Treatment of metabolic disorders in canine animals

Abstract

The present invention relates to one or more SGLT2 inhibitors or pharmaceutically acceptable forms thereof for use in the treatment and/or prevention of a metabolic disorder in a canine animal, preferably wherein the metabolic disorder is one or more selected from the group consisting of: ketoacidosis, pre-diabetes, insulin dependent diabetes mellitus, insulin resistance diabetes, insulin resistance, obesity, hyperglycemia, hyperglycemia induced cataract formation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, low grade systemic inflammation, hepatic lipidosis, inflammation of the pancreas, metabolic disorder consequences, such as hypertension, renal dysfunction and/or muscoskeletal disorders, and/or Syndrome X (metabolic syndrome), wherein preferably the development of hyperglycemia induced cataract formation is prevented or remission is achieved and/or wherein preferably the development of metabolic disorder consequences, such as hypertension, renal dysfunction and/or muscoskeletal disorders, is prevented or progression is slowed or remission is achieved.

Claims

1. A method of treatment of a metabolic disorder in a canine animal comprising administering to the canine animal a composition comprising one or more active agents, where the one or more active agents consist of one or more SGLT2 inhibitors or pharmaceutically acceptable forms thereof; wherein: the one or more SGLT-2 inhibitors or pharmaceutically acceptable forms thereof comprises 1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene or a pharmaceutically acceptable form thereof represented by the following formula: ##STR00029## the metabolic disorder is one or more selected from the group consisting of ketoacidosis, pre-diabetes, insulin dependent diabetes mellitus, insulin resistance diabetes, insulin resistance, obesity, hyperglycemia, hyperglycemia induced cataract formation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, low grade systemic inflammation, hepatic lipidosis, inflammation of the pancreas, metabolic disorder consequences comprising hypertension, renal dysfunction and/or musculoskeletal disorders, and Syndrome X (metabolic syndrome); and the one or more SGLT-2 inhibitors or pharmaceutically acceptable forms thereof is administered at a dose of no greater than 2.0 mg/kg bodyweight.

2. The method of claim 1, wherein the development of hyperglycemia induced cataract formation is prevented or remission is achieved and/or the development of metabolic disorder consequences prevented or progression is slowed or remission is achieved.

3. The method of claim 1, wherein the metabolic disorder is selected from the group consisting of clinical conditions associated with pre-diabetes, insulin dependent diabetes mellitus and insulin resistance.

4. The method of claim 3, wherein the clinical conditions are one or more conditions selected from the group consisting of ketoacidosis, insulin resistance, obesity, hyperglycemia, hyperglycemia induced cataract formation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, low grade systemic inflammation, hepatic lipidosis, inflammation of the pancreas, metabolic disorder consequences comprising hypertension, renal dysfunction and/or musculoskeletal disorders, and Syndrome X (metabolic syndrome).

5. The method of claim 1, wherein the canine animal is suffering from diabetes.

6. The method of claim 1, wherein the canine animal is suffering from pre-diabetes or insulin dependent diabetes.

7. The method of claim 1, wherein the canine animal is a dog.

8. The method of claim 1, wherein the pharmaceutically acceptable form thereof is a crystalline complex between the one or more SGLT-2 inhibitors and one or more amino acids.

9. The method of claim 8, wherein the one or more amino acids comprise proline.

10. The method of claim 8, wherein the one or more amino acids comprise L-proline.

11. The method of claim 1, wherein the one or more SGLT-2 inhibitors or pharmaceutically acceptable forms thereof is administered orally or parenterally.

12. The method of claim 1, wherein the one or more SGLT-2 inhibitors or pharmaceutically acceptable forms thereof is administered once per day.

13. The method of claim 1, wherein the one or more SGLT-2 inhibitors or pharmaceutically acceptable forms thereof is administered at a dose of 0.01 to 1.0 mg/kg bodyweight.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1 Urinary glucose excretion in Beagle dogs after single oral dosing of Compound A. The urine was individually sampled 0-24 h and 24-48 h after administration. After 24 h, urine obtained by insertion of a catheter into the bladder was added to the freely sampled urine. In controls and the low dose (0.01 mg/kg) virtually no glucose was detectable in urine. Indicated p-values above bars are versus control (*, p<0.05).

(2) FIG. 2 Sigmoidal dose response for the urinary glucose excretion in the time period 0-24 h after administration of Compound A as shown. ED50 for glucose excretion by Compound A in Beagle dogs is 0.1 mg/kg (95% CI 0.02-0.52 mg/kg).

