido, alkanoylamino, oxo, acyl, arylcarbonylamino, amino, nitro, cyano, thiol, and /or alkylthio and/or any alkyl substituent.

The term "alkanoyl", as used here either alone or as part of other terms, refers to an alkyl bonded to a carbonyl group.

The term "halogen" or "halo", as used here either alone or as part of other terms, refers to chlorine, bromine, fluorine and iodine; with chlorine and fluorine being more preferred.

The term "metal ion" refers to alkali metal ions such as sodium, potassium or lithium, and alkaline earth metal ions such as magnesium and calcium, as well as zinc and aluminum.

The term "aryl" or "Aryl", used here either alone or as part of other terms, unless otherwise specified, refers to monocyclic and bicyclic aromatic groups having from 6 to 10 carbon atoms in the ring (such as phenyl or naphthyl , including 1-naphthyl and 2-naphthyl), which may optionally include 1-3 additional rings combined with a carbon ring or a heterocyclic ring (such as, for example, an aryl, cycloalkyl, heteroaryl or cycloheteroalkyl ring, for example

and may optionally be substituted on suitable carbon atoms with 1, 2 or 3 groups including hydrogen, halo, haloalkyl, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkyl-alkyl, cycloheteroalkyl, cycloheteroalkylalkyl, aryl, heteroaryl , arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, alkoxycarbonyl, arylcarbonyl, arylalkenyl, aminocarbonylaryl, arylthioarylsulfinyl, arylazo, heteroarylalkyl, heteroarylalkenyl, heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro, cyano, amino, substituted amines, where amine contains 1-2 substituents (to which includes alkyl, aryl or any other aryl compound mentioned in the definitions), thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonyl ylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl , arylsulfonylamino and arylsulfonaminocarbonyl and/or any alkyl substituent mentioned here.

The terms "lower alkoxy", "alkoxy", "aryloxy" or "aralkoxy" as used herein, either alone or as part of other terms, unless otherwise specified, refer to any alkyl, aralkyl or aryl groups mentioned above, associated with an oxygen atom.

The terms "lower alkylthio", "alkylthio", "arylthio" or "aralkylthio" as used herein, either alone or as part of other terms, unless otherwise specified, refer to any alkyl, aralkyl or aryl groups mentioned above, associated with a sulfur atom.

The term "polyhaloalkyl" as used herein refers to an "alkyl" group as defined above having 2 to 9, preferably 2 to 5 halo substituents such as F or Cl, preferably F such as CF ₃ CH ₂ , CF ₃ or CF ₃ CF ₂ CH ₂ .

The term "polyhaloalkyloxy" as used herein refers to an "alkoxy" or "alkyloxy" group as defined above having 2 to 9, preferably 2 to 5 halo substituents, such as F or Cl, preferably F, such as CF ₃ CH ₂ O, CF ₃ O or CF ₃ CF ₂ CH ₂ O.

The term "prodrug esters" as used herein refers to esters and carbonates obtained by reacting one or more hydroxyl compounds of formula I with an alkyl, alkoxy or aryl-substituted acylating agent using methods to prepare acetates, pivalates, methyl carbonates. , benzoates and the like, well known to those skilled in the art. In addition, esters based on carboxylic and phosphoric acids, such as methyl, ethyl, benzyl, etc., are known from the prior art.

Examples of such esters are:

When the compounds of formula I are in the form of acids, they can form pharmaceutically acceptable salts such as alkali metal salts such as lithium, sodium or potassium; alkaline earth metal salts such as calcium or magnesium, as well as zinc and aluminum, and other cations such as ammonium, chlorine, diethanolamine, lysine (D or L), ethylenediamine, tert-butylamine, tert-octylamine, tri-(hydroxymethyl) aminomethane (TRIS), N-methyl glucosamine (NMG), triethanolamine and dehydroabietylamine.

The present invention also contemplates all stereoisomers of the compounds, whether in admixture or in purified or partially purified form. The compound of the present invention may have asymmetric centers at any of the carbon atoms, including any R substituent. Therefore, the compound of formula I may exist in the form of enantiomers or diastereomers or mixtures thereof. In the preparation of compounds, racemates, enantiomers or diastereomers may be used as starting compounds. Once obtained, enantiomers and diastereomers can be separated using conventional methods such as chromatography or fractional crystallization.

If necessary, the compounds of formula I can be used in combination with one or more other antidiabetic agents or one or more other drugs, which can be administered either orally in the same dosage form, as well as in a separate oral dosage form or by injection.

Such antidiabetic agents which may be used in conjunction with SGLT2 inhibitors of formula I include 1,2,3 or more antidiabetic agents or antihyperglycemic agents, including insulin secretagogues or insulin sensitizers, or other antidiabetic agents, preferably having a different mechanism of action than SGLT2 inhibitors. and including biguanides, sulfonylurea derivatives, glucosidase inhibitors, PPARy agonists such as thiazolidinediones, aP2 inhibitors, PPARα/γ dual agonists, dipeptidyl peptidase IV (DP4) inhibitors and/or meglitinides, as well as insulin, glucagon-like peptide 1, PTP1 inhibitors B, glycogen phosphorylase inhibitors and/or glucose-6-phosphatase inhibitors.

Other drugs that may be used in conjunction with SGLT2 inhibitors of formula I include anti-obesity agents, antihypertensive agents, antiplatelet agents, anti-atherosclerotic agents and/or lipid-lowering agents.

The SGLT2 inhibitor of formula I can also be used in conjunction with drugs to treat complications of diabetes. Such drugs include PKC inhibitors and/or AGE inhibitors.

It is believed that the use of the compounds of formula I in combination with 1,2,3 or more other antidiabetic agents leads to a more significant antihyperglycemic effect than when using the same drugs alone and greater than the total additive antihyperglycemic effect due to these drugs.

Such an antidiabetic agent may be an oral antihyperglycemic agent, preferably from the biguanide group, such as metformin or phenformin or salts thereof, preferably metformin HCl.

When a drug from the biguanide group is used as an additional antidiabetic agent, the compound of formula I is administered in a weight ratio to this drug equal to 0.01:1-100:1, preferably 0.1:1-5:1.

Also, the additional anti-diabetic agent may preferably be an agent from the group of sulfonylurea derivatives such as glubiride (also known as glibenclamide), glimepiride (described in US Pat. No. 4,379,785), glipizide, gliclazide or chlorpropamide, or any known agent from this group or other an antihyperglycemic agent that acts on ATP-dependent β-cell channels; glubiride and glipizide are preferred and may be used in the same or different oral dosage forms.

The compound of formula I is then introduced in a weight ratio to the sulfonylurea derivative of 0.01:1-100:1, preferably 0.2-10:1.

In addition, the additional anti-diabetic agent may be an agent from the group of glucosidase inhibitors such as acarbose (described in US Pat. No. 4,904,769) or miglitol (described in US Pat. No. 4,639,436), which can be used in the same or different oral dosage forms.

The compound of formula I is then administered in a weight ratio to the glucosidase inhibitor of 0.01:1-100:1, preferably 0.5:1-50:1.

