EA043941B1 - TREATMENT OF METABOLIC DISORDERS IN FELIDES - Google Patents

Description

Field of technology to which the invention relates

The invention relates to veterinary medicine, in particular to the treatment and/or prevention of metabolic disorders in felines.

Prerequisites for creating the invention

Felines, such as cats, are susceptible to various metabolic disorders. A number of metabolic disorders are known in felines, including hyperglycemia, insulin resistance, diabetes (eg, type 1 or type 2 diabetes mellitus or prediabetes), hepatic lipidosis, obesity, hyperinsulinemia, impaired glucose tolerance, ketosis (eg. ketoacidosis), dyslipidemia, dysadipokinemia, obesity, subclinical inflammation or systemic inflammation, in particular low-grade systemic inflammation, which also includes inflammation of adipose tissue, syndrome X (metabolic syndrome), atherosclerosis and/or inflammation of the pancreas. There are various correlations between these disorders. Among these disorders in cats, diabetes mellitus, particularly prediabetes and type 2 diabetes mellitus, as well as hyperglycemia, insulin resistance, hepatic lipidosis and obesity, are of increasing importance. This may be at least partly due to changes in the living conditions and feeding behavior of pets over recent years.

Diabetes mellitus is characterized by disturbances in the metabolism of carbohydrates, proteins and triglycerides based on a relative or absolute lack of insulin. This is a relatively common endocrinopathy in felines such as cats. The incidence in cats has increased approximately 5-12 times over the past four decades, approximately 0.5-1.2%. Several risk factors have been identified: age, obesity, sterilization and gender. Males, neutered, old and obese (>10 years) are likely to have the greatest risk of developing diabetes.

In accordance with the existing classification, diabetes mellitus is divided into three classes:

(1) type 1 diabetes, which results from loss of function of insulin-secreting cells, such as through immunological destruction of beta cells or the presence of autoantibodies to insulin (human juvenile diabetes);

(2) type 2 diabetes, which results from the failure of insulin-stimulated cells to respond properly to an insulin stimulus; it is also associated, for example, with amyloid accumulation in beta cells; Type 2 diabetes usually develops over a long period of so-called prediabetes;

(3) secondary diabetes mellitus, which may occur due to diabetogenic drugs (for example, long-acting glucosteriods, megestrol acetate, etc.) or other primary diseases such as pancreatitis, pancreatic adenocarcinoma, Cushing's syndrome, hypo- or hyperthyroidism , tumors that produce growth hormone, leading to acromegaly.

Type 2 diabetes in particular is a growing problem for cat populations throughout the developed world. Changes in the lifestyles of cat owners are affecting their cats - they are increasingly indoors, have reduced activity levels and eat high-calorie diets, leading to obesity and a susceptibility to type 2 diabetes. As these trends continue, the incidence of Diabetes mellitus in cats increases accordingly.

Several oral hypoglycemic agents have been approved for the treatment of diabetes mellitus in humans, particularly type 2 diabetes mellitus. These drugs act, for example, by stimulating pancreatic insulin secretion in a glucose-independent or glucose-dependent manner (sulfonylureas/meglitinides, or DPP IV inhibitors, respectively), by increasing tissue sensitivity to insulin (biguanides, thiazolidinediones), or by slowing postprandial absorption of glucose in the intestine (alpha-glucosidase inhibitors).

Other approaches to treat diabetes and reduce hyperglycemia in humans are also being considered, including inhibition of the renal sodium-dependent glucose cotransporter SGLT2. SGLT2 in the kidney regulates glucose levels by mediating the reabsorption of glucose back into plasma after blood filtration. Thus, inhibition of SGLT2 induces glycosuria and may lead to decreased blood glucose levels. For example, the compound 1-cyano-2-(4-cyclopropylbenzyl)-4-(beta-Dglucopyranos-1-yl)benzene is described as an SGLT2 inhibitor in WO 2007/128749. A wide variety of additional SGLT2 inhibitors are also known. WO 2011/117295, which relates to the treatment of predominantly carnivorous non-human animals with dipeptidyl peptidase IV (DPP-IV) inhibitors, highlights various SGLT2 inhibitors among numerous other types of compounds in the context of the use of combination therapy with DPP-IV inhibitors.

Inhibition of SGLT2 has not previously been indicated for the treatment of metabolic disorders in felines such as cats. In felines, medications for treating metabolic disorders are much less developed than in humans. Unfortunately, even if treatment or prevention is effective in humans, for example, or other non-feline animals,

- 1 043941 it is not possible to conclude that the same approach would be effective, safe, and also advisable in felines such as the cat.

Felines differ significantly from humans or, for example, dogs in terms of their metabolism.

Being strictly carnivores, felines are not well adapted to carbohydrates in their diet. For example, feline liver does not show glucokinase activity (Tanaka et al., Vet Res Commun 2005, 29 (6): 477-485). In most mammals, such as dogs or humans, hepatic glucokinases act as a glucose sensor that allows hepatic metabolism to adequately respond to changes in plasma glucose concentrations. In addition, insulin release from the cat pancreas has been found to be less sensitive to glucose as a stimulus compared to most other species (Curry et al., Comp Biochem Physiol 1982. 72A(2): 333-338).

Another adaptation to a strictly carnivorous diet relates to the utilization of proteins and fats for energy production, i.e. gluconeogenesis. In omnivores, gluconeogenesis occurs primarily in a state of fasting. In contrast, in obligate carnivores such as cats, gluconeogenesis is found to be constitutively active in the liver, regardless of nutritional status, and after food consumption is even higher than in the fasted state (Hoenig et al. Am J Physiol, 2011, 301(6):R1798-1807, Verbrugghe et al., Crit Rev Food Sci Nutr. 2012;52(2): 172-182).

Accordingly, the pathophysiology of metabolic disorders in felines, and consequently their response to treatment for such disorders, differs from that of other species.

As a diabetic complication, such as diabetes, vision problems and cataracts are common in dogs but are rare in felines.

Oral diabetes medications that are known as human drugs, such as glipizide (a sulfonylurea), work in a small proportion of cats, but these medications may be completely ineffective if the pancreas is not working. Moreover, in some studies, glipizide and other oral hypoglycemic drugs have been shown to cause side effects such as vomiting and jaundice, and damage the pancreas further, resulting in a decreased chance of diabetes remission for cats. They have also been demonstrated to cause liver damage. Even lower efficacy has been shown for other groups of compounds, e.g. meglitinides, biguanides, thiazolidinediones and aglucosidase inhibitors (Palm C.A. et al., Vet Clin Small Anim 2013, 43: 407-415).

Insulin injections are currently considered the gold standard for treating diabetic cats. However, cats are known to be unpredictable in their response to exogenous insulin. No single type of insulin is generally effective in maintaining glycemic control, even when administered twice daily. Even with strict owner compliance, control is often lax and common secondary problems arise. Many owners find it impossible to achieve acceptable levels of compliance because synchronizing food intake and insulin administration is not possible in most cases. Ultimately, many cats with diabetes are euthanized due to the disease.

Factors governing patient and owner compliance are also quite different. In cats, for example, oral administration is even more desirable than in humans.

Treatment that allows for strict adherence and therefore improved glycemic control compared with currently available insulin-based treatments will help reduce disease progression and delay or prevent the onset of complications in many animals.

Additionally, even when diabetic cats are treated aggressively with insulin and clinical remission is achieved, this also does not necessarily normalize insulin secretion, pancreatic beta cell function, and/or insulin resistance. Cats remain prone to new onset diabetes. It would be desirable to have a method of treating diabetes mellitus in felines that better improves, for example, insulin resistance and pancreatic beta cell function Reusch S.E. et al., Schweizer Archiv fuer Tierheilkunde 2011, 153811): 495-500).

Thus, there is still a great need for effective, safe and acceptable treatments for metabolic disorders, including diabetes, in felines.

Disclosure of the Invention Summary of the Invention

The inventors of the present invention have surprisingly found that inhibition of SGLT2 is effective and safe in the treatment and/or prevention of metabolic disorders in felines.

The present invention thus provides the use of one or more SGLT2 inhibitors or pharmaceutically acceptable crystalline forms thereof in the treatment and/or prevention of a metabolic disorder in a feline, wherein said one or more SGLT2 inhibitors is a glucopyranosyl-substituted derivative of the common benzene formulas (1)

R ²

where R ¹ is cyano, Cl or methyl;

R ² is H, methyl, methoxy or hydroxyl; And

R ³ is cyclopropyl, hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, 3-methyl-but-1-yl, cyclobutyl, cyclopentyl , cyclohexyl, 1-hydroxycyclopropyl, 1-hydroxycyclobutyl, 1-hydroxycyclopentyl, 1-hydroxycyclohexyl, ethynyl, ethoxy, difluoromethyl, trifluoromethyl, pentafluoroethyl, 2-hydroxyethyl, hydroxymethyl, 3hydroxypropyl, 2-hydroxy-2-methyl-prop-1-yl, 3 -hydroxy-3-methyl-but-1-yl, 1-hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1 -hydroxy-1-trifluoromethyl-ethyl, 2-methoxy-ethyl, 2-ethoxy-ethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxy-ethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethylsulfinyl, ethylsulfonyl, trimethylsilyl, (R)tetrahydrofuran- 3-yloxy or (S)-tetrahydrofuran-3-yloxy or cyano, wherein one or more hydroxyl groups of the β-E-glucopyranosyl group are optionally further acylated with groups selected from ( _C1.18 alkyl)carbonyl, ( _C1.18 alkyl )oxycarbonyl, phenylcarbonyl and phenyl-(C1. ₃ alkyl)carbonyl, wherein such one or more SGLT2 inhibitors or pharmaceutically acceptable crystalline forms thereof are administered at a dose of 0.01 to 5.0 mg/kg body weight per day.

Additional aspects of the invention are defined below, as well as in the claims.

A pharmaceutically acceptable form of one or more SGLT2 inhibitors may be a crystalline complex between one or more SGLT2 inhibitors and one or more amino acids such as proline.

In accordance with the invention, one or more SGLT2 inhibitors, or a pharmaceutically acceptable form thereof, can be provided, for example, for oral or parenteral administration, preferably for oral administration.

One or more SGLT2 inhibitors, or a pharmaceutically acceptable form thereof, may be administered in dosages of 0.1 to 3.0 mg/kg body weight per day, preferably 0.2 to 2.0 mg/kg body weight per day, more preferably from 0.1 to 1 mg/kg body weight per day. Thus, one or more SGLT2 inhibitors, or a pharmaceutically acceptable form thereof, can be formulated to administer 0.1 to 3.0 mg/kg body weight per day, preferably 0.2 to 2.0 mg/kg body weight per day, more preferably from 0.1 to 1 mg/kg body weight per day.

One or more SGLT2 inhibitors, or a pharmaceutically acceptable form thereof, are preferably administered only once daily.

The present invention also provides a pharmaceutical composition that includes one or more SGLT2 inhibitors, or a pharmaceutically acceptable form thereof, for use in accordance with the invention as disclosed herein.

In the examples provided herein, the therapeutic and/or prophylactic benefits of SGLT2 inhibition in accordance with the present invention are demonstrated experimentally. The experimental data disclosed in this application are intended to illustrate the invention, but are not intended to have a limiting effect on the scope of protection as defined herein below and in the claims.

In particular, the present inventors have unexpectedly discovered that use of one or more SGLT2 inhibitors according to the present invention advantageously results in a reduction in insulin resistance in treated insulin resistant felines. That is, likewise, use of one or more SGLT2 inhibitors according to the present invention advantageously results in increased insulin sensitivity in treated insulin-resistant felines. Insulin sensitivity can be calculated using various simplified indices, for example, during the glucose tolerance test in the form of the modified Belfiore index (1 /log(ΔAUC-glucose * ΔAU C-insulin)).

The invention thus makes it possible to improve the treatment and/or prevention of diabetes, in particular type 2 diabetes mellitus, in felines.

Use of one or more SGLT2 inhibitors according to the present invention

- 3 043941 advantageously leads to reduced excursions in insulin levels, for example, as measured by an intravenous glucose tolerance test (ivGTT), or after consumption of any form of glucose, for example, consumption of a high carbohydrate meal (acute postprandial increase in insulin levels) or following a stress-induced increase in blood glucose levels. In particular, the use of one or more SGLT2 inhibitors according to the invention advantageously also results in reduced second phase insulin secretion, for example as measured by the ivGTT process, or after consumption of any other form of glucose, for example after a meal.

The use of one or more SGLT2 inhibitors in accordance with the present invention advantageously also results in decreased levels of non-esterified fatty acids in the blood plasma or improved elimination of non-esterified fatty acids from the bloodstream, for example, as measured in the ivGTT process, or after consumption of any other form increasing insulin levels in the blood.

The use of one or more SGLT2 inhibitors according to the present invention thus generally results in improved glucose tolerance, i.e. reduces glucose intolerance.

The glucose deviation in the insulin tolerance test (ivITT) of a feline treated according to the invention is also preferably improved compared to an untreated animal.

The use of one or more SGLT2 inhibitors in accordance with the present invention also advantageously results in a reduction in body fat levels, blood leptin levels and/or respiratory gas exchange ratio (RER). The invention is also associated with anti-obesity effects and may in particular advantageously prevent body weight gain and/or result in weight loss in felines. In one aspect, the invention thus allows for the control of obesity and/or obesity-related metabolic disorders in felines.

