EA040378B1 - USE OF SGLT2 INHIBITOR IN TREATMENT AND/OR PREVENTION OF METABOLIC DISORDER IN CANINE ANIMAL - Google Patents

Description

The field of technology to which the invention belongs

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

Prerequisites for the creation of the invention

Canines, such as dogs, are subject to various metabolic disorders. A number of metabolic disorders are known in canines, including hyperglycemia, insulin resistance, diabetes, hepatic lipidosis, obesity, hyperinsulinemia, impaired glucose tolerance, ketosis (particularly ketoacidosis), dyslipidemia, dysadipokinemia, obesity, subclinical inflammation, or systemic inflammation, in particular mild systemic inflammation, which also includes inflammation of the adipose tissue, syndrome X (metabolic syndrome) and/or inflammation of the pancreas. There are various correlations between these disorders. Among these disorders in dogs, diabetes mellitus, in particular prediabetes and insulin-dependent diabetes mellitus, as well as hyperglycemia, insulin resistance and obesity, are gaining in importance. This may be at least partly due to changing living conditions and eating habits and the fact that pets are living longer due to improvements in preventive veterinary care in 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 canines such as dogs. The incidence of diabetes in dogs has increased by about 1.0% in recent decades. Several risk factors have been identified: age, obesity, sterilization, sex and breed.

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

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

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

(3) secondary diabetes mellitus, which may be due to diabetogenic drugs (eg, long-acting glucosteroids, 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.

Canine diabetes is not easy to classify, although there are clear similarities and differences between the human and canine disease. There is no evidence for a canine equivalent of type 2 diabetes, despite the fact that obesity is as much of a problem in pet dogs as it is in their owners.

The disease can be broadly classified into diabetes associated with insulin deficiency and non-insulin-dependent diabetes (Catchpole et al., Diabetologia 2005. 48: 1948-1956). Insulin deficiency is the most common type. In contrast to the human situation, it is not usually found in young dogs, but rather, perhaps similar to latent/late autoimmune adult type 1 diabetes (LADA), which is characterized by progressive destruction of beta cells in an autoimmune pattern.

Autoimmunity in dogs, however, is controversial. Because antibodies have only been found in a subgroup of dogs with canine diabetes and this is discussed as a consequence rather than a cause of the disease (Catchpole et al., Diabetologia 2005. 48: 1948-1956).

In addition, diabetes associated with diestrus/gestational insulin resistance is common in intact female dogs.

Several oral hypoglycemic agents have been approved for the treatment of diabetes 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. glucose in the intestine (alpha-glucosidase inhibitors).

Some oral hypoglycemic agents have been used but have either been ineffective in diabetic dogs, such as sulfonylureas, or have shown some effects on glycemic control but have been unfavorable due to the high prevalence of side effects, such as alpha-glucosidase inhibitors ( Nelson et al., J Small Anim Pract 2000, 41, 486-490).

Other approaches to treat diabetes and reduce hyperglycemia are also being considered, including inhibition of the renal sodium-dependent glucose cotransporter SGLT2. SGLT2 in the kidney

- 1 040378 regulates glucose levels by mediating the reabsorption of glucose back into plasma after blood filtration. Thus, inhibition of SGLT2 induces glycosuria and can lead to a decrease in blood glucose levels. For example, the compound 1-cyano-2-(4-cyclopropyl-benzyl)-4-(beta-D-glucopyranos-1-yl)benzene is described as an SGLT2 inhibitor in WO 2007/128749. A wide variety of additional SGLT2 inhibitors is also known. In WO 2011/117295, which relates to the treatment of predominantly carnivorous non-human animals with dipeptidyl peptidase IV (DPP-IV) inhibitors, various SGLT2 inhibitors stand out 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 considered for the treatment of metabolic disorders in canines such as dogs. In representatives of canine drugs for the treatment of metabolic disorders are much less developed than in humans. Unfortunately, even if treatment or prophylaxis is effective, for example, in humans or other non-canine animals, it is not possible to conclude that the same approach would be effective, safe, and also beneficial in canine animals such as dog.

Canids differ significantly from humans or, for example, from other carnivores such as cats in terms of their metabolism.

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

Dogs show obesity and all the signs of the metabolic syndrome, similar to those, for example, for humans, as well as for cats. Unlike these species, this syndrome does not progress to type 2 diabetes in canines. The pathophysiological hallmark of type 2 diabetes mellitus in humans, as well as in felines, amyloid deposition in the pancreatic islet, is absent in dogs (Verkest, Vet J, available at doi.org/10.1016/j.tvjl.2013.09.057).

A diabetic complication, such as vision problems, is common in dogs but rare in felines. Although retinopathy is often found in diabetic humans, it is rare in dogs, but vision problems occur due to keratopathy and, in particular, cataracts. They account for up to 80% in diabetic dogs (Beam et al. Vet. Ophtalmol. 1999. 2, 169-172).

Insulin injections are now considered the gold standard for treating diabetic dogs. However, no single type of insulin is generally effective in maintaining glycemic control even when administered twice daily. Even regulated diabetics can eventually reach a point where their blood glucose is no longer controlled and insulin must be adjusted in either dose or type. In addition, even with strict adherence by the owner, control is often weak and general secondary problems occur. Many owners find it impossible to achieve acceptable levels of compliance because synchronization of food intake and insulin administration is not possible in most cases. Ultimately, many diabetic dogs are euthanized because of the disease.

The factors governing patient and owner compliance are also very different. In dogs, oral administration, for example, is even more desirable than in humans.

Treatment that would allow for strict adherence to, and therefore improve glycemic control over, current insulin-based therapies would help attenuate disease progression and delay or prevent complications in many animals.

No satisfactory treatment is currently available for metabolic disorders such as obesity, insulin resistance, hyperglycemia, hyperinsulinemia, impaired glucose tolerance, hepatic lipidosis, dyslipidemia, dysadipokinemia, subclinical inflammation, or systemic inflammation, particularly mild systemic inflammation. inflammation, which also includes adipose tissue, and associated disorders such as syndrome X (metabolic syndrome). In addition, these metabolic disorders may be associated with or induced by hypo- or hyperthyroidism, elevated cortisone levels (hyperadrenocorticism, Cushing's disease) and/or excess growth hormone (acromegaly). These metabolic disorders can be clinically manifested, for example, by elevated blood pressure, cardiomyopathy, renal dysfunction, and/or musculoskeletal disorders in canines.

Thus, there is still a particular need for effective, safe and acceptable methods for the treatment of metabolic disorders, including diabetes, in canines.

Disclosure of invention

Summary of essence

The present inventors have surprisingly found that the use of an SGLT2 inhibitor is effective and safe in the treatment and/or prevention of metabolic disorders in canines.

The present invention thus provides for the use of one or more inhibitors

- 2 040378

SGLT2 or pharmaceutically acceptable crystalline forms thereof in the treatment and/or prevention of a metabolic disorder in a canine in a canine animal in need of such treatment and/or prevention, wherein one or more SGLT2 inhibitors is selected from the group consisting of the following compounds or their pharmaceutically acceptable crystalline forms:

glucopyranosyl-substituted benzene derivative of general formula (1)

where R ¹ denotes cyano, Cl or methyl;

R ² is H, methyl, methoxy or hydroxy; 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, 1hydroxycyclopropyl, 1-hydroxycyclobutyl, 1-hydroxycyclopentyl, 1-hydroxycyclohexyl, ethynyl, ethoxy, difluoromethyl, trifluoromethyl, pentafluoroethyl, 2-hydroxyethyl, hydroxymethyl, 3-hydroxypropyl, 2-hydroxy-2-methylprop-1-yl, 3 -hydroxy-3-methyl-but-1-yl, 1-hydroxy-1-methylethyl, 2,2,2-trifluoro-1hydroxy-1-methylethyl, 2,2,2-trifluoro-1-hydroxy-1-trifluoromethyl -ethyl, 2-methoxyethyl, 2-ethoxyethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxyethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethylsulfinyl, ethylsulfonyl, trimethylsilyl, (R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran -3-yloxy or cyano;

or a derivative thereof, wherein one or more hydroxyl groups of the eD-glucopyranosyl group are acylated with groups selected from (C _1-18 -alkyl)carbonyl, (C _1-18 -alkyl)oxycarbonyl, phenylcarbonyl, and phenyl-(C _{1- 3} -alkyl)carbonyl;

dapagliflozin represented by formula (3)

canagliflozin represented by formula (4) empagliflozin represented by formula (5) ertugliflozin represented by formula (9) a compound represented by formula (18) wherein R ³ is selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)-tetrahydrofuran-3- yloxy or (S)-tetrahydrofuran-3-yloxy;

wherein one or more SGLT2 inhibitors or pharmaceutically acceptable crystalline forms thereof are administered at a dose of 0.1 to 4.0 mg/kg body weight.