A compound of formula I can be used in combination with a PPAR agonist such as the oral antihypoglycemic drug thiazolidinedione or another insulin sensitizer (which has an insulin sensitizing effect in NIDDM patients) such as troglitazone (Rezulin® from Warner-Lambert, described in US Pat. 4,572,912), rosiglitazone (SKB), pioglitazone (Takeda), MCC-555 from Mitsubishi (described in US Pat. No. 5,594,016), GL-262570 from Glaxo-Welcome, englitazone (CP-68722 from Pfizer), or darglitazone (CP-86325). Pfizer, isaglitazone (MIT/J&J), JTT-501 (JPNT/P&U), L-895645 (Merk), R-119702 (Sakyo/WL), NN-2344 (Dr. Reddy/NN), or YM-440 ( Yamanouchi), preferably rosiglitazone and pioglitazone.

The compound of formula I is then introduced in a weight ratio to thiazolidinedione equal to 0.01:1-100:1, preferably 0.2:1-10:1.

Sulfonylureas and thiazolidinedione in amounts less than 150 mg of an oral anti-diabetic agent may be combined in one tablet with a compound of formula I.

The compound of formula I can also be used in combination with an antihyperglycemic agent such as insulin or a glucagon-like peptide 1(GLP-1) such as GLP-1(1-36)amide, GLP-1(7-36)amide, GLP-1 (7-37) (as described in Habener US Pat. No. 5,614,492, incorporated herein by reference), as well as AC2993 (Amylen) and LY-315902 (Lilly), which can be injected, intranasally, transdermally or buccally.

If used, metformin, sulfonylurea derivatives such as glubiride, glimepiride, glipiride, glipizide, chlorpromamide and gliclazide, and glucosidase inhibitors such as acarbose and miglitol, or insulin (injectable, pulmonary, buccal or oral) may be used according to formulations above, and in the amounts and doses indicated in the doctor's desk reference.

If used, metformin and its salts can be used in amounts ranging from 500 to 2000 mg per day, which can be administered in single or divided doses from one to four times a day.

When used, antidiabetic agents from the thiazolidinedione group can be used in amounts ranging from 0.01 to 2000 mg per day, which can be administered either in single or divided doses from one to four times a day.

If used, insulin can be used in accordance with the formulations, quantities and dosages indicated in the doctor's desk reference.

When used, GLP-1 peptides can be administered as oral, buccal, intranasal, or parenteral dosage forms as described in US Pat.

Among other things, an antidiabetic agent that can be used in conjunction with a compound of formula I can be a PPAR α/γ dual agonist such as AR-H039242 (Astra/Zeneca), GW-409544 (Glaxo-Wel1come), KRP297 (Kyorin Merck), as well as those described by Murakami et al. "A novel insulin sensitizer works as a coligand for peroxisome proliferation-activated receptor alpha (PPAR alpha) and for PPAR gamma. Effect of PPAR activation on abnormal hepatic lipid metabolism in obese Zucker rats ", Diabetes 47, 1841-1847 (1998), as well as the agents disclosed in application No. 60/155,400 of September 22, 1999, mentioned here by reference, and administered at the dosages indicated therein; preferred are the compounds mentioned in the above application as the most suitable.

Also, another anti-diabetic agent may be an aP2 inhibitor such as described in US Patent Application No. 09/391,053, filed September 7, 1999, and US Patent Application No. 60/127,745, filed April 5, 1999, used at the dosages indicated therein. Preferred are the compounds mentioned in the above applications as the most suitable.

In addition, another anti-diabetic agent may be a DP4 inhibitor such as described in WO 99/38501, WO 99/46272, WO 99/67279 (PROBIODRUG), WO 99/67278 (PROBIODRUG), WO 99/61431 (PROBIODRUG), NVP-DPP728A (1-[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S)-pyrrolidine)(Novartis) (preferred), as described by Hunges et al, Biochemistry, 38 (36), 11597-11603, 1999, TSL-225 (tryptophyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (described in Yamada et al, Biorg. & Med. Chem. Lett.8 (1998) 1537-1540, 2-cyanopyrrolidides and 4-cyanopyrrolidides as described in Ashworth et al., Bioorg. & Med. Chem. Lett., Vol. 6, No. 22, pp. 1163-1166 and 2745-2748 (1996) used at the dosages indicated in the above references.

A compound from the meglitinide group which may be used in conjunction with a compound of formula I may be repaglinide, nateglinide (Novartis) or KAD1229 (PF/Kissei); rapaglinide is preferred.

The SGLT2 inhibitor of formula I is then administered in a weight ratio of 0.01:1-100:1, preferably 0.2:1-10:1, to meglitinide, PPAR agonist, PPAR α/γ dual agonist, aP2 inhibitor or DP4 inhibitor.

Lipid-lowering agents which may be used in conjunction with compounds of formula I may include 1,2,3 or more of the MRI inhibitors, HMG-CoA reductase inhibitors, squalene synthetase inhibitors, fibric acid derivatives, ALAT inhibitors, lipoxygenase inhibitors, absorption inhibitors cholesterol, inhibitors of intestinal Na ⁺ /bile acid co-transporter, regulators of LDL receptor activity, bile acid sequestrants and/or nicotinic acid and its derivatives.

MTP inhibitors that may be used in accordance with the present invention include the MTP inhibitors described in US Pat. No. 5,595,872; US Pat. No. 5,739,135; US Pat. No. 5,712,279; See also US Pat. No. 5,962,440. Preferred is each of the MTP inhibitors described in the above documents. All of the aforementioned US patents and applications are incorporated herein by reference.

The lipid-lowering agent may be an agent from the group of HMG-CoA reductase inhibitors, which includes, but is not limited to, mevastatin and related compounds described in US Pat. No. 3,983,140; lovastatin (mevinolin) and related compounds described in US patent No. 4,231,938; pravastatin and related compounds described in US patent No. 4,346,227; simvastatin and related compounds described in US Pat. Nos. 4,448,784 and 4,450,171. Lipid-lowering agents can also be compounds described in US patent applications No. 60/211,594 and 60/211,595. Other HMG-CoA reductase inhibitors that can be used in accordance with the present invention include (but are not limited to) fluvastatin described in US patent No. 5,354,772; cerivastatin as described in US Pat. Nos. 5,006,530 and 5,177,080; atorvastatin described in US Pat. Nos. 4,681,893, 5,273,995, 5,385,929 and 5,686,104; atavastatin (nivastatin from Nissan/Sankyo (NK-104)), described in US patent No. 5,011,930; visastatin from Shionogi-Astra/Zeneca (ZD-4522) described in US Pat. No. 5,260,440 and statin-related compounds described in US Pat. No. 5,753,675; pyrazole analogs of mevalonolactone derivatives described in US patent No. 4,613,610, indene analogs of mevalonolactone derivatives described in PCT application WO 86/03488; 6-[2-(substituted-pyrrol-1-yl)-alkyl)puran-2-ones and their derivatives described in US patent No. 4,647,576; SC-45355 dichloroacetate from Searl (3-substituted pentanedioic acid derivative); imidazole analogs of mevalonolactone derivatives as described in PCT application WO 86/07 054; derivatives of 3-carboxy-2-hydroxy-propane-phosphonic acid, described in French patent No. 2,596,393; derivatives of 2,3-disubstituted pyrrole, furan and thiophene, described in the application for European patent No. 0,221,025; naphthyl analogues of mevalonolactone, described in US patent No. 4,686,237; octahydrognaphthalenes as described in US Pat.