Effects of applications in accordance with the present invention (i.e., the above-mentioned preferred effects on insulin resistance/sensitivity, changes in insulin levels, second phase insulin secretion, glucose tolerance, elimination of non-esterified fatty acids, body fat, blood leptin levels, values RER and/or body weight) are also advantageous in that they allow for subclinical treatment, such as the treatment of prediabetic conditions in felines. They thus provide the opportunity to prevent or delay the onset of diabetes mellitus in felines. In particular, they provide the ability to prevent or delay the development of certain metabolic disorders, symptoms or conditions as described herein (such as hyperglycemia, impaired glucose tolerance, insulin resistance, abnormal changes in insulin or glucose levels, high levels of non-esterified fatty acids). acids or leptin in the blood, obesity and/or loss of pancreatic beta cell function) in diabetes mellitus, in particular type 2 diabetes mellitus, in felines.

An additional advantage of the present invention is that the use of one or more SGLT2 inhibitors is effective only against metabolic disorders, i.e. If the use of one or more SGLT2 inhibitors is desired in a feline, provide monotherapy (ie, stand-alone therapy; ie, no other drugs are administered to the animal to treat or prevent the same metabolic disorder). The invention also allows for combination therapy using another drug (eg, an additional insulin-sensitizing drug or insulin itself).

An additional advantage of the present invention is that it has surprisingly been found that the use of one or more SGLT2 inhibitors is effective only against metabolic disorders, i.e. If the use of one or more SGLT2 inhibitors is desired in a feline, provides monotherapy (ie, stand-alone therapy; ie, when no other drugs are administered to the animal to treat or prevent the same metabolic disorder). This invention also provides the possibility of replacing insulin therapy in felines, or combination therapy using insulin or another drug (eg, a hypoglycemic drug). Such a combination advantageously results in a reduction in the dose and/or frequency with which insulin or other drug (eg, a hypoglycemic drug) is administered compared to feline therapy using insulin or other drug. Most preferably, the feline can gradually reduce the dose of insulin or other medication. Thus, clinical remission is achieved.

Thus, the use of one or more SGLT2 inhibitors according to the present invention provides improved treatment and/or prevention of metabolic diseases,

- 4 043941 as disclosed in this application, including diabetes and/or prediabetes in felines.

Effects of administration of one or more SGLT2 inhibitors in accordance with the present invention (e.g., the beneficial effects described above on insulin resistance/sensitivity, insulin excursion, second phase insulin secretion, glucose tolerance, elimination of non-esterified fatty acids, body fat, leptin levels blood, RER values, body weight and/or hyperglycemia) may refer to the same or a comparison feline before administration of one or more SGLT2 inhibitors in accordance with the present invention, and/or refers to a comparison feline that has not received said treatment (eg placebo group). In any case, when a comparison is made, it can be made after a certain period of treatment, which is, for example, 1, 2, 3, 4, 5, 6 or 7 days; 10 days, 14 days; 2, 3, 4, 5, 6, 7 or 8 weeks; 1, 2, 3 or 4 months. Preferably the treatment period is 4 weeks. Alternatively, the treatment period may be 6 or 8 weeks. Alternatively, the treatment period may be 8 weeks or more, for example 8-16 weeks.

An additional advantage of the present invention is that one or more SGLT2 inhibitors can be effectively administered orally to a feline. In addition, one or more SGLT2 inhibitors in accordance with the present invention can be administered only once a day. These benefits allow for better compliance between the treated animal and its owner. This results in better glycemic control of disorders (such as diabetes) for which felines are treated with insulin. In general, the use of one or more SGLT2 inhibitors in accordance with the present invention thus helps to attenuate (ie, delay or prevent) the development of metabolic disorders and delay or prevent the onset of metabolic disorders (eg, diabetes) and its complications in representatives of felines.

Accordingly, the present invention also provides pharmaceutical compositions that include one or more SGLT2 inhibitors in accordance with the invention for use in the treatment and/or prevention of metabolic disorders in felines.

The invention also provides methods for treating and/or preventing metabolic disorders in felines, which include administering to the feline in need of such treatment and/or prevention an effective dose of one or more SGLT2 inhibitors as described herein.

Preferably, the use of one or more SGLT2 inhibitors in accordance with the present invention does not cause hypoglycemia.

Effects of administration of one or more SGLT2 inhibitors according to the present invention (e.g., the above-mentioned beneficial effects on insulin resistance/sensitivity, insulin excursion, second phase insulin secretion, glucose tolerance, elimination of non-esterified fatty acids, body fat, leptin levels blood, RER values, body weight and/or hyperglycemia) may refer to the same or a comparison feline before administration of one or more SGLT2 inhibitors in accordance with the present invention, and/or may refer to a comparison feline that received standard treatment when using insulin (eg, control group) or received no treatment.

An additional advantage of the present invention is that one or more SGLT2 inhibitors can be effectively administered to a feline orally, for example, in liquid form. In addition, one or more SGLT2 inhibitors in accordance with the present invention can be administered only once a day. These advantages allow for optimal dosing and compliance by the animal being treated and the owner.

In general, the use of one or more SGLT2 inhibitors in accordance with the present invention may thus attenuate, delay or prevent the development of a metabolic disorder, for example, the metabolic disorders disclosed herein, or may slow or prevent the onset of a metabolic disorder and its complications in felines.

Definitions

All values and concentrations presented in this application are subject to inherent variability, which is acceptable in biological science, within ±10%. The term approximately also refers to this acceptable variation.

Treatment effects disclosed herein (such as increase, decrease, excess, prolongation, increase, decrease, improvement, delay, pathological levels or any other change or deviation relative to the reference/control) may be observed at a statistical significance of p< 0.05, preferably <0.01.

When reference is made herein to a deviation (e.g. increase, decrease, excess, extension, increase, decrease, improvement, delay, pathological levels or any other change or deviation relative to control), the deviation may be, for example, 5% or more, in particular 10% or more, more preferably 15% or more, more preferably 20% or more, more preferably 30% or more, more preferably 40% or more, or, in

- 5 043941 in particular, 50% or more, in relation to the corresponding control values, unless otherwise stated. Typically the deviation will be at least 10%, i.e. 10% or more. The deviation may also be 20%. The deviation can also be 30%. The deviation can also be 40%. An appropriate control value can be obtained from a group of control animals that are treated with placebo instead of one or more SGLT2 inhibitors, or a group of animals that are not treated.

In this application, for example, an insulin deviation or a glucose deviation refers to a change in concentration or level in the blood over a period of time. The magnitude of deviations, such as insulin deviations or glucose deviations, can be expressed as area under the curve (AUC) values.

As used herein, the terms active substance or active ingredient include one or more SGLT2 inhibitors, or any pharmaceutically acceptable forms thereof (eg, prodrug or crystalline form) for use in accordance with the present invention. When combined with one or an additional active compound, the term active ingredient or active substance may also include the additional active compound.

In this application, the expression associated with, in particular, covers the expression caused by something.

In this application, ivGTT refers to an intravenous glucose tolerance test. ivGTT may typically use 0.8 g dextrose per kg body weight.

In this application, ivITT refers to an intravenous insulin tolerance test. In ivITT, 0.05 units may typically be used. insulin per kg body weight.

SGLT2 inhibitors.

SGLT2 inhibitors for use in accordance with the invention include, but are not limited to, glucopyranosyl-substituted benzene derivatives, for example, as described in WO 01/27128, WO 03/099836, WO 2005/092877, WO 2006/034489, WO 2006/ 064033, WO 2006/117359, WO 2006/117360, WO 2007/025943, WO 2007/028814, WO 2007/031548, WO 2007/093610, WO 2007/128749, WO 2008/0 49923, WO 2008/055870, WO 2008/ 055940, WO 2009/022020 or WO 2009/022008.

In addition, one or more SGLT2 inhibitors for use in accordance with the invention may be selected from the group consisting of the following compounds or pharmaceutically acceptable forms thereof:

(1) glucopyranosyl-substituted benzene derivative of general formula (1)

R ²

where R ¹ is cyano, Cl or methyl (most preferably cyano);

R ² is H, methyl, methoxy or hydroxy (most preferably H) and

R ³ is cyclopropyl, hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, 3-methyl-but-1-yl, cyclobutyl, cyclopentyl, cyclohexyl , 1-hydroxycyclopropyl, 1-hydroxycyclobutyl, 1-hydroxycyclopentyl, 1-hydroxycyclohexyl, ethynyl, ethoxy, difluoromethyl, trifluoromethyl, pentafluoroethyl, 2-hydroxyethyl, hydroxymethyl, 3-hydroxypropyl, 2-hydroxy-2-methyl-prop-1-yl, 3 -hydroxy-3-methyl-but-1-yl, 1-hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1-hydroxy -1-trifluoromethyl-ethyl, 2-methoxy-ethyl, 2-ethoxy-ethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxy-ethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethylsulfinyl, ethylsulfonyl, trimethylsilyl, (R)-tetrahydrofuran-3 -yloxy or (8)-tetrahydrofuran-3-yloxy or cyano;

where R ³ is preferably selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)tetrahydrofuran-3-yloxy or (B)-tetrahydrofuran-3-yloxy; and most preferably R ³ is cyclopropyl, or a derivative thereof, wherein one or more hydroxyl groups of the β-D-glucopyranosyl group are acylated with groups selected from (C _{1 .18} alkyl)carbonyl, (C ₁ . ₁₈ alkyl)oxycarbonyl, phenylcarbonyl and phenyl-(C ₁ .zalkyl)carbonyl.

Preferred SGLT2 inhibitors are glucopyranosyl-substituted benzene derivatives. Optionally, one or more hydroxyl groups of the glucopyranosyl group in such one or more SGLT2 inhibitors can be acylated with groups selected from ( _C1.18 alkyl)carbonyl, ( _C1.18 alkyl)oxycarbonyl, phenylcarbonyl and _phenyl- ( _C1 . alkyl)carbonyl.

More preferred are the glucopyranosyl-substituted benzonitrile derivatives of formula (1) as disclosed hereinabove. Even more preferred are glucopyranosyl-substituted benzonitrile derivatives of formula (18):

- 6 043941

where R ³ denotes cyclopropyl, hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, iso-butyl, tert-butyl, 3-methyl-but-1-yl, cyclobutyl, cyclopentyl, cyclohexyl, 1-hydroxy-cyclopropyl, 1-hydroxycyclobutyl, 1-hydroxycyclopentyl, 1-hydroxycyclohexyl, ethynyl, ethoxy, difluoromethyl, trifluoromethyl, pentafluoroethyl, 2-hydroxyethyl, hydroxymethyl, 3-hydroxypropyl, 2-hydroxy-2-methyl-prop- 1 -yl, 3-hydroxy-3-methyl-but-1 -yl, 1 -hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1 hydroxy-1-methyl-ethyl, 2,2,2 -trifluoro-1-hydroxy-1-trifluoromethyl-ethyl, 2-methoxy-ethyl, 2-ethoxy-ethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxy-ethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethylsulfinyl, ethylsulfonyl, trimethylsilyl, ( R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3-yloxy or cyano (wherein R ³ is preferably selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)-tetrahydrofuran-3-yloxy or (S)- tetrahydrofuran-3-yloxy; and R ³ is most preferably cyclopropyl, or a derivative thereof, wherein one or more hydroxyl groups of the β-E-glucopyranosyl group are acylated with groups selected from (C1. ₁₈ alkyl)carbonyl, (C1. ₁₈ alkyl)oxycarbonyl, phenylcarbonyl and phenyl-(C1_ ₃ alkyl)carbonyl.

Metabolic disorders.

A metabolic disorder may be diabetes, prediabetes, obesity, and/or any disorder, disease, condition, or symptom associated with one or more of these disorders. In particular, the metabolic disorder may be hyperglycemia, insulin resistance, diabetes and/or hepatic lipidosis. Additional associated metabolic disorders include hyperinsulinemia, impaired glucose tolerance, ketosis (particularly ketoacidosis), hyperlipidemia, elevated levels of fatty acids and/or glycerol in the blood, syndrome X (metabolic syndrome), atherosclerosis, pancreatic inflammation, adipose tissue inflammation and /or loss of pancreatic beta cell function.

In some embodiments, the metabolic disorder is diabetes. As used herein, diabetes may be prediabetes, type 1 diabetes mellitus, or type 2 diabetes mellitus. In particular, diabetes may be type 2 diabetes mellitus. In some embodiments, diabetes may be associated with obesity.

In some embodiments, the metabolic disorder is hyperglycemia. As used herein, hyperglycemia may be associated with diabetes, such as type 2 diabetes mellitus. In some embodiments, hyperglycemia may be associated with obesity. Hyperglycemia can be chronic.

In some embodiments, the metabolic disorder is insulin resistance. As used herein, insulin resistance may be associated with diabetes, such as type 2 diabetes mellitus. In some embodiments, insulin resistance may be associated with obesity.

In some embodiments, the metabolic disorder is impaired glucose tolerance (IGT). As used herein, impaired glucose tolerance may be associated with diabetes, such as type 2 diabetes. In some embodiments, impaired glucose tolerance may be associated with obesity.