(3) FIG. 3 shows an X-ray powder diffraction pattern of a representative batch of a crystalline complex of compound A with L-proline (1:1)

(4) FIG. 4 shows a DSC/TG diagram of a representative batch of a crystalline complex of compound A with L-proline (1:1)

EXAMPLES

(5) The following examples show the beneficial therapeutic effects on glycaemic control and/or insulin resistance, etc., of using one or more SGLT2 inhibitors in canine animals, according to the present invention. These examples are intended to illustrate the invention in more detail without any limitation of the scope of the claims.

Example 1

Pharmacokinetics (PK) of Compound A Single Oral Dosing in Dogs

(6) Compound A was administered to overnight fasted dogs. The groups (n=4 per group) received a single administration of either oral vehicle (DI water) containing the SGLT2 inhibitor Compound A at a dose of 1 mg/kg and 10 mg/kg or intravenous vehicle (saline) containing the SGLT2 inhibitor Compound A at a dose of 1 mg/kg. PK measurements were taken until day 3 after a single administration of compound A or its vehicle.

(7) TABLE-US-00002 TABLE 2 Pharmacokinetic data, single dose (i.v. 1 mg/kg, oral 1 and 10 mg/kg) i.v. p.o. p.o. Parameter 1 mg/kg 1 mg/kg 10 mg/kg t.sub.max [hour] mean 1.9 0.6 C.sub.max [nmol/L] mean 4845 51525 CL [mL/min/kg] mean 0.63 CL/F [mL/min/kg] mean 0.64 0.71 F [%] mean 101 92 AUC.sub.0.fwdarw.∞ [nmol .Math. h/l] mean 67025 67675 616750 T.sub.1/2 [hour] mean 13.4 13.9 14.5

Example 2

The Effect of Compound A on Urinary and Blood Glucose After Single Dosing in Dogs

(8) Beagle dogs were fasted overnight and received a single oral administration of Compound A at doses of 0 mg/kg b.w., 0.01 mg/kg b.w., 0.1 mg/kg b.w., or 1 mg/kg b.w. (n=3 per group) followed by a rinsing with water (1 mL/kg b.w.)

(9) Compound A was moistened with a small volume of a 1% (w/v) aqueous Polysorbat 80 (Tween80, Polyoxyethylene Sorbitan Monooleate, ICN Biomedicals) solution and then dissolved by slowly adding a large volume of a 0.5% (w/v) aqueous hydroxyethylcellulose (Natrosol 250 HX, Boehringer Ingelheim) solution and stirring at room temperature for about 15 minutes. The final concentration of Polysorbat 80 was 0.015%. Compound A was applied in a volume of 2 mL/kg. b.w.

(10) The animals were kept individually in metabolic cages and received food 2 h after administration. They had free access to water during the experiment. Urine was collected in the time intervals 0-8 h, 8-24 h, 24-32 h, and 32-48 h after administration. A catheter (Eickemeyer) was inserted into the bladder to completely collect the 24 h urine. This urine was combined with the urine that had been excreted in the 8-24 h period. A volume of 5 mL of a 10% solution of sodium azide in saline had been added to each urine collection tube before sampling. Volume of urine was determined and samples were frozen for subsequent determination of glucose concentration.

(11) During the experiment, blood samples were drawn from a forearm vein. Blood was collected in EDTA tubes prior to administration of vehicle or Compound A, and subsequently at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h, 32 h, and 48 h time points post-dose. Plasma was prepared following blood collection and frozen for determination of glucose concentration. A prominent increase of urinary glucose concentration and volume was evident at the two higher doses already 8 h after administration (FIG. 1). Neither dose of compound A induced hypoglycemia, or altered the blood glucose level in dogs as compared to normal reference values.

(12) In respect to urinary glucose excretion it is thus estimated that the ED.sub.50 is 0.1 mg/kg (FIG. 2).