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

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

- 3 040378

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

One or more SGLT2 inhibitors, or a pharmaceutically acceptable crystalline form thereof, may be administered at 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 0.1 to 1 mg/kg of body weight per day. Thus, one or more SGLT2 inhibitors, or a pharmaceutically acceptable crystalline form thereof, may 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 of body weight per day.

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

In the examples provided in this application, the therapeutic and/or prophylactic benefits based on the inhibition of SGLT2 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 those that have a restrictive effect on the scope of protection as defined in this application below and in the claims.

In particular, the present inventors have unexpectedly found that the use of one or more SGLT2 inhibitors according to the present invention advantageously results in a reduction in hyperglycemia and/or in addition to an improved, for example, 9- or 24-hour glycemic profile in hyperglycemic (e.g., diabetic) ) dogs. Thus, this may lead to a decrease in the level of insulin needed to treat diabetic canines.

Since absorption and onset of action (glucosuria) are very rapid and noticeable (examples 1 and 2), treatment in canines with a newly diagnosed metabolic disorder allows the optimal dose to be established within a short period of time (eg, 7-14 days).

The invention demonstrates a significant improvement and optimized treatment of hyperglycemia and thus provides the possibility of preventing or delaying the development of the disease or inducing remission of complications associated with hyperglycemia, in particular the occurrence of diabetic cataracts, in canines.

An additional advantage of the present invention is that the use of one or more SGLT2 inhibitors is only effective against metabolic disorders, that is, if the desired use of one or more SGLT2 inhibitors in a canine provides a monotherapy - that is, a separate therapy; in other words, when no other drugs are administered to the animal to treat or prevent the same metabolic disorder - with the sole exception of non-insulin dependent diabetes.

In addition, this invention also provides the possibility of combination therapy using insulin. Such a combination advantageously results in a reduction in the dose and/or frequency at which insulin is administered as compared to monotherapy in a representative canine using insulin.

Advantageously, the use of one or more SGLT2 inhibitors in accordance with the present invention does not cause hypoglycemia (Example 2).

An additional advantage, in particular, is that the use of one or more SGLT2 inhibitors in accordance with the present invention leads to a reduction in insulin resistance in insulin resistant canines that have been treated. In other words, the use of one or more SGLT2 inhibitors according to the present invention advantageously results in increased insulin sensitivity in insulin resistant treated canines.

Thus, the use of one or more SGLT2 inhibitors in accordance with the present invention provides improved treatment and/or prevention of metabolic diseases as disclosed in this application, including diabetes, in canines.

The effects of the use of one or more SGLT2 inhibitors of the present invention may refer to the same or comparable canine prior to administration of one or more SGLT2 inhibitors of the present invention, and/or refer to a comparable canine that has not received said treatment (e.g. 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, eg 8-16 weeks, ie. 8, 9, 10, 11, 12, 13, 14, 15 or 16 weeks.

An additional advantage of the present invention is that one or more SGLT2 inhibitors can be effectively administered orally to a canine. In addition, one or more SGLT2 inhibitors in accordance with the present invention can be administered only once a day. These advantages allow for better compliance with the regimen of the animal being treated and its owner. This results in better glycemic control of disorders (eg diabetes) for which canines are treated with insulin. In general, the use of one or more SGLT2 inhibitors in accordance with the present invention thus helps to attenuate (i.e., delay or prevent) the development of metabolic disorders and delay or prevent the onset of metabolic disorders (e.g., diabetes) and its complications in canine individuals. .

The effects of using one or more SGLT2 inhibitors of the present invention (e.g., the beneficial effects on hyperglycemia mentioned above) may refer to the same or a comparable canine before administration of one or more SGLT2 inhibitors of the present invention, and/or may refer to to a comparable canine that received standard treatment with insulin (eg, a control group), or to one that received no treatment.

An additional advantage of the present invention is that one or more SGLT2 inhibitors can be effectively administered orally to a canine, eg in liquid form. In addition, one or more SGLT2 inhibitors in accordance with the present invention may be administered only once a day. These advantages allow for optimal dosing and compliance in both the treated animal 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, such as the metabolic disorders disclosed herein, or may delay the development or prevent the onset of a metabolic disorder and its complications. in canines, for example, high blood pressure, renal dysfunction and/or musculoskeletal disorders are prevented or retarded or remission achieved.

Definitions

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

The effects of the treatment disclosed in this application (such as improvement, attenuation or late onset of a disorder, disease or condition, or improvement, attenuation, increase or delay in any effect, indicator, marker level or other parameter related to the disorder, disease or condition) , can be observed at statistical significance p<0.05, preferably <0.01.

When reference is made in this application to a deviation (e.g., increase, decrease, excess, prolongation, increase, decrease, improvement, delay, abnormal levels, or any other change or deviation from a control), the deviation may be, for example, 5% or more, in particular 10% or more, in particular 15% or more, in particular 20% or more, in particular 30% or more, in particular 40% or more, or in particular 50% or more than the corresponding control values, unless otherwise indicated. Typically, the deviation will be at least 10%, ie 10% or more. The deviation can 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 a placebo instead of one or more SGLT2 inhibitors, or from a group of animals that have not been treated.

In this application, for example, insulin deviation or glucose deviation refers to a change in blood concentration or level 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 crystalline form thereof, for use in accordance with the present invention. In the case of combination with one or more active compound, the term active ingredient or active substance may also include the additional active compound.

In this application, the expression clinical(s) condition(s) refers to pathological(them) condition(s) or physiological changes that are identifiable, ie. visible and measurable, such as blood counts, and those that are associated with and/or cause a disorder and/or disease.

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. In ivGTT can

- 5 040378 typically 0.8 g of dextrose per kg of body weight is used.

In this application, ivITT refers to an intravenous insulin tolerance test. In ivITT, 0.05 units can typically be used. insulin per kg of 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 2008/049923, Wo 2008/055870, Wo 2008, Wo 2008, Wo 2008, Wo 2008, Wo 2008, Wo 2008 /055940, WO 2009/022020 or WO 2009/022008.

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

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

^R2

OH where R ¹ is cyano, C1 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, t-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-methylprop-1 -yl, 3 -hydroxy-3-methyl-but-1-yl, 1-hydroxy-1-methylethyl, 2,2,2-trifluoro-1 hydroxy-1-methylethyl, 2,2,2-trifluoro-1-hydroxy-1 - trifluoromethylethyl, 2-methoxyethyl, 2-is xyethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxyethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethyl sulfinyl, ethyl sulfonyl, trimethylsilyl, (R)-τeτpahydropoφypan-3-ylokci or (3)-tetrahydrofuran -3-yloxy or cyano;

where R ³ is preferably selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)tetrahydrofuran-3-yloxy or (3)-tetrahydrofuran-3-yloxy; and most preferably R is cyclopropyl, or a derivative thereof, wherein one or more of the hydroxyl groups of the β-D-glucopyranosyl group are acylated with groups selected from ( _{C1_18} alkyl)carbonyl, ( _{C1_18} alkyl)oxycarbonyl, phenylcarbonyl, and phenyl -( _{C1_3} -alkyl)carbonyl;

(3) dapagliflozin represented by formula (3)

(4) canagliflozin represented by formula (4)

(5) empagliflozin represented by formula (5)

he (9) ertugliflozin represented by formula (9)

and the compound represented by formula (18)

He

-6040378 where R ³ is selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3-yloxy.

The term dapagliflozin, as used in this application, refers to dapagliflozin of the above structure, as well as its pharmaceutically acceptable forms, including its hydrates and solvates, as well as its crystalline forms. The compound and methods for its synthesis are described, for example, in WO 03/099836. Preferred hydrates, solvates and crystalline forms are described, for example, in patent applications WO 2008/116179 and WO 2008/002824.