In some embodiments, the metabolic disorder is hyperinsulinemia. In this application, hyperinsulinemia may be associated with diabetes, for example, type 2 diabetes mellitus. In some embodiments, hyperinsulinemia may be associated with obesity.

In some embodiments, the metabolic disorder is one or more of hyperglycemia, insulin resistance, and hepatic lipidosis. In some embodiments, the metabolic disorder is selected from hyperglycemia and insulin resistance.

In some embodiments, the metabolic disorder is one or more of hyperinsulinemia, impaired glucose tolerance, hyperglycemia, and insulin resistance.

In some embodiments, the feline is obese. For example, in accordance with the invention, one or more metabolic disorders selected from hyperglycemia, insulin resistance and hepatic lipidosis can be treated and/or prevented in an obese feline. In addition, for example, hyperinsulinemia and/or impaired glucose tolerance can be treated and/or prevented in an obese feline. In addition, one or more disorders selected from ketosis (particularly ketoacidosis), hyperlipidemia, elevated levels of fatty acids and/or glycerol, syndrome X (metabolic syndrome), atherosclerosis, pancreatic inflammation, adipose tissue inflammation, and loss of beta function pancreatic cells may be treated and/or prevented in an obese feline.

In some embodiments, the feline suffers from diabetes, for example, type 2 diabetes mellitus. For example, in accordance with the invention, one or more metabolic disorders selected from the group of hyperglycemia, insulin resistance and hepatic lipidosis may be treated and/or prevented in a feline that suffers from diabetes, e.g., type 2 diabetes mellitus. Additionally, for example, hyperinsulinemia and/or impaired glucose tolerance may be treated and/or prevented in a feline that suffers from diabetes, e.g. for diabetes mellitus type 2. In addition, one or more disorders selected from ketosis (particularly ketoacidosis), hyperlipidemia, elevated levels of fatty acids and/or glycerol, syndrome X (metabolic syndrome), atherosclerosis, pancreatic inflammation, adipose inflammation tissue and loss of pancreatic beta cell function may be treated and/or prevented in a feline that suffers from diabetes, such as type 2 diabetes mellitus.

In some embodiments, the feline is obese and suffers from diabetes, such as type 2 diabetes mellitus. In some embodiments, the feline suffers from diabetes, such as type 2 diabetes mellitus, but not obesity. In some embodiments, the feline is obese but not diabetic.

The present invention also provides the use of one or more SGLT2 inhibitors to treat and/or prevent pancreatic beta cell degeneration. For example, by increasing pancreatic beta cell mass, and/or improving and/or restoring the functionality (ie, insulin secretion) of pancreatic beta cells in a feline.

Ketosis is a state of increased levels of ketone bodies in the body. Ketoacidosis can be described as a type of metabolic acidosis that is caused by high concentrations of ketone bodies produced by the breakdown of fatty acids and the deamination of amino acids. Two common ketones that are produced in the human body are acetoacetic acid and β-hydroxybutyrate. There are mainly three ketones found in cats: acetoacetic acid, beta-hydroxybutyrate and pyruvic acid. Ketoacidosis may cause an odor to the subject's breath. This is due to acetone, a direct by-product of the spontaneous decomposition of acetoacetic acid.

Ketoacidosis is an extreme and uncontrolled form of ketosis. Ketosis is also a normal response to prolonged fasting. In ketoacidosis, the body is unable to adequately regulate ketone production, particularly through the production of acetyl-CoA, thus causing such a severe accumulation of ketoacids that the blood pH drops significantly, i.e. Excess ketone bodies can significantly acidify the blood. In extreme cases, ketoacidosis can be fatal.

Ketoacidosis can occur when the body produces high levels of ketone bodies by metabolizing fatty acids (ketosis), and insulin does not slow down this production sufficiently (for example, due to insulin resistance/reduced insulin sensitivity). The presence of high blood sugar levels (hyperglycemia), caused by a lack of insulin, can lead to a further increase in blood acidity. In healthy individuals, this usually does not occur because the pancreas produces insulin in response to rising ketone/blood sugar levels.

Ketoacidosis is most common in advanced diabetes mellitus, when the liver breaks down fat and proteins in response to the perceived need for respiratory substrate.

Prediabetes in felines is characterized by hyperinsulinemia, end-organ insulin resistance, impaired glucose tolerance, including, for example, an altered insulin response to glycemic challenge, and also, for example, stress-induced. Prediabetes is also often associated with obesity. Prediabetes may also be associated with intermittent hyperglycemia.

Type 2 diabetes in felines is characterized by both decreased insulin production and insulin resistance in target organs. Reduced insulin production may, for example, be due to amyloid accumulation in beta cells, glucose toxicity, and/or infection of the pancreas. The defect in beta cell function is usually progressive and, in some felines, results in complete loss of insulin secretion. Genetic factors, glucose steroids, progesterone or lack of exercise, and obesity are possible causes of insulin resistance. For example, in healthy cats, insulin sensitivity decreases by 50% after weight increases >40%. Cats with diabetes are thought to have primarily type 2 diabetes, based on the fact that most diabetic cats have islet amyloid, which has been called the hallmark of type 2 diabetes.

Only a significant minority of cats are thought to have secondary diabetes mellitus.

Clinical signs of diabetes mellitus observed in felines include:

- 8 043941 include polydipsia, polyuria, weight loss and/or polyphagia. In cats, anorexia is more often described as polyphagia. Pathognomonic for diabetes mellitus in cats is a plantigrade posture (weakness of the hind legs, metatarsus touching the ground when the cat walks). It is caused by diabetic neuropathy.

In addition, particularly relevant clinical signs of diabetes mellitus in felines in the context of the present invention are hyperglycemia and glycosuria. Hyperglycemia in a feline (eg, cat) is defined as plasma glucose values above normal values (3.9-8.3 mmol/L or 70-150 mg/dL), for example, 8 mmol/L or more or 150 mg/dL or more plasma glucose. Glucosuria in a feline (eg cat) is defined as urinary glucose levels above normal values (0-2 mmol/L or 36 mg/dL). The renal threshold is reached at a blood glucose concentration of about 11 to 17 mmol/L or 200 to 300 mg/dL.

The diagnosis of diabetes mellitus in felines may alternatively be based on three criteria, such as the following:

(1) measuring fasting blood glucose > 250 mg/dL;

(2) glycosuria, as defined above; and (3) one or more of the following: polyuria, polydipsia, polyphagia, weight loss despite good appetite, or ketonuria (without evidence of severe ketoacidosis).

In addition to and to support the above diagnostic criteria, additional tests may include hematology, blood chemistry, x-ray and/or abdominal ultrasound.

Preferably, the use of one or more SGLT2 inhibitors in accordance with the present invention allows the maintenance and/or establishment of normal or near-normal blood glucose concentrations. However, unlike human therapy, this is not always considered necessary for diabetic animals, and therefore does not always represent a treatment goal in accordance with the present invention. According to the invention, the blood glucose concentration can also be maintained at a level of, for example, 5.5 to 16.6 mmol/l or 100 to 300 mg/dl. For felines this will often be satisfactory.

One of the goals of treating prediabetes or diabetes in felines in accordance with the present invention may be for owners to eliminate visible signs (eg, lethargy, polyuria, polydipsia, weight loss, polyphagia, etc.) that occur secondary to hyperglycemia in untreated animals. Additional treatment targets or treatment effects may be one or more of any of the beneficial effects of the invention disclosed herein, including, but not limited to, any one or more of improved glucose tolerance, increased insulin sensitivity, decreased resistance to insulin, improved glucose excursion in ivITT, improved insulin excursion in ivGTT or oral glucose tolerance test (OGTT), decreased second-phase insulin secretion, decreased body fat, body weight and/or blood leptin, decreased ratio respiratory gas exchange (RER) and/or lack of weight gain if the animal is obese.

Diabetic remission is used in cats when normal (or near-normal) blood glucose concentrations are achieved, clinical signs improve, and insulin can be discontinued or omitted for at least four consecutive weeks. However, pancreatic beta cell viability may not be fully restored. The use of one or more SGLT2 inhibitors, and thereby reducing blood glucose concentrations, as well as improving insulin resistance and pancreatic beta cell function, is believed to be critical for achieving and maintaining diabetes remission in felines.

Insulin resistance can be described as a condition in which the normal amount of insulin is insufficient to produce a normal insulin response from fat, muscle and liver cells. Insulin resistance in fat cells reduces the effect of insulin and leads to increased hydrolysis of storage triglycerides in the absence of measures that either increase insulin sensitivity or provide additional insulin. Increased mobilization of storage lipids in these cells increases the level of free fatty acids in the blood plasma. Insulin resistance in muscle cells reduces glucose uptake (and thus local storage of glucose as glycogen), while insulin resistance in liver cells results in impaired glycogen synthesis and an inability to suppress glucose production. Elevated blood fatty acid levels, decreased muscle glucose uptake, and increased hepatic glucose production may contribute to elevated blood glucose levels (hyperglycemia).

Surrogate measures of insulin sensitivity can be calculated according to QUICKI (Quantitative Insulin Sensitivity Control Index: I/log (glucose*insulin)) for baseline blood levels. For dynamic studies, for example, the definition of the modified Belfiore index of the glucose tolerance test (I/log((AAUCglucose*(ΔAUC-insulin))) can be used.

Insulin resistance may be present in association with obesity, visceral obesity, hypertension and dyslipidemia associated with increased levels of triglycerides, small compact low-density lipoprotein (sdLDL) particles and reduced HDL cholesterol levels. Regarding visceral obesity, a wealth of evidence in humans suggests two strong links to insulin resistance. First, unlike subcutaneous adipose tissue, visceral fat cells produce significant amounts of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 and -6, etc. In numerous experimental models, these proinflammatory cytokines profoundly disrupted normal insulin action in fat and muscle cells, and they may represent a major contributor to the whole-body insulin resistance observed in human patients with visceral obesity. Similarly, in felines, excessive fat deposits contribute to a low degree of systemic inflammation. The cause of the vast majority of cases of insulin resistance remains unknown. There is a clear hereditary component. However, there are some reasons to suspect that insulin resistance is associated with high carbohydrate feed. Inflammation also appears to be involved in insulin resistance.

Hyperinsulinemia can be described as a condition in which there are excess levels, i.e. those that are approximately 35 pmol/L above the baseline level or approximately 200 pmol/L, such as under glycemic load (eg, ivGTT or stress), of insulin circulating in the blood. As mentioned, it is usually present in cases of insulin resistance, and may be a consequence of this in felines.

Impaired glucose tolerance can be described as a condition in which the response after a glycemic challenge, such as after a meal or after an exercise test (glucose tolerance test) or after a stress-induced increase in blood glucose concentration, the glycemic peak glucose deviation is greater high and/or the duration of the glucose deviation is prolonged.

Dyslipidemia or hyperlipidemia is the presence of elevated or abnormal levels of lipids and/or lipoproteins in the blood. Abnormal lipid and lipoprotein profiles are considered a highly modifiable risk factor for cholesterol-related cardiovascular disease. Glycerol is a precursor for the synthesis of triacylglycerols (triglycerides) and phospholipids in the liver and adipose tissue. When the body uses stored fat as an energy source, glycerol and fatty acids enter the bloodstream after hydrolysis of triglycerides. The glycerol component can be converted to glucose in the liver and provides energy for cellular metabolism. Normal levels of free fatty acids in the blood of a pet (such as a cat) are triglyceride concentrations of 50 to 100 mg/dL (0.6 to 1.2 mmol/L). Normal blood cholesterol levels are, for example, 70 to 150 mg/dL for a cat.

Dysadipokinemia can be characterized as a condition in which the level of biologically active substances circulating in the blood plasma, which are produced in adipose tissue and act in an autocrine/paracrine or endocrine manner, is abnormal. For example, increased leptin levels and/or decreased adiponectin levels.

Subclinical inflammation or systemic inflammation, in particular low-grade systemic inflammation, is characterized by increased expression and secretion of pro-inflammatory cytokines such as tumor necrosis factor-alpha and/or lower expression and secretion of anti-inflammatory cytokines such as interleukin-10 and/or their corresponding receptors.

Obesity can be described as a medical condition in which excess fat accumulates to the extent that it can have an adverse effect on a person's health, resulting in a decrease in life expectancy. Obese cats, for example, have a body condition score (BCS) that is greater than 6 (out of 9).

Metabolic disorders that are treated and/or prevented in accordance with the invention include syndrome X (metabolic syndrome). This disorder can be described as a combination of medical disorders that increase the risk of developing cardiovascular disease and diabetes. Metabolic syndrome is also known as metabolic syndrome X (metabolic syndrome), syndrome X (metabolic syndrome), insulin resistance syndrome, Riven syndrome and CHAOS (as an acronym for coronary heart disease, hypertension, atherosclerosis, obesity and stroke).

The precise mechanisms behind the complex pathways of metabolic syndrome are not yet fully understood. The pathophysiology is extremely complex and only partially understood. Most patients are older, obese, sedentary, and have some degree of insulin resistance. The most important factors in order are: (1) overweight and obesity, (2) genetics, (3) aging and (4) sedentary lifestyle, i.e. low physical activity and excess calorie intake.