Example 3

Treatment of Insulin Dependent Diabetes in Dogs

(13) Treating Dogs With Insulin Dependent Diabetes

(14) Insulin with the Compound A according to the invention or a combination of active substances according to the invention, in addition to producing an acute improvement in the glucose metabolic situation, prevents deterioration in the metabolic situation in the long term and reduces the insulin dose needed to treat the diabetic canine. This can be observed if dogs are treated for a shorter or longer period, e.g. 2-4 weeks or 3 months to 1 year, with the pharmaceutical composition according to the invention and are compared to the metabolic situation prior to treatment or with dogs that have been treated with e.g. insulin alone. There is evidence of therapeutic success if daily mean blood glucose and fructosamine level are reduced as compared to pre-treatment level. Further evidence of therapeutic success is obtained if a significantly smaller percentage of the dogs treated with a pharmaceutical composition according to the invention, compared with dogs who have been treated with other medications, undergo transient deterioration in the glucose metabolic position (e.g. hyper- or hypoglycaemia).

Example 4

Improvement of Insulin Resistance Diabetes in Female Dogs With Dioestrus/Gestational Diabetes

(15) Insulin resistance diabetes is a frequently found form of diabetes in intact female canine animals. Therapy with Compound A may be provided with the objective of preventing the transition to manifest diabetes. In studies over a shorter or longer period (e.g. 2-4 weeks or 1-2 years) the success of the treatment is examined by determining the fasting glucose values and/or the glucose values after a meal or after a loading test (intravenous glucose tolerance test or food tolerance test after a defined meal) during the study throughout the different phases of the menstrual cycle and/or after the end of the period of therapy for the study and comparing them with the values before the start of the study and/or with those of a placebo group. In addition, the fructosamine value can be determined before and after therapy and compared with the initial value and/or compared with dogs that have been treated with other medications or placebo. A significant drop in the fasting or non-fasting glucose and/or fructosamine levels demonstrates the efficacy of the treatment of insulin resistance—diabetes and preventing manifest diabetes in female dogs with a history of dioestrus/gestational diabetes.

Example 5

Treatment of Hyperglycaemia

(16) In clinical studies in dogs with metabolic disorders running for different lengths of time (e.g. 2 weeks to 12 months) the success of the treatment is checked using the measurement of baseline blood glucose and/or blood fructosamine.

(17) The improvement of glycaemic control can furthermore be determined establishing diurnal blood glucose curves, e.g. a 9 or 24 hour blood glucose curve starting prior to medication and repeated measurements post dosing.

(18) A significant fall in these values during or at the end of the study, compared with the initial value or compared with a placebo group, or a group given a different therapy, proves the efficacy of a pharmaceutical composition according to the invention in the reduction of hyperglycaemia in dogs.

(19) Alternatively, the effect of compound A on hyperglycaemia can be shown in dogs subject to a continuous glucose infusion (hyperglycaemic clamp). The normalization of the hyperglycaemia can be evaluated as compared to no treatment and/or to a combined treatment with insulin and/or a treatment with insulin alone.

Example 6

Prevention or Treatment of Hyperglycaemia Associated Complications

(20) The treatment of hyperglycaemic or insulin dependent or insulin resistance diabetic dogs with Compound A according to the invention or a combination of active substances according to the invention prevents or reduces hyperglycaemia associated complications, e.g. cataract formation.

(21) Evidence of the therapeutic success is compared with dogs that have been treated with other antidiabetic medicaments or with placebo. The success of the treatment is determined e.g. by ophthalmological eye examination of the development or the progression or the regression of cataract formation. And/or the time to development of a cataract and/or progression of the cataract maturation may be determined and be compared to dogs who have been treated with other antidiabetic medicaments or with placebo.

Example 7

Treatment of Insulin Resistance

(22) In clinical studies in insulin resistant dogs running for different lengths of time (e.g. 4 weeks to 12 months) the success of the treatment is checked using the measurement of baseline blood glucose, blood fructosamine and blood insulin and/or c-peptide level and the corresponding relation between the parameter in the individual dog.

(23) Also the glucose and blood lipids (e.g. NEFA) and/or insulin values after a meal or after a loading test (glucose tolerance test or insulin tolerance test) during or after the end of the period of therapy for the study can be compared with the values before the start of the study and/or with those of insulin resistant dogs who have been treated with other medications or placebo.

