Application of mGlu5 inhibitor in treating or preventing non-alcoholic fatty liver disease and / or obesity and improving leptin sensitivity

Abstract

The invention relates to application of an mGlu5 inhibitor in improvement of non-alcoholic fatty liver disease and / or obesity and improvement of leptin sensitivity. Specifically, the invention provides application of an mGlu5 inhibitor or pharmaceutically acceptable salt thereof in preparation of a medicine for treating or preventing the following diseases in a subject: non-alcoholic fatty liver disease, obesity and / or hyperleptinemia; and the use of an mGlu5 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for improving leptin sensitivity in a subject.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of mGlu5 inhibitors in treating or preventing non-alcoholic fatty liver disease and / or obesity, and improving leptin sensitivity. Background Art

[0002] With its increasing incidence, non-alcoholic fatty liver disease (NAFLD) has become a global health concern, threatening the lives and health of nearly 30% of adults worldwide. If left untreated, NAFLD can irreversibly progress to non-alcoholic fatty liver disease (NAFLD), and eventually develop into cirrhosis and liver cancer.

[0003] The incidence of obesity has been increasing in recent years. Currently, there are nearly 2 billion overweight people in the world, of which 650 million are obese. [1] According to the World Health Organization, about 3.4 million people die each year from complications of obesity. [2] .

[0004] mGlu5 is a metabotropic glutamate receptor that is primarily distributed in the central nervous system and plays an important role in central nervous system disorders such as anxiety, depression, autism, and pain.

[0005] Basimglurant (2-Chloro-4-[1-(4-fluoro-phenyl)-2,5-dimethyl-1H-imidazol-4-ylethynyl]-pyridine) is a highly effective and safe mGlu5 negative allosteric modulator that has been reported to be used to treat neurological diseases such as fragile X syndrome, major depressive disorder, and trigeminal neuralgia. [4] .

[0006]

[0007] CTEP is a long-acting, orally bioavailable allosteric antagonist of the mGlu5 receptor that has been reported to be used to treat metastatic melanoma, etc.

[0008]

[0009] AZD 2066 is a selective, orally active and blood-brain permeable mGluR5 antagonist that has been reported to be used to treat neuropathic pain, major depression and gastroesophageal reflux disease.

[0010] Summary of the Invention

[0011] The present invention provides use of an mGlu5 inhibitor or a pharmaceutically acceptable salt thereof in improving non-alcoholic fatty liver disease and / or obesity and improving leptin sensitivity.

[0012] In a first aspect of the present invention, the present invention provides use of an mGlu5 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing the following conditions in a subject: non-alcoholic fatty liver disease, obesity and / or hyperleptinemia.

[0013] In some embodiments, the subject has an intact leptin signaling pathway.

[0014] In some embodiments, the mGlu5 inhibitor may include one or more of the group consisting of: Basimglurant, CTEP, AZD 2066:

[0015]

[0016] In some embodiments, the mGlu5 inhibitor can be Basimglurant.

[0017] In some embodiments, the subject is selected from a rodent, a primate.

[0018] In a second aspect of the present invention, the present invention provides use of an mGlu5 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for improving leptin sensitivity in a subject.

[0019] In some embodiments, the subject has an intact leptin signaling pathway.

[0020] In some embodiments, the mGlu5 inhibitor may include one or more of the group consisting of: Basimglurant, CTEP, AZD 2066.

[0021] In some embodiments, the mGlu5 inhibitor can be Basimglurant.

[0022] In some embodiments, the subject is selected from a rodent, a primate.

[0023] In a third aspect, the present invention provides a use of a pharmaceutical composition in the preparation of a medicament for treating or preventing the following conditions in a subject: non-alcoholic fatty liver disease, obesity and / or hyperleptinemia; wherein the pharmaceutical composition comprises one or more mGlu5 inhibitors selected from the group consisting of: basimglurant, CTEP, AZD 2066 and a pharmaceutically acceptable carrier or excipient; preferably, the mGlu5 inhibitor is basimglurant.

[0024] In a fourth aspect, the present invention provides a use of a pharmaceutical composition in the preparation of a medicament for improving leptin sensitivity in a subject; wherein the pharmaceutical composition comprises one or more mGlu5 inhibitors selected from the group consisting of: basimglurant, CTEP, AZD 2066 and a pharmaceutically acceptable carrier or excipient; preferably, the mGlu5 inhibitor is basimglurant. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The illustrative examples of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0026] Figure 1 A schematic diagram of the experimental technical route is shown.