Another risk factor is diabetes mellitus. At least in humans, the large majority (~75%) of patients with type 2 diabetes or impaired glucose tolerance (IGT) have metabolic syndrome.

The pathophysiology is typically characterized by the development of visceral fat, after which adipocytes (fat cells) of the visceral fat increase plasma levels of TNF-alpha and alter the levels of several other substances (eg, adiponectin, resistin, PAI-1). TNF-alpha has been shown not only to induce the production of inflammatory cytokines, but possibly triggers a cellular signaling system by interacting with the TNF-alpha receptor, which can lead to insulin resistance.

Current first-line therapy is lifestyle changes (eg, caloric restriction and physical activity). However, drug treatment is often necessary. Individual disorders that contribute to metabolic syndrome may be treated separately. Diuretics and ACE inhibitors may be used to treat hypertension. Cholesterol drugs can be used to lower LDL cholesterol and triglyceride levels if they are elevated and raise HDL levels if they are low. Such treatment may be combined with the use of one or more SGLT2 inhibitors according to the present invention.

Metabolic disorders that are treated and/or prevented in accordance with the invention include inflammation of the pancreas (pancreatitis). This disorder can occur either in an acute form or in a chronic form. Chronic pancreatitis may occur with or without steatorrhea and/or with or without diabetes mellitus.

Pancreatitis can be caused by hypertriglyceridemia (particularly when triglyceride values exceed 1500 mg/dL (16 mmol/L), hypercalcemia, viral infection, injury, vasculitis (i.e. inflammation of the small blood vessels in the pancreas) and may be an autoimmune disorder pancreatitis.

Metabolic disorders, in particular dyslipidemia and elevated serum triglyceride levels, are risk factors for the development of pancreatitis, and thus can be considered in conjunction with pancreatitis in accordance with the present invention. Accordingly, the present invention also relates to the prevention of pancreatitis.

Metabolic disorders that are treated and/or prevented in accordance with the invention include inflammation of adipose tissue (panniculitis), which is a group of disorders characterized by inflammation of adipose tissue.

Panniculitis can occur in any adipose tissue (cutaneous and/or visceral). It can be diagnosed based on a deep skin biopsy, and can be further classified using histological characteristics based on the location of the inflammatory cells (within the fat lobules or in the septa that separate them) and based on the presence or absence of vasculitis. Panniculitis can also be classified based on the presence or absence of systemic symptoms.

Metabolic diseases, in particular pancreatitis, are risk factors for the development of panniculitis and thus can be considered in accordance with the present invention in combination with panniculitis. Accordingly, the present invention also relates to the prevention of panniculitis.

Representatives of cats.

As used herein, a feline is a member of the family Felidae (i.e., felines). Such an animal may therefore belong to either the subfamily Felinae or the subfamily Pantherinae. The term feline includes the term cat, such as a domestic cat. The term domestic cat covers the terms Felis catus and Felis silvestris catus.

Pharmaceutically acceptable forms.

In this application, references to SGLT2 inhibitors and/or their use in accordance with the invention cover pharmaceutically acceptable forms of SGLT2 inhibitors, unless otherwise specified.

In accordance with the invention, any pharmaceutically acceptable form of the SGLT2 inhibitor can be used, for example, formula (1), preferably formula (18). For example, a crystalline form may be used. Prodrug forms may also be covered by the present invention.

Prodrugs may include, for example, esters and/or hydrates. The term prodrug is also intended to include any covalently linked carrier that releases the active compound of the invention in vivo when the prodrug is administered to a mammalian subject. Prodrug forms of the compound of the present invention can be prepared by modifying

- 11 043941 functional groups present in a compound according to the invention such that the modifications are cleaved, either by a conventional method or in vivo, to form the parent compound according to the invention.

Crystalline forms for use in accordance with the present invention include a complex of an SGLT2 inhibitor with one or more amino acids (see, for example, WO 2014/016381). The amino acid for such use may be a naturally occurring amino acid. The amino acid may be a proteogenic amino acid (including L-hydroxyproline) or a non-proteogenic amino acid. The amino acid may be a D- or L-amino acid. In some preferred embodiments, the amino acid is proline (L-proline and/or D-proline, preferably L-proline). Thus, this application discloses a crystalline complex between one or more naturally occurring amino acids and an SGLT2 inhibitor, for example, a crystalline complex between one or more naturally occurring amino acids and a glucopyranosyl-substituted benzene derivative of an SGLT2 inhibitor, preferably an SGLT2 inhibitor of formula (1), more preferably formula (18 ).

This application further discloses the use of one or more crystalline complexes, as defined herein above or below, to prepare a pharmaceutical composition that is useful for the treatment and/or prevention of diseases or conditions that may be affected by sodium-dependent inhibition. glucose cotransporter SGLT, preferably SGLT2. This application further discloses the use of one or more crystalline complexes, as defined above or below herein, to prepare a pharmaceutical composition for inhibiting the sodium-dependent glucose cotransporter SGLT2.

A crystalline complex between one or more naturally occurring amino acids (eg, proline, preferably L-proline) and an SGLT2 inhibitor is a preferred pharmaceutically acceptable form of the SGLT2 inhibitor for use in accordance with the present invention. In particular, a crystalline complex between one or more naturally occurring amino acids (e.g., proline, preferably L-proline) and a glucopyranosyl-substituted benzene derivative of an SGLT2 inhibitor, preferably an SGLT2 inhibitor of formula (1), more preferably formula (18), is a preferred pharmaceutically acceptable form an SGLT2 inhibitor for use in accordance with the present invention. This application further discloses a method for preparing one or more crystalline complexes as defined herein above and below, said method comprising the following steps:

(a) preparing a solution of an SGLT2 inhibitor (eg, a glucopyranosyl-substituted benzene derivative or an SGLT2 inhibitor of formula (1), preferably formula (18)) and one or more natural amino acids in a solvent or mixture of solvents;

(b) keeping the solution to precipitate a crystalline complex from it;

(c) removing the precipitate from the solution; and (d) optionally drying the precipitate until any excess of said solvent or solvent mixture is removed.

Certain pharmaceutical activity is a basic requirement that must be fulfilled by a pharmaceutically active agent before it is approved as a drug in the market. However, there are a number of additional requirements that a pharmaceutically active agent must meet. These requirements are based on various parameters that are related to the nature of the active substance itself. Without limitation, examples of such parameters are the stability of the active agent under various environmental conditions, its stability during the manufacturing process of a pharmaceutical drug, and the stability of the active agent in the final dosage compositions. The pharmaceutically active substance used to prepare pharmaceutical compositions must be as pure as possible, and its long-term storage stability must be guaranteed under various environmental conditions. This is very important in order to prevent the use of pharmaceutical compositions which contain, in addition to the actual active substance, for example, their breakdown products. In such cases, the content of the active substance in the drug may be less than indicated.

Uniform distribution of the drug in the composition is a critical factor, in particular when the drug must be administered in low doses. In order to ensure uniform distribution, the particle size of the active substance can be reduced to an acceptable level, for example by grinding. Since disintegration of the pharmaceutically active substance as a side effect of grinding (or micronization) should be avoided as far as possible, despite the harsh conditions required during the process, it is essential that the active substance be highly stable during the grinding process. Only when the active substance is sufficiently stable during the grinding process is it possible to obtain a homogeneous pharmaceutical preparation that always contains a given amount of active substance in a reproducible manner.

- 12 043941

Another problem that may arise during the grinding process to obtain the desired pharmaceutical is the energy input caused by the process and the stress on the surface of the crystals. This can in some cases lead to polymorphic changes, amorphization or changes in the crystal lattice. Since the pharmaceutical quality of a pharmaceutical formulation requires that the active substance always have the same crystalline morphology, the stability and properties of the crystalline active substance are also subject to stringent requirements from this point of view.

The stability of the pharmaceutically active substance is also important in pharmaceutical compositions to determine the shelf life of a particular drug; The shelf life is the period of time during which a drug can be administered without any risk. Therefore, the high stability of the drug in the above pharmaceutical compositions under various storage conditions is an additional advantage for the patient and the manufacturer.

Moisture absorption reduces the pharmaceutically active substance content as a result of increased weight caused by water absorption. Pharmaceutical compositions that tend to absorb moisture should be protected from moisture during storage, for example by adding suitable desiccant agents or by storing the drug in an environment where it is protected from moisture. It is therefore preferred that the pharmaceutically active substance be at best slightly hygroscopic.

In addition, the presence of a well-defined crystalline form allows the drug substance to be purified by recrystallization.

In addition to the requirements stated above, it should generally be borne in mind that any change in the solid state of a pharmaceutical composition that is capable of improving its physical and chemical stability provides a significant advantage over less stable forms of the same drug.

A crystalline complex between a natural amino acid and an SGLT2 inhibitor (eg, a glucopyranosyl-substituted benzene derivative or an SGLT2 inhibitor of formula (1) or formula (18)) satisfies the important requirements set forth in this application above.

Preferably, the naturally occurring amino acid is present in either its (D) or (L) enantiomeric form, most preferably as the (L) enantiomer.

Further preferred are those crystalline complexes of the present invention that are formed between an SGLT2 inhibitor (eg, Formula (1), preferably Formula (18)) and one natural amino acid, most preferably between the compound and the (L) enantiomer of the natural amino acid.

Preferred amino acids in accordance with this invention are selected from the group consisting of phenylalanine and proline, in particular (L)-proline and (L)-phenylalanine.

According to a preferred embodiment, the crystalline complex is characterized in that the natural amino acid is proline, in particular (L)-proline.

Preferably, the molar ratio of SGLT2 inhibitor (eg, formula (1), preferably formula (18)) and natural amino acid is in the range of from about 2:1 to about 1:3; more preferably from about 1.5:1 to about 1:1.5, even more preferably from about 1.2:1 to about 1:1.2, most preferably about 1:1. In this case, this implementation option is called a complex (1:1) or 1:1 complex.

Thus, a preferred crystalline complex according to the present invention is a 1:1 complex between said SGLT2 inhibitor (eg, one of formula (1), preferably formula (18)) and proline; in particular, the specified SGLT2 inhibitor and Lproline.

According to a preferred embodiment, the crystalline complex, in particular the 1:1 complex of said SGLT2 inhibitor with L-proline, is a hydrate.

Preferably, the molar ratio of crystalline complex to water is in the range of from about 1:0 to 1:3; more preferably from about 1:0 to 1:2, even more preferably from about 1:0.5 to 1:1.5, most preferably from about 1:0.8 to 1:1.2, in particular about 1 :1.

The crystalline complex of said SGLT2 inhibitor with proline, in particular L-proline and water, can be identified and distinguished from other crystalline forms according to its characteristic X-ray powder diffraction (XRPD) patterns.

For example, a crystalline complex of compound A with L-proline is preferably characterized by a powder X-ray diffraction pattern that includes peaks at 20.28, 21.14 and 21.64° 2Θ (±0.1° 2Θ), wherein said powder x-ray diffraction pattern is obtained by using CuKa1 radiation.

- 13 043941

Compound A has the following chemical formula (2):

In particular, said powder X-ray diffraction pattern includes peaks at 4.99, 20.28, 21.14, 21.64 and 23.23° 2Θ (±0.1° 2Θ).

In particular, the specified powder X-ray diffraction pattern includes peaks at 4.99, 17.61, 17.77, 20.28, 21.14, 21.64, 23.23 and 27.66° 2Θ (±0.1°2Θ ), wherein said powder X-ray diffraction pattern is obtained using CuKal radiation.

Even more specifically, said powder X-ray diffraction pattern includes peaks at

4.99, 15.12, 17.61, 17.77, 18.17, 20.28, 21.14, 21.64, 23.23 and 27.66° 2Θ (±0.1° 2Θ), wherein said powder X-ray diffraction pattern is obtained using CuKa1 radiation.

Even more specifically, the crystalline complex of compound A and L-proline is characterized by a powder X-ray diffraction pattern obtained using CuKa1 radiation, which includes peaks at degrees 2Θ (±0.1° 2Θ), as presented in table. 1.

Table 1

Powder X-ray diffraction pattern of the crystalline complex of compound A and L-proline _________ (only peaks up to 30° in 2Θ are shown): _________

2 Θ [°] d-parameter [A] Intensity 1/1« [%] 4.99 17.68 39 7.01 12.61 6 8.25 10.70 AND 9.95 8.88 12 13.15 6.73 thirty 13.33 6.64 10 14.08 6.28 4 15.12 5.85 32 16.40 5.40 12 16.49 5.37 13 17.11 5.18 6 17.61 5.03 32 17.77 4.99 35 18.17 4.88 32 18.32 4.84 28 18.72 4.74 8 19.16 4.63 thirty 19.96 4.45 26 20.28 4.37 56 20.60 4.31 7 21.14 4.20 84 21.64 4.10 100 22.33 3.98 15 23.23 3.83 41 24.06 3.70 4 24.51 3.63 15 24.93 3.57 26 25.89 3.44 23 26.21 3.40 AND 26.84 3.32 8 27.66 3.22 38 27.96 3.19 9 28.26 3.16 5 28.44 3.14 6 28.75 3.10 6 29.18 3.06 19

Even more specifically, this crystalline complex is characterized by a powder X-ray diffraction pattern obtained using CuKa1 radiation, which includes peaks at degrees 2Θ (±0.1° 2Θ, as shown in Fig. 11).

