Benzimidazole compounds for treating metabolic disorders

Abstract

The present invention relates to benzimidazole compounds of formula I, or pharmaceutically acceptable salts, crystals, hydrates, prodrugs, metabolites or solvates thereof, for the treatment of obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. [Formula 1] JPEG2023522062000017.jpg4863

Description

[Technical Field]

[0001] The present invention relates to benzimidazole compounds for the treatment of obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. [Background technology]

[0002] Obesity, diabetes, and nonalcoholic fatty liver disease (NAFLD) have become major public health problems worldwide, and the scale of these epidemics continues to grow annually. Obesity is a chronic disease that represents a major risk factor for the development of several comorbid conditions, including diabetes and NAFLD (Wang C and Liao JK (2013). A Mouse Model of Diet-Induced Obesity and Insulin Resistance. Methods Mol Biol. 821: 421-433). The World Health Organization (WHO) estimated that in 2016, there were more than 1.9 billion overweight adults worldwide, of whom 650 million were obese. The global prevalence of NAFLD is estimated at 25% of the global population. However, even higher prevalence rates (42%-70%) have been observed among patients with diabetes. Furthermore, it is predicted that one in 11 adults will suffer from diabetes.

[0003] NAFLD is a complex disease that can be classified into nonalcoholic fatty liver disease (NAFL) and nonalcoholic steatohepatitis (NASH) depending on its severity. NAFL consists of fatty liver without significant hepatocellular injury, while NASH is characterized by a combination of steatosis, hepatocellular injury, and inflammation, with or without fibrosis. NAFLD can further progress to cirrhosis, hepatocellular carcinoma, and other complications for which liver transplantation is the only treatment option. Currently, there are no approved medications on the market for the treatment of NAFLD. Given the high level of unmet clinical need in this area, there is significant interest in developing new therapies to treat obesity, diabetes, NAFLD, and NASH.

[0004] Melatonin (N-acetyl-5-methoxytryptamine) is a naturally occurring hormone that influences the circadian regulation of glucose and insulin levels, synchronizing metabolism with the daily feeding and fasting cycle (Cipolla-Neto J et al. (2014). Melatonin, energy metabolism, and obesity: a review. J Pineal Res. 56(4):371-81). Genetic variants in the melatonin receptor and altered melatonin signaling have been associated with elevated fasting plasma glucose levels, impaired insulin secretion, and increased risk of type 2 diabetes (Bouatia-Naji N et al. (2009). A variant near MTNR1B is associated with increased fasting plasma glucose levels and type 2 diabetes risk. Nat Genet. 41(1):89-94; Tuomi T et al. (2016). Increased Melatonin Signaling Is a Risk Factor for Type 2 Diabetes. Cell Metab. 23(6):1067-1077). Studies in mice genetically ablated for MT1 (MT1- / -) and MT2 (MT2- / -) have demonstrated differential effects of these receptors on glucose and insulin metabolism. MT1- / - mice exhibit a robust metabolic phenotype with higher cumulative weight gain, hyperglycemia, and pronounced insulin resistance.In contrast, MT2- / - mice exhibited decreased hepatic insulin sensitivity but increased insulin release (Owino S et al. (2018). Nocturnal activation of melatonin receptor type 1 signaling modulates diurnal insulin sensitivity via regulation of PI3K activity. J Pineal Res. 64(3); Tuomi T et al. (2016). Increased Melatonin Signaling Is a Risk Factor for Type 2 Diabetes. Cell Metab. 23(6):1067-1077). Furthermore, the regulation of MT1 and MT2 shows important differences upon melatonin stimulation. MT2 activation desensitizes cAMP responses, whereas MT1 activation leads to cAMP hypersensitivity during the subsequent drug withdrawal period (Witt-Enderby PA et al. (1998). Physiological exposure to melatonin supersensitizes the cyclic adenosine 3',5'-monophosphate-dependent signal transduction cascade in Chinese hamster ovary cells expressing the human mt1 melatonin receptor. Endocrinology.139(7):3064-71; Karamitri A et al. (2019). Melatonin in type 2 diabetes mellitus and obesity. Nat Rev Endocrinol.15(2):105-125). These findings suggest that drugs modulating MT1 / MT2 receptors may have a positive impact on the treatment of obesity, diabetes, and liver metabolic diseases; however, drugs targeting specific melatonin receptors (MT1 and MT2) in peripheral tissues have yet to be developed. Summary of the Invention

[0005] The present invention relates to benzimidazole compounds of formula I, or pharmaceutically acceptable salts, crystals, hydrates, prodrugs, metabolites or solvates thereof, for the treatment of obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. [ka]

[0006] The benzimidazole derivative compounds of the present invention also include their pharmaceutically acceptable salts, crystals, hydrates, and solvates, as long as they significantly impart the activity of the compounds.The benzimidazole derivative compounds of the present invention also include their prodrugs and metabolites, as well as slightly modified free base molecules (e.g., by incorporating substituents).These modified free base molecules, as known to those skilled in the art, result in equivalent compounds and do not significantly alter the pharmacophore of the molecules for the treatment of obesity, diabetes, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis.