[0027] Figure 2 The effect of basimglurant on body weight and food intake in high-fat diet-induced obese mice was shown. A vehicle or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered to high-fat diet-induced obese mice for 2 weeks (n=5 per group).

[0028] A. Schematic diagram of basimglurant administration to high-fat diet-induced obese mice.

[0029] B. Body weight curves (g) of mice during administration of basimglurant or vehicle.

[0030] C. Cumulative total food intake (g) of mice during basimglurant or vehicle administration.

[0031] Gray circles represent the solvent-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM, and unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01.

[0032] Figure 3Basimglurant reduces lipid accumulation in adipose tissue of high-fat diet-induced obese mice. High-fat diet-induced obese mice were intraperitoneally administered with a vehicle or basimglurant (3 mg / kg / 2 days) for 2 weeks (vehicle group n=5, basimglurant group n=4).

[0033] A. Representative H&E staining results of white adipose tissue (WAT) and brown adipose tissue (BAT) in high-fat diet-induced obese mice after administration of basimglurant or vehicle.

[0034] B. Statistical results of the average area of WAT lipid droplets (μm 2 ).

[0035] C. Statistical results of the average area of BAT lipid droplets (μm 2 ).

[0036] D. Subcutaneous white adipose tissue (sWAT) mass (g).

[0037] E. Gonadal white adipose tissue (gWAT) mass (g).

[0038] F. Subcutaneous white adipose tissue mass / body weight ratio (sWAT / Weight, %).

[0039] G. Gonadal white adipose tissue mass / body weight ratio (gWAT / Weight, %).

[0040] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are presented as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 25 μm.

[0041] Figure 4 Basimglurant was shown to improve ectopic fat accumulation in the liver of high-fat diet-induced obese mice. High-fat diet-induced obese mice were intraperitoneally administered with a vehicle or basimglurant (3 mg / kg / 2 days) for 2 weeks.

[0042] A. Representative H&E and Oil Red O staining results of liver tissues from high-fat diet-induced obese mice after administration of basimglurant or vehicle.

[0043] B. Statistical results of the average area of lipid droplets in H&E-stained liver sections (μm 2 )(n=5 per group).

[0044] C. Statistical results of the average area of lipid droplets in liver Oil Red O-stained sections (μm 2 )(n=5 per group).

[0045] D. Mouse liver tissue weight (g) (n=5 per group).

[0046] E. Liver tissue mass / body weight ratio (%) (n=5 per group) F. Triglyceride content in mouse liver (mg / g) (solvent group n=4, basimglurant group n=3).

[0047] G. Total cholesterol content (mg / g) in mouse liver (solvent group n=4, basimglurant group n=3).

[0048] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 50 μm.

[0049] Figure 5 Basimglurant showed no significant effect on the body weight and food intake of healthy mice fed a normal diet. Vehicle or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered to healthy wild-type C57BL / 6J mice fed a normal diet for 2 weeks (n=5 per group).

[0050] A. Schematic diagram of basimglurant administration to healthy mice fed a normal chow diet.

[0051] B. Body weight curves (g) of mice during administration of basimglurant or vehicle.

[0052] C. Cumulative total food intake (g) of mice during basimglurant or vehicle administration.

[0053] Gray circles represent the solvent-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM, and unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01.

[0054] Figure 6 Basimglurant did not significantly affect lipid accumulation in adipose tissue of chow-fed healthy mice. A vehicle or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered to chow-fed healthy mice for 2 weeks (n=5 per group).

[0055] A. Representative H&E staining results of WAT and BAT in chow-fed healthy mice after administration of basimglurant or vehicle.

[0056] B. Statistical results of the average area of WAT lipid droplets (μm 2 ).

[0057] C. Statistical results of the average area of BAT lipid droplets (μm 2 ).

[0058] D. sWAT mass (g). E. gWAT mass (g).

[0059] F. sWAT mass / body weight ratio (sWAT / Weight, %).

[0060] G. gWAT mass / body weight ratio (gWAT / Weight, %).

[0061] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 50 μm.

[0062] Figure 7 Basimglurant showed no significant effect on ectopic fat accumulation in the liver of healthy mice fed a normal diet. The vehicle or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered to healthy mice fed a normal diet for 2 weeks (n=5 per group).

[0063] A. Representative H&E staining results of liver tissue from chow-fed healthy mice after administration of basimglurant or vehicle.