The term canagliflozin, as used in this application, refers to canagliflozin of the above structure, as well as its pharmaceutically acceptable forms, including its hydrates and solvates, as well as its crystalline forms. The compound and methods for its synthesis are described, for example, in WO 2005/012326 and WO 2009/035969. Preferred hydrates, solvates and crystalline forms are described, for example, in patent application WO 2008/069327.

The term empagliflozin, as used in this application, refers to empagliflozin of the above structure, as well as its pharmaceutically acceptable forms, including its hydrates and solvates, as well as its crystalline forms. The compound and methods for its synthesis are described, for example, in WO 2005/092877, WO 2006/120208 and WO 2011/039108. A preferred crystal form is described, for example, in patent applications WO 2006/117359 and WO 2011/039107.

The term ertugliflozin, as used in this application, refers to ertugliflozin of the above structure, as well as its pharmaceutically acceptable forms, including its hydrates and solvates, as well as its crystalline forms. The connection is described, for example, in WO 2010/023594.

Preferred SGLT2 inhibitors are glucopyranosyl-substituted benzene derivatives. Optionally, one or more of the hydroxyl groups of the glucopyranosyl group in such one or more SGLT2 inhibitors may be acylated with groups selected from (C _1-18 alkyl)carbonyl, (C _1-18 alkyl)oxycarbonyl, phenylcarbonyl, and phenyl-(C _1-3 -alkyl)carbonyl.

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

where R ³ is selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3-yloxy; And

R ³ is most preferably cyclopropyl.

Thus, in preferred embodiments, the SGLT2 inhibitor according to the present invention is a glucopyranosyl-substituted benzene derivative of an SGLT2 inhibitor, preferably an SGLT2 inhibitor of formula (1), more preferably formula (18) in each case as defined herein above.

metabolic disorders

The 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, impaired glucose tolerance, insulin resistance, insulin-dependent diabetes and/or hepatic lipidosis. Additional relevant metabolic disorders include hyperinsulinemia, impaired glucose tolerance, ketosis (particularly ketoacidosis), hyperlipidemia, dyslipidemia, elevated blood levels of fatty acids and/or glycerol, syndrome X (metabolic syndrome), and/or inflammation of the pancreas, systemic mild inflammation, inflammation of adipose tissue.

In some embodiments, the metabolic disorder is diabetes. In this application, diabetes can be pre-diabetes, insulin-dependent diabetes, and non-insulin-dependent diabetes. In particular, the diabetes may be insulin-dependent diabetes.

In some embodiments, the metabolic disorder is hyperglycemia. In this application, hyperglycemia may be associated with diabetes, for example, insulin dependent diabetes or non-insulin dependent diabetes. In some embodiments, hyperglycemia may be associated with obesity. Hyperglycemia can be chronic.

In some embodiments, the metabolic disorder is insulin resistance. In this application, insulin resistance may be associated with diabetes, such as non-insulin dependent diabetes. In some embodiments, insulin resistance may be associated with obesity.

In some embodiments, the metabolic disorder is impaired glucose tolerance (IGT). In this application, impaired glucose tolerance may be associated with

- 7 040378 diabetes, for example, with insulin-dependent diabetes or non-insulin-dependent 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, such as non-insulin dependent diabetes. 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 canine 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 canine. In addition, for example, hyperinsulinemia and/or impaired glucose tolerance may be treated and/or prevented in an obese canine. In addition, one or more disorders selected from ketosis (particularly ketoacidosis), hyperlipidemia, elevated levels of fatty acids and/or glycerol, syndrome X (metabolic syndrome), inflammation of the pancreas, inflammation of adipose tissue, may be treated and/or prevention in an obese canine.

In some embodiments, the canine is not obese. The metabolic disorder may be associated with and/or caused by, for example, hypo- or hyperthyroidism, hypercortisolism (hyperadrenocorticism, Cushing's disease) and/or excess growth hormone (acromegaly). 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 a non-obese canine. In addition, for example, hyperinsulinemia and/or impaired glucose tolerance may be treated and/or prevented in a non-obese canine. In addition, one or more disorders selected from ketosis (particularly ketoacidosis), hyperlipidemia, elevated fatty acid and/or glycerol levels, syndrome X (metabolic syndrome), pancreatic inflammation, and/or adipose tissue inflammation may be treated and /or prevention in a non-obese canine.

In some embodiments, the canine is suffering from diabetes, such as insulin dependent diabetes or non-insulin dependent diabetes. For example, in accordance with the invention, one or more metabolic disorders selected from the group of hyperglycemia, impaired glucose tolerance, and hepatic lipidosis can be treated and/or prevented in a canine that suffers from diabetes, for example, insulin-dependent diabetes or non-insulin-dependent diabetes. . In addition, for example, hyperinsulinemia and/or insulin resistance may be treated and/or prevented in a canine that suffers from diabetes, such as non-insulin dependent diabetes. In addition, one or more disorders selected from ketosis (particularly ketoacidosis), hyperlipidemia, elevated levels of fatty acids and/or glycerol, syndrome X (metabolic syndrome), inflammation of the pancreas, inflammation of adipose tissue, may be treated and/or prevention in a canine that suffers from diabetes, such as non-insulin dependent diabetes.

In some embodiments, the canine is not diabetic but is obese.

In some embodiments, the canine is not obese, but diabetic.

The present invention also provides the use of one or more SGLT2 inhibitors for the treatment and/or prevention of complications associated with hyperglycemia. For example, by improving daily blood glucose control, the development or progression of, or regression of, cataract formation in a canine can thus be delayed or prevented.

Ketosis is a state of elevated 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 formed from the breakdown of fatty acids and from the deamination of amino acids. Two common ketones that form in the human body are acetoacetic acid and β-hydroxybutyrate. In dogs, three ketones are mainly found: acetoacetic acid, beta-hydroxybutyrate, and pyruvic acid. Ketoacidosis can produce an odor on 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

- 8 040378 regulate the production of ketone, in particular by the production of acetyl-CoA, thus causing such a serious accumulation of keto acids that the pH value of the blood decreases significantly, that is, an excess of 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 sufficiently slow this production (eg, due to insulin resistance/insulin sensitivity or lack of insulin). The presence of high blood sugar levels (hyperglycemia) caused by a lack of insulin can lead to further acidification of the blood. In healthy individuals, this usually does not occur because the pancreas produces insulin in response to rising blood ketones/sugar levels.

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

Prediabetes in canines is characterized by hyperinsulinemia, insulin resistance in target organs, impaired glucose tolerance, including, for example, an altered insulin response to glycemic challenge, and also, for example, induced by stress. Prediabetes is also often associated with obesity. Prediabetes can also be associated with intermittent hyperglycemia.

Non-insulin-dependent diabetes in canines is characterized by both increased insulin production and resistance to insulin in target organs, and as a result, hyperglycemia. This is often seen in intact diabetic female canines and is mainly associated with progesterone, which acts as an internal insulin antagonist. Thus, it is most often associated with either menstrual cycles, such as diestrus, or pregnancy - gestational. Genetic factors, glucose steroids, lack of exercise and obesity are possible additional causes of insulin resistance.

Clinical signs of diabetes that are seen in canines include polydipsia, polyuria, weight loss and/or polyphagia. In contrast, in other animals such as cats, anorexia has been described more frequently than polyphagia.

In addition, particularly relevant clinical signs of diabetes mellitus in canines in the context of the present invention are hyperglycemia and glycosuria. Hyperglycemia in a canine animal (e.g., dogs) is defined as a plasma glucose value above normal values (3.5-7 mmol/L or 60-120 mg/dL), such as 8 mmol/L or more or 150 mg /dL or more plasma glucose. Glycosuria in a canine animal (eg, dogs) is defined as urinary glucose levels above normal (0-2 mmol/L or 36 mg/dL). The renal threshold is reached at a blood glucose concentration of about 8-11 mmol/L or 150 to 200 mg/dL.

Diagnosis of diabetes mellitus in canines may alternatively be based on three criteria, such as the following:

(1) measurement of 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-rays, 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 in diabetic animals, and therefore is not always the goal of treatment in accordance with the present invention. In accordance with the invention, the concentration of glucose in the blood can also be maintained at a level, for example, from 5.5 to 16.6 mmol/l or from 100 to 300 mg/dl. For canines, this will often be satisfactory.