[0007] Another aspect of the present invention is directed to pharmaceutically acceptable acid addition salts of compounds of formula I, such as inorganic addition salts such as, for example, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; pharmaceutically acceptable organic addition salts of compounds of formula I, such as, for example, acetate, propionate, hexanoate, heptanoate, glycolate, pyruvate, lactate, malonate, malate, maleate, fumarate, tartrate, citrate, succinate, mesylate, acetonitrile, besylate, tosylate, xinafoate, benzoate, p-toluenesulfonate, cinnamate, p-chlorobenzenesulfonate, 2-naphthalenesulfonate, p-toluenesulfonate, camphorsulfonate, trimethyl acetate, t-butyl acetate, lauryl sulfate, gluconate, glutarate, hydroxynaphthoate, salicylate, stearate, muconate, mandelate, 2-hydroxyethanesulfonic acid; alkali addition salts of metals including sodium, potassium, lithium, calcium, magnesium, and bismuth; bromides; pharmaceutically acceptable salts of primary, secondary, or tertiary amines with organic bases; salts of amino acids including arginine, lysine, and histidine; and salts of caffeine, procaine, hydrobromide, choline, betaine, ethylenediamine, glucosamine, theobromine, purines, and morpholine.

[0008] The present invention also relates to pharmaceutical compositions comprising a compound of Formula I and a pharmaceutically acceptable excipient for treating obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.

[0009] The selection of excipients to be used and the preparation of pharmaceutical compositions are generally made taking into account the type of administration route, the physical and chemical compatibility of the excipients with the active ingredient, the method of preparation of the pharmaceutical form (dosage form) and its impact on efficacy. These excipients are known in the art, described in the literature (e.g., Handbook of Pharmaceutical Manufacturing Formulations - Vol. 1 a 6-2004 - Sarfaraz K. Niazi - CRC Press and Remington's Pharmaceutical Sciences, Mack Publishing), and are widely used by technical experts in this field.

[0010] For therapeutic use and administration, the benzimidazole compounds of the present invention can be formulated using conventional techniques and appropriate excipients into compositions suitable for oral, parenteral, nasal, rectal, transmucosal and transdermal administration.

[0011] There is no particular limitation on the dosage form containing the benzimidazole compound of the present invention.For example, as a solid for oral administration, suitable dosage forms may be tablets, pills, dragees, capsules, granules, powders, pellets, freeze-dried preparations and similar forms.As a liquid for oral administration, suitable dosage forms may be solutions, dispersions, suspensions, emulsions, oils, syrups and the like.Liquid dosage forms may be used for injection, such as intravenous, intramuscular, subcutaneous, or intradermal administration.

[0012] Other examples of dosage forms include liposomes and nanoparticles, and any other mode of administration known to those skilled in the art. The dosage forms can provide immediate release, controlled release, and delayed release of the benzimidazole compounds of the present invention.

[0013] In another embodiment, the present invention relates to the use of a compound of formula I in the manufacture of a pharmaceutical composition for the treatment of obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.

[0014] Another embodiment of the invention is a dosage form as described above, which contains, for example, 0.01-5000 mg of a benzimidazole compound of the invention, and which can be administered one or more times daily during treatment.

[0015] In certain embodiments, the composition may contain at least one other active ingredient, in addition to the benzimidazole compound, used in the treatment of obesity, diabetes, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis, such as metformin, insulin and its derivatives, sulfonylureas, SGLT-2 inhibitors, DPP4 inhibitors, GLP-1 agonists, meglitinides, thiazolidinediones, α-glucosidase inhibitors, FXR agonists, PPAR agonists, ASK1 inhibitors, CCR2 and CCR5 antagonists, caspase inhibitors, insulin sensitizers, and cholic acid-arachidic acid complexes.

[0016] In another aspect, the present invention relates to methods of treating obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis with a benzimidazole compound of Formula I, or a pharmaceutically acceptable salt, crystal, hydrate, solvate, prodrug, or metabolite thereof. [ka] [Brief explanation of the drawings]

[0017] [Figure 1]Figures 1A-D show the effects of the compound of Formula I on body weight (BW) and food intake (FI) in male SD rats maintained on an HFD. The benzimidazole compound of Formula I, the comparator compound of Formula II, and dapagliflozin were administered to animals fed an HFD. Animals fed normal chow and an HFD were administered vehicle. Data are presented as adjusted means-SEM for n = 9-10 animals (BW and BW gain) and n = 5 cages (FI and daily average FI) per group. Data were analyzed by ANCOVA using BW on day 1 (A and C) and average FI during the baseline period (B and D) as covariates, excluding the chow vehicle group. Treatment groups were compared to HFD vehicle by separate Williams tests (compounds I and II) and multiple t-tests (dapagliflozin), with p < 0.05 considered statistically significant. Statistical significance is indicated as "a," "b," "c," "d," and "e" for the HFD groups treated with dapagliflozin, 10 mg / kg Compound II, 30 mg / kg Compound II, 10 mg / kg Compound I, and 30 mg / kg Compound I, respectively. # (p=0.051) and & (p=0.098) indicate a trend toward significance compared to HFD vehicle.