In addition, phosphinic acid compounds having the ability to inhibit HMG-CoA reductase and suitable for use in accordance with the present invention are described in GB 2205837.

Suitable squalene synthetase inhibitors for use in accordance with the present invention include, but are not limited to, the α-phosphonosulfonates described in US Pat. No. 5,712,396; the inhibitors described in Biller et al, J. Med. Chem., 1988, Vol.31, No. 10, pp. 1869-1871, including isoprenoid (phosphinyl-methyl) phosphonates, as well as other known squalene synthetase inhibitors, such as those described in US Pat. M., and Poulter, CD., Current Pharmaceutical Design, 2, 1-40 (1996).

In addition, additional squalene synthetase inhibitors suitable for use in accordance with the present invention include the terpenoid pyrophosphates described by P. Otiz de Mintelliano et al, J. Med. Chem., 1977, 20 , 243-249; farnesyl diphosphate A analogs and presqualene pyrophosphate analogs (PSQ-PP) described by Corey and Volante, J. Am. Chem. Soc, 1976, 98, 1291-1293; phosphinylphosphonates as described by McClard, RW et al, JACS, 1987, 109, 5544; and cyclopropanes as described in Capson, TL, June 1987, Dept. Med. Chem. U of Utach, abstract, table, pp. 16,17, 40-43, 48-51, summary.

Other lipid-lowering agents suitable for use in accordance with the present invention include (but are not limited to) fibric acid derivatives such as fenofibrate, gemfibrozil, clofibrate, bezafibrate, ciprofibrate, clinofibrate, and the like; probucol and related compounds described in US patent No. 3,674,836; (robucol and gemfibrozil are preferred); bile acid sequestrants such as cholestyramine, colestipol and DEAE-Sephadex (Secholex®, Policexide®), and lipostabil (Phone-Poulenc); Eisai E-5050 (N-substituted ethanolamine derivative); imanixil (NOE-402); tetrahydrolipstatin (THL); istigmastanylphosphorylcholine (SPC, Roche); aminocyclodextrin (Tanabe Seiyoki); Ajinomoto AJ-814 (azulene derivative); melinamide (Sumitomo); Sandoz 58-035, American Cyanamid CL-277,082 and CL-283, 546 (disubstituted urea derivatives); a nicotinic acid; acipimox; atsifran; neomycin; p-aminosalicylic acid; aspirin, poly(diallylmethylamine) derivatives described in US patent No. 4,759,923; poly(diallyldimethylammonium chloride) quaternary amine; and ions such as those described in US Pat. No. 4,027,009, as well as other known serum cholesterol lowering agents.

Another lipid-lowering agent may be an ACAT inhibitor such as described, for example, in Drugs of the Future 24, 9-15 (1999), (Avasimibe); "The ACAT inhibitor, C1-1011, is an effective agent for preventing and slowing the development of fatty streaks in the aorta in hamsters", Nicolosi et al, Atherosclerosis (Shannon, Irel). (1998), 137(1), 77-85; "Pharmacological profile of FCE 27677: a novel ACAT inhibitor with potential hypolipidemic activity due to selective suppression of hepatic secretion of ApoB100-bearing lipoprotein", Ghiselli, Giancarlo, Cardiovasc. Drug Rev. (1998), 16(1), 16-30; "RP 73163: Bioavailable alkylsulfinyl-diphenylimidazole ACAT inhibitor", Smith, C, et al, Bioorg. Med. Chem. Lett. (1996), 6(1), 47-50; "ASAT inhibitors: physiological mechanisms of lipid-lowering and anti-atherosclerotic activity in experimental animals", Krause et al, Editor(s): Ruffolo, Robert R., Jr.; Hollinger, Mannfred A., Inflammation: mediator pathways (1995), 173-98, Publisher: CRC, Boca Raton, Fla.; "ASAT inhibitors: potential anti-atherosclerotic agents", Sliskovic et al, Curr. Med. Chem. (1994), 1(3), 204-25; "Acyl-CoA inhibitors: cholesterol O-acyl transferase (ACAT) as hypocholesterolemic agents. b. The first water-soluble ACAT inhibitor with lipid-regulating activity." Acyl-CoA inhibitors: cholesterol acyltransferase (ACAT)7. Development of a series of substituted N-phenyl-N'-[(1-phenylcyclopentyl)methyl]urea derivatives with high hypocholesterolemic activity", Stout et al, Chemtracts: Org. Chem. (1995), 8(6), 395-62, or TS-962 (Taisho Pharmaceutical Co. Ltd).

The lipid-lowering agent may also be an LD2 receptor upregulator such as MD-700 (Taisho Pharmaceutical Co. Ltd) or LY295427 (Eli Lilly).

The lipid-lowering agent may also be a cholesterol absorption inhibitor, preferably SCH48461 from Shering-Plough, as well as those described in Atherosclerosis 115, 45-63 (1995) and J. Med. Chem. 41, 973 (1998).

The lipid-lowering agent can also be an inhibitor of the intestinal Na ⁺ /bile acid co-transporter such as described in Drugs of the Future, 24, 425-430 (1999).

Preferred lipid-lowering agents are pravastatin, lovastatin, simvastatin, atorvastatin, fluvastatin, cerivastatin, atavastatin and rosuvastatin.

The US patents cited above are incorporated herein by reference. The dosage of the drugs used corresponds to the dosage given in the doctor's desk reference or in the above patents.

Compounds of formula I, declared in accordance with the present invention, will be used in a weight ratio to lipid-lowering agents (if used), equal to 500:1-1:100.

The dosage used must be carefully selected in accordance with the age, weight and physical condition of the patient, as well as the route of administration, dosage form and mode of administration and the desired result.

The dosages and formulations of the lipid-lowering agents are as set forth in the various patents and applications cited above.

Dosages and formulations of other lipid-lowering agents, if used, are in accordance with the latest edition of the Physician's Desk Reference.

When administered orally, a satisfactory result can be obtained by using an MTP inhibitor in an amount ranging from 0.01 mg/kg to 500 mg, preferably from 0.1 mg to 100 mg, administered one to four times a day.

Preferred oral dosage forms, such as tablets or capsules, contain an MTP inhibitor in an amount that ranges from 1 to about 500 mg, preferably from 2 to about 400 mg, more preferably from 5 to about 250 mg, from one to four once a day.

When administered orally, a satisfactory result can be obtained with the use of HMG-CoA reductase inhibitors, for example, pravastatin, lovastatin, simvastatin, atorvastatin, fluvastatin or cerivastatin at dosages indicated in the physician's desk reference, for example, in an amount ranging from 1 to 2000 mg , preferably from 4 to 200 mg.

The squalene synthetase inhibitor may be administered in an amount ranging from 10 mg to 2000 mg and preferably from 25 mg to 200 mg.

Preferred oral dosage forms such as tablets or capsules contain the HMG-CoA reductase inhibitor in an amount that varies from 0.1 to 100 mg, preferably from 5 to 80 mg, and more preferably from 10 mg to 40 mg.

A preferred oral dosage form, such as tablets or capsules, contains the squalene synthetase inhibitor in an amount that ranges from 10 to 500 mg, preferably 25 to 200 mg.