In addition, said crystalline complex of compound A with L-proline has a melting point above 89°C, in particular in the range from about 89°C to about 115°C, more preferably in the range from about 89°C to about 110°C (determined by 14 043941 using DSC; evaluated as the temperature of the onset of decomposition; heating rate 10 K/min).

This crystalline complex can be observed to melt upon dehydration. The resulting DSC curve is shown in Fig. 12.

This crystalline complex of compound A with L-proline shows weight loss by thermogravimetric determination (TG). The observed weight loss indicates that the crystalline form contains water, which may be bound by adsorption and/or may be part of the crystal lattice, i.e. the crystalline form may be present as a crystalline hydrate. The water content in crystalline form ranges from 0 to about 10% by weight, in particular from 0 to about 5% by weight, even more preferably from about 1.5 to about 5% by weight. The dotted line in Fig. 2 depicts a weight loss of 2.8 to 3.8% water. From the observed weight loss, a stoichiometry close to that of the monohydrate can be estimated.

Said crystalline complex has the following advantageous physicochemical properties, which are useful in the preparation of a pharmaceutical composition. In particular, the crystalline complex has high physical and chemical stability under various environmental conditions and during drug production. For example, the crystals can be produced in a form and particle size that are particularly useful in the process of preparing solid pharmaceuticals. In addition, the crystals have high mechanical strength, which makes it possible to grind the crystals. In addition, the crystalline complex does not exhibit a high tendency to absorb moisture and is chemically stable, that is, the crystalline complex makes it possible to obtain a solid pharmaceutical composition with a long shelf life. On the other hand, the crystalline complex has advantageously high solubility over a wide pH range, which is advantageous for the preparation of solid pharmaceutical preparations for oral administration.

Powder X-ray diffraction patterns can be obtained using a STOE STADI P diffractometer in transmission mode, equipped with an OED and a Cu anode as an X-ray source (CuKa1 radiation, λ=1.54056 A, 40 kV, 40 mA). In table 1, the values of 2Θ [°] denote the diffraction angle in degrees, and the values of d [A] denote the specified distances in A between the grating planes. The intensity shown in FIG. 11, given in impulses per second.

In order to take into account experimental error, the 2Θ quantities described above should be considered accurate to ± 0.1° 2Θ, in particular ± 0.05° 2Θ. That is, when assessing whether a given crystal sample of Compound A is in crystalline form in accordance with the 2Θ values described above, the 2Θ value that was experimentally observed for the sample should be considered identical to the characteristic value described above if it is within ±0 ,1° 2Θ characteristic value, in particular if it is within ± 0.05° 2Θ characteristic value.

The melting point is determined by DSC (differential scanning calorimetry) using a DSC 821 (Mettler Toledo). Weight loss is determined using thermal gravimetry (TG) using the TGA 851 (Mettler Toledo).

Also disclosed herein is a method for preparing a crystalline complex as defined in this application above and in this application below, wherein said method includes the following steps:

(a) providing a solution of an SGLT2 inhibitor as described herein (eg, Compound A or another SGLT2 inhibitor described herein) and one or more naturally occurring amino acids in a solvent or mixture of solvents;

(b) keeping the solution to precipitate a crystalline complex from it;

(c) removing the precipitate from the solution; and (d) optionally drying the precipitate until any excess of said solvent or solvent mixture is removed.

In accordance with step (a), a solution of an SGLT2 inhibitor (eg, Compound A or another SGLT2 inhibitor described herein) and one or more natural amino acids in a solvent or mixture of solvents is prepared. Preferably, the solution is saturated or at least nearly saturated or even supersaturated with respect to the crystalline complex. In step (a), the SGLT2 inhibitor may be dissolved in a solution that includes one or more natural amino acids, or one or more natural amino acids can be dissolved in a solution that includes the SGLT2 inhibitor. In an alternative procedure, the SGLT2 inhibitor is dissolved in a solvent or mixture of solvents to form a first solution, and one or more natural amino acids are dissolved in a solution or mixture of solvents to form a second solution. Thereafter, said first solution and said second solution are combined to obtain a solution in accordance with step (a).

Preferably, the molar ratio of natural amino acid to SGLT2 inhibitor (eg, Compound A or any other SGLT2 inhibitor described herein) in solution corresponds to the molar ratio of natural amino acid to SGLT2 inhibitor in the crystalline complex that is prepared. Thus, the preferred molar ratio is in the range from about 1:2 to 3:1; most preferably about 1:1.

Suitable solvents are preferably selected from the group consisting of C ₁ . ₄ α-alkanols, water, ethyl acetates, acetonitriles, acetones, diethyl ether, tetrahydrofuran and mixtures of two or more of these solvents.

More preferred solvents are selected from the group consisting of methanol, ethanol, isopropanol, water and a mixture of two or more of these solvents, in particular a mixture of one or more of these organic solvents with water.

Particularly preferred solvents are selected from the group consisting of ethanol, isopropanol, water and a mixture of ethanol and/or isopropyl alcohol and water.

In the case of a mixture of water and one or more C ₁ . ₄ α-alkanols, in particular methanol, ethanol and/or isopropanol, most preferably ethanol, the preferred volume ratio of water to alkanol being in the range from about 99:1 to 1:99; more preferably from about 50:1 to 1:80; even more preferably from about 10:1 to 1:60.

Preferably, step (a) is carried out at approximately room temperature (about 20°C) or at elevated temperature to approximately the boiling point of the solvent or solvent mixture used.

In accordance with a preferred embodiment, the starting material of an SGLT2 inhibitor (eg, Compound A or any other SGLT2 inhibitor described herein) and/or one or more natural amino acids and/or a solvent and mixture of solvents contains an amount of H ₂ O that is is at least necessary for the formation of SGLT2 inhibitor hydrate; in particular at least 1 mole, preferably at least 1.5 mole of water per mole of SGLT2 inhibitor. Even more preferably, the amount of water is at least 2 moles of water per mole of SGLT2 inhibitor. This means that either the starting material of the SGLT2 inhibitor (eg, compound A), or one or more natural amino acids, or the specified solvent, or mixtures of solvents, or the specified compounds and/or solvents in combination contain an amount of H2O such as specified above. For example, if the SGLT2 inhibitor starting material (eg, Compound A) or natural amino acid in step (a) contains a sufficient amount of water as described above, then the water content of the solvent(s) is not required.

In order to reduce the solubility of the crystalline complex according to the present invention in solution, one or more antisolvents may be added in step (a) and/or step (b), preferably during step (a) or at the beginning of step (b) . Water is an example of an acceptable antisolvent. The amount of antisolvent is preferably chosen to provide a supersaturated or saturated solution with respect to the crystalline complex.

In step (b), the solution is kept for a period of time sufficient to obtain a precipitate, i.e. crystalline complex. The temperature of the solution in step (b) is approximately the same or lower than in step (a). During aging, the temperature of the solution is preferably reduced, preferably to a temperature in the range of 20 to 0°C or even lower. Step (b) can be carried out with or without stirring. As is known to one skilled in the art, by using the time period and temperature differences in step (b), the size, shape and quality of the resulting crystals can be controlled. In addition, crystallization can be induced by methods that are known in the art, for example, by mechanical means such as scratching or rubbing the contact surface of the reaction vessel, for example, using a glass rod. Optionally, the (nearly) saturated or supersaturated solution may be inoculated with seed crystals.

In step (c), the solvent(s) may be removed from the precipitate by known methods such as, for example, filtration, suction, decantation and centrifugation.

In step (d), excess solvent(s) is removed from the precipitate by methods known to one skilled in the art, such as, for example, by reducing the partial pressure of the solvent(s), preferably in vacuum and/or by heating above a temperature of approximately 20°C, preferably in the temperature range below 100°C, even more preferably below 85°C.

Compound A can be synthesized by methods as specifically and/or generally described or set forth in international application WO 2007/128749, which is incorporated herein by reference in its entirety, and/or the examples disclosed herein below. . The biological properties of compound A can also be studied as described in WO 2007/128749.

The crystalline complex as described herein is preferably used as the active drug substance in substantially pure form, i.e. substantially free from other SGLT2 inhibitor crystalline forms (eg Compound A). However, the present invention also embraces the crystalline complex in admixture with other crystalline form or forms. If the active drug substance is a mixture of crystalline forms, it is preferred that the substance contains at least 50% by weight, even more preferably at least 90% by weight, most preferably at least , 95% by weight of the crystalline complex as described herein.

In view of its ability to inhibit SGLT activity, the crystalline complex according to the invention is suitable for use in the treatment and/or prophylactic treatment of conditions or diseases that may be affected by inhibition of SGLT, in particular SGLT-2 activity, in particular metabolic disorders , as described in this application. The crystalline complex according to the invention is also suitable for the preparation of pharmaceutical compositions for the treatment and/or prophylactic treatment of conditions or diseases that may be affected by inhibition of SGLT, in particular SGLT-2 activity, in particular metabolic disorders, as described in this application. A crystalline complex as described herein (particularly Compound A with a natural amino acid, eg proline, particularly L-proline) is also suitable for use in the treatment of felines.

Pharmaceutical compositions and formulations.

SGLT2 inhibitors for use in accordance with the invention can be obtained in the form of pharmaceutical compositions. They can be obtained in the form of solid or liquid compositions. In any case, they are preferably prepared for oral administration, preferably in liquid form for oral administration. SGLT2 inhibitors can, however, also be prepared, for example, for parenteral administration.

Solid dosage forms include tablets, granular forms, and other solid forms such as suppositories. Among solid forms, tablets and granular forms are preferred.

Pharmaceutical compositions within the meaning of the present invention may contain an SGLT2 inhibitor in accordance with the present invention and one or more excipients. Any excipient that allows or maintains the intended therapeutic effect may be used. Such fillers are available to one skilled in the art. Useful fillers are, for example, anti-blocking agents (used to reduce adhesion between powder (pellets) and punch faces and thereby prevent sticking to tableting punches), binding agents (a solution of binding agents or dry binding agents that hold ingredients together), coatings ( to protect tablet ingredients from damage by air moisture and make large or bad-tasting tablets easier to swallow), disintegrants (to allow tablets to break down when reconstituted), fillers, diluents, flavorings, colors, glidants (flow regulators to to promote powder flow by reducing particle friction and cohesion), lubricants (to prevent ingredients from sticking together and sticking to tablet punches or capsule filling apparatus), preservatives, sorbents, sweeteners, etc.

Dosage forms in accordance with the invention, for example solid compositions, may contain carriers and/or disintegrating agents selected from the group of sugars and sugar alcohols, for example mannitol, lactose, starch, cellulose, microcrystalline cellulose and cellulose derivatives, for example methylcellulose, etc. like them.

Manufacturing procedures for formulations suitable for felines are known to those skilled in the art and for solid compositions include, for example, direct compression, dry granulation and wet granulation. In the direct compression process, the active ingredient and all other excipients are placed together in a compression device, which is directly used to compress tablets from this material. The resulting tablets may then be coated in order to protect them from physical and/or chemical attack, for example with a material known in the art.

A unit administration form, for example a unit liquid dosage form or a unit solid composition, for example a tablet, may contain from 0.1 to 10 mg, or, for example, from 0.3 to 1 mg, from 1 to 3 mg, from 3 up to 10 mg; or 5 to 2500 mg, or, for example, 5 to 2000 mg, 5 to 1500 mg, 10 to 1500 mg, 10 to 1000 mg, or 10 to 500 mg of an SGLT2 inhibitor for use in accordance with the present invention. As one skilled in the art will appreciate, the content of the SGLT2 inhibitor in a solid composition or any formulation as described herein for administration to a feline may be increased or decreased as appropriate depending on the body weight of the feline. who is being treated.

In one embodiment, the pharmaceutical composition for use in accordance with the present invention is for oral or parenteral administration, preferably for oral administration. In particular, oral administration is facilitated by the use of excipients that modify the odor and/or tactile properties of the pharmaceutical composition for the intended patient, for example, as described herein.

When an SGLT2 inhibitor for use in accordance with the present invention is formulated for oral administration, it is preferred that the excipients impart properties

- 17 043941 for example, taste and/or chewability, which make the composition suitable for administration to a feline animal.

Additionally, liquid compositions are preferred. Liquid preparations may be, for example, solutions, syrups or suspensions. They may be administered directly to the cat or may be mixed with the cat's food and/or drink (eg, drinking water, etc.). One of the advantages of a liquid dosage form (similar to a granular formulation) is that such a dosage form allows for precise dosing. For example, an SGLT2 inhibitor can be dosed precisely according to the feline's body weight. Typical formulations of liquid compositions are known to those skilled in the art.

Dosing and administration.