[0018] [Figure 2]Figures 2A-C show the effects of the compound of Formula I on plasma glucose (A), insulin (B), and HOMA-IR (C) in male SD rats maintained on an HFD. Compound I, Compound II, and dapagliflozin were administered to animals fed an HFD. Animals fed normal chow and an HFD were administered vehicle. Parameters were assessed on day 51. Data are presented as adjusted mean ± SEM (n = 9-10). Statistical analysis was performed using a general linear model (glucose and HOMA-IR) and robust regression (insulin) of log-transformed data (insulin), with treatment as a factor and bleeding order and BW on day 1 (except for comparison with the chow vehicle group) as covariates. Treatment groups were compared to HFD vehicle by separate Williams tests (Compound II and Compound I) and multiple t-tests (dapagliflozin), with statistical significance indicated by **p<0.01 and ***p<0.001. Comparisons with diet vehicle were performed by multiple t-test (HFD vehicle and dapagliflozin groups) and Dunnett's test (Compound I and Compound II), and statistically significant differences are indicated by † (p<0.05) and †† (p<0.01).

[0019] [Figure 3]Figures 3A–H show the effects of compound of formula I on hepatic triglycerides (A–B), glycogen (C–D), and histopathology (E–H) in male SD rats maintained on an HFD. Parameters were evaluated after chronic oral treatment with 10 and 30 mg / kg of compound I, 5 mg / kg of dapagliflozin, and vehicle in animals receiving an HFD. Vehicle was also administered to animals receiving a normal diet. Hepatic triglycerides were quantified using a Cobas C111 clinical analyzer (TRIGL 04657594 190). Glycogen quantification of liver samples was performed using a commercially available kit (Thermo BioVision, Glycogen Assay Kit, K646-100). Data are presented as raw data including mean ± SEM (A–D) and mean ± SEM (E–H), n = 9–10. Statistical analysis of liver triglycerides (AB) and glycogen (CD) was performed using a general linear model of log-transformed data (triglycerides only), with treatment and cohort as factors and day 1 termination sequence and BW as covariates. Treatment groups were compared to HFD vehicle by separate Williams tests (compound I) and multiple t-tests (dapagliflozin), with statistical significance indicated by * (p<0.05), ** (p<0.01), and *** (p<0.001). Comparisons with diet vehicle were performed by multiple t-tests (HFD vehicle and dapagliflozin groups) and Dunnett's test (compound I), with statistical significance indicated by † (p<0.05), †† (p<0.01), and ††† (p<0.001). Analysis of liver histopathology data (EH) was performed using the exact Wilcoxon rank-sum test. Statistically significant differences are indicated by * (p<0.05) when compared with HFD vehicle, and by † (p<0.05), †† (p<0.01) and ††† (p<0.001) when compared with chow vehicle. DETAILED DESCRIPTION OF THE INVENTION

[0020] Research has led to the discovery of certain benzimidazole (also known as N-[2-(5-chloro-2,6-dimethoxy-benzimidazol-1-yl)-ethyl]-acetamide) derivatives represented by formula I, or pharmaceutically acceptable salts, crystals, hydrates, prodrugs, metabolites or solvates thereof: [ka]

[0021] 1 H NMR (400 MHz CDCl 3) δ 1.91 (s, 3H), 3.52 - 3.57(m, 2H), 3.91 (s, 3H), 4.10 - 4.14( m, 5H), 5.62 (br s, 1H), 6.80 (s, 1H), 7.52 (s, 1H). 13 C NMR (125 MHz, CDCl3) δ ppm 23.16; 39.00; 41.16; 56.94; 57.21; 93.02; 116.73; 119.08; 133.20; 133.68; 150.85; 157.61; 170.81. MS (ESI) m / z calculated C 13H 16ClN 3O 3: 297.0880; found [M+H] + 298.0977.

[0022] This compound was designed to act preferentially in peripheral tissues by i) acting as a melatoninergic agonist with higher potency at the MT1 receptor than at the MT2 receptor (Table 1) and ii) exhibiting higher peripheral exposure than central exposure, i.e., a low brain-to-plasma ratio (Table 2). The benzimidazole derivatives of Formula I according to the present invention are potent, peripherally preferred melatonin receptor agonists with moderate selectivity for the MT1 receptor, and modulate weight gain and insulin resistance, hepatic triglyceride levels, and histologically determined steatosis. Therefore, the benzimidazole derivatives of Formula I according to the present invention add significant value to the treatment of obesity, diabetes, and NAFLD / NASH.

[0023] Accordingly, the present invention relates to a benzimidazole compound of formula I, or a pharmaceutically acceptable salt, crystal, hydrate, prodrug, metabolite or solvate thereof, for the treatment of obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. [ka]

[0024] The benzimidazole derivative compounds of the present invention also include their pharmaceutically acceptable salts, crystals, hydrates, and solvates, as long as they significantly impart the activity of the compounds.The benzimidazole derivative compounds of the present invention also include their prodrugs and metabolites, as well as slightly modified free base molecules (e.g., by incorporating substituents).These modified free base molecules, as known to those skilled in the art, result in equivalent compounds and do not significantly alter the pharmacophore of the molecules for the treatment of obesity, diabetes, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis.

[0025] Without excluding other options, suitable salts of the benzimidazole compounds of the present invention include their pharmaceutically acceptable acid addition salts, such as inorganic addition salts, for example, hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, etc.; pharmaceutically acceptable organic addition salts, such as acetates, propionates, hexanoates, heptanoates, glycolates, pyruvates, lactates, malonates, malates, maleates, fumarates, tartrates, citrates, succinates, mesylates, acetonitriles, besylates, tosylates, xinafoates, benzoates, p-toluenesulfonates, cinnamates, p-chlorobenzenesulfonates, 2-naphthalenesulfonates, p-toluenesulfonates, 2-isopropylmethylcelluloses ... salts of benzophenonesulfonates, camphorsulfonates, trimethyl acetate, t-butyl acetate, lauryl sulfate, gluconates, glutarates, hydroxynaphthoates, salicylates, stearates, muconates, mandelates, 2-hydroxyethanesulfonic acid; alkali addition salts of metals including sodium, potassium, lithium, calcium, magnesium, and bismuth; bromides; pharmaceutically acceptable salts of primary, secondary, or tertiary amines with organic bases; salts of amino acids including arginine, lysine, and histidine; and salts of caffeine, procaine, hydrobromide, choline, betaine, ethylenediamine, glucosamine, theobromine, purines, and morpholine.