Another lipid-lowering agent may also be a lipoxygenase inhibitor, including a 15-lipoxygenase (15-LO) inhibitor belonging to the class of benzimidazole derivatives such as described in WO 97/12615; LO-15 inhibitors as described in WO 97/12613; the isothiazolones described in WO 96/38144; and 15-LO inhibitors as described by Sendobry et al in "Attenuation of diet-induced atherosclerosis in rabbits with a highly selective 15-lipoxygenase inhibitor having significant antioxidant activity", Brit. J. Pharmacology (1997) 120, 1199-1206, and Cornicelli et al, "15-lipoxygenase and its inhibition: a new therapeutic possibility in the treatment of vascular disease", Current Pharmaceutical Design, 1999, 5, 11-20.

The compounds of formula I and the lipid-lowering agents may be used together in the same oral dosage form or as separate dosage forms taken simultaneously.

The compositions described above may be administered in the dosage forms described above in single or divided doses from one to four times a day. It is preferable to start treatment with a combination of low dosages with a gradual increase in dosage to a high one.

Preferred lipid-lowering agents are pravastatin, simvastatin, lovastatin, atorvastatin, fluvastatin, cerivastatin, atavastatin and rosuvastatin.

In addition, other drugs that can be used in conjunction with the compounds of formula I are 1,2,3 or more drugs for the treatment of obesity, including beta-3-adrenergic receptor agonists, lipase inhibitors, serotonin reuptake inhibitors (and dopamine), thyroid beta receptor drugs, anorectics, NPY antagonists, leptin analogs and/or MC4 agonists.

The beta-3 adrenergic receptor agonist which may be used in conjunction with a compound of formula I may be AJ9677 (Takeda/Dainippon), L750355 (Merck) or CP331648 (Pfizer) or any other known beta-3 adrenergic receptor agonist, such as those described US Pat. Nos. 5,541,204, 5,770,615, 5,491,134, 5,776,983 and 5,488,064 AJ9677, L750.335 and CP331648 are preferred.

Lipase inhibitors that can be used in conjunction with the compounds of formula I are orlistat and ATL-962 (Alizyme); orlistat is preferred.

Serotonin (and dopamine) reuptake inhibitors which may be used in conjunction with compounds of formula I are sibutramine, topiramate (Johnson & Johnson) or axokine (Regeneron); sibutramine and topiramate are preferred.

The thyroid beta receptor agent which may be used in conjunction with compounds of formula I may be a thyroid receptor ligand such as described in WO 97/21993 (U. Cal SF), WO 99/00353 (KaroBio) and GB 98/284425 (KaroBio); the compounds disclosed in KaroBio applications are preferred.

Anorectic agents that can be used in conjunction with the compounds of formula I are dexamphetamine, phentermine, phenylpropanolamine or mazindol; dexamphetamine is preferred.

Compounds of formula I and agents for the treatment of obesity can be used together both in the same dosage form and in the form of separate dosage forms taken simultaneously, the dosage and mode of administration is well known to specialists, and is also indicated in the doctor's desk reference.

Examples of antiplatelet drugs that can be used in conjunction with the compounds of the present invention are abciximab, ticlopidine, eptifibatide, dipyridamole, aspirin, anagrelide, tirofiban and/or clopidogrel.

Antihypertensive drugs that can be used in conjunction with the compounds of the present invention include ACE inhibitors, calcium antagonists, alpha-blockers, diuretics, centrally acting drugs, angiotensin-II antagonists, beta-blockers, and vasopeptidase inhibitors.

Examples of ACE inhibitors are lisinopril, enalapril, quinapril, benazepril, fosinopril, ramipril, captopril, enalaprilat, moexipril, tandolapril and perindopril; examples of calcium antagonists are amlodipine, diltiazem, nifedipine, verapamil, felodipine, nisoldipine, isradipine and nicardipine; examples of alpha blockers are terazosin, doxazosin and prazosin; examples of diuretics are hydrochlorothiazide, torasemide, furosemide, spironolactone and indapamide; examples of centrally acting drugs are clonidine and guanfacine; examples of angiotensin-II inhibitors are losartan, valsartan, irbesartan, candesartan and telmisartan; examples of beta-blockers are metoprolol, propanolol, atenolol, carvedilol and sotalol; and examples of vasopeptidase inhibitors are omapatrilat and hemopatrilat.

The method of the present invention involves the use of a pharmaceutical composition comprising a compound of formula I, either with or without another antidiabetic, lipid-lowering or other drug, and a pharmaceutically acceptable carrier or diluent. The pharmaceutical composition can be prepared using conventional solid or liquid carriers or diluents and pharmaceutical additives appropriate to the desired route of administration. The compounds can be administered to mammals, including humans, monkeys, dogs, etc., both orally, for example in the form of tablets, capsules, granules or powders, and parenterally, in the form of injections, as well as intranasally or via transdermal patches. The dose for an adult is preferably in the range of 10 to 2000 mg per day, and may be administered as a single dose or divided into several doses from 1 to 4 times a day.

A typical injectable dosage form is prepared by placing a compound of formula I under aseptic conditions in an ampoule which is then aseptically freeze-dried and sealed. Before administration, the contents of the ampoule are dissolved in 2 ml of saline to obtain an injectable preparation.

The inhibitory activity of a compound of formula I can be determined using the assays below.

Study of SGLT2 activity

The human SGLT2 mRNA sequence (GenBank #M95549) was cloned by reverse transcription and amplification of human kidney mRNA using standard molecular biology techniques. The cDNA sequence was stably transfected into CHO cells and the clones were assayed for SGLT2 activity as described in Ryan et al. (1994). Evaluation of SGLT2 inhibition activity in cell lines selected by clonal selection was performed as described by Ryan et al., with the following modifications. Cells were grown in 96-well plates for 2-4 days to 75,000 or 30,000 cells per well in F-12 nutrient mixture (Ham's F-12) supplemented with 10% fetal bovine serum, 300 μg/ml geneticin and penicillin-streptomycin . In the monolayer state, the cells were washed twice with 10 mM Hepes/Tris solution, pH 7.4, 137 mM N-methyl-D-glucamine, 5.4 mM KCl, 2.8 mM CaCl g, 1.2 mM MgSO ₄ . Cells were then incubated with 10 µM [ ¹⁴ C]AMG and 10 µM inhibitor (final DMSO=0.5%) in 10 mM Herpes/Tris solution, pH 7.4, 137 mM NaCl, 5.4 mM KCl, 2.8 mM CaCl g, 1.2 mM MgSO ₄ at 37°C for 1.5 h. Label uptake studies were performed in ice-cold IX PBS containing 0.5 mmol phlorizin, and cells were lysed in 0.1% NaOH. After addition of MicroScint scintillation fluid, the cells were allowed to mix for 1 hour and then [ ¹⁴ C]AMG was counted on a TopCount counter. The control was carried out both in the presence of NaCl and in its absence. To determine the EC ₅₀ index, 10-fold inhibitor concentrations were used at 2 log intervals in the appropriate response zone in triplicate on the plates. Ryan MJ, Johnson G, Kirk J, Fuerstenberg SM, Zager RA, and Torok-Storb B. 1994. HK-2: An immortal proximal tubular epithelial cell line of healthy adult kidney. Kidney International 45: 48-57.