Hyperglycemia provokes cataract, which has an early manifestation, rapid development and bilateral symmetry. Clouding of the lens in the eye leads to deterioration or complete loss of vision.

Insulin resistance can be described as a condition in which the normal amount of insulin is insufficient to produce a normal response to insulin 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. An increase in the 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

- 9 040378 local storage of glucose as glycogen), while insulin resistance in liver cells leads to impaired glycogen synthesis and an inability to suppress glucose production. Elevated levels of fatty acids in the blood, reduced glucose uptake by the muscles, and increased production of hepatic glucose can contribute to elevated blood glucose (hyperglycemia). Obese dogs have been found to be insulin resistant, i.e., a fivefold decrease in insulin sensitivity compared to lean animals.

Insulin resistance may be associated with obesity, visceral obesity, hypertension, and dyslipidemia associated with elevated triglycerides, low-density lipoprotein (LDL) small compact particles, and reduced levels of LDL cholesterol. With regard to visceral obesity, a lot of evidence in humans suggests two strong links with 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. It has been shown in numerous experimental models that these pro-inflammatory cytokines profoundly disrupt the normal action of insulin in fat and muscle cells, and they may represent a major factor in the onset of whole body insulin resistance seen in patients with visceral obesity. Similarly, in canines, excessive body fat contributes to low-grade systemic inflammation. The cause of the vast majority of cases of insulin resistance remains unknown. There is a clear hereditary component. However, there is some reason to suspect that insulin resistance is associated with high carbohydrate diets. Inflammation is also likely to be involved in the onset of insulin resistance.

Hyperinsulinemia can be characterized as a condition in which there are excessive levels, i.e., those that are about 35 pmol/L above baseline or about 200 pmol/L, of insulin circulating in the blood, for example, under glycemic load (for example, at ivGTT or stress). As already mentioned, it is commonly present in cases of insulin resistance, and may be a consequence of this in canines.

Impaired glucose tolerance can be described as a condition in which, in response to a glycemic load, for example, after a meal or after a stress test (glucose tolerance test) or after a stress-induced increase in blood glucose concentration, the glycemic peak of glucose deviation is higher 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 to be a highly modifiable risk factor for cholesterol-mediated 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 dog) are triglyceride concentrations of 50 to 150 mg/dl. Normal blood cholesterol levels are, for example, 130 to 300 mg/dL for a dog.

Dysadipokinemia can be characterized as a condition in which circulating plasma levels of biologically active substances that are produced in adipose tissue and act in an autocrine/paracrine or endocrine manner are abnormal, for example, an increase in leptin and/or a decrease in adiponectin.

Subclinical or systemic inflammation, in particular mild 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 respective receptors.

Obesity can be described as a medical condition in which excess body fat accumulates to the extent that it can adversely affect a person's health, resulting in a reduced life expectancy. Obese dogs, for example, have a body condition score (BCS) that is over 7 (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 overt clinical consequences - for example, high blood pressure, cardiomyopathy, renal dysfunction, and/or musculoskeletal disorders in canines.

Metabolic syndrome is also known as metabolic syndrome X (metabolic syndrome), syndrome X (metabolic syndrome), insulin resistance syndrome,

- 10 040378

Riven and CHAOS (as an abbreviation for coronary heart disease, hypertension, atherosclerosis, obesity, and stroke).

The exact mechanisms of the complex pathways of the metabolic syndrome are not yet fully known. The pathophysiology is extremely complex and only partially elucidated. 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.

The pathophysiology is typically characterized by the development of visceral fat, after which adipocytes (fat cells) of visceral fat increase plasma levels of TNF-alpha and alter levels of a number of 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 modification (eg, caloric restriction and physical activity). However, medical treatment is often necessary. Accordingly, the present invention also provides for the prevention of clinically relevant consequences of metabolic disorders, eg, high blood pressure, cardiomyopathy, renal dysfunction, and/or musculoskeletal disorders in canines.

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, trauma, vasculitis (i.e., inflammation of the small blood vessels in the pancreas) and can be autoimmune pancreatitis.

Metabolic disorders, in particular dyslipidemia and elevated serum triglyceride levels, are risk factors for the development of pancreatitis and thus may 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 (skin and/or visceral). It can be diagnosed on the basis of 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 on the basis of 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.

Canine representatives

In this application, the representative canine may be a member of the Canidae (ie, canine) family. Such an animal could thus belong either to the subfamily Canini (referring to wolves) or to the subfamily Vulpini (referring to foxes). The term canine encompasses the term dog, such as a domestic dog. The term domestic dog covers the terms Canis lupus familiaris and Canis lupus dingo.

Pharmaceutically acceptable forms

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

Any pharmaceutically acceptable crystalline form of the SGLT2 inhibitor can be used according to the invention, for example formula (1), preferably formula (18).

Crystal 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). For example, a crystalline complex of 1-cyano-2-(4-cyclopropylbenzyl)-4-(P-O-glucopyranos-1-yl)benzene (formula (2); Compound A) with proline (eg Lproline) is preferred.

Thus, this application discloses a crystalline complex between one or more natural 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 of formula (18 ).

A crystalline complex between one or more natural amino acids (eg, proline, preferably L-proline) and an SGLT2 inhibitor is the 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 the preferred pharmaceutically acceptable form of the inhibitor. SGLT2 for use in accordance with the present invention.

In addition, this application discloses a method for obtaining one or more crystalline complexes, as defined in this application above and below, where this method includes the following steps:

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

(b) holding the solution to precipitate a crystalline complex therefrom;

(c) removing the precipitate from the solution; and (d) drying the precipitate, optionally, to remove any excess of said solvent or mixture of solvents.

Certain pharmaceutical activity is known to be a basic requirement that must be met when using a pharmaceutically active agent before it is approved as a drug on the market. However, there are a number of additional requirements that a pharmaceutically active agent must meet. These requirements are based on various parameters which are related to the nature of the active substance itself. Without limitation, examples of such parameters are the stability of an active agent under various environmental conditions, its stability during the manufacture of a pharmaceutical product, and the stability of an active agent in final drug formulations. The pharmaceutically active substance used to prepare pharmaceutical compositions should be as pure as possible, and its long-term storage stability should 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 degradation products. In such cases, the content of the active substance in the medicinal product may be less than indicated.

Uniform distribution of the drug in the composition is a critical factor, in particular when the drug is to be administered at 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 degradation of the pharmaceutically active substance as a side effect of milling (or micronization) should be avoided as far as possible, despite the stringent conditions required during the process, it is essential that the active agent be very stable during the milling process. Only when the active substance is sufficiently stable during the milling process can a homogeneous pharmaceutical preparation be obtained which always contains the desired amount of the active substance in a reproducible manner.

Another problem that may arise during the grinding process to obtain the desired pharmaceutical preparation is the energy input caused by this process and the stress on the surface of the crystals. This may in some cases lead to polymorphic changes, amorphization or to a change 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 the 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 content of the pharmaceutically active substance as a result of the 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 desiccants or by storing the drug in an environment where it is protected from moisture. It is therefore preferred that the pharmaceutically active substance is at best slightly hygroscopic.

In addition, the presence of a well-defined crystalline form allows purification of the medicinal substance by recrystallization.

In addition to the requirements above, it should generally be understood 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 (e.g., a glucopyranosyl-substituted benzene derivative or an SGLT2 inhibitor of formula (1) or formula (18) satisfies the important requirements stated 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.

Also preferred are those crystalline complexes according to the present invention which form between an SGLT2 inhibitor (e.g., 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 according to the 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, when the molar ratio of the SGLT2 inhibitor (e.g., formula (1), preferably formula (18) and natural amino acid is in the range from about 2:1 to about 1:3; more preferably from about 1.5:1 to about 1:1.5, even more preferably about 1.2:1 to about 1:1.2, most preferably about 1:1 In this case, such an implementation is referred to as 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 (e.g. such of formula (1), preferably formula (18) and proline; in particular said 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, when the molar ratio of the crystalline complex and water is in the range 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.