Example 8

Treatment of Pre-Diabetes in Dogs

(24) The efficacy of SGLT2 inhibition in accordance with the invention in the treatment of pre-diabetes characterised by pathological fasting glucose and/or impaired glucose tolerance and/or insulin resistance can be tested using clinical studies. In studies over a shorter or longer period (e.g. 2-4 weeks or 1-2 years) the success of the treatment is examined by determining the fasting glucose values and/or the glucose values after a meal or after a loading test (intravenous glucose tolerance test or food tolerance test after a defined meal or insulin tolerance test) after the end of the period of therapy for the study and comparing them with the values before the start of the study and/or with those of a placebo group. In addition, the fructosamine value can be determined before and after therapy and compared with the initial value and/or the placebo value. A significant drop in the fasting or non-fasting glucose and/or fructosamine levels demonstrates the efficacy of the treatment of pre-diabetes

Example 9

Effects on Body Weight, Body Composition, Dyslipidemia and Dysadipokinemia

(25) Treating dogs with metabolic disorders such as obesity, dyslipidaemia, dysadipokinemia, hepatic lipidosis, subclinical inflammation or systemic inflammation, in particular low grade systemic inflammation, which also comprises adipose tissue, and associated disorders, such as Syndrome X (metabolic syndrome), and/or insulin resistance, hyperglycaemia, hyperinsulinaemia, impaired glucose tolerance is also in pursuit of the goal of preventing the transition or slowing the progression to e.g. clinically manifest consequences of the metabolic disorders e.g. hypertension, cardiomyopathy, renal dysfunction and/or musculoskeletal disorders in canine animals.

(26) The efficacy of a treatment can be investigated in a comparative clinical study in which dogs are treated over a lengthy period (e.g. 3-12 months) with either Compound A or a combination of active substances or with placebo or with a non-drug therapy (e.g. diet) or other medicaments. Prior, during and at the end of the therapy the parameter can be determined: body weight (scale) and body composition e.g. with dual-energy x-ray absorptiometry. In plasma or serum lipid (e.g. Triglycerides, Cholesterol, NEFA) and adipokine (e.g. adiponectin, leptin) profiles as well as inflammatory markers (e.g. c-reactive protein, monocyte chemoattractant protein-I) can be measured. Insulin and glucose level can be determined basal as well as e.g. after a loading test. Renal parameter can be determined in blood and urinary samples (e.g. urea, creatinine, urinary albumin). Additionally, the blood pressure and/or also evidences of cardiomyopathy can be investigated with echocardiographic doppler ultrasound measurements. An improvement in musculoskeletal disorders (e.g. osteoarthritis) can be quantified e.g. with activity, lameness, and pain scores.

Example 10

Preparation of 1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene (compound A)

(27) The following example of synthesis serves to illustrate a method of preparing 1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene (compound A). A method of preparing its crystalline complex with L-proline is also described. It is to be regarded only as a possible method described by way of example, without restriction of the scope of the invention. The terms “room temperature” and “ambient temperature” are used interchangeably and denote temperatures of about 20° C. The following abbreviations are used:

(28) DMF dimethylformamide

(29) NMP N-methyl-2-pyrrolidone

(30) THF tetrahydrofuran

Preparation of 4-bromo-3-hydroxymethyl-1-iodo-benzene

(31) ##STR00020##

(32) Oxalyl chloride (13.0 mL) is added to an ice-cold solution of 2-bromo-5-iodo-benzoic acid (49.5 g) in CH.sub.2Cl.sub.2 (200 mL). DMF (0.2 mL) is added and the solution is stirred at room temperature for 6 h. Then, the solution is concentrated under reduced pressure and the residue is dissolved in THF (100 mL). The resulting solution is cooled in an ice-bath and LiBH.sub.4 (3.4 g) is added in portions. The cooling bath is removed and the mixture is stirred at room temperature for 1 h. The reaction mixture is diluted with THF and treated with 0.1 M hydrochloric acid. Then, the organic layer is separated and the aqueous layer is extracted with ethyl acetate. The combined organic layers are dried (Na.sub.2SO.sub.4) and the solvent is evaporated under reduced pressure to give the crude product.

(33) Yield: 47.0 g (99% of theory)

Preparation of 4-bromo-3-chloromethyl-1-iodo-benzene

(34) ##STR00021##

(35) Thionyl chloride (13 mL) is added to a suspension of 4-bromo-3-hydroxymethyl-1-iodo-benzene (47.0 g) in dichloromethane (100 mL) containing DMF (0.1 mL). The mixture is stirred at ambient temperature for 3 h. Then, the solvent and the excess reagent is removed under reduced pressure. The residue is triturated with methanol and dried.