The following Examples illustrate the present invention. Temperatures, unless otherwise specified, are expressed in centigrade units.

Example

A. 5-bromo-2-chloro-4'-ethoxybenzophenone

To a stirred suspension of commercial 5-bromo-2-chlorobenzoic acid (250 g, 1.06 mol) in 450 ml of CH ₂ Cl ₂ containing oxalyl chloride (1.1 mol) was added 1.5 ml of DMF. After the intensive evolution of gas ceased, the reaction mixture was stirred overnight, after which the volatile compounds were removed in vacuo using a rotary evaporator. After dissolving the crude 5-bromo-2-chlorobenzoyl chloride in 200 ml CH ₂ Cl ₂ , the yellow solution was placed in a 2 liter three-necked flask equipped with an external mixer and an internal thermometer. This stirred mixture was cooled to -3° after which phenetol (130 g, 1.08 mol) was added.

Next, AlCl ₃ (140 g, 1.07 mol) was added with a solid funnel over 30 minutes to prevent the temperature from rising above 4°. A significant amount of gaseous HCl, which began to evolve after the addition of 60% AlCl ₃ , was captured by passing the gas through a stirred concentrated NaOH solution. High Performance Liquid Chromatography (HPLC) indicated 95% completion of the reaction within 10 minutes. after the completion of the addition of AlCl ₃ . Next, the mixture was stirred at 4°C for 1 h, after which the reaction mixture was cooled with ice. Subsequently, the suspension was diluted with H ₂ O (1 l) and was extracted three times with CH ₂ Cl ₂ . The combined organic extracts were washed twice with 1H. HCl, once H ₂ O, twice 1M NaOH and twice brine, then dried over Na ₂ SO ₄ . After removal of volatiles, using HPLC, a precipitate was obtained, which is a mixture of ortho/para isomers in a ratio of 1:7.

Recrystallization twice with 400 ml absolute ethanol gave 230 g (64%) of 5-bromo-2-chloro-4'-ethoxybenzophenone.

B. 5-bromo-2-chloro-4'-ethoxydiphenylmethane

To a stirred solution of Et ₃ SiH (400 ml, 2.51 mol) and 5-bromo-2-chloro-4'-ethoxybenzophenone (390 g, 1.15 mol) in 900 ml of a mixture of 1,2-dichloroethane/MeCN in a ratio of 1:2 at temperature of 10° was added BF ₃ ⋅Et ₂ O (150 ml, 1.58 mol) provided that the temperature did not exceed 20°C. Moderate heat evolution occurred during the addition.

The mixture was stirred over night at a temperature of 20°C, after which HPLC revealed the completion of the reaction by 90%. After adding 40 ml of Et ₃ SiH and 15 ml of BF ₃ ⋅Et ₂ O, the reaction mixture was heated to 50°C for 3 hours (it was noted that high temperatures enhance the formation of the Ritter reaction product - N-acetyl 5-bromo-2-chloro- 4'-ethoxydiphenylmethylamine). To cool the reaction mixture, 120 g KOH in 300 ml H ₂ O was added. After stirring for 2 hours, the layers were separated. The aqueous layer was extracted twice with CH ₂ Cl ₂ ; the combined organic layers were washed once with 300 ml of 2M KOH, twice with H ₂ O containing 10% brine to enhance separation of the layers, and twice with brine just before drying over Na ₂ SO ₄ . After removal of the volatiles, the residue was recrystallized from absolute ethanol to give 230 g of 5-bromo-2-chloro-4'-ethoxydiphenylmethane as a white solid.

C. 2,3,4,6-tetra-O-Trimethylsilyl-β-D-glucolactone

To a solution of gluconolactone (239 g, 1.34 mol) and N-methylmorpholine (1180 ml, 10.73 mol) stirred at -5°C in 2.4 L of THF under Ar was added trimethylsilyl chloride (1022 ml, 8.05 mol) through a dropping funnel in such a way that so that the temperature does not exceed 5°C. Then the reaction mixture was stirred for 1 h, after which it was heated to 35°C for 5 hours and stirred overnight, resulting in a decrease in the temperature of the mixture to 20°C. The mixture was then diluted with 3.6 L of toluene and cooled to 0-5°C before carefully adding 7 L of H ₂ O so that the temperature did not exceed 10°C. Attention should be paid to intense heat evolution during the addition of the first portion of H ₂ O. After mixing, the phases were separated and separated. The organic phases were washed with aq. NaH ₂ PO ₄ (2 L), H ₂ O (1 L) and brine (1 L). The organic layer was then concentrated under vacuum using a rotary evaporator; the resulting light yellow oil was treated twice with 250 ml of toluene and re-concentrated to give 616 g of the title compound.

184 ml of 2.5 M n-BuLi in hexane drop by drop, so that the temperature does not rise above -70°C. After stirring for 30 minutes, the solution was cannulated into a stirred at -78°C solution of part C of 2,3,4,6-tetra-O-trimethylsilyl-β-D-glucolactone (236 g, 0.51 mol) in 1.1 l of toluene, so that the temperature remains below -70°C. After stirring the solution for 30 min. at a temperature of -78°C it was cooled by adding 1 l of MeOH containing methanesulfonic acid (41.8 ml, 0.64 mol). The reaction mixture was stirred overnight and the temperature of the mixture rose to 20°C. HPLC revealed two new peaks corresponding to the weight of the expected O-methyl glucoside; the ratio varied from 95:5 to 80:20. The desired product corresponds to the main peak with a shorter retention time. It should be noted that a longer reaction time or the addition of more than 50% methanesulfonic acid tends to convert all of the isomeric product to the desired O-methyl glucoside. After completion of the reaction, the mixture was cooled by adding NaHCO ₃ (37 g, 0.37 mol) in 200 ml H ₂ O. If the pH is not weakly basic, add more NaHCO ₃ before doubling the H ₂ O layers and extracting three times with EtOAc. The combined EtOAc fractions were washed with brine and dried over Na ₂ SO ₄ . After concentration using a rotary evaporator, the residue was dissolved in hot toluene (150 ml). The resulting solution was poured into a liter of stirred hexane. The precipitate was collected by vacuum filtration; the resulting filter cake was washed twice with 500 ml of hexane and then air-dried to give 171 g of the desired compound as a white solid.

Et3SiH ( _65.27 g, _0.56 mol), and then BF ₃ ⋅Et ₂ O (59.75 g, 0.42 mol) so that the temperature remains between -5° and -10°C. The temperature of the stirred solution was raised to 0° C. over 5 hours. When high performance liquid chromatography (HPLC) showed completion of the reaction, the reaction mixture was cooled by adding saturated aqueous NaHCO ₃ (310 ml). Organic volatiles were removed over vacuum using a rotary evaporator. The precipitate was partitioned between EtOAc and H ₂ O using 2 L of each. After phase separation, the aqueous layer was extracted twice with 2 L of EtOAc. The combined organic phases were washed with H ₂ O (2 L) and brine (2 L), then dried over MgSO ₄ and concentrated using a rotary evaporator to obtain 104.6 g of a yellow solidified foam. After dissolving the resulting precipitate in CH ₂ Cl ₂ (750 ml), pyridine (200 g, 2.53 mol) and Ac ₂ O (261.1 g, 2.56 mol) were added in one portion. After heat evolution was completed, which raised the temperature of the mixture from 28°C to 47°C, DMAP (1.56 g, 13 mmol) was added. After 1.5 h the reaction mixture was cooled by adding H ₂ About (1.8 l), HPLC showed the completion of the reaction. The mixture was extracted twice with CH ₂ Cl ₂ (total volume 2.7 l); the combined organic layers were washed twice with 1N. HCl (1.8 L), twice with brine (1.8 L), then dried over MgSO ₄ . After concentration with a rotary evaporator, the residue was recrystallized from absolute ethanol (750 ml) to give 8×9.5 g of the desired tetraacetylated β-C-glucoside as a white solid. The initial liquid contained the corresponding α-C-glucoside and the more polar furanose isomer.