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 the drug manufacturing process. For example, crystals can be obtained in a form and particle size that are particularly suitable in the process for the preparation of solid pharmaceutical preparations. In addition, the crystals have high mechanical strength, which makes it possible to grind the crystals. In addition, the crystalline complex does not show 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 STOESTADI P-diffractometer in transmission mode, equipped with a displacement sensitive sensor (OED) and a Cu anode as an X-ray source (CuK _a1 radiation, λ=1.54056 A,

- 13 040378 kV, 40 mA). The 2Θ[°] values indicate the diffraction angle in degrees, and the d[A] values indicate the specified distances in A between the grating planes. The intensity shown in Fig. 3 is given in pulses per second.

In order to take into account the experimental error, the values of the 2Θ quantity described above should be considered accurate up to ± 0.1° 2Θ, in particular, ± 0.05° 2Θ. In other words, when evaluating whether a given sample of compound A crystals in the crystalline form according to the 2Θ values described above, the 2Θ value that is experimentally observed for the sample should be considered identical with 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 by thermal gravimetry (TG) using a TGA 851 (Mettler Toledo).

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

(a) obtaining a solution of an SGLT2 inhibitor as described in this application and one or more natural amino acids in a solvent or mixture of solvents;

(b) holding the solution to precipitate a crystalline complex therefrom;

(c) removing the precipitate from the solution; and (d) drying the precipitate, optionally, to remove any excess of said solvent or mixture of solvents.

According to step (a), a solution of the SGLT2 inhibitor and one or more natural amino acids is prepared in a solvent or mixture of solvents. 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 naturally occurring amino acids, or one or more naturally occurring amino acids may be dissolved in a solution that includes the SGLT2 inhibitor. According to an alternative procedure, the SGLT2 inhibitor is dissolved in a solvent or solvent mixture to form a first solution, and one or more natural amino acids are dissolved in a solution or solvent mixture to form a second solution. Thereafter, said first solution and said second solution are combined to form a solution according to step (a).

Preferably, the molar ratio of natural amino acid and SGLT2 inhibitor in solution corresponds to the molar ratio of natural amino acid and SGLT2 inhibitor in the crystalline complex that is obtained. 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 C1.4-alkanols, water, ethyl acetates, acetonitriles, acetones, diethyl ether, tetrahydrofuran and a mixture 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 mixtures of ethanol and/or isopropyl alcohol with water.

In the case of a mixture of water and one or more _{C1_4} alkanols, in particular methanol, ethanol and/or isopropanol, most preferably ethanol, the preferred volume ratio of water and alkanol is 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 about room temperature (about 20° C.) or at an elevated temperature up to about the boiling point of the solvent or solvent mixture used.

In accordance with a preferred embodiment, the starting material of the SGLT2 inhibitor and/or one or more natural amino acids and/or the solvent and solvent mixture contains an amount of H ₂ O which is at least necessary to form a hydrate of the SGLT2 inhibitor; in particular at least 1 mol, preferably at least 1.5 mol of water per mol 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, or one or more natural amino acids, or the indicated solvent, or mixtures of solvents, or the indicated compounds and/or solvents in combination, contain the amount of H 2 O as indicated above. For example, if the starting material of the SGLT2 inhibitor or natural amino acid in step (a) contains a sufficient amount of water, as indicated above, then the water content of the solvent(s) is not necessary.

In order to reduce the solubility of the crystalline complex according to the given

- 14 040378 solution, in step (a) and/or step (b), one or more anti-solvents can be added, preferably during step (a) or at the beginning of step (b). Water is an example of a suitable anti-solvent. The amount of anti-solvent is preferably chosen so as to obtain a supersaturated or saturated solution with respect to the crystalline complex.

In step (b), the solution is held for a period of time sufficient to produce a precipitate, i.e., a crystalline complex. The temperature of the solution in step (b) is about the same or lower than in step (a). While holding, the temperature of the solution is preferably lowered, preferably to a temperature in the range of 20 to 0° C. or even lower. Step (b) can be carried out with or without agitation. As known to the person skilled in the art, the size, shape and quality of the resulting crystals can be controlled by the time period and temperature difference in step (b). In addition, crystallization can be induced using 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 with a glass rod. Optionally, a (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 filtration, decantation and centrifugation.

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

The crystalline complex as described herein is preferably used as the active drug in substantially pure form, ie substantially free of other SGLT2 inhibitor crystalline forms. However, the present invention also encompasses a crystalline complex in admixture with another crystalline form or forms. If the active drug substance is a mixture of crystalline forms, then 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. weight of the crystalline complex, as described in this application.

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 which may be affected by SGLT inhibition, in particular SGLT-2 activity, in particular metabolic disorders, as described in this application. A crystalline complex as described herein (particularly with a naturally occurring amino acid, eg proline, in particular L-proline) is also suitable for use in the treatment of canines.

Pharmaceutical compositions and formulations

One or more SGLTS inhibitors for use in accordance with the invention may be formulated into pharmaceutical compositions. It can be obtained in the form of solid or liquid compositions. In any case, it is preferably formulated for oral administration, preferably in liquid form for oral administration. One or more SGLTS 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 one or more SGLT2 inhibitors in accordance with the present invention and one or more excipients. Any inert excipient that allows or maintains the intended therapeutic effect can be used. Such fillers are available to the person skilled in the art. Useful excipients are, for example, release agents (used to reduce adhesion between powder (granules) and punch faces and thereby prevent sticking to tablet punches), binders (binder solution or dry binders that hold ingredients together), coatings (to protect tablet ingredients from damage by moisture in the air and make large or unpalatable tablets easier to swallow), disintegrating agents (to allow tablets to break when reconstituted), fillers, diluents, flavors, colors, glidants (flow regulators for in order to promote the flow of the powder by reducing interparticle friction and cohesion), lubricants (to prevent ingredients from sticking together and sticking to

- 15 040378 tablet punches or capsule filling machine), preservatives, sorbents, sweeteners, etc.

Dosage forms according to 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 and the like. them.

Manufacturing procedures for formulations suitable for canines are known to those skilled in the art, and for solid formulations include, for example, direct compression, dry granulation, and wet granulation. In a direct compression process, the active ingredient and all other excipients are placed together in a compression device which is directly used to compress tablets of 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, e.g. a liquid unit dosage form or a unit solid composition, e.g. a tablet, may contain from 0.01 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 will be appreciated by one of skill in the art, the amount of SGLT2 inhibitor in a solid composition or any formulation as described herein for administration to a canine can be increased or decreased as appropriate, depending on the body weight of the canine. which 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 excipients that alter 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, such as palatability and/or chewability, that make the composition suitable for administration to a canine.

In addition, liquid compositions are preferred. Liquid preparations may be, for example, solutions, syrups or suspensions. They may be administered directly to the dog, or may be mixed with food and/or drink (eg, drinking water, etc.) for the dog. One of the advantages of a liquid dosage form (similar to a formulation in granular form) is that such a dosage form provides accurate dosing. For example, the SGLT2 inhibitor can be dosed exactly according to the body weight of the canine. Typical formulations of liquid compositions are known to those skilled in the art.

Dosage and administration

The practitioner of skill in the art can determine suitable dosages for use in accordance with the present invention. Preferred dosage units include mg/kg, that is, mg of the SGLT2 inhibitor per unit body weight of the canine. The SGLT2 inhibitor according to the invention may, for example, be administered at a dose of 0.1 to 4.0 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 of body weight per day.

The practitioner of skill in the art is able to prepare the SGLT2 inhibitor of the present invention for administration at the desired dose.

Preferably, according to the invention, the SGLT2 inhibitor is administered no more than three times a day, more preferably no more than twice a day, most preferably only once a day. The frequency of administration may be adapted to the dog's typical feeding frequency.

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

Preferably, after administration and the time required for the SGLT2 inhibitor to reach the bloodstream, such levels are maintained 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. SGLT2 inhibitor can be injected directly into the oral cavity of animals

- 16 040378 (for example, with a syringe, preferably a syringe graduated according to body weight) or together with food or drink for the animal (for example, drinking water or the like), in each case, preferably in liquid form. However, the SGLT2 inhibitor may also be administered, eg parenterally, or by any other route of administration, eg rectally.

An SGLT2 inhibitor may be used alone or in combination with another drug. In some embodiments, one or more SGLTS inhibitors are used in combination with one or more other oral hypoglycemic agents. 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 co-administration drug with the SGLT2 inhibitor is not administered concurrently 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, such as 12 months or more.