[0064] B. Mouse liver tissue mass (g).

[0065] C. Liver tissue mass / body weight ratio (%).

[0066] D. Triglyceride content in mouse liver (mg / g).

[0067] E. Total cholesterol content in mouse liver (mg / g).

[0068] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 50 μm.

[0069] Figure 8Basimglurant has been shown to have a modest anti-obesity effect in wild-type mice fed a high-fat diet for two weeks. Healthy mice were switched from a standard diet to a high-fat diet for two weeks to simulate the early stages of obesity. During this period, a vehicle or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered. Another group continued to be fed a standard diet and served as a negative control (n=5 per group).

[0070] A. Schematic diagram of diet induction and basimglurant administration in wild-type mice. B. Body weight curves (g) of mice during basimglurant or vehicle administration.

[0071] C. Body weight changes (%) of mice during administration of basimglurant or vehicle.

[0072] D. Cumulative total food intake (g) of mice during basimglurant or vehicle administration.

[0073] In Figure C, dark gray represents the normal diet group, light gray represents the high-fat diet-solvent administration group, and blue represents the high-fat diet-Basimglurant administration group.

[0074] In Panels B and D, gray circles represent the vehicle-treated group, and blue squares represent the basimglurant-treated group. Data are presented as mean ± SEM, and analyzed by one-way ANOVA with Tukey's multiple comparison correction. *P < 0.05, **P < 0.01.

[0075] Figure 9 Basimglurant has been shown to have a modest effect in treating fatty liver disease and reducing fat accumulation in wild-type mice fed a high-fat diet for two weeks. Healthy mice were switched from a standard diet to a high-fat diet for two weeks to simulate the early stages of obesity. During this period, a vehicle or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered. Another group continued to be fed a standard diet and served as a negative control (n=5 per group).

[0076] A. Representative H&E staining results of liver tissue, WAT, and BAT in mice after basimglurant and diet induction administration.

[0077] B. Liver tissue mass / body weight ratio (%).

[0078] C. sWAT mass / body weight ratio (sWAT / Weight, %).

[0079] D. gWAT mass / body weight ratio (gWAT / Weight, %).

[0080] Dark gray circles represent the standard diet group, light gray circles represent the high-fat diet-solvent group, and blue squares represent the high-fat diet-basimglurant group. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA with Tukey's multiple comparison correction. *P < 0.05, **P < 0.01.

[0081] Figure 10 It was shown that basimglurant had no significant effect on body weight and food intake in ob / ob mice and db / db mice.

[0082] A. Changes in body weight (%) of ob / ob mice during administration of basimglurant or vehicle Ob / ob mice were intraperitoneally administered with vehicle or basimglurant (3 mg / kg / day) for 3 weeks (n=5 per group).

[0083] B. Cumulative total food intake (g) of ob / ob mice during administration of basimglurant or vehicle Ob / ob mice were intraperitoneally administered with vehicle or basimglurant (3 mg / kg / day) for 3 weeks (n=5 per group).

[0084] C. Changes in body weight (%) of db / db mice during administration of basimglurant or solvent. Solvent or basimglurant (3 mg / kg / day) was intraperitoneally administered to db / db mice for 3 weeks (n=5 per group).

[0085] D. Cumulative total food intake (g) of db / db mice during administration of basimglurant or solvent. Solvent or basimglurant (3 mg / kg / day) was intraperitoneally administered to db / db mice for 3 weeks (n=5 per group).

[0086] Gray circles represent the solvent-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM, and unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01.

[0087] Figure 11 It was shown that Basimglurant did not significantly affect lipid accumulation in adipose tissue of ob / ob mice. Ob / ob mice were intraperitoneally administered with a vehicle or Basimglurant (3 mg / kg / day) for 3 weeks (n=5 per group).

[0088] A. Representative H&E staining results of WAT and BAT in ob / ob mice after administration of basimglurant or vehicle.

[0089] B. Statistical results of the average area of WAT lipid droplets in ob / ob mice (μm 2 ).

[0090] C. Statistical results of the average area of BAT lipid droplets in ob / ob mice (μm 2 ).

[0091] D. sWAT mass / body weight ratio (sWAT / Weight, %) of ob / ob mice.

[0092] E. gWAT mass / body weight ratio (gWAT / Weight, %) of ob / ob mice.

[0093] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are presented as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 25 μm.