[0026] The present invention also relates to pharmaceutical compositions comprising a compound of Formula I and a pharmaceutically acceptable excipient for treating obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.

[0027] The selection of excipients to be used and the preparation of pharmaceutical compositions are generally made taking into account the type of administration route, the physical and chemical compatibility of the excipients with the active ingredient, the method of preparation of the pharmaceutical form (dosage form) and its impact on efficacy. These excipients are known in the art, described in the literature (e.g., Handbook of Pharmaceutical Manufacturing Formulations - Vol. 1 a 6-2004 - Sarfaraz K. Niazi - CRC Press and Remington's Pharmaceutical Sciences, Mack Publishing), and are widely used by technical experts in this field.

[0028] Pharmaceutical excipients are typically classified and subclassified based on the function they serve in pharmaceutical compositions and / or manufacturing techniques. They may be referred to as diluents, binders, disintegrants and deflocculating agents, lubricants, suspending agents, thickeners, solvents, surfactants, slip agents, anti-agglomerating and flow agents, coating agents, plasticizers, sweeteners, tonicity agents, colorants, preservatives, antioxidants, pH control and adjusters, complexing agents used to mask flavor, improve solubility, promote formulation stability, and modify bioavailability, as well as chelating agents, flavors, and flavoring agents.

[0029] Diluents are pharmaceutical excipients found in solid dosage forms such as tablets, capsules, pills, pellets, powders, and granules to increase the volume and weight of the formulation. They may also be used in liquid and semi-solid pharmaceutical forms for the same purpose. Examples of diluents suitable for preparing the pharmaceutical compositions of the present invention include, but are not limited to, calcium carbonate, calcium carbonate, calcium phosphate, calcium sulfate, microcrystalline cellulose, powdered cellulose, dextrin, glucose, fructose, kaolin, anhydrous and / or monohydrate lactose, maltose, sorbitol, various starches (corn, wheat, potato, tapioca), pregelatinized starch, sucrose, and sugar.

[0030] Binders are pharmaceutical excipients included in formulations to facilitate the aggregation of powders into granules during the mixing (and granulation) stage, using water as the granulation fluid or hydroalcoholic mixtures and other solvents. Binders can also be used in dry mixing processes where no liquid is required. Examples of binders suitable for preparing the pharmaceutical compositions of the present invention include, but are not limited to, acacia gum, alginic acid, ammonium methacrylate copolymers, carbomer copolymers and homopolymers and interpolymers, starch (corn, wheat, potato, tapioca), microcrystalline cellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, dextrin, maltodextrin, maltose, sucrose, gelatin, glucose, guar gum, and povidone.

[0031] Disintegrants and anti-agglomerating agents are pharmaceutical excipients that can accelerate the disintegration and dissolution of formulations when in contact with biological fluids. Examples of disintegrants and anti-agglomerating agents suitable for preparing the pharmaceutical compositions of the present invention include, but are not limited to, alginic acid, starch, sodium alginate, croscaramelose sodium, sodium glycolate, sodium carboxymethylcellulose, microcrystalline cellulose, and crospovidone.

[0032] Lubricant is an excipient that reduces the friction between particles in the formulation, and also reduces the friction between particles and the wall of the device used for their preparation.The example of lubricant suitable for preparing the pharmaceutical composition of the present invention includes but is not limited to calcium stearate, magnesium stearate, zinc stearate, mineral oil, polyethylene glycol, sodium lauryl sulfate, sodium stearyl fumarate, starch, stearic acid, talc, type I hydrogenated vegetable oil.

[0033] Suspending agents and thickening agents are excipients used in formulations to ensure the stability of dispersed systems (such as suspensions and emulsions), to reduce the settling rate of particles or the fluidity of liquid formulations.Examples of suspending agents and thickening agents suitable for preparing the pharmaceutical composition of the present invention include, but are not limited to, acacia gum, agar, alginic acid, aluminum monostearate, bentonite, carbomer, copolymer carbomer, homopolymer carbomer, interpolymer carbomer, calcium and sodium carboxymethylcellulose, carrageenan, microcrystalline cellulose, dextrin, guar gum, gellan gum, hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose, magnesium aluminum silicate, pectin, polyethylene oxide, polyvinyl alcohol, povidone, propylene glycol alginate, sodium alginate, silicon dioxide, colloidal silicon dioxide, starch (corn, wheat, potato, tapioca), tragacanth gum, and xanthan gum.

[0034] Solvents are excipients used to dissolve other substances in preparing liquid, semi-solid, and solid compositions, and in the latter case to ensure easier mixing and / or provide a uniform concentration of active pharmaceutical ingredients or other excipients. Examples of solvents suitable for preparing pharmaceutical compositions of the present invention include, but are not limited to, water, ethanol, isopropanol, vegetable oils (corn, cotton, sesame, soybean), mineral oil, glycerin, sorbitol, and oleic acid.