In some embodiments, one or more SGLTS inhibitors are used in conjunction with insulin, preferably in simultaneous, sequential, and/or intermittent combinations. Such co-administration may also be in the form of a fixed dose combination (FDC), for example, a combination consisting of one or more SGLTS inhibitors and insulin, which are combined into a single dosage form, which is manufactured and distributed in certain fixed doses. In some embodiments of the present invention, an 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 canine is not treated with said compound. In some embodiments, the SGLT2 inhibitor is not used in combination with a DPP-IV inhibitor, i.e., the canine is not being treated with a DPPIV inhibitor.

In some embodiments, the SGLT2 inhibitor is used as monotherapy, i.e., as a separate therapy, in other words, no other drug is administered to the dog to treat or prevent the same metabolic disorder, i.e., the metabolic disorder for which an SGLT2 inhibitor is administered. For example, no other drug is administered to a dog to treat or prevent the same metabolic disorder for a period of at least 2, 3, or 4 weeks before and after administration of an SGLT2 inhibitor.

Brief description of the figures

Fig. Figure 1 Urinary glucose excretion in beagle dogs after a single oral dose of Compound A. A urine sample was taken individually 0-24 hours and 24-48 hours after ingestion. After 24 hours, urine obtained by inserting a catheter into the bladder was added to the freely collected urine. In the controls and at the low dose (0.01 mg/kg), almost no glucose was detected in the urine. The indicated p values are above the margin of error compared to the control (*, p<0.05).

Fig. 2 is the sigmoid dose effect for urinary glucose excretion over a period of time 0-24 hours after administration of Compound A, as shown. The ED ₅₀ for glucose excretion with compound A in beagle dogs is 0.1 mg/kg (95% CI 0.02-0.52 mg/kg).

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

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

Examples

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

Example 1 Pharmacokinetics (PK) of a single oral dose of Compound A in dogs.

Compound A was administered to dogs after an overnight fasting period. Groups (n=4 per group) received either oral vehicle (deionized water) containing an SGLT2 inhibitor in the form of Compound A at 1 mg/kg and 10 mg/kg or intravenous vehicle (saline) containing an SGLT2 inhibitor in the form of compound A at a dose of 1 mg/kg. PK measurements were made up to day 3 after a single administration of Compound A or its vehicle.

- 17 040378

Table 2.

data, single dose mg/kg, and 10 mg/kg)

Parameter intravenously 1 mg/kg orally 1 mg/kg orally 10 mg/kg tmax [h] average value 1.9 0.6 Stax [Nmol/L] average value 4845 51525 CL [ml/min/kg] average value 0.63 CL/F [ml/min/kg] average value 0.64 0.71 F [%] average value 101 92 AUCo >, [nmol-hour/l] average value 67025 67675 616750 Τι/g [h] average 13.4 13.9 14.5 meaning

Example 2 Effect of Compound A on Urine and Blood Glucose Concentrations Following Single Doses in Dogs.

Beagle dogs were kept without food overnight and given a single oral dose of Compound A at 0 mg/kg by weight, 0.01 mg/kg by weight, 0.1 mg/kg by weight, or 1 mg/kg by weight (n=3 per group), followed by washing with water (1 ml/kg by weight).

Compound A was wetted with a small volume of 1% (w/v) aqueous solution of Polysorbate 80 (Tween 80, polyoxyethylene sorbitol monooleate, ICN Biomedicals) and then dissolved by slowly adding a large volume of 0.5% (w/v) aqueous solution of hydroxyethyl cellulose (Natrosol 250 HX, Boehringer Ingelheim) and stirred at room temperature for about 15 minutes. The final concentration of Polysorbate 80 was 0.015%. Compound A was used in a volume of 2 ml/kg by weight.

Animals were kept separately in metabolic cages and fed 2 hours after administration. They had free access to water during the experiment. Urine was collected at 0-8 hours, 8-24 hours, 24-32 hours and 32-48 hours after administration. A catheter (Eickemeyer) was inserted into the bladder for complete round-the-clock urine collection. This urine was mixed with urine that was excreted between 8 and 24 hours. A 5 ml volume of 10% sodium azide solution in saline was added to each urine collection tube until a sample was obtained. The volume of urine was determined and the samples were frozen for subsequent determination of the glucose concentration.

During the experiment, blood samples were taken from the vein of the forearm. Blood was collected in EDTA tubes prior to vehicle or Compound A administration and then collected at time points at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h, 32 h, and 48 hours after drug administration. Upon receipt, blood plasma was collected and frozen to determine the glucose concentration.

A significant increase in urinary glucose concentration and volume was evident at the two high doses 8 hours after administration (FIG. 1).

None of the doses of Compound A caused hypoglycemia or abnormal blood glucose levels in dogs compared to normal controls.

The level of excretion of glucose in the urine, as it was established in this way, showed that the value of ED50 was 0.1 mg/kg (Fig. 2).

Example 3 Treatment of Insulin-Dependent Diabetes in Dogs.

Treatment of dogs with insulin dependent diabetes.

Insulin in combination with Compound A according to the present invention or the combination of active substances according to the present invention, in addition to obtaining a dramatic improvement in glucose metabolism, prevents metabolic deterioration in the long term and reduces the dose of insulin needed to treat a canine that suffers from diabetes. This can be observed if dogs are treated for a shorter or longer period of time, such as 2-4 weeks or 3 months to 1 year, using a pharmaceutical composition according to the present invention, and compared with the metabolic rate prior to treatment. or with dogs that have been treated with, for example, insulin alone. There is evidence of therapeutic success if the average daily glucose level and the level of fructosamine in the blood are reduced compared to the level before treatment. Further evidence of therapeutic success is obtained if a significantly lower percentage of dogs treated with the pharmaceutical composition of the present invention, compared to dogs treated with other drugs, experience a transient impairment in glucose metabolism (e.g. hyper or hypoglycemia).

Example 4 Improvement in non-insulin dependent diabetes in female dogs with diestrus/pregnancy diabetes.

Non-insulin-dependent diabetes is often found in intact female canids.

- 18 040378

Compound A therapy may be provided to prevent progression to overt diabetes. In studies over short or long periods of time (for example, 2-4 weeks or 1-2 years), treatment success was studied by determining fasting blood glucose values, and/or glucose values after a meal, or after an exercise test (intravenous test glucose tolerance or food tolerance test after an appropriate meal) during the study of different stages of the menstrual cycle and / or after the end of the therapy period to study and compare them with the values before the start of the study and / or with the values obtained from the placebo group. Additionally, the value of fructosamine can be determined before and after therapy and/or can be compared with the initial value for dogs that were treated with other drugs or placebo. Significant reductions in fasting or non-fasting glucose and fructosamine levels demonstrate efficacy in treating non-insulin dependent diabetes or preventing the onset of diabetes in female dogs with a history of diestrus/pregnancy-associated diabetes.

Example 5 Treatment of Hyperglycemia.

In clinical studies in dogs with metabolic disorders that were treated for varying lengths of time (eg, 2 weeks to 12 months), treatment success was tested using baseline blood glucose and/or blood fructosamine measurements.

Improvement in blood glucose control can also be measured by 24-hour blood glucose curves, such as a 9 or 24-hour blood glucose curve that starts before drug administration and contains multiple measurements after drug administration.

A significant reduction in these values during or at the end of the study compared to baseline, or compared to the placebo group or other therapy group, proves the effectiveness of the pharmaceutical composition according to the invention in reducing hyperglycemia in dogs.

Alternatively, the effect of compound A on hyperglycemia can be demonstrated in dogs that have been subjected to a prolonged infusion of glucose (hyperglycemic clamp). Normalization of hyperglycemia can be assessed in comparison to no treatment and/or combination therapy with insulin and/or treatment with insulin alone.

Example 6 Prevention or treatment of complications associated with hyperglycemia.

Treatment of dogs that suffer from hyperglycemia, or insulin-dependent diabetes or non-insulin-dependent diabetes, with Compound A according to the present invention, or using a combination of active substances in accordance with the present invention, prevents or reduces complications associated with hyperglycemia, such as cataract formation.