A person skilled in the art can determine suitable dosages for use in accordance with the present invention. Preferred dosage units include mg/kg, i.e. mg of SGLT2 inhibitor per unit body weight of the feline. The SGLT2 inhibitor according to the invention can, for example, be administered at a dose of 0.01-5 mg/kg per day, for example 0.01-4 mg/kg, for example 0.01-3 mg/kg, for example 0 .01-2 mg/kg, for example, 0.01-1.5 mg/kg, for example, 0.01-1 mg/kg, for example, 0.01-0.75 mg/kg, for example, 0.01 -0.5 mg/kg, for example 0.01-0.4 mg/kg, for example 0.01-0.4 mg/kg per day; or 0.1 to 3.0 mg/kg per day, preferably 0.2 to 2.0 mg/kg per day, more preferably 0.1 to 1 mg/kg per day. In another preferred embodiment, the dose is 0.020.5 mg/kg per day, more preferably 0.03-0.4 mg/kg per day, for example 0.03-0.3 mg/kg per day.

A practitioner of the art is able to prepare an SGLT2 inhibitor in accordance with the present invention for administration at the desired dose.

Preferably, in accordance with the invention, the SGLT2 inhibitor is administered no more than three times per day, more preferably no more than twice per day, most preferably only once per day. The frequency of administration can be adapted to the cat's typical feeding frequency.

According to the invention, the SGLT2 inhibitor can be administered in such a way as to achieve the appropriate concentration of SGLT2 inhibitors in the blood plasma (for example, the maximum concentration in the blood plasma or the concentration in the blood plasma after a certain period of time, for example, 4, 8, 12 or 24 hours after oral administration, preferably approximately 8 hours after oral administration). For example, for Compound A, the plasma concentration (e.g., peak plasma concentration or plasma concentration after a specified period of time after oral administration) may be in the range of 2 to 4000 nM, e.g., 20 to 3000, or e.g. from 40 to 2000 nM.

Preferably, following administration and the time required for the SGLT2 inhibitor to reach the bloodstream, such levels remain in the blood for a time interval of at least 12 hours, more preferably at least 18 hours, most preferably at least , 24 hours

According to the invention, the SGLT2 inhibitor is preferably administered orally in liquid or solid form. The SGLT2 inhibitor can be administered directly into the oral cavity of animals (for example, using a syringe, preferably a syringe graduated according to body weight) or along with the animal's food or drink (for example, drinking water or the like), in each case, preferably in liquid form. However, the SGLT2 inhibitor can also be administered, for example, parenterally or by any other route of administration, for example rectally.

An SGLT2 inhibitor may be used alone or in combination with another drug. In some embodiments, one or more SGLT2 inhibitors, preferably Compound A, is used in combination with one or more other oral hypoglycemic drugs. When an SGLT2 inhibitor is used in combination with an additional drug, the SGLT2 inhibitor and any additional drugs may be administered simultaneously, sequentially (in any order), and/or according to specific time intervals of the drug regimen. In such embodiments, when the additional drug for combination administration with the SGLT2 inhibitor is not administered concomitantly with the SGLT2 inhibitor, the SGLT2 inhibitor and any additional drug are preferably administered over a period of at least 2 weeks, 1 month, 2 months , 4 months, 6 months or longer, for example, for 12 months or more.

In some embodiments of the present invention, the SGLT2 inhibitor (whether used alone or in combination with another drug) is not used in combination with 1-[(3-cyanopyridin-2-yl)methyl]-3-methyl- 7-(2-butyn-1-yl)-8-[3-(R)aminopiperidin-1-yl]xanthine or a pharmaceutically acceptable salt thereof, i.e. the feline is not treated with the compound. In some embodiments of the present invention, the SGLT2 inhibitor is not used in combination with a DPP-IV inhibitor, i.e. The feline is not being treated with a DPP-IV inhibitor.

In some embodiments of the present invention, the SGLT2 inhibitor is used as monotherapy, i.e. as a separate therapy, i.e. no other drug is administered to a cat to treat or prevent the same metabolic disorder, i.e. metabolic

- 18 043941 disorder for which an SGLT2 inhibitor is administered. For example, no other drug is administered to a cat for the treatment or prevention of the same metabolic disorder for a period of time of at least 2, 3, or 4 weeks before and after administration of an SGLT2 inhibitor.

Brief description of the figures

Fig. 1 shows the correlation between plasma levels of Compound A and urinary glucose excretion normalized to creatinine (g/creat). This represents an unambiguous logarithmic-linear relationship.

Fig. 2 shows the profiles of blood glucose and insulin secretion in the intravenous glucose tolerance test (ivGTT) of normal weight cats according to Hoenig (Mol Cell Endocrinol 2002, 197(1-2): 221-229) (iv GTT [lg/ kg]) and in cats with insulin resistance and obesity before (dashed line - before the study, before) and after 4 weeks of treatment with compound A (solid line). The increased and prolonged second phase secretion in insulin resistant and obese cats used in this study was significantly improved by treatment with Compound A.

Fig. The 3 values show the areas under the curve (AUC) for blood insulin and the sensitivity index for surrogate insulin (the relationship of insulin and blood glucose as expressed by the modified Belfiore index) in cats with insulin resistance during an intravenous glucose tolerance test ( ivGTT) before (before) and after and after 4 weeks of treatment (after) using compound A or its vehicle (control). Treatment with Compound A resulted in a significant reduction in insulin AUC (Panel A) as well as significantly improved insulin sensitivity (Panel B).

Fig. 4 shows the time course of blood glucose concentrations [mmol/L] after insulin administration to cats with insulin resistance during the intravenous insulin tolerance test (ivITT) before (dashed line - pre-test, before) and after 4 weeks of treatment when using compound A (solid line when using compound A or its vehicle (control). In animals that were not treated (control), insulin sensitivity (IS) decreased throughout the study period (panel A). In comparison, treatment using compound A was accompanied by a significant improvement in IS (panel B).

Fig. 5 shows time course of blood non-esterified fatty acid (NEFA) levels [mEq/L] following insulin administration to insulin-resistant cats in the ivITT process before (dashed line - pre-study, before) and after 4 weeks of treatment with Compound A (solid line) when using compound A or its vehicle (control). In untreated animals (control), NEFA elimination was significantly impaired (Panel A), while it was significantly improved by treatment with Compound A (Panel B).

Fig. 6 shows blood leptin concentrations that decreased significantly over the course of the study in treated cats.

Fig. 7 shows a decrease in respiratory exchange rate (RER) (indicative of increased lipid utilization) in treated animals, as measured by indirect calorimetry.

Fig. 8 shows that blood levels of β-hydroxybutyrate (β-HB/BHB) increased after 4 weeks of treatment with Compound A.

Fig. 9 shows a positive correlation between the change in blood leptin concentration and the change in RER value before treatment and 4 weeks after treatment with Compound A or vehicle (control).

Fig. 10 shows a negative correlation between blood levels of β-hydroxybutyrate (βHB/HB) and the RER value 4 weeks after treatment with Compound A.

Fig. 11 shows a powder X-ray diffraction pattern of a representative batch of a crystalline complex of Compound A with L-proline (1:1).

Fig. 12 shows a DSC/TG diagram of a representative batch of a crystalline complex of Compound A with L-proline (1:1).

Fig. 13 shows the average blood glucose value from the 9-hour glucose curve on the day of the visit.

Fig. 14 shows the level of serum fructosamine on the day of the visit.

Fig. 15 Preliminary data from four cats demonstrate that fasting insulin concentrations increased relative to a concomitant decrease in mean glucose (from the 9-hour glucose curve) on day 7 compared with day 1. Subsequently, insulin concentrations reached a plateau level. which can be explained by the almost normalized glucose concentration. This demonstrates the normal physiological situation in fasted animals: when glucose levels are within the normal range (fasted state), no increase in insulin concentrations is expected to be present anymore. These preliminary

- 19 043941 data obtained from fasting insulin readings from four cats support the stated indications of loss of pancreatic beta cell function and remission of a metabolic disorder, preferably remission of diabetes, as this demonstrates an increase in insulin concentrations and a decrease in glucose concentrations to normal values and thus , reflects a return to normal physiological response.

Examples

The following examples show beneficial therapeutic effects on glycemic control and/or insulin resistance, etc., by administering one or more SGLT2 inhibitors to felines in accordance with the present invention. These examples are intended to illustrate the invention in more detail without in any way limiting the scope of the claims.

Example 1: Pharmacokinetics (PK)/pharmacodynamics (PD) of a single oral dose of Compound A in cats.

Compound A was administered to cats after an overnight fasting period. Groups (n=3 per group) received a single oral dose of either vehicle (water) alone or vehicle containing the SGLT2 inhibitor in the form of Compound A at doses of 0.01, 0.1 and 1 mg/kg. PK/PD measurements were performed up to day 4 after a single administration of Compound A or its vehicle.

table 2

Pharmacokinetic data, single dose (0.01/0.1/1.0 mg/kg)

Parameter 0.01 mg/kg 0.1 mg/kg 1.0 mg/kg tmax [hour] average value 1 1.3 1 Stach [NMOL/L] average value 9 77 1173 AUCo >, [nmol-hour/l] average value thirty 358 5379 Τι/g [hour] average value 1.2 2.9 5.4

Pharmacodynamic data.

Significant increases in urinary glucose concentrations were evident at doses >0.01 mg/kg as early as 8 hours after administration (group means: controls, 1.4 mmol/L; 0.01 mg/kg, 1.4 mmol/L ; 0.1 mg/kg -46.1 mmol/l; 1 mg/kg - 239.3 mmol/l) and was constant for 24 hours.

None of the three doses of Compound A changed blood glucose levels in cats compared to normal reference values.

None of the three doses of Compound A altered kidney function in cats. The increase in urinary glucose excretion is apparently dependent on the dose and plasma concentration of the compound (log-linear correlation), as shown in FIG. 1.

Example 2: Effect of Compound A on urinary and blood glucose concentrations after repeated dosing in cats.

Compound A was administered to cats after an overnight fasting period. Groups (n=3 per group) received either vehicle alone (PillPocket®) or vehicle containing an SGLT2 inhibitor (dry compound) once daily orally at 1 and 3 mg/kg for three subsequent days. Glucose concentrations in urine and blood were measured.

Significant increases in urinary glucose concentrations were evident at both doses as early as 8 hours after administration. Maximum urinary concentrations did not increase further after repeated dosing and were similar to those at doses of 1 and 3 mg/kg (mean values 281 mmol/L and 209 mmol/L, respectively).

None of the doses of Compound A changed blood glucose levels in cats compared to normal reference values.

With regard to urinary glucose excretion, it was assessed using the ED5 ₀ indicator, which was < 1 mg/kg.

Example 3: Effect of Compound A on urinary and blood glucose concentrations after repeated dosing in cats.

Compound A was administered to normoglycemic obese cats that had ad libitum access to food. Groups (n=6 per group) received either vehicle alone (gelatin capsules) or vehicle containing an SGLT2 inhibitor (dry compound) once daily orally at a dose of 1 mg/kg for 4 weeks. Glucose concentrations in urine and blood were measured.

Urinary glucose concentrations increased significantly at the end of the study - controls - 0.6 mmol/l; 1 mg/kg - 489 mmol/l.

No changes in blood glucose levels were observed.

Example 4: Treating Prediabetes: Preventing Type 2 Diabetes in Cats.

The effectiveness of SGLT2 inhibition according to the present invention in the treatment of prediabetes characterized by abnormal fasting glucose levels and/or impaired glucose tolerance and/or insulin resistance can be tested through clinical studies. In studies over a shorter or longer period of time (eg, 2-4 weeks or 1-2 years), the success of treatment was tested by determining fasting glucose concentrations and/or postprandial glucose concentrations, or after an exercise test (oral glucose tolerance test or food tolerance test after eating a specific meal) after the end of the study treatment period and compare them with the values before the start of the study and/or with those for the placebo group. In addition, the fructosamine concentration value can be determined before and after therapy and compared with the initial value and/or with the value for the placebo group. Significant reductions in fasting or postprandial glucose levels and/or fructosamine levels demonstrate the effectiveness of treatment for prediabetes. In addition, the significant reduction in the number of patients who develop the manifestation of type 2 diabetes mellitus when treated using the pharmaceutical composition in accordance with this invention, compared with another form of treatment, demonstrates its effectiveness in preventing the transition from prediabetes to the manifestation of diabetes.

Case Study 5 Treatment of Prediabetes: Improvement of Insulin Resistance in Cats.

The example below shows the beneficial effect of Compound A on insulin resistance in obese cats. Compound A was administered to normoglycemic, insulin-resistant, obese cats that had ad libitum access to food. Groups (N=6 per group) received either vehicle alone (gelatin capsules) or vehicle containing an SGLT2 inhibitor (dry compound) once daily orally at a dose of 1 mg/kg for 4 weeks. The following experiments were conducted before the start of treatment and at the end of the 4-week treatment period approximately 24 hours after the last compound/vehicle administration.

An intravenous glucose tolerance test (ivITT, 0.8 g/kg dextrose) was performed on cats after an overnight fasting period. Blood was taken with a catheter through the jugular vein. Blood samples were taken at -5, 0, 5, 10, 15, 30, 45, 60, 90, 120, 180 min after glucose application.

Fluctuations in glucose and insulin concentrations were quantified by calculating baseline glucose-adjusted AUC. An intravenous insulin tolerance test (ivITT, 0.05 U/kg regular insulin) was performed on cats after an overnight fasting period. Blood was taken with a catheter through the jugular vein. Blood samples were taken at -5, 0, 5, 10, 15, 30, 45, 60, 90, 120, 180 min after glucose application.