[0035] Surfactants, also known as surface tension modifiers, are excipients with various functions and are used as emulsifiers, humectants, and / or solubilizers. Examples of surfactants suitable for preparing the pharmaceutical compositions of the present invention include benzalkonium chloride, benzethonium chloride, cetylpyridinium chloride, nonoxynol 9, octoxynol 9, polyoxyl 50 stearate, polyoxyl 10 oleyl ether, polyoxyl 20 cetostearyl ether, polyoxalate 35 lysine oil, hydrogenated polyoxalate 40 lysine oil, polyoxyl 40 stearate, polyoxyl lauryl ether, polyoxyl stearyl ether, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, sodium cetoesteryl sulfate, sodium lauryl sulfate, sorbitan monolaurate, sorbitan monooleate, sorbitan Monostearate, sorbitan sesquioleate, sorbitan trioleate, cetyl alcohol, oleyl alcohol, poloxamer, propylene glycol monostearate, carbomer copolymers and interpolymers, cholesterol, monoethanolamine, diethanolamine, triethanolamine, diethylene glycol stearate, sodium docusate, ethylene glycol stearate, glyceryl distearate, glyceryl monolinoleate, glyceryl monooleate, glyceryl monostearate, lanolin alcohol, lecithin, mono and diglycerides, sodium stearate, stearic acid, emulsifying wax.

[0036] Flow agents, anti-agglomerating agents, and slip agents are excipients used in formulations to promote flow and reduce clumping in powder flow and solid conduction funnels during processing. Examples of flow agents, anti-agglomerating agents, and slip agents suitable for preparing the pharmaceutical compositions of the present invention include, but are not limited to, calcium silicate, magnesium silicate, colloidal silicon dioxide, and talc.

[0037] Coating agents are excipients that can be used for various functions, such as masking unpleasant tastes and odors, controlling the rate of drug release, improving appearance, facilitating swallowing, and controlling drug release in the gastrointestinal tract (e.g., enteric coating). Examples of coating agents suitable for preparing the pharmaceutical composition of the present invention include, but are not limited to, ammonium methacrylate copolymer, sodium carboxymethylcellulose, cellulose acetate phthalate, cellulose acetate, copovidone, ethylcellulose and its aqueous dispersion, gelatin, pharmaceutical varnish, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose acetate succinate, hydroxypropyl methylcellulose phthalate, maltodextrin, methacrylic acid copolymer and its dispersion, methylcellulose, polyethylene glycol, vinyl acetate phthalate, shellac, modified pregelatinized starch, sucrose, titanium dioxide, carnauba wax, and microcrystalline wax.

[0038] Plasticizers are excipients added to other drugs to increase their plasticity and resilience (elasticity). They are important components for imparting desired physical properties to polymer systems. Examples of plasticizers suitable for preparing the pharmaceutical compositions of the present invention include, but are not limited to, acetyl tributyl citrate, acetyl triethyl citrate, ricin oil, diacetylated monoglyceride, dibutyl sebacate, sorbitol, dextrin, diethyl phthalate, glycerin, polyethylene glycol, polyethylene glycol monomethyl ether, propylene glycol, benzyl benzoate, triacetin, tributyl citrate, triethyl citrate, and chlorobutanol.

[0039] For parenteral administration, isotonic solutions, i.e., solutions with an osmotic pressure similar to that of the tissues they contact, are generally used to avoid hemolysis and reduce pain and discomfort during administration. Examples of tonicity agents frequently used to ensure the isotonicity of pharmaceutical compositions of the present invention include, but are not limited to, dextrose, glycerin, mannitol, sodium chloride, and potassium chloride.

[0040] Sweetener is the agent used to mask unpleasant taste and sweeten oral preparations.The example of the sweetener suitable for preparing pharmaceutical compositions of the present invention includes but is not limited to acesulfame potassium, aspartame, acesulfame aspartame salt, glucose, dextrose, fructose, galactose, maltitol, maltose mannitol, saccharin, saccharin calcium, saccharin sodium, sorbitol, sucralose, saccharin, sugar, tagatose.

[0041] The scope of compositions covered by the present invention also includes pharmaceutical colorants that are included in dosage forms to give each pharmaceutical a distinct appearance and allow easy differentiation of specific formulations among formulations with similar physical properties. Examples of pharmaceutical colorants that may be used in the compositions of the present invention include red ferric oxide, yellow ferric oxide, mixed ferric oxide, caramel, titanium dioxide, FD&C colorants, and D&C colorants.

[0042] Depending on the route of administration and the physical and chemical properties inherent to the compounds of the present invention, pharmaceutical compositions prepared with these compounds may contain additional substances capable of stabilizing and preserving the components and preventing and / or avoiding premature degradation. These additional excipients may act as antioxidants, preservatives, and pH adjusters and regulators. Examples of excipients suitable for preparing pharmaceutical compositions of the present invention and used with these properties include, but are not limited to, ascorbic acid, sorbic acid, sodium metabisulfite, α-tocopherol, methylparaben, propylparaben, butylparaben, sodium sulfite, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), phenol, benzyl alcohol, benzalkonium chloride, benzethonium chloride, cetylpyridinium chloride, benzoic acid, sodium benzoate, sodium propionate, boric acid, and pH adjusters. The latter include organic and inorganic acids, bases, and buffers, commonly used in pharmaceutical compositions.