(36) Yield: 41.0 g (82% of theory)

Preparation of 4-bromo-1-iodo-3-phenoxymethyl-benzene

(37) ##STR00022##

(38) Phenol (13 g) dissolved in 4 M KOH solution (60 mL) is added to 4-bromo-3-chloromethyl-1-iodo-benzene (41.0 g) dissolved in acetone (50 mL). NaI (0.5 g) is added and the resulting mixture is stirred at 50° C. overnight. Then, water is added and the resulting mixture is extracted with ethyl acetate. The combined extracts are dried (Na.sub.2SO.sub.4) and the solvent is evaporated under reduced pressure. The residue is purified by chromatography on silica gel (cyclohexane/ethyl acetate 19:1).

(39) Yield: 38.0 g (79% of theory)

Preparation of 1-bromo-4-(1-methoxy-D-glueopyranos-1-yl)-2-(phenoxymethyl)-benzene

(40) ##STR00023##

(41) A 2 M solution of iPrMgCl in THF (11 mL) is added to dry LiCl (0.47 g) suspended in THF (11 mL). The mixture is stirred at room temperature until all the LiCl is dissolved. This solution is added dropwise to a solution of 4-bromo-1-iodo-3-phenoxymethyl-benzene (8.0 g) in tetrahydrofuran (40 mL) cooled to −60° C. under argon atmosphere. The solution is warmed to −40° C. and then 2,3,4,6-tetrakis-O-(trimethylsilyl)-D-glucopyranone (10.7 g, 90% pure) in tetrahydrofuran (5 mL) is added. The resulting solution is warmed to −5° C. in the cooling bath and stirred for another 30 min at this temperature. Aqueous NH.sub.4Cl solution is added and the resultant mixture is extracted with ethyl acetate. The combined organic extracts are dried over sodium sulfate and the solvent is removed under reduced pressure. The residue is dissolved in methanol (80 mL) and treated with methanesulfonic acid (0.6 mL) to produce the more stable anomer solely. After stirring the reaction solution at 35-40° C. overnight, the solution is neutralized with solid NaHCO.sub.3 and the methanol is removed under reduced pressure. The remainder is diluted with aqueous NaHCO.sub.3 solution and the resulting mixture is extracted with ethyl acetate. The combined extracts are dried over sodium sulfate and the solvent is evaporated to yield the crude product that is submitted to reduction without further purification.

(42) Yield: 7.8 g (93% of theory)

Preparation of 1-bromo-4-(2,3,4,6-tetra-O-acetyl-D-glueopyranos-1-yl)-2-(phenoxymethyl)-benzene

(43) ##STR00024##

(44) Boron trifluoride diethyletherate (4.9 mL) is added to a solution of 1-bromo-4-(1-methoxy-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene (8.7 g) and triethylsilane (9.1 mL) in dichloromethane (35 mL) and acetonitrile (50 mL) cooled to −20° C. at such a rate that the temperature maintains below −10° C. The resultant solution is warmed to 0° C. over a period of 1.5 h and then treated with aqueous sodium hydrogen carbonate solution. The resulting mixture is stirred for 0.5 h, the organic solvent is removed and the residue is extracted with ethyl acetate. The combined organic layers are dried over sodium sulfate and the solvent is removed. The residue is taken up in dichloromethane (50 mL) and pyridine (9.4 mL), acetic anhydride (9.3 mL) and 4-dimethylaminopyridine (0.5 g) are added in succession to the solution. The solution is stirred for 1.5 h at ambient temperature and then diluted with dichloromethane. This solution is washed twice with 1 M hydrochloric acid and dried over sodium sulfate. After the solvent is removed, the residue is recrystallized from ethanol to furnish the product as a colourless solid.

(45) Yield: 6.78 g (60% of theory)

(46) Mass spectrum (ESI.sup.+): m/z=610/612 (Br) [M+NH.sub.4].sup.+

Preparation of 2-(phenoxymethyl)-4-(2,3,4,6-tetra-O-acetyl-D-glueopyranos-1-yl)-benzonitrile

(47) ##STR00025##

(48) A flask charged with zinc cyanide (1.0 g), zinc (30 mg), Pd.sub.2(dibenzylideneacetone).sub.3*CHCl.sub.3 (141 mg) and tri-tert-butylphosphonium tetrafluoroborate (111 mg) is flushed with argon. Then a solution of 1-bromo-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene (5.4 g) in NMP (12 mL) is added and the resulting mixture is stirred at room temperature for 18 h. After dilution with ethyl acetate, the mixture is filtered and the filtrate is washed with aqueous sodium hydrogen carbonate solution. The organic phase is dried (sodium sulfate) and the solvent is removed. The residue is recrystallized from ethanol.