Fluctuations in glucose and non-esterified fatty acids (NEFA) were quantified by calculating baseline AUC adjusted for glucose and NEFA.

The significance of differences in means between groups was assessed by two-way repeated measures (time and treatment) ANOVA and using post hoc multiple comparisons with control or matched baseline measures.

Fluctuations in glucose levels during the ivGTT did not change over the study period or due to treatment. Insulin fluctuations did not change during the study period in control cats, but decreased significantly in treated cats compared to baseline values (p<0.05).

As shown in FIG. 2, compared with non-obese cats, the obese cats used in the present study had a decreased first-phase insulin secretion profile and an increased and longer-lasting second phase. As shown in panel B of FIG. 2, treatment with Compound A resulted in a significant improvement in the second phase insulin secretion profile.

Insulin sensitivity was significantly increased in treated cats compared to baseline values (p<0.05). This was demonstrated by calculating the ratio between glucose and insulin using the modified Belfiore index (1/log(ΔAUCgluc*ΔAUCins).

Area under the curve values for blood insulin and the ratio of blood insulin to blood glucose as presented by the modified Belfiore index for insulin sensitivity in cats with insulin resistance during the intravenous glucose tolerance test (ivGTT) before (pre) and after (post) 4 weeks of treatment with Compound A or its vehicle (control) are shown in FIG. 3.

Fluctuations in glucose concentrations during ivITT worsened significantly throughout the study period in control animals (p<0.05) (see Fig. 4, panel A). This was similar to the elimination of NEFA (see Fig. 5, panel A). In contrast, in cats treated with Compound A, the glucose curve did not change throughout the study period (see FIG. 4, panel B), and NEFA elimination was significantly improved by treatment with Compound A (p< 0.01; see Fig. 5, panel B).

These data indicate that in obese cats, insulin resistance was significantly improved after 4 weeks of treatment with Compound A. Since insulin resistance is a characteristic feature of prediabetes, these data strongly suggest that

- 21 043941 that compound A is capable of treating prediabetes in felines.

In clinical studies in diabetic cats treated for varying periods of time (eg, 2 weeks to 12 months), success in improving insulin resistance can be tested by measuring baseline blood glucose, blood fructosamine, and blood insulin levels. , followed by monitoring the development of these levels in individual cats throughout the study period. In addition, postprandial or post-exercise glucose and insulin values (glucose tolerance test or insulin tolerance test) after the end of the study treatment period can be compared with values before the start of the study and/or with those of diabetic cats that were treated with other drugs.

Example 6 Treatment of type 2 diabetes in cats.

Treatment of cats with type 2 diabetes mellitus with the pharmaceutical composition of the invention, in addition to producing a dramatic improvement in glucose metabolism, prevents long-term metabolic deterioration. This can be observed if cats are treated for a shorter or longer period of time, for example, 2-4 weeks or 3 months to 1 year, using the pharmaceutical composition in accordance with the present invention, and compared with metabolism before treatment or with cats that have been treated with insulin or other sugar-lowering medications. There is evidence of therapeutic success if mean daily blood glucose and fructosamine levels are reduced compared to pre-treatment levels. Further evidence of therapeutic success is obtained if a significantly lower percentage of cats treated with the pharmaceutical composition of the present invention, compared with cats treated with other drugs, undergo transient deterioration in glucose metabolism (eg, hyper- or hypoglycemia).

Example 7: Improvement of pancreatic beta cell function.

In clinical studies in diabetic cats treated for varying periods of time (eg, 4 weeks to 12 months), treatment success was tested by measuring baseline blood glucose, blood fructosamine, and blood insulin levels and the corresponding relationship between the parameters for individual cat. In addition, for example, arginine stimulation can be used to test the ability of pancreatic beta cells to secrete insulin.

A significant increase in blood insulin levels (either before treatment or after stimulation with arginine) during or at the end of the study compared with the baseline value or compared with the placebo group or group receiving other therapy demonstrates the effectiveness of the pharmaceutical composition in accordance with this invention, in improving the function of pancreatic beta cells in diabetic cats (Fig. 15).

Example 8. Remission of diabetes.

In clinical studies in diabetic cats treated over a longer period of time (eg, 3 months to 1 year), treatment success was tested by measuring baseline blood glucose, blood fructosamine, and blood insulin levels and the corresponding relationship between parameters for an individual cat. There is evidence of therapeutic success if laboratory values decrease from pre-treatment levels without the need for insulin injections (Fig. 15).

When Compound A is used in combination, for example, with insulin or other drugs that effectively reduce hyperglycemia, the feline may be able to discontinue the insulin or other drug and still maintain glycemic control within normal limits.

Most preferably, administration of Compound A is discontinued to the feline.

Example 9: Reducing hyperglycemia.

In clinical studies in diabetic cats that were treated for varying periods of time (eg, 1 day to 12 months), the success of treatment in cats with hyperglycemia was tested by measuring blood glucose levels or blood fructosamine levels. A significant reduction in these values during the study or at the end of the study compared to the baseline value or compared to the placebo group or group receiving other therapy demonstrates the effectiveness of the pharmaceutical composition in accordance with the invention in reducing hyperglycemia in cats.

Example 10: Body composition and body fat reduction.

The following example shows the beneficial effects of Compound A in obese cats. Compound A was administered to obese cats with easy access to food. Groups (N=6 per group) received either vehicle alone (gelatin capsules) or vehicle containing an SGLT2 inhibitor (dry compound) at a dose of 1 mg/kg orally once daily for 4 weeks.

- 22 043941

The following experiments were conducted before the start of treatment and at the end of the 4-week treatment period approximately 24 hours after the last compound/vehicle administration. As shown in FIG. 6, blood leptin concentrations decreased significantly during the study period in treated cats (mean values: before: 2482 pmol/L, after: 2213 pmol/L, p < 0.05).

Indirect calorimetry shows the effect of treatment on energy metabolism. The respiratory exchange ratio (RER; the ratio between the amount of exhaled CO ₂ and inhaled O2, see Fig. 7) shows significantly increased fatty acid metabolism (lipid utilization) in treated animals (mean RER values: 0.749 pre-treatment, 0.728 post-treatment; p <0.01).

Increased lipid utilization was also reflected in increased blood β-hydroxybutyrate (β-HB/BHB) concentrations, as shown in FIG. 8. The increase in β-hydroxybutyrate concentrations did not exceed normal control values.

These changes in relevant data throughout the study show a significant correlation and suggest that the treatment demonstrates beneficial effects on body composition.

In summary, the data show a positive correlation between the change in blood leptin concentrations and the change in RER before and after 4 weeks of treatment with Compound A (Figure 9), as well as a negative correlation between blood levels of β-hydroxybutyrate (β-HB/BHB). and RER (Fig. 10).

Liver parameters were unchanged and no ketones were detected in the urine. Thus, the shift in lipid and carbohydrate metabolism is within normal physiological ranges.

As a consequence, 4 weeks of treatment in obese cats clearly showed that dyslipidokineemia was improved, and furthermore, the shift in metabolic substrate utilization from glucose to lipids represents a clear advantage in the treatment of obese cats. These data strongly suggest that Compound A can treat prediabetes in felines.

Example 11: Pilot Clinical Study of Compound A in Client-Owned Diabetic Cats.

The following data were obtained from 4 diabetic cats that were purportedly treated with 1 mg/kg orally administered Compound A once daily for 28 days. Diagnosis of diabetes mellitus was made based on a blood glucose >250 mg/dL (13.9 mmol/L) at screening, either glycosuria or serum fructosamine >400 μmol/L, and the persistence of at least one clinical condition/ symptom consistent with diabetes [lethargy, polyuria, polydipsia, polyphagia, weight loss and/or plantigrade hind legs (diabetic polyneuropathy)].

The results showed that the mean (FIG. 13) blood glucose, as shown in the 9-hour blood glucose curve, decreased significantly in all 4 cats from baseline at the end of the study. The reduction was present as early as day 7 and was unexpectedly found to be to a degree that was comparable to long-term insulin therapy. In comparison, a comparable reduction in mean blood glucose levels was not observed in 14 cats treated with Vetsulin® before day 14 (NADA 141-236, Freedom of Information Summary, Vetsulin). Serum fructosamine levels confirmed this good glycemic control and also decreased to below 350 μmol/L (excellent control according to laboratory interpretation rules) in all cats on day 28 (Figure 14). In contrast, the mean serum fructosamine level for cats treated with Vetsulin was 546 on day 30, and remained elevated to 462 on day 60 (NADA 141-236, Freedom of Information Summary, Vetsulin) .

All cats showed improvement in at least one clinical condition/symptom, and 3 of 4 cats showed improvement in at least 3 owner-rated clinical conditions/symptoms. The investigator assessed that all cats had improved overall diabetes control. Urinary glucose excretion was reduced in all cats by the end of the study. No hypoglycemia (defined as blood glucose less than 70 mg/dL) was reported.

In conclusion, these data indicate that Compound A can be used to treat diabetic cats using once-daily oral therapy comparable to twice-daily long-term insulin therapy.

Example 12 Preparation of 1-cyano-2-(4-cyclopropylbenzyl)-4-(β-D-glucopyranos-1-yl)benzene (compound A).

The following synthetic example serves to illustrate the method for preparing 1-cyano-2-(4cyclopropylbenzyl)-4-(β-D-glucopyranos-1-yl)benzene (compound A). A method for preparing its crystalline complex with L-proline is also described. This should be considered only as a possible method described by way of example, without limiting the scope of the present invention. The terms room temperature and ambient temperature are used interchangeably and indicate a temperature of approximately 20°C. The following abbreviations are used:

- 23 043941

DMF - dimethylformamide;

NMP - N-methyl-2-pyrrolidone;

THF - etrahydrofuran.

Preparation of 4-bromo-3-hydroxymethyl-1-iodobenzene

Oxalyl chloride (13.0 ml) was added to an ice-cold solution of 2-bromo-5-iodobenzoic acid (49.5 g) in CH ₂ Cl ₂ (200 ml). DMF (0.2 ml) was added and the solution was stirred at room temperature for 6 hours. The solution was then concentrated under reduced pressure and the residue was dissolved in THF (100 ml). The resulting solution was cooled in an ice bath and LiBH4 (3.4 g) was added in portions. The cooling bath was removed and the mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with THF and treated with 0.1 M hydrochloric acid. Then, the organic layer was separated, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were dried (Na ₂ SO ₄ ) and the solvent was evaporated under reduced pressure to obtain the raw material.

Yield: 47.0 g (99% of theory).

Preparation of 4-bromo-3-chloromethyl-1-iodobenzene

Thionyl chloride (13 ml) was added to a suspension of 4-bromo-3-hydroxymethyl-1-iodobenzene (47.0 g) in dichloromethane (100 ml) containing DMF (0.1 ml). The mixture was stirred at room temperature for 3 hours. Then the solvent and excess reagent were removed under reduced pressure. The residue was triturated with methanol and dried.

Yield: 41.0 g (82% of theoretical) Preparation of 4-bromo-1-iodo-3-phenoxymethyl-benzene

Phenol (13 g) dissolved in 4 M KOH (60 ml) was added to 4-bromo-3-chloromethyl-1iodobenzene (41.0 g) dissolved in acetone (50 ml). NaI (0.5 g) was added and the resulting mixture was stirred at 50°C overnight. Water was then added and the resulting mixture was extracted with ethyl acetate. The combined extracts were dried (Na ₂ SO ₄ ) and the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography (cyclohexane/ethyl acetate 19:1).

Yield: 38.0 g (79% of theory).

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

M solution of iPrMgCl in THF (11 ml) was added to dry LiCl (0.47 g) suspended in THF (11 ml). The mixture was stirred at room temperature until all of the LiCl had dissolved. This solution was added dropwise to a solution of 4-bromo-1-iodo-3-phenoxymethylbenzene (8.0 g) in tetrahydrofuran (40 ml) cooled to -60°C under argon. The solution was heated to -40°C and then 2,3,4,6-tetrakis-O-(trimethylsilyl)-D-glucαpyranone (10.7 g, 90% purity) in tetrahydrofuran (5 ml) was added. The resulting solution was heated to -5°C in a cooling bath and stirred for an additional 30 minutes at this temperature. An aqueous solution of NH4Cl was added and the resulting mixture was extracted with ethyl acetate. The combined organic extracts were dried over sodium sulfate and the solvent was removed under reduced pressure. The residue was dissolved in methanol (80 ml) and treated with methanesulfonic acid (0.6 ml) to obtain an exceptionally more stable anomer. After stirring the reaction solution at 35-40°C overnight, the solution was neutralized with solid NaHCO ₃ and the methanol was removed under reduced pressure. The residue was diluted with aqueous NaHCO ₃ solution and the resulting mixture was extracted with ethyl acetate. The combined extracts were dried over sodium sulfate and the solvent was evaporated to obtain the raw material, which was reduced without further purification.

Yield: 7.8 g (93% of theory).