[0043] Pharmaceutical compositions containing the compounds of the present invention may contain and be prepared with (a) complexing agents to mask flavors, improve solubility, promote formulation solubility, and / or modify bioavailability; and (b) fragrances and flavoring agents used to modify and mask unpleasant odors and flavors and to impart pleasant odors and flavors. Several substances and preparations are commercially available for such uses, and their use is limited to approved drugs and officially approved drugs that are compatible with the ingredients in the composition. For therapeutic use and administration, the benzimidazole compounds of the present invention can be formulated into compositions suitable for oral, parenteral, nasal, rectal, transmucosal, and transdermal administration using conventional techniques and appropriate excipients.

[0044] There is no particular limitation on the dosage form containing the benzimidazole compound of the present invention.For example, as a solid for oral administration, suitable dosage forms may be tablets, pills, dragees, capsules, granules, powders, pellets, freeze-dried preparations and similar forms.As a liquid for oral administration, suitable dosage forms may be solutions, dispersions, suspensions, emulsions, oils, syrups and the like.Liquid dosage forms may be used for injection, such as intravenous, intramuscular, subcutaneous, or intradermal administration.

[0045] Other examples of dosage forms include liposomes and nanoparticles, and any other mode of administration known to those skilled in the art. The dosage forms can provide immediate release, controlled release, and delayed release of the benzimidazole compounds of the present invention.

[0046] Another embodiment of the invention is a dosage form as described above, which contains, for example, 0.01-5000 mg of a benzimidazole compound of the invention, and which can be administered one or more times daily during treatment.

[0047] In certain embodiments, the composition may contain at least one other active ingredient, in addition to the benzimidazole compound, used in the treatment of obesity, diabetes, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis, such as metformin, insulin and its derivatives, sulfonylureas, SGLT-2 inhibitors, DPP4 inhibitors, GLP-1 agonists, meglitinides, thiazolidinediones, α-glucosidase inhibitors, FXR agonists, PPAR agonists, ASK1 inhibitors, CCR2 and CCR5 antagonists, caspase inhibitors, insulin sensitizers, and cholic acid-arachidic acid complexes.

[0048] In other embodiments, the present invention relates to the use of a compound of formula I in i) the preparation of a pharmaceutical composition or ii) the treatment of obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.

[0049] In another aspect, the present invention relates to methods of treating obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis with a benzimidazole compound of Formula I, or a pharmaceutically acceptable salt, crystal, hydrate, solvate, prodrug, or metabolite thereof. [ka]

[0050] Non-exhaustive examples illustrating the effects of the benzimidazole derivatives of formula I of the present invention are given below. Example

[0051] Example 1: Effect of benzimidazole derivatives of formula I

[0052] The effects of the benzimidazole derivatives of formula I were evaluated in Sprague Dawley (SD) rats maintained on a high-fat diet (HFD), which represents a diet-induced obesity (DIO) model that exhibits changes similar to those seen in humans with obesity, diabetes, and nonalcoholic fatty liver disease (NAFLD), including insulin resistance and hepatic triglyceride accumulation (van Herck MA et al. (2017). Animal Models of Nonalcoholic Fatty Liver Disease—A Starter's Guide. Nutrients. 9(10). pii: E1072; Wang C and Liao JK (2013). A Mouse Model of Diet-Induced Obesity and Insulin Resistance. Methods Mol Biol. 821: 421-433). The effects of the benzimidazole derivative of formula I (a more potent agonist at MT1 than at MT2) were compared with those of N-[3-(5-chloro-2-ethoxy-6-methoxy-benzimidazol-1-yl)-propyl]-acetamide (structure represented by formula II below; a melatoninergic agonist with higher potency at MT2 than at the MT1 receptor; i.e., the opposite of the benzimidazole derivative of formula I according to the present invention), and dapagliflozin (an SGLT2 inhibitor that normalizes blood glucose levels and reduces body weight in a DIO model) (Millar P et al. (2017). Metabolic and neuroprotective effects of dapagliflozin and liraglutide in diabetic mice. J Endocrinol. 234(3):255-267; Devenny JJ et al. (2012). Weight loss induced by chronic dapagliflozin treatment is attenuated by compensatory hyperphagia in diet-induced obesity). (DIO) rats. Obesity. 20: 1645-1652.). [ka]

[0053] 1 H NMR (400 MHz, CDCl 3) δ 1.48 (t, J = 7.0 Hz, 3H), 1.94 - 2.01( m, 5H), 3.24 (q, J = 4.80 Hz, 2H), 3.92 (s, 3H), 3.99 (t, J = 6.8 Hz, 13 C NMR (125 MHz, CDCl 3) δ ppm 14.74; 23.28; 28.73; 36.91; 39.70; 57.11; 66.48; 93.25; 116.79; 119.10; 132.35; 134.10; 150.67; 157.06; 170.22. MS (ESI) m / z calculated C 15H 20ClN 3O 3: 325.1193; found [M + H]+ 326.1292

[0054] In the HFD model, animals were initially maintained on an HFD for 4 weeks to induce significant body weight (BW) gain before treatment initiation. HFD animals were then treated by oral administration of 10 and 30 mg / kg of the compound of formula I, 10 and 30 mg / kg of the compound of formula II, and 5 mg / kg of dapagliflozin once daily. Two vehicle-treated control groups were also included, receiving normal chow (chow vehicle) and HFD (HFD vehicle).