[0094] Figure 12 Basimglurant did not significantly affect lipid accumulation in adipose tissue of db / db mice. The vehicle or basimglurant (3 mg / kg / day) was intraperitoneally administered to db / db mice for 3 weeks.

[0095] A. Representative H&E staining results of WAT and BAT in db / db mice after administration of basimglurant or vehicle (n=5 per group).

[0096] B. Statistical results of the average area of WAT lipid droplets in db / db mice (μm 2 )(n=5 per group).

[0097] C. Statistical results of the average area of BAT lipid droplets in db / db mice (μm 2 )(n=5 per group).

[0098] D. sWAT mass / body weight ratio (sWAT / Weight, %) of db / db mice (vehicle group n=5, basimglurant group n=4).

[0099] E. gWAT mass / body weight ratio (gWAT / Weight, %) of db / db mice (vehicle group n=5, basimglurant group n=4).

[0100] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are presented as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 25 μm.

[0101] Figure 13 Basimglurant showed no significant effect on ectopic fat accumulation in the liver of ob / ob mice. Ob / ob mice were intraperitoneally administered with a vehicle or basimglurant (3 mg / kg / day) for 3 weeks (n=5 per group).

[0102] A. Representative H&E staining results of liver tissues from ob / ob mice after administration of basimglurant or vehicle.

[0103] B. Statistical results of the average area of lipid droplets in H&E-stained sections of ob / ob mouse liver (μm 2 ).

[0104] C. Liver tissue mass / body weight ratio (%) of ob / ob mice.

[0105] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 50 μm.

[0106] Figure 14 Basimglurant showed no significant effect on ectopic fat accumulation in the liver of db / db mice. The vehicle or basimglurant (3 mg / kg / day) was intraperitoneally administered to db / db mice for 3 weeks.

[0107] A. Representative H&E staining results of liver tissues of db / db mice after administration of basimglurant or vehicle (n=5 per group).

[0108] B. Statistical results of the average area of lipid droplets in H&E-stained sections of db / db mouse liver (μm 2 )(n=5 per group).

[0109] C. Liver tissue mass / body weight ratio (%) of db / db mice (solvent group n=5, basimglurant group n=4).

[0110] Gray circles represent the vehicle-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM. Unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. Scale bar, 50 μm.

[0111] Figure 15 Basimglurant administration improves hyperleptinemia. Leptin levels in mouse serum after different treatments.

[0112] A. A solvent or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered to normal chow-fed healthy mice for 2 weeks (solvent group n=5, basimglurant group n=4).

[0113] B. A solvent or basimglurant (3 mg / kg / 2 days) was chronically intraperitoneally administered to high-fat diet-induced obese mice for 2 weeks (solvent group n=5, basimglurant group n=3).

[0114] C. A solvent or basimglurant (3 mg / kg / day) was intraperitoneally administered to high-fat diet-induced obese mice for 4 days (n=6 per group).

[0115] D. The diet of healthy mice was switched from normal diet to high-fat diet for 2 weeks to simulate the early stage of obesity. During this period, solvent or basimglurant (3 mg / kg / 2 days) was intraperitoneally administered. Another group continued to be fed with normal diet and served as a negative control (normal diet group n=5, high-fat diet-solvent group n=5, high-fat diet-basimglurant group n=3).

[0116] Gray circles represent the solvent-administered group, and blue squares represent the basimglurant-administered group. Data are expressed as mean ± SEM, and unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01.

[0117] Figure 16 Basimglurant was shown to improve leptin sensitivity in mice fed a high-fat diet.

[0118] A. Schematic diagram of the leptin sensitivity test. Obese mice induced by a high-fat diet were intraperitoneally administered with either vehicle or basimglurant (3 mg / kg / day) for 4 days. Sixteen hours after the last administration, saline or leptin (1 mg / kg) was intraperitoneally administered, and the mice were dissected 40 minutes later (n=3 per group).

[0119] B. Immunoblotting was used to detect the p-STAT3 protein level in the mouse hypothalamus.

[0120] Gray represents the solvent-administered group, and blue represents the basimglurant-administered group. Data are expressed as mean ± SEM, and unpaired two-tailed t-test was used. *P < 0.05, **P < 0.01. DETAILED DESCRIPTION

[0121] The section headings are for organizational purposes only and are not to be construed as limiting the subject matter described to specific aspects or embodiments.