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

Boron diethyletherate trifluoride (4.9 ml) was 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 is maintained below -10°C. The resulting solution was heated to 0°C for a period of 1.5 hours and then treated with an aqueous sodium hydrogen carbonate solution. The resulting mixture was stirred for 0.5 hour, the organic solvent was removed, and the residue was extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and the solvent was removed. The residue was taken up in dichloromethane (50 ml) and pyridine (9.4 ml), acetic anhydride (9.3 ml) and 4-dimethylaminopyridine (0.5 g) were added successively to the solution. The solution was stirred for 1.5 hours at room temperature and then diluted with dichloromethane. This solution was washed twice with 1 M hydrochloric acid and dried over sodium sulfate. The solvent was then removed and the residue was recrystallized from ethanol to obtain the product as a colorless solid.

Yield: 6.78 g (60% of theory).

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

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

cyanide

Through a flask loaded with zinc (1.0 g), zinc (30 mg),

Pd ₂ (dibenzylideneacetone) ₃ ·CHCl ₃ (141 mg) and tri-tert-butylphosphonium tetrafluoroborate (111 mg) were purged with argon. A solution of 1-bromo-4-(2,3,4,6-tetra-O-acetyl-P-glucopyranos-1-yl)-2(phenoxymethyl)benzene (5.4 g) in NMP (12 ml) was then added. , and the resulting mixture was stirred at room temperature for 18 hours. After dilution with ethyl acetate, the mixture was filtered and the filtrate was washed with an aqueous sodium hydrogen carbonate solution. The organic phase was dried (sodium sulfate) and the solvent was removed. The residue was recrystallized from ethanol.

Yield: 4.10 g (84% of theory).

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

Alternatively, the compound described above was synthesized using 1-bromo-4-(2,3,4,6-tetra-O-acetyl-P-glucopyranos-1-yl)-2-(phenoxymethyl)benzene as the starting material 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

A 33% solution of hydrobromic acid in acetic acid (15 ml) was 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 was stirred at 55°C for 6 hours and then cooled in an ice bath. The reaction mixture was neutralized with a cooled aqueous potassium carbonate solution, and the resulting mixture was extracted with ethyl acetate. The combined organic extracts were dried over sodium sulfate and the solvent was removed under reduced pressure. The residue was taken up in ethyl acetate/cyclohexane (1:5) and the precipitate was separated by filtration and dried at 50°C to give pure product.

Yield: 0.52 g (75% of theory)

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

Preparation of 4-cyclopropylphenylboronic acid

I

HE

A 2.5 M solution of n-butyllithium in hexane (14.5 ml) was added dropwise to 1-bromo-4-cyclopropylbenzene (5.92 g) dissolved in THF (14 ml) and toluene (50 ml) and cooled to -70°C. The resulting solution was stirred at -70°C for 30 minutes before adding triisopropiborate (8.5 ml). The solution was heated to -20°C and then treated with 4 M aqueous hydrochloric acid (15.5 ml). The reaction mixture was then warmed to room temperature and the organic phase was separated. The aqueous phase was extracted with ethyl acetate and the combined organic phases were dried (sodium sulfate). The solvent was evaporated and the residue was washed with a mixture of ether and cyclohexane to give the product as a colorless solid.

Yield: 2.92 g (60% of theory).

Mass spectrum (ESI ^- ): m/z=207 (Cl) [M+HCOO] ^- .

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

O

2-bromomethyl-4-(2,3,4,6-tetra-O-acetyl-E-glucopyranos-1yl)benzonitrile (1.60 g), 4-cyclopropyl-phenylboronic acid (1.0 d), potassium carbonate (1.85 g) and a degassed 3:1 mixture of acetone and water (22 ml). The mixture was stirred at room temperature for 5 minutes, pre-cooled in an ice bath. Palladium dichloride (30 mg) was then added and the reaction mixture was stirred for 16 hours at room temperature. The mixture was then diluted with brine and extracted with ethyl acetate. The combined extracts were dried over sodium sulfate and the solvent was removed under reduced pressure. The residue was dissolved in methanol (20 ml) and treated with 4 M aqueous potassium hydroxide solution (4 ml). The resulting solution was stirred at room temperature for 1 hour and then neutralized with 1 M hydrochloric acid. The methanol was evaporated and the residue was diluted with brine and extracted with ethyl acetate. The collected organic extracts were dried over sodium sulfate and the solvent was removed. The residue was subjected to chromatography on silica gel (dichloromethane/methanol 1:0 -> 8:1).

Yield: 0.91 g (76% of theory).

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

Preparation of a crystalline complex (1:1) of compound A with L-proline

L-proline (0.34 g) dissolved in 2.1 ml of ethanol and water (10:1 v/v) was added to a solution of 1-cyano-2-(4-cyclopropylbenzyl)-4-(β-D- glucopyranos-1-yl)benzene (1.17 g, prepared as described above) dissolved in 2 ml of ethanol. The resulting solution was left to settle at room temperature. After approximately 16 hours, the crystalline complex was isolated as white crystals by filtration. If necessary, crystallization can be initiated, for example, by scratching with a glass rod or metal spatula, or by introducing seed crystals. The remaining solvent was removed by storing the crystals at slightly elevated temperature (30 to 50°C) under vacuum for approximately 4 hours to obtain 1.27 g of a crystalline 1:1 complex of L-proline and 1-cyano-2-(4- cyclopropylbenzyl)-4(e-D-glucopyranose-1-yl)benzene.

Several batches of the crystalline complex were prepared according to the production method described above. The X-ray powder diffraction profiles were consistent. Melting points were determined by DSC and estimated as the onset temperature of decomposition. Examples of melting points were approximately 89, 90, 92, 101 and 110°C. Powder X-ray diffraction pattern, as presented in Table. 1 and as shown in FIG. 11, DSC and TG diagrams in Fig. 12 corresponds to a batch with a melting point of approximately 90°C.

- 26 043941

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

Example 13. Recipes.

Several examples of formulations are described in which the term active substance refers to an SGLT2 inhibitor or a pharmaceutically acceptable form thereof, eg a prodrug or crystalline form, for use in accordance with the invention. When combined with one or more additional active agents, the term active agent may also include the additional active agent.

Tablets containing 100 mg of active substance

Compound:

the tablet contains:

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

Method of obtaining:

The active substance, lactose and starch were mixed together and uniformly wetted using an aqueous solution of polyvinylpyrrolidone. The moistened composition was then screened (sieve size 2.0 mm) and dried in a multi-tier dryer at 50° C., after which it was screened again (sieve size 1.5 mm) and lubricant was added. The final mixture was compressed to form tablets.

Tablet weight: 220 mg

Diameter: 10 mm, two-plane, faceted on both sides and with a notch on one side.

Tablets containing 150 mg of active substance

Composition: 1 tablet contains:

active substance 150.0 mg lactose powder 89.0 mg corn starch 40.0 mg colloidal silicon dioxide 10.0 mg polyvinylpyrrolidone 10.0 mg Magnesium stearate 1.0 mg

300.0 mg

Receipt:

The active substance, mixed with lactose, corn starch and silica, was moistened with a 20% aqueous solution of polyvinylpyrrolidone and passed through a 1.5 mm sieve. The granules, dried at 45°C, were passed through the same sieve again and mixed with the specified amount of magnesium stearate. Tablets were pressed from the mixture.

Tablet weight: 300 mg

Matrix: 10 mm, flat

Hard gelatin capsules containing 150 mg of active substance Composition:

capsule contains:

active substance 150.0 mg corn starch (dried) lactose (powdered) approximately 180.0 mg approximately 87.0 mg Magnesium stearate 3.0 mg approximately 420.0 mg

Receipt:

The active substance was mixed with the excipients, passed through a 0.75 mm sieve and homogeneously mixed using suitable equipment. The resulting mixture was packed into hard gelatin capsules with size 1.

Filled capsule: approximately 320 mg

Capsule shell: hard gelatin capsule size 1.

- 27 043941

Suppositories containing 150 mg of active substance Composition:

1 suppository contains: active substance 150.0 mg polyethylene glycol 1500 550.0 mg polyethylene glycol 6000 460.0 mg polyoxyethylene sorbitol 840.0 mg monostearate

2000.0 mg

Receipt:

After the suppository mass was melted, the active substance was homogeneously distributed in this mixture, and the molten mixture was poured into cooled molds.

Ampoules containing 10 mg of active substance

Compound:

active substance

0.01 N hydrochloric acid/NaCl double distilled water

10.0 mg in the required amount up to 2.0 ml

Receipt:

The active substance was dissolved in the required amount of 0.01 N HCl, the solution was made isotonic using a known salt, filtered sterilely and transferred into 2 ml ampoules.

Ampoules containing 50 mg of active substance

Compound:

active substance

0.01 N hydrochloric acid/NaCl double distilled water

50.0 mg in the required amount up to 2.0 ml

Receipt:

The active substance was dissolved in the required amount of 0.01 N HCl, the solution was made isotonic using a known salt, filtered sterilely and transferred into 10 ml ampoules.

Links

1) Curry et al., Comp Biochem Physiol. 1982. 72A(2): 333-338;

2) EP 1213296;

3) EP 1354888;

4) EP 1344780;

5) EP 1489089;

6) Hoenig, Mol Cell Endocrinol 2002, 197(1-2): 221-229;

7) Hoenig et al., Am J Physiol, 2011, 301(6):R1798-1807;

8) NADA 141-236 Freedom of Information Vetsulin;

9) Palm C.A. et al., Vet Clin Small Anim 2013, 43: 407-415;

10) Reusch S.E. et al., Schweizer Archiv filer Tierheilkunde 2011, 153811): 495-500;

11) Tanaka et al., Vet Res Commun. 2005, 29(6):477-485;

12) Verbrugghe et al., Crit Rev Food Sci Nutr. 2012;52(2): 172-182;

13) WO 01/27128;

14) WO 03/099836;

15) WO 2004/007517;

16) WO 2004/080990;

17) WO 2005/012326;

18) WO 2005/092877;

19) WO 2006/034489;

20) WO 2006/064033;

21) WO 2006/117359;

22) WO 2006/117360;

23) WO 2006/120208;

24) WO 2007/025943;

25) WO 2007/028814;

26) WO 2007/031548;

27) WO 2007/093610;

28) WO 2007/114475;

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Claims (4)

29) WO 2007/128749; 30) WO 2007/140191; 31) WO 2008/002824; 32) WO 2008/013280; 33) WO 2008/042688; 34) WO 2008/049923; 35) WO 2008/055870; 36) WO 2008/055940; 37) WO 2008/069327; 38) WO 2008/116179; 39) WO 2009/014970; 40) WO 2009/022008; 41) WO 2009/022020; 42) WO 2009/035969; 43) WO 2010/023594; 44) WO 2011/039107; 45) WO 2011/039108; 46) WO 2011/117295; 47) WO 2014/016381. CLAIM 1. The use of one or more SGLT2 inhibitors or pharmaceutically acceptable crystalline forms thereof for the treatment and/or prevention of a metabolic disorder in a feline animal, wherein said one or more SGLT2 inhibitors is a glucopyranosyl-substituted benzene derivative of general formula (1) where R ¹ is cyano, Cl or methyl; R ² is H, methyl, methoxy or hydroxyl; And R ³ is cyclopropyl, hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, 3-methyl-but-1-yl, cyclobutyl, cyclopentyl, cyclohexyl , 1-hydroxycyclopropyl, 1-hydroxycyclobutyl, 1-hydroxycyclopentyl, 1-hydroxycyclohexyl, ethynyl, ethoxy, difluoromethyl, trifluoromethyl, pentafluoroethyl, 2-hydroxyethyl, hydroxymethyl, 3hydroxypropyl, 2-hydroxy-2-methyl-prop-1-yl, 3-hydroxy -3-methyl-but-1-yl, 1-hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1-hydroxy -1-trifluoromethyl-ethyl, 2-methoxy-ethyl, 2-ethoxy-ethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxy-ethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethylsulfinyl, ethylsulfonyl, trimethylsilyl, (R)tetrahydrofuran-3- yloxy or (S)-tetrahydrofuran-3-yloxy or cyano, wherein one or more hydroxyl groups of the eD-glucopyranosyl group are optionally further acylated with groups selected from (C ₁ . ₁₈ alkyl)carbonyl, (C ₁ . ₁₈ alkyl) oxycarbonyl , phenylcarbonyl and phenyl-(C ₁ . ₃ alkyl)carbonyl, wherein such one or more SGLT2 inhibitors or pharmaceutically acceptable crystalline forms thereof are administered at a dose of 0.01 to 5.0 mg/kg body weight per day. 2. Use according to claim 1, wherein the metabolic disorder is one or more selected from the group consisting of: ketoacidosis, prediabetes, type 1 or type 2 diabetes mellitus, insulin resistance, obesity, hyperglycemia, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, low-grade systemic inflammation, hepatic lipidosis, atherosclerosis, pancreatic inflammation, neuropathy, and/or syndrome X (metabolic syndrome), and/or loss of pancreatic beta cell function, and/or where remission of a metabolic disorder is achieved and/or maintained, preferably remission of diabetes. 3. Use according to any one of claims 1 and 2, wherein the metabolic disorder is prediabetes, type 1 diabetes mellitus or type 2 diabetes mellitus and/or clinical conditions associated with prediabetes, type 1 diabetes mellitus or type 2 diabetes mellitus, preferably prediabetes or type 2 diabetes mellitus, and/or clinical conditions associated with prediabetes or type 2 diabetes mellitus. 4. Use according to claim 3, wherein said clinical condition is one or more of -