Evidence of therapy success is compared with dogs treated with other antidiabetic drugs or placebo. The success of the treatment was determined, for example, using an ophthalmological examination of the eye for the development or progression or regression of cataract formation. Alternatively or additionally, the time of cataract formation and/or the progression of cataract maturation can be determined and compared to dogs that have been treated with other antidiabetic drugs or placebo.

Example 7 Treatment of insulin resistance.

In clinical studies using insulin resistant dogs that were treated for varying lengths of time (e.g., 2 weeks to 12 months), treatment success was tested using baseline blood glucose, blood fructosamine, blood insulin, and/or or the level of C-peptide and the corresponding relationship between the parameters in a particular dog.

Also, glucose values and blood lipid spectrum (for example, the level of non-esterified fatty acids) and / or insulin levels after a meal and after a stress test (glucose tolerance test or insulin tolerance test) during or after the end of the treatment period for study can be compared with the values pre-study and/or insulin resistant dogs treated with other drugs or placebo.

Example 8 Treatment of Prediabetes in Dogs.

The efficacy of the SGLT2 inhibitors of the present invention in the treatment of prediabetes, which is characterized by abnormal fasting blood glucose and/or impaired glucose tolerance and/or insulin resistance, can be tested in clinical trials. In studies over a short or long period of time (for example, 2-4 weeks or 1-2 years), treatment success was studied by determining the values of fasting blood glucose and / or glucose values after a meal or after an exercise test (intravenous test for glucose tolerance or food tolerance test after an appropriate meal or insulin tolerance test) after the end of the treatment period to study and compare them with the values before the start of the study and/or with the values of the placebo group. Additionally, the value of fructosamine can be determined before and after therapy and compared with the initial value and/or with the value for the placebo group. A significant decrease in glucose and/or fructosamine levels on an empty stomach or not on an empty stomach demonstrates the effectiveness of the treatment of prediabetes.

Example 9 Effects on body weight, body composition, dyslipidemia and dysadipokinemia.

Treatment of dogs with metabolic disorders such as obesity, dyslipidemia, dysadipokinemia, hepatic lipidosis, subclinical inflammation or systemic inflammation, in particular, mild systemic inflammation, which also includes inflammation of the adipose tissue, and associated disorders such as syndrome X ( metabolic syndrome) and/or insulin resistance, hyperglycemia, hyperinsulinemia, impaired glucose tolerance, also aims to prevent transition or slow progression towards, for example, clinically apparent consequences of metabolic disorders, for example, high blood pressure, cardiomyopathy, renal dysfunction and/or musculoskeletal disorders in canines.

The efficacy of treatment can be investigated in a comparative clinical trial in which dogs were treated over an extended period of time (eg 3-12 months) with either Compound A, or a combination of active substances, or placebo, or drug-free therapy (eg, with diet) or other medications. Before, during and after the end of therapy, the following parameters can be determined: body weight (size) and body composition, for example, using dual-energy x-ray absorptiometry. Profiles of lipids (eg, triglycerides, cholesterol, non-esterified fatty acids) and adipokines (eg, adiponectin, leptin) in plasma or serum can be measured, as well as inflammatory markers (eg, C-reactive protein, monocyte chemoattractant protein 1). Insulin and glucose levels can be determined both before and after, for example, a stress test. Renal parameters can be determined in blood and urine samples (eg, urea, creatinine, urinary albumin). Additionally, blood pressure and/or also evidence of cardiomyopathy can be studied using echocardiographic Doppler ultrasound measurements. Improvement in musculoskeletal disorders (eg, osteoarthritis) can be quantified by, for example, activity levels, lameness, and pain scores.

Example 10 Preparation of 1-cyano-2-(4-cyclopropylbenzyl)-4-(β-O-glucopyranos-1-yl)benzene (compound A).

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

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

Vg

I

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 added in portions of LiBH ₄ (3.4 g). 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 0.1M 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 give a crude product. 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. The solvent and excess reagent were then removed under reduced pressure. The residue was triturated with methanol and dried. Yield: 41.0 g (82% of theory).

Preparation of 4-bromo-1-iodine-3-phenoxymethylbenzene

- 20 040378

Phenol (13 g) dissolved in 4 M KOH solution (60 ml) was added to 4-bromo-3-chloromethyl-1-iodobenzene (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-O-glucopyranose-1-yl)-2-(phenoxymethyl)benzene

A 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 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)-O-glucapyranone (10.7 g, 90% pure) in tetrahydrofuran (5 ml) was added. The resulting solution was heated to -5°C in a cooling bath and stirred for an additional 30 min at this temperature. Was added an aqueous solution of NH ₄ Cl, and the resulting mixture was extracted with ethyl acetate. The combined organic extracts were dried over sodium sulfate and the solvent removed under reduced pressure. The residue was dissolved in methanol (80 ml) and treated with methanesulfonic acid (0.6 ml) to give 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 NaHCO3 and the resulting mixture was extracted with ethyl acetate. The combined extracts were dried over sodium sulfate and the solvent evaporated to give a crude product 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-O-glucopyranos-1-yl)-2-(phenoxymethyl)benzene

Boron diethyl etherate trifluoride (4.9 ml) was added to a solution of 1-bromo-4-(1-methoxy-O-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. over a period of 1.5 hours and then treated with aqueous sodium hydrogen carbonate solution. The resulting mixture was stirred for 0.5 h, 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 h 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, the residue was recrystallized from ethanol to give the product as a colorless solid. Yield: 6.78 g (60% of theory). Mass spectrum (ESI ⁺⁾ : m/z = 610/612 (Br) [M+NH4] ⁺ .

Obtaining 2-(phenoxymethyl)-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1 -yl)benzonitrile

Argon was passed through a flask loaded with zinc cyanide (1.0 g), zinc (30 mg), Pb ₂ (dibenzylideneacetone) ₃ ·CHCl ₃ (141 mg) and tri-tert-butylphosphonium tetrafluoroborate (111 mg). Then a solution of 1-bromo-4-(2,3,4,6-tetra-O-acetyl-O-glucopyranose-1-yl)-2-(phenoxymethyl)ben-21 040378 sol (5.4 g) in NMP (12 ml) 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 solution of sodium bicarbonate. 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+NH4] ⁺ .

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

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

A 33% solution of hydrobromic acid in acetic acid (15 ml) was added to a solution of 2phenyloxymethyl-4-(2,3,4,6-tetra-O-acetyl-O-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 h and then cooled in an ice bath. The reaction mixture was neutralized with a chilled 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 removed under reduced pressure. The residue was taken up in ethyl acetate/cyclohexane (1:5) and the precipitate was filtered off and dried at 50° C. to give the pure product. Yield: 0.52 g (75% of theory). Mass spectrum (ESI ⁺ ): m/z = 543/545 (Br) [M+NH ₄ ] ⁺ .

Obtaining 4-cyclopropylphenylboronic acid

A 2.5 M solution of n-butyl lithium 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 down to -70°C. The resulting solution was stirred at -70°C for 30 min before adding triisopropyl borate (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 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 (ESF): m/z = 207 (Cl) [M+HCOO] ^- .

Preparation of 1-cyano-2-(4-cyclopropylbenzyl)-4-(v-O-glucopyranos-1-yl)benzene

2-bromomethyl-4-(2,3,4,6-tetra-O-acetyl-O-glucopyranos-1yl)benzonitrile (1.60 g), 4-cyclopropyl-phenylboronic acid (1.0 d), potassium carbonate (1.85 g) and degassed 3:1 mixture of acetone and water (22 ml). The mixture was stirred at room temperature for 5 min before being cooled in an ice bath. Then palladium dichloride (30 mg) was 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 silica gel chromatography (dichloromethane/methanol 1:0>8:1). Yield: 0.91 g (76% of theory). Mass spectrum (ESI ⁺ ): m/z = 413 [M+NH4] ⁺ .

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

L-proline (0.34 g) dissolved in 2.1 ml of a mixture of ethanol and water (10:1 v/v) was added to a solution of 1-cyano-2-(4-cyclopropylbenzyl)-4-(v-O- glucopyranos-1-yl)benzene (1.17 g, prepared as above) dissolved in 2 ml ethanol. The resulting solution was left to settle at room temperature. Approximately 16 hours later, 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 seeding. 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-(3D-glucopyranose-1 -yl)benzene.