Alternatively, the D portion of the O-methyl glucoside may be first acetylated and then reduced to give the desired tetraacetylated C-aryl glycoside, according to the following procedure.

A solution of part D of O-methylglucoside (3.0 g, 6.8 mmol) in toluene (45 ml) containing diisopropylethylamine (6.9 ml, 40 mmol) was cooled to 0°C, after which acetic anhydride (3.35 ml, 35.5 mmol) was added and DMAP (84 mg, 0.6-8 mmol). The temperature of the solution rose to 20°C for 6 h, after which thin layer chromatography revealed the completion of the formation of tetraacetate. The reaction mixture was cooled by adding 50 ml of 20% H ₃ RO ₄ . After separation of the layers, the aqueous phase was extracted twice with toluene. The combined organic phases were washed once with 50 ml H ₂ O and then concentrated under vacuum. The resulting oil was dissolved in 20 ml of toluene and concentrated to obtain a thick oil (4.15 g), which was used further without further purification.

A solution of the above crude oil (4.15 g, 6.8 mol) in MeCN (60 ml) containing one equivalent of H ₂ O (123 mg, 6.8 mol) was cooled to 0°C, after which Et ₃ SiH (3.27 ml, 20.5 mmol) and then BF ₃ ⋅Et ₂ O (1.73 ml, 13.7 mmol). After stirring for 1 hour, the temperature of the solution rose to 20°C. After 4 hours, a single analysis was performed using high performance liquid chromatography (HPLC), which showed no progressive reaction over 60%, after which 2 ml of Et ₃ SiH and 1 ml of BF ₃ ⋅Et ₂ O were added. Two hours later, HPLC revealed the absence of source material. After adding aq. NaHCO ₃ to cool the reaction mixture, the latter was stirred for 30 minutes, after which it was extracted three times with EtOAc. The combined organic layers were washed once with aq. NaHCO ₃ and brine, then dried over Na ₂ SO ₄ . The oil obtained after concentration under vacuum was dissolved in 70 ml of hot 25% EtOAc/hexane. During cooling, 2.45 g of the desired tetraacetylated β-C-arylglucoside crystallized and was successively isolated by filtration.

To a solution of tetraacetylated β-C-glucoside (27.2 g, 49 mmol) obtained in accordance with point E) in 480 ml of a mixture of THF/MeOH/H ₂ O in a ratio of 2:3:1 was added LiOH⋅ H ₂ O (2.3 g, 57 mmol). After stirring overnight, the volatiles were removed using a rotary evaporator. The precipitate after dissolution in EtOAc (300 ml) was washed once with brine (150 ml), once with brine (50 ml) containing 10 ml of 5% aq. KHSO ₄ , and again with saline (50 ml), then dried over Na ₂ SO ₄ . The volatiles were removed using a rotary evaporator and the resulting oil in a minimum amount of CH ₂ Cl ₂ was frothed under vacuum to obtain 20.4 g of the desired C-aryl glucoside as a white glassy substance containing 0.11 mol% EtOAc.

HPLC retention time: 7.08 min, 94% purity, YMC S5 C-18 4.6×50 mm column, 2.5 ml/min, detection at 220 nm; 8 min - 0-100% B gradient maintained for 5 min at 100% B. Solvent A: 10% MeOH/H ₂ O + 0.2% H ₃ PO ₄ . Solvent B: 90% MeOH/H ₂ O + 0.2% H ₃ PO ₄ .

¹ H NMR (500 MHz, CD ₃ OD) δ 7.33 (d, 1H, J=6 Hz), 7.31 (d, 1H, J=2.2 Hz), 7.31 (dd, 1H, J=6 Hz, J=2.2 Hz), 7.07 (d, 2H, J=8.8 Hz), 6.78 (d, 2H, J=8.8 Hz), 4.07-3.90 (m, 7H), 3.85 (d, 1H, J=10.6 Hz), 3.69 ( dd, 1H, J=5.3, 10.6 Hz), 3.42-3.25 (m, 4Н) Hz), 1.34 (t, 3H, J=7 Hz).

¹³ C NMR (125 MHz, CD ₃ OD) δ 158.8, 140.0, 139.9, 134.4, 132.9, 131.9, 130.8, 130.1, 128.2, 115.5, 82.9, 82.2, 79.7, 76.4, 71.9, 64.5, 63.1, 39.2, 15.2.

Anal Calcd for C ₂₁ H ₂₅ ClO ₆ LC-MS [M+Na ⁺ ] 431; found 431.

The preparation of intermediates is described below, as well as their spectral characteristics.

White solid. HPLC: column Zorbax SB C18, 4.6×75 mm, 0-100% B for 8 min, exposure 4 min, A=10% MeOH / 90% H ₂ O / 0.2% H ₃ PO ₄ , V \u003d 10% H ₂ O / 90% MeOH / 0.2% H ₃ PO ₄ , room. pace. = 7.60 min, 100% purity.

LC-MS: Phenomex Luna 5u column, 4.6×50 mm, 0-100% B for 4 min, hold 1 min, A=10% CH ₃ CN / 90% H ₂ O / 10 mM NH ₄ Ac , B=10% H ₂ O / 90% CH ₃ CN / 10 mM NH ₄ Ac, room. pace. = 2.00 min, calculated for C ₂₂ H ₃₁ ClNO ₇ (M + NH4 ₎ ⁺ 456.2, found 456.3.

¹ H NMR (400 MHz, CD ₃ OD) δ 7.77 (s, 1H), 7.22-7.27 (m, 2H), 7.02 (d, J=8.8, 2H), 6 .74 (d, J=9.3, 2H), 5.01 (s, 1H), 4.84 (s, 1H), 3.75-4.06 (m, 8H), 3.47-3 , 60 (m, 2H), 3.17 (t, J=8.3 Hz, 1H), 3.00 (m, 1H), 2.89 (s, 3H), 1.33 (t, J= 6.6, 1H). ¹³ C NMR (100 MHz, CD ₃ OD) δ 157.3, 138.6, 136.6, 134.4, 131.4, 130.0, 129.7, 129.3, 126.7, 114.4, 101.0, 74.7, 72.5, 70.1, 63.4, 62.0, 49.2, 38.5, 14.9.