(49) Yield: 4.10 g (84% of theory)

(50) Mass spectrum (ESI.sup.+): m/z=557 [M+NH.sub.4].sup.+

(51) Alternatively, the compound described above is synthesized starting from 1-bromo-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene using copper(I) cyanide (2 equivalents) in NMP at 210° C.

Preparation of 2-bromomethyl-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile

(52) ##STR00026##

(53) A 33% solution of hydrobromic acid in acetic acid (15 mL) is added to a solution of 2-phenyloxymethyl-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile (0.71 g) and acetic anhydride (0.12 mL) in acetic acid (10 ml). The resulting solution is stirred at 55° C. for 6 h and then cooled in an ice-bath. The reaction mixture is neutralized with chilled aqueous potassium carbonate solution, and the resultant mixture is extracted with ethyl acetate. The combined organic extracts are dried over sodium sulfate and the solvent is removed under reduced pressure. The residue is taken up in ethyl acetate/cyclohexane (1:5), and the precipitate is separated by filtration and dried at 50° C. to give the pure product.

(54) Yield: 0.52 g (75% of theory)

(55) Mass spectrum (ESI.sup.+): m/z=543/545 (Br) [M+NH.sub.4].sup.+

Preparation of 4-cyclopropyl-phenylboronic acid

(56) ##STR00027##

(57) 2.5 M solution of nButyllithium in hexane (14.5 mL) is added dropwise to 1-bromo-4-cyclopropyl-benzene (5.92 g) dissolved in THF (14 mL) and toluene (50 mL) and chilled to −70° C. The resultant solution is stirred at −70° C. for 30 min before triisopropyl borate (8.5 mL) is added. The solution is warmed to −20° C. and then treated with 4 M aqueous hydrochloric acid (15.5 mL). The reaction mixture is further warmed to room temperature and then the organic phase is separated. The aqueous phase is extracted with ethyl acetate and the combined organic phases are dried (sodium sulfate). The solvent is evaporated and the residue is washed with a mixture of ether and cyclohexane to give the product as a colourless solid.

(58) Yield: 2.92 g (60% of theory)

(59) Mass spectrum (ESI.sup.−): m/z=207 (Cl) [M+HCOO].sup.−

Preparation of 1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene

(60) ##STR00028##

(61) An Ar filled flask is charged with 2-bromomethyl-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile (1.60 g), 4-cyclopropyl-phenylboronic acid (1.0 g), potassium carbonate (1.85 g) and a degassed 3:1 mixture of acetone and water (22 mL). The mixture is stirred at room temperature for 5 min, before it is cooled in an ice-bath. Then palladium dichloride (30 mg) is added and the reaction mixture is stirred for 16 h at ambient temperature. The mixture is then diluted with brine and extracted with ethyl acetate. The combined extracts are dried over sodium sulfate and the solvent is removed under reduced pressure. The residue is dissolved in methanol (20 mL) and treated with 4 M aqueous potassium hydroxide solution (4 mL). The resulting solution is stirred at ambient temperature for 1 h and then neutralized with 1 M hydrochloric acid. The methanol is evaporated, and the residue is diluted with brine and extracted with ethyl acetate. The organic extracts collected are dried over sodium sulfate, and the solvent is removed. The residue is chromatographed on silica gel (dichloromethane/methanol 1:0.fwdarw.8:1).

(62) Yield: 0.91 g (76% of theory)

(63) Mass spectrum (ESI.sup.+): m/z=413 [M+NH.sub.4].sup.+

Preparation of a Crystalline Complex (1:1) of Compound A With L-proline

(64) L-proline (0.34 g) dissolved in 2.1 mL of a mixture of ethanol and water (volume ratio 10:1) is added to a solution of 1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene (1.17 g, obtained as described above) dissolved in 2 mL ethanol. The resulting solution is allowed to stand at ambient temperature. After about 16 h the crystalline complex is isolated as white crystals by filtration. If necessary the crystallisation may be initiated by scratching with a glass rod or metal spatula for example or by inoculating with seed crystals. Residual solvent is removed by storing the crystals at slightly elevated temperature (30 to 50° C.) under vacuum for about 4 h to yield 1.27 g of the crystalline 1:1 complex of L-proline and 1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene.