Received several batches of the crystalline complex in accordance with the above method of obtaining. The X-ray powder diffraction profiles coincided. Melting points were determined using DSC and were evaluated as the temperature of the start of decomposition. Examples of melting points were approximately 89°C, 90°C, 92°C, 101°C and 110°C. Powder x-ray diffraction pattern, as presented in table. 1 and as shown in FIG. And, the DSC and TG diagrams in FIG. 12 correspond to a batch with a melting point of approximately 90°C.

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

Example 11 Recipes.

Several formulation examples are described in which the term active substance denotes an SGLT2 inhibitor or a pharmaceutically acceptable form thereof, eg prodrug or crystalline form, for use in accordance with the invention. In the case of combination with one or more additional active substances, the term active substance may also include the additional active substance.

Tablets containing 100 mg of the active substance.

Compound:

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

Receipt method.

The active substance, lactose and starch were mixed together and uniformly wetted using an aqueous solution of polyvinylpyrrolidone. Thereafter, the moistened composition was screened (sieve size 2.0 mm) and dried using a stacked dryer at 50° C., after which it was screened again (sieve size 1.5 mm) and lubricant was added. The final mixture was subjected to compression to obtain tablets.

Tablet weight: 220 mg.

Diameter: 10 mm, biplanar, faceted on both sides and notched on one side.

Tablets containing 150 mg of the active substance.

Compound:

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, cornstarch 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 indicated amount of magnesium stearate. Tablets were compressed from the mixture.

Tablet weight: 300 mg.

Matrix: 10 mm, flat.

Hard gelatin capsules containing 150 mg of the active substance.

-23040378 capsule contains: active substance corn starch (dried) lactose (powdered) magnesium stearate

Compound:

150.0 mg approximately 180.0 mg approximately 87.0 mg

3.0 mg approximately 420.0 mg

Receipt.

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

Filled capsule: approximately 320 mg.

Capsule shell: hard gelatin capsule size 1.

Suppositories containing 150 mg of the active substance.

Compound:

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

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

Ampoules containing 10 mg of the active substance.

Compound:

active substance 10.0 mg

0.01 N hydrochloric acid in the required amount / NaCl bidistilled water 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, sterile filtered and transferred into 2 ml ampoules.

Ampoules containing 50 mg of the active substance.

Composition: active substance 50.0 mg

0.01 N hydrochloric acid in the required amount / NaCl bidistilled water 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, sterile filtered and transferred into 10 ml ampoules.

-24040378

Links

1) Beam et al. Vet. Ophthalmol. 1999. 2, 169-172

2) Catchpole et al., Diabetologia 2005. 48: 1948-1956

3) EP 1 213 296

4) EP 1 354 888

5) EP 1 344 780

6) EP 1 489 089

7) Nelson et al. J small Anim Pract 2000, 41, 486-490

8) Verkest, Vet J, in press doi.org/10.1016/j.tvj 1.2013.09.057

9) Wang et al. J Diabet. Compl. in press, doi:0.1016/j.jdiacomp.2013.11.002

10) WO 01/27128

11) WO 03/099836

12) W02004/007517

13) WO2004/080990

14) WO2005/012326

15) WO2005/092877

16) WO2006/034489

17) WO2006/064033

18) WO 2006/117359

19) WO2006/117360

20) WO2006/120208

21) WO2007/025943

22) WO2007/028814

23) WO2007/031548

24) WO2007/093610

25) WO2007/114475

26) WO2007/128749

27) WO2007/140191

28) WO2008/002824

29) WO2008/013280

30) WO2008/042688

31) WO2008/049923

32) WO2008/055870

33) WO2008/055940

34) W02008/069327

35) WO2008/116179

36) WO2009/014970

37) WO2009/022008

38) WO2009/022020

39) W02009/035969

40) WO2010/023594

41) WO2011/039107

42) W02011/039108

43) WO2011/117295

441 WO 2014/016381

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Claims (6)

CLAIM 1. 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 canine animal in need of such treatment and/or prevention, wherein the one or more SGLT2 inhibitors is selected from the group consisting of the following compounds or their pharmaceutically acceptable crystalline forms: glucopyranosyl-substituted benzene derivative of general formula (1) where R ¹ denotes cyano, Cl or methyl; R ² is H, methyl, methoxy or hydroxy; 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, 1hydroxycyclopropyl, 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-methylethyl, 2,2,2-trifluoro-1hydroxy-1-methylethyl, 2,2,2-trifluoro-1-hydroxy-1 -trifluoromethylethyl, 2-methoxyethyl, 2-ethoxyethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy, 2-methyloxyethyloxy, methylsulfanyl, methylsulfinyl, methylsulfonyl, ethylsulfinyl, ethylsulfonyl, trimethylsilyl, (R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3 -yloxy or cyano; or a derivative thereof, wherein one or more hydroxyl groups of the eD-glucopyranosyl group are acylated with groups selected from (C _1-18 -alkyl)carbonyl, (C _1-18 -alkyl)oxycarbonyl, phenylcarbonyl, and phenyl-(C _{1- 3} -αkyl)carbonyl; dapagliflozin represented by formula (3) canagliflozin represented by formula (4) empagliflozin represented by formula (5) ertugliflozin represented by formula (9) a compound represented by formula (18) wherein R ³ is selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)-tetrahydrofuran-3-yloxy, or (S)-tetrahydrofuran-3-yloxy; wherein one or more SGLT2 inhibitors or pharmaceutically acceptable crystalline forms thereof are administered at a dose of 0.1 to 4.0 mg/kg body weight. 2. Use according to claim 1, wherein the one or more SGLT2 inhibitors is selected from the group consisting of a glucopyranosyl-substituted benzene derivative of the general formula (1) - 26 040378 where R ³ is preferably selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3-yloxy; and most preferably R ³ is cyclopropyl; canagliflozin represented by formula (4) (5) empagliflozin represented by formula (5) ertugliflozin represented by formula (9) a compound represented by formula (18) wherein R ³ is selected from cyclopropyl, ethyl, ethynyl, ethoxy, (R)-tetrahydrofuran-3-yloxy, or (S )-tetrahydrofuran-3-yloxy. 3. Use according to any one of claims 1-2, wherein the metabolic disorder is one or more selected from the group consisting of ketoacidosis, prediabetes, insulin dependent diabetes, non-insulin dependent diabetes, insulin resistance, obesity, hyperglycemia, education cataracts caused by hyperglycemia, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, mild systemic inflammation, hepatic lipidosis, inflammation of the pancreas, consequences of metabolic disorders such as high blood pressure, renal dysfunction and/or musculoskeletal disorders, and/or syndrome X (metabolic syndrome), preferably prediabetes, insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, insulin resistance, where hyperglycemia-induced cataract formation is preferably prevented or remission is achieved and/or where Preferably, the development of the consequences of a metabolic disorder, such as high blood pressure, renal dysfunction and/or musculoskeletal disorders, is prevented or progression is slowed down, or remission is achieved. 4. Use according to any one of claims 1 to 3, wherein the metabolic disorder is selected from clinical conditions associated with prediabetes, insulin dependent diabetes mellitus and/or insulin resistance. 5. Use according to claim 4, wherein said clinical condition is one or more of ketoacidosis, insulin resistance, obesity, hyperglycemia, hyperglycemia-induced cataract formation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, mild systemic inflammation, hepatic lipidosis, pancreatic inflammation, metabolic sequelae such as high blood pressure, renal dysfunction, and/or musculoskeletal disorders, and/or syndrome X (metabolic syndrome ), where the formation of cataracts caused by hyperglycemia is preferably prevented or remission is achieved, and/or where the development of the consequences of a metabolic disorder such as high blood pressure, renal dysfunction and/or musculoskeletal disorders is preferably prevented or the progression is slowed, or to remission is achieved. 6. Use according to any one of claims 1 to 5, wherein said metabolic disorder is ketoacidosis, insulin resistance, obesity, hyperglycemia, hyperglycemia-induced cataract formation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation , systemic inflammation, mild systemic inflammation, hepatic lipidosis, inflammation of the pancreas, consequences of metabolic disorders such as high blood pressure, renal dysfunction, and/or musculoskeletal disorders, and/or syndrome X (metabolic syndrome), where cataract formation, called- -