White solid, t _pl = 120°C (not corrected). HPLC t _R =3.9 min, 100% purity. HPLC retention time on a Phenomex Luna C-18 column, 4.6 mm x 50 mm, eluent with a 4 min gradient 0-100% solvent B; while solvent A: 10: 90: 0.1 CH ₂ OH = H ₂ O - TFA and solvent B: 90:10: 0.1 CH ₃ OH = H ₂ O - TFA.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.35 (d, J=8.4, 1H), 7.19 (dd, J=1.8, 8.4, 1H), 7.07 (d , J=1.8, 1H), 7.05 (d, J=8.8, 2H), 6.82 (d, J=8.8, 2H), 5.28 (t, J=9, 2, 1H), 5.20 (t, J=9.2, 1H), 5.05 (t, J=9.2, 1H), 4.31 (d, J=9.7, 1H), 4.26 (dd, J=4.8, 12.8, 1H), 4.14 (dd, J=2.2, 12.4, 1H), 3.95-4.07 (m, 4H) , 3.80 (m, 1H), 2.08 (s, 3H), 2.04 (s, 3H), 1.99 (s, 3H), 1.71 (s, 3H), 1.40 ( t, J=7.0, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 170.7, 170.3, 169.45, 168.7, 157.5, 139.1, 135.1, 134.6, 131.0, 129.79, 126.0, 114.5, 79.5, 76.1, 74 .1, 72.5, 68.5, 63.4, 62.3, 38.2 , 20.7, 20.6, 20.3, 14.8. HRMS calculated for C ₂₉ H ₃₃ ClNaO ₁₀ (M+Na) ⁺ : 599, 1660; found: 599, 1649. Calculated for C ₂₉ H ₃₃ ClO ₁₀ : C, 60, 36; H, 5.76. Found: C, 60.43; H, 5.56.

For the main anomer: HPLC t _R =3.45 min, 100% purity. HPLC retention time on a Phenomex Luna C-18 column, 4.6 mm x 50 mm, eluent with a 4 min gradient 0-100% solvent B; while solvent A: 10: 90: 0.1 CH ₃ OH - H ₂ O - TFA; solvent B: 90: 10: 0.1 CH ₃ OH - H ₂ O - TFA.

¹ H NMR (400 MHz, CD ₃ OD) δ 7.54 (d, J=2.2, 1H), 7.45 (dd, J=2.2, 8.4, 1H), 7.35 ( d, J=8.4, 1H), 7.08 (d, J=8.8, 2H), 6.79 (d, J=8.8, 2H), 4.08 (d, J=15 .0, 1H), 3.99 (d, J=15.0, 1H), 3.98 (q, J=7.0, 2H), 3.92 (dd, J=2.2, 11, 8, 1H), 3.80 (dd, J=5.3, 11.9, 1H), 3.74 (t, J=9.2, 1H), 3.57 (m, 1H), 3, 41 (d, J=8.8, 1H), 3.08 (d, J=9.7, 1H), 3.06 (s, 3H), 1.35 (t, J=7.0, 3H ). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 158.2, 139.0, 138.5, 134.2, 132.4, 131.2, 130.1, 129.2, 127.6, 114.8, 101.8, 78.0, 75.3, 74.5, 71.1, 63.8, 62.1, 49.0, 38.7, 14.5 . Calculated for C ₂₂ H ₂₇ ClO ₇ : C, 60, 20; H, 6.20; Cl, 8.07. Found: C, 60, 05; H, 6.21; Cl, 8.01.

Transparent oil. HPLC: Zorbax SB C 18 column, 4.6 x 75 mm, 0-100% B for 8 min, hold 4 min, A=10% MeOH / 90% H ₂ O / 0.2% H ₃ PO ₄ , B \u003d 10% H ₂ O / 90% MeOH / 0.2% H ₃ PO ₄ , room. pace. = 8.85 min, 98.1% purity.

LC-MS: Phenomex Luna 5u column, 4.6 x 50 mm, 0-100% B for 4 min, hold 1 min, A=10% CH ₃ CN / 90% H ₂ 0 / 10 mM NH ₄ Ac, B=10% H ₂ O / 90% CH ₃ CN / 10 mM NH ₄ Ac, room. pace. = 3.55 min. Calcd for C ₃₀ H ₃₉ ClNO ₁₁ (M+NH ₄ ) + 624.2, found 624.3. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.38 (d, J=8.3, 1H) , 7.27 (dd, J=1.7, 8.8, 1H), 7.21 (d, J=2.2, 1H), 7.03 (d, J=8.8, 2H), 6.81 (d, J=8.2, 2H), 5.55 (t, J=9.9, 1H), 5.21 (t, J=9.6, 1H), 4.92 (d , J=10.5, 1H), 4.34 (dd, J=5.0, 12.0, 1H), 4.20 (dd, J=2.8, 12.1, 1H), 3, 97-4.08 (m, 5H), 3.09 (s, 3H), 2.10 (s, 3H), 2.09 (s, 3H), 1.94 (s, 3H), 1.76 (s, 3H), 1.39 (t, J=7.2, 3H). ¹³ C NMR (100 MHz, CD ₃ OD) δ 170.7, 170.2, 170.1, 169.6, 169.0, 157.5, 139.0, 135.1, 134.7, 131.1, 129.8, 129.7, 129.3, 126.0, 11 4.6, 100.1 80.8, 73.9, 71.3, 68.9 , 68.8, 63.4, 62.1, 52.1, 49.5, 38.4, 20.7, 20.6, 20.3, 14.9.

5-bromo-2-chloro-4'-ethoxydiphenylmethane

White solid; m.p. 37°C (not corrected). HPLC t _R =4.43 min, 100% purity. HPLC retention time on a Phenomex Luna C-18, 4.6 mm x 50 mm column, 4 min gradient eluent, 0-100% solvent B; solvent A: 10: 90: 0.1 CH ₃ OH - H ₂ O - TFA; solvent B: 90: 10: 0.1 CH ₃ OH -H ₂ O - TFA.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.20-7.28 (m, 3H), 7.08 (d, J=8.8, 2H), 4.00 (q, J=7.0 , 2H), 3.96 (s, 2H), 1.40 (t, J=7.0, 3H). ^13C NMR (125 MHz, _CDCl3 ) δ 157.6, 141.3, 133.5, 133.1, 130.9, 130.5, 130.4, 130.0, 120.4, 114.6, 63.4, 38.2, 14.9. Calculated for C ₁₅ H ₁₄ BrClO: C, 55, 32; H, 4.33. Found: C, 55.22; H, 4.25.

5-lithium-2-chloro-4'-ethoxydiphenylmethane

Data could not be obtained, product is thermally unstable above -75°C, rapid decomposition occurs,

or

5-bromo-2-chloro-4'-ethoxybenzophenone

White solid; m.p. = 60°C (not corrected). HPLC t _R =3.9 min, 100% purity. HPLC retention time using Phenomex Luna C-18 column, 4.6 mm x 50 mm, eluent with 4 min gradient 0-100% solvent B; solvent A: 10: 90: 0.1 CH ₃ OH - H ₂ O - TFA; solvent B: 90: 10: 0.1 CH ₃ OH - H ₂ O - TFA.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.76 (d, J=8.8, 2H), 7.53 (dd, J=2.2, 8.4, 1H), 7.48 (d , J=2.2, 1H), 7.32 (d, J=8.8, 1H), 6.93 (d, J=8.8, 2H), 4.11 (q, J=7, 0.2H), 1.45 (t, J=7.0, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 192.0, 163.9, 140.7, 133.7, 132.6, 131.4, 130.1, 128.6, 120.5, 114.5, 63.9, 14.6.