[0122] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present invention belongs. Methods and materials similar or equivalent to the methods and materials described herein can also be used in the practice of the present invention, and some suitable methods and materials will be introduced below. The entire contents of all publications, patent applications, patents and other references mentioned in the text are incorporated herein by reference. In the event of a conflict, this specification (including definitions) shall prevail. In addition, the materials, methods and examples described herein are illustrative only and are not intended to be limiting.

[0123] All publications and patents mentioned herein are incorporated by reference in their entirety, just as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of a conflict, the present specification (including any definitions therein) will control. However, the reference to any reference, article, publication, patent, patent disclosure, or patent application cited herein is not, and should not be construed as, an admission or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country / region in the world.

[0124] In this specification, any concentration range, percentage range, ratio range or integer range should be understood to include any integer value within the range, and where appropriate, include its fraction (e.g., one tenth and one hundredth of an integer), unless otherwise stated. The term "approximately", when immediately before a number or numerical value, refers to the plus or minus 10% of that number or numerical range. The term "about", when immediately after a number or numerical value, refers to the plus or minus 10% of that number or numerical range. It should be understood that the terms "one" and "a" when used herein, unless otherwise stated, refer to one or more of the listed components. The use of options (e.g., "or") should be understood to refer to any one, both, or any combination of the options. The term "and / or" should be understood to refer to any one or both of the options. As used herein, the terms "include" and "comprising" are synonymous.

[0125] In the present invention, the term "pharmaceutically acceptable salt" refers to an acid addition salt prepared by reacting a compound of the present invention with a pharmaceutically acceptable acid, or a salt formed by reacting a compound having an acidic group with a basic compound. The acid is preferably selected from an inorganic acid (such as hydrochloric acid, sulfuric acid, phosphoric acid or hydrobromic acid), and an organic acid (such as oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, lysine, histidine, citric acid or benzoic acid); the basic compound is preferably selected from sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate or potassium bicarbonate. The above-mentioned pharmaceutically acceptable salts are easy to separate and can be purified by conventional separation methods, such as solvent extraction, dilution, recrystallization, column chromatography and preparative thin layer chromatography.

[0126] As used herein, the term "treat" or "treatment" refers to ameliorating one or more symptoms of a disease or condition. The term "prevent" refers to delaying, retarding, or interrupting one or more symptoms of a disease or condition.

[0127] The terms "nonalcoholic fatty liver disease" and "NAFLD" are used interchangeably to refer to a heterogeneous group of diseases characterized by diffuse fatty liver on imaging or marked hepatocellular steatosis with predominantly macrovesicular or bullous changes on pathology, excluding alcohol and other clearly liver-damaging factors.

[0128] In this application, the terms "hepatic steatosis" and "ectopic hepatic fat accumulation" are used interchangeably and both refer to the accumulation of fat in hepatocytes, a primary symptom of non-alcoholic fatty liver disease. Histologically, "hepatic steatosis" or "ectopic hepatic fat accumulation" manifests as an increase in the number and size of vacuoles in H&E staining and an increase in the number and size of red lipid droplets in Oil Red O staining. Biochemically, it manifests as an increase in triglyceride (TAG) content in the liver.

[0129] In the present invention, the term "obesity" refers to a subject's weight exceeding the standard weight. For mouse subjects, an obese subject refers to a mouse weighing more than 45 g. For human subjects, an obese subject refers to a subject with a body mass index (BMI) greater than or equal to 24 kg / m 2 Subjects with a BMI greater than or equal to 28 kg / m 2 of the subjects.

[0130] In the present invention, the term "weight loss" is different from "diet weight". "Weight loss" refers to a reduction in the weight of a subject. "Weight loss" means that the subject's adipose tissue mass and lipid content are reduced as confirmed by histological examination. Generally, in obese subjects, subjects after "weight loss" are often accompanied by weight loss (i.e., "weight loss"). However, the result or conclusion of the subject's weight loss cannot be obtained only through the reduction of the subject's weight (i.e., "weight loss") without histological examination, and the changes in the subject's adipose tissue cannot be concluded based solely on the changes in the subject's weight.

[0131] In this application, "wild-type" mice refer to the C57BL / 6J strain. Except for ob / ob and db / db mice, or unless otherwise indicated, all mice used were wild-type C57BL / 6J. "High-fat diet-induced obese mice," "healthy mice," "healthy mice fed a normal diet," and "wild-type mice fed a high-fat diet for two weeks" are all of the same wild-type C57BL / 6J strain. These four mice differ in their feeding regimens.

[0132] In the present invention, "high-fat diet", "high-fat feed" and "HFD