(65) Several batches of the crystalline complex according to the above preparation are obtained. The X-ray powder diffraction patterns coincide. The melting points are determined via DSC and evaluated as onset-temperature. Examples of melting points are approximately 89° C., 90° C., 92° C., 101° C. and 110° C. The X-ray powder diffraction pattern as contained in Table 1 and as depicted in FIG. 11 and the DSC and TG diagram in FIG. 12 correspond to a batch with a melting point of approximately 90° C.

(66) The X-ray powder diffraction pattern of the crystalline complex of the compound A and L-proline (peaks up to 30° in 2Θ) is provided above in Table 1.

Example 11

Formulations

(67) Some examples of formulations are described in which the term “active substance” denotes an SGLT2 inhibitor or pharmaceutically acceptable form thereof, e.g. a prodrug or a crystalline form, for use according to the invention. In the case of a combination with one or additional active substances, the term “active substance” may also include the additional active substance.

(68) Tablets Containing 100 mg of Active Substance

(69) Composition:

(70) 1 tablet contains:

(71) TABLE-US-00003 active substance 100.0 mg lactose 80.0 mg corn starch 34.0 mg polyvinylpyrrolidone 4.0 mg magnesium stearate 2.0 mg 220.0 mg

(72) Method of Preparation:

(73) The active substance, lactose and starch are mixed together and uniformly moistened with an aqueous solution of the polyvinylpyrrolidone. After the moist composition has been screened (2.0 mm mesh size) and dried in a rack-type drier at 50° C. it is screened again (1.5 mm mesh size) and the lubricant is added. The finished mixture is compressed to form tablets.

(74) Weight of tablet: 220 mg

(75) Diameter: 10 mm, biplanar, facetted on both sides and notched on one side.

(76) Tablets Containing 150 mg of Active Substance

(77) Composition:

(78) 1 tablet contains:

(79) TABLE-US-00004 active substance 150.0 mg powdered lactose 89.0 mg corn starch 40.0 mg colloidal silica 10.0 mg polyvinylpyrrolidone 10.0 mg magnesium stearate 1.0 mg 300.0 mg

(80) Preparation:

(81) The active substance mixed with lactose, corn starch and silica is moistened with a 20% aqueous polyvinylpyrrolidone solution and passed through a screen with a mesh size of 1.5 mm. The granules, dried at 45° C., are passed through the same screen again and mixed with the specified amount of magnesium stearate. Tablets are pressed from the mixture. Weight of tablet: 300 mg die: 10 mm, flat

(82) Hard Gelatine Capsules Containing 150 mg of Active Substance

(83) Composition:

(84) 1 capsule contains:

(85) TABLE-US-00005 active substance 150.0 mg com starch (dried) approx. 180.0 mg lactose (powdered) approx. 87.0 mg magnesium stearate 3.0 mg approx. 420.0 mg

(86) Preparation:

(87) The active substance is mixed with the excipients, passed through a screen with a mesh size of 0.75 mm and homogeneously mixed using a suitable apparatus. The finished mixture is packed into size 1 hard gelatine capsules. Capsule filling: approx. 320 mg Capsule shell: size 1 hard gelatine capsule.

(88) Suppositories containing 150 mg of active substance

(89) Composition:

(90) 1 suppository contains:

(91) TABLE-US-00006 active substance 150.0 mg polyethyleneglycol 1500 550.0 mg polyethyleneglycol 6000 460.0 mg polyoxyethylene sorbitan monostearate 840.0 mg 2,000.0 mg

(92) Preparation:

(93) After the suppository mass has been melted the active substance is homogeneously distributed therein and the melt is poured into chilled moulds.

(94) Ampoules Containing 10 mg Active Substance

(95) Composition:

(96) TABLE-US-00007 active substance 10.0 mg 0.01N hydrochloric acid/NaCl q.s. double-distilled water ad 2.0 ml

(97) Preparation:

(98) The active substance is dissolved in the necessary amount of 0.01 N HCl, made isotonic with common salt, filtered sterile and transferred into 2 ml ampoules.

(99) Ampoules Containing 50 mg of Active Substance

(100) Composition:

(101) TABLE-US-00008 active substance 50.0 mg 0.01N hydrochloric acid/NaCl q.s. double-distilled water ad 10.0 ml

(102) Preparation:

(103) The active substance is dissolved in the necessary amount of 0.01 N HCl, made isotonic with common salt, filtered sterile and transferred into 10 ml ampoules.

REFERENCES

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