[0055] BW and food intake (FI) were assessed throughout the study, and the results are shown in Figure 1. Animals on the HFD vehicle steadily increased in body weight from an initial average of 459.9 g on day 1 to 612.4 g on day 57. Interestingly, compared to the HFD, treatment with the compound of formula I (10 and 30 mg / kg po) significantly reduced body weight (BW) (p<0.05) from days 13 to 32 (p<0.05), and treatment with dapagliflozin (5 mg / kg po) significantly reduced BW (p<0.05) from days 2 to 48, 50 to 54, and 56 (Figure 1A). Additionally, a trend toward decreased BW gain from days 1 to 57 compared with HFD vehicle (Figure 1C) was observed in animals treated with 10 and 30 mg / kg po of compound I (p = 0.098) and 5 mg / kg po of dapagliflozin (p = 0.051). In contrast, treatment with compound of formula II (10 and 30 mg / kg) did not significantly affect daily BW gain. Compound I and compound II did not affect daily food intake compared with the HFD vehicle group, except for a single change observed in the latter (Figures 1B and 1D). In contrast, dapagliflozin significantly increased daily FI from days 8 to 56 (p < 0.05) and increased the mean daily FI (p < 0.001) compared with HFD vehicle.

[0056] The degree of insulin resistance and glucose intolerance was assessed using the homeostatic model assessment for insulin resistance (HOMA-IR) method (van Dijk TH et al. (2013). A novel approach to monitor glucose metabolism using stable isotopically labeled glucose in longitudinal studies in mice. Lab Anim. 47(2):79-88.). HOMA-IR was calculated based on plasma insulin and glucose concentrations after a 4-h fast on day 51. HFD-vehicle animals showed significant increases in plasma glucose (+17.6%, p<0.01), insulin (+56.4%, p<0.05), and HOMA-IR (+79.3%, p<0.01) compared with the chow-vehicle group (Figure 2). As expected, dapagliflozin significantly reduced plasma glucose (-21.6%, p<0.001), plasma insulin (-56.7%, p<0.001), and plasma HOMA-IR (-66.7%, p<0.001) compared with HFD vehicle. Notably, the compound of formula I also significantly reduced plasma glucose (-16.4%, p<0.01) at an oral dose of 10 mg / kg. At the maximum dose (30 mg / kg po), compound I of the present invention significantly reduced both plasma glucose (-17%, p<0.01) and HOMA-IR (-36%, p<0.01) compared with HFD vehicle (Figures 2A and 2C). In contrast, the comparative compound (compound II) did not change plasma glucose, insulin, or HOMA-IR compared with HFD vehicle.

[0057] After the study ended (days 65 / 66), the effects on body composition and weights of selected tissues (brown fat pad, epididymal fat pad, liver, and gastrocnemius muscle) were assessed. Tissue weights were analyzed in two ways: i) day 1 BW (body weight) was used as a covariate to correct for differences in BW at the start of the study, or ii) final BW (days 65 / 66) was used as a covariate to correct for changes due to weight loss (Table 3). Dapagliflozin resulted in a significant increase in liver weight compared to HFD vehicle when data were corrected for final BW (+10.8%, p<0.01). In contrast, in analyses corrected for initial BW, a trend toward decreased liver weight was observed in both groups of animals treated with the benzimidazole derivative of formula I (p=0.12). Apart from these changes, no significant effects on the weights of selected tissues were observed compared to the HFD vehicle group, regardless of correction. Body composition was determined using a FoodScan™ near-infrared analyzer to assess moisture, protein, fat content, and final carcass weight (Table 4). No effect of the benzimidazole derivative of formula I according to the present invention or comparative compound II was observed on any of the assessed parameters compared to the HFD vehicle control. Dapagliflozin significantly reduced final carcass weight compared to the HFD vehicle (p<0.05), but body composition was unchanged.

[0058] Since only the benzimidazole derivative of formula I according to the present invention affected BW gain, liver weight, plasma glucose levels, and HOMA-IR in the HFD model, while the comparative benzimidazole compound II did not, the specific effects of compound I on the animal liver were evaluated in comparison with dapagliflozin and the control group. HFD vehicle animals showed significantly increased liver triglyceride content and concentration (+473% and +409%, respectively) compared with the dietary vehicle control (Figures 3A and 3B). Interestingly, compared with HFD vehicle, treatment with compound of formula I (10 and 30 mg / kg po) significantly reduced liver triglyceride content (-41.3% and -55.1%, respectively, p<0.05), and 30 mg / kg po of compound I significantly reduced liver triglyceride concentration by 51.6% (p<0.01). In contrast, dapagliflozin treatment did not significantly reduce liver triglyceride content or concentration compared to HFD vehicle controls. Liver glycogen content and concentration were similar in both diet vehicle and HFD vehicle animals (Figures 3C and 3D). The compound of formula I (10 and 30 mg / kg po) significantly (p<0.05) reduced liver glycogen content (36.6% and 36.6%, respectively) and concentration (42.8% and 40.8%, respectively) compared to HFD vehicle. No effect of dapagliflozin on liver glycogen content or concentration was observed compared to HFD vehicle controls.