HRMS calculated for C ₁₅ H ₁₃ BrClO ₂ (M+H) ⁺ : 338, 9787; found 338, 9785. Calculated for C ₁₅ H ₁₂ BrClO ₂ : C, 53, 05; H, 3.56. Found: C, 53.11; H, 3.28.

Spectral instruments and conditions

NMR spectra ( ¹ H, ¹³ C) were recorded on JEOL JNM-EPC-500, JEOL GSX400, and Bruker 400 spectrometers. Chemical shifts are indicated in ppm. (ppm), in the field of weak fields relative to the internal standard, tetramethylsilane; coupling constants (J values) are given in Hz (Hz). Elemental analysis was performed at the Discovery Analytical Science Department. Melting points were determined on a Hoover Uni-melt apparatus and were not corrected. LC/MS was recorded on a Shimadzu LC-10AT instrument equipped with a SIL-10A injector, the SDD-10AV detector was typically operated at 200 nm and using a Micromass ZMD mass spectrometer. Unless otherwise indicated, LC/MS or HPLC retention times were determined on a Phenomex Luna C-18 column, 4.6 mm x 50 mm, eluent with a 4 min gradient of 0-100% solvent B; while solvent A: 10: 90: 0.1 CH ₃ OH - H ₂ O - TFA and solvent B: 90: 10: 0.1 CH ₃ OH - H ₂ O - TFA.

biological data

Compound of Formula I (Example)

is an active and selective inhibitor of the main luminal glucose transporter in the renal proximal tubule (SGLT2).

- At doses as low as 0.1 mg/kg, the compound of formula I stimulated significant urinary glucose excretion in normal rats, and at doses as low as 0.01 mg/kg, the compound of formula I stimulated urinary glucose excretion in rats ZDF suffering from glucosuria due to diabetes. These data suggest that the compound of formula I acts on the diabetic kidney to lower the threshold glucose in the kidney. The property of the compounds of formula I to stimulate glucose release in normal rats is stronger than in the case of sergliflozin, 9T-1095, or florizin administered subcutaneously.

- In male ZDF rats, administration of a compound of formula I for 15 days resulted in significantly lower acceleration and plasma glucose levels compared to vehicle-treated rats.

- In a clamp study of hyperinsulinemia-normoglycemia, a significant increase in glucose infusion and glucose utilization rate was accompanied by a significant decrease in HGP in Formula I treated rats compared to vehicle treated rats.

- Preclinical pharmacology of the compound of formula I suggests that this agent can correct environmental hyperglycemia and improve metabolic status in diabetic ZDF rats. Given the absence of side effects in the gastrointestinal tract or hyperglycemia, these data suggest that the compound of formula I is a possible therapeutic candidate for the treatment of diabetes.

Increased selectivity for SGLT2 compared to SGLT1

The in vitro SGLT (EC50) potency of a compound of formula I and, in separate experiments, Example 12 of W0 01/27128 was evaluated by monitoring the inhibition of accumulation of radiolabeled α-methyl-D-glucopyranoside (AMG) by Chinese hamster ovary cells stably expressing SGLT2 and human SGLT1.

Average EC ₅₀ values for SGLT2 and SGLT1 for each compound are presented in Table 1 below.

As shown in Table 1, the compound of formula I exhibits a mean EC ₅₀ against SGLT2 of 1.1 nM compared to an average EC ₅₀ for Example 12 of W0 '128 of 1.0 nM. With respect to SGLT1, the compound of formula I and Example 12 from WO '128 showed mean EC ₅₀ values of 1390 nM and 762 nM, respectively.

These results show that the compound of formula I is highly selective (1.263 times) for SGLT2 over SGLT1 and about 1.7 times more selective for SGLT2 over SGLT1 relative to Example 12 of WO 01/27128.

Increased ability to lower blood glucose levels 5 hours after oral administration in rats with STZ-induced diabetes (short-term animal model)

The in vivo efficacy of the compound of formula I and Example 12 of WO 01/27128 was evaluated using a rat model of STZ-induced diabetes. Following oral administration of a compound of formula I as a single dose of 0.03 or 0.1 mg/kg, blood glucose levels were measured every hour for 5 hours, during which the rats were deprived of food. In separate studies, a single dose of 0.03 or 0.1 mg/kg of Example 12 from WO 01/27128 was administered.

The results are presented in Table 2 below. The 5 hour values in Table 2 represent the mean percent reduction in blood glucose levels from baseline for this cohort of rats. Since the rats did not have access to food during this 5 hour period, the blood glucose values of both treated and untreated rats decreased. To correct for this effect, the percent change in glucose at hour 5 for the vehicle control group was subtracted from the percent change in glucose at 5 hours post-dose.

The mean percent change at 5 hours in the drug groups was compared to the mean percent change at 5 hours from the vehicle group using a T-test. All p values were less than or equal to 0.0006, which was considered statistically significant.

As shown in Table 2, the mean reduction in blood glucose levels compared to control was higher for the Formula I compound than for Example 12 of WO 01/27128 (32.9% vs. 26.2% at the 0.03 mg/kg dose; 42.7% vs. 35.9% at a dose of 0.1 mg/kg). This corresponds to a 25% ((32.9-26.2)/26.2) and 19% ((42.7-35.9)/35.9) increase in the reduction in blood glucose levels under the action of the compound of formula I compared to Example 12 at 0.03 and 0.1 mg/kg, respectively.

Increased ability to lower plasma glucose over a 15-day period after oral administration in rats with STZ-induced diabetes (long-term animal model)

The in vivo efficacy of the compound of formula I and Example 12 of WO 01/27128 was also studied in a head-to-head comparison study in a rat model with STZ-induced diabetes (Zucker's diabetic obese rat, a model genetically predisposed to developing diabetes) during a two-week subchronic study.

10. In this direct comparative study, the compound of formula I and Example 12 were administered daily at a dose of 0.5 mg/kg. Plasma glucose concentration was measured at baseline (ie, baseline plasma glucose prior to the 1st dose of drug) and after days 8 and 15 of treatment following an 18 hour fast. The results are shown in Table 3 below.

Mean fasting plasma glucose values were compared between vehicle and drug treated rats on days 8 and 15. The T-tests performed compared fasting plasma glucose values in the drug-treated and vehicle-treated groups. The p values were less than or equal to 0.002, which was considered statistically significant.

As shown in Table 3, Example 12 of WO 01/27128 reduced fasting glucose levels from about 304 to 196 mg/dl (35.4% reduction; 33.6% reduction compared to control) at day 15, while the compound of formula I reduced fasting glucose levels from about 304 to 138 mg/dl (54.5% decrease; 53.2% decrease compared to control) over the same period.

13. These data demonstrate that the compound of formula I is 58% ((53.2-33.6)/33.5)) more effective than Example 12 in lowering plasma glucose compared to control over a 15 day period.

Claims (2)

A pharmaceutical composition having an inhibitory effect on SGLT2, comprising a compound of the formula and a pharmaceutically acceptable carrier.