[0059] Liver NAFLD was assessed by histopathological analysis (Figure 3E-H). Samples were sectioned, stained, and scored according to the previously described method (Kleiner DE et al. (2005). Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology. 41(6):1313-21; Nishida T et al. (2013). Spontaneous onset of nonalcoholic steatohepatitis and hepatocellular carcinoma in a mouse model of metabolic syndrome. Lab Invest. 93(2):230-41). Parameters analyzed were steatosis (0-3), intralobular inflammation (0-3), and hepatocyte ballooning (0-2). A total "NAFLD Activity Score" (NAS) score was calculated as the sum of all parameters (0-8). Livers of diet-vehicle animals had no pathology scores (zero) for steatosis, intralobular inflammation, hepatocyte ballooning, or total NAS score. In the HFD-vehicle group, pathology scores for steatosis, intralobular inflammation, and total NAS were significantly (p<0.05) increased, whereas hepatocyte ballooning was not significantly affected. Notably, the benzimidazole derivative of Formula I according to the present invention (30 mg / kg po) significantly (p<0.05) reduced steatosis by 57% compared to the HFD group (Figure 3F). Intralobular inflammation was also reduced in animals treated with the benzimidazole derivative of Formula I according to the present invention at a dose of 30 mg / kg, and was no longer significantly different from diet-vehicle controls (Figure 3G). A trend toward significance was also observed for total NAS values ​​between this group and HFD-vehicle animals (Figure 3E, p=0.126). In contrast, no effects on the evaluated parameters were observed in rats treated with dapagliflozin compared with the HFD vehicle group. [Table 1]

[0060] Cellular functional assays were performed in agonist mode. EC50: half maximal effective concentration. IA: intrinsic activity. [Table 2]

[0061] Data are averaged from 2-3 animals at each time point. Compounds were administered orally to Wistar (100 mg / kg) and Sprague Dawley (SD) rats (10 and 30 mg / kg), and samples (plasma and brain) were collected at selected time points. ND: Undetermined due to brain concentrations below the lower limit of quantitation (5 ng / g). [Table 3]

[0062] Data are presented as mean ± SEM (n = 9-10). Mean values ​​were adjusted for differences in BW between treatment groups on Day 1 and Day 65 / 66 (end of study). SEM was calculated from the residuals of the statistical model. The compound of Formula I, the compound of Formula II, dapagliflozin, and vehicle were administered to animals fed a normal diet and an HFD, and parameters were assessed at termination. Data were analyzed by ANCOVA with treatment and cohort as factors and Day 1 BW and final carcass weight as covariates (used to adjust the data for Day 1 and Day 65 / 66 BW, respectively). Treatment groups were compared with HFD vehicle by individual Williams' tests (Compound I and Compound II) and multiple t-tests (dapagliflozin), and statistical significance is indicated by ** (p < 0.01). # (p = 0.12) indicates a significant trend compared to HFD vehicle. [Table 4]

[0063] Data are presented as adjusted mean ± SEM (n = 9-10). SEM was calculated from the residuals of the statistical model. Analysis was performed by robust regression of the data with treatment as a factor and BW on day 1 as a covariate. Treatment groups were compared with HFD vehicle by separate Williams tests (compound I and compound II) and multiple t-tests (dapagliflozin), with statistical significance indicated by * (p < 0.05). Comparisons with diet vehicle were performed by multiple t-tests (HFD vehicle and dapagliflozin groups) and Dunnett's test (compound I and compound II), with statistical significance indicated by † (p < 0.05) and †† (p < 0.01).

Claims

1. A pharmaceutical composition for treating obesity, diabetes, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis, comprising: A pharmaceutical composition comprising a benzimidazole compound of Formula I or a pharmaceutically acceptable salt, crystal, hydrate, or solvate thereof, and a pharmaceutically acceptable excipient. 【Chemistry 1】

2. 10. The pharmaceutical composition of claim 1, comprising 0.01-5000 mg of the benzimidazole compound of formula I.

3. Pharmaceutically acceptable salts of the benzimidazole compounds of Formula I include inorganic addition salts including hydrochloride, hydrobromide, sulfate, nitrate, phosphate; acetate, propionate, hexanoate, heptanoate, glycolate, pyruvate, lactate, malonate, malate, maleate, fumarate, tartrate, citrate, succinate, mesylate, acetonitrile, besylate, tosylate, xinafoate, benzoate, p-toluenesulfonate, cinnamate, p-chlorobenzenesulfonate, 2-naphthalenesulfonate, p-toluenesulfonate, camphorsulfonate, trimethyl acetate, t-butyl acetate, laurate ...

2. The pharmaceutical composition of claim 1, wherein the salt is selected from the group consisting of organic addition salts including methyl, methyl gluconate, glutarate, hydroxynaphthoate, salicylate, stearate, muconate, mandelate, and 2-hydroxyethanesulfonic acid; alkali addition salts of metals including sodium, potassium, lithium, calcium, magnesium, and bismuth; bromides; pharmaceutically acceptable salts of primary, secondary, or tertiary amines with organic bases; salts of amino acids including arginine, lysine, and histidine; and salts of caffeine, procaine, hydrobromide, choline, betaine, ethylenediamine, glucosamine, theobromine, purine, and morpholine.

4. Use of a benzimidazole compound of formula I, a pharmaceutically acceptable salt, crystal, hydrate, or solvate thereof in the manufacture of a pharmaceutical composition for treating obesity, diabetes, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis. 【Chemistry 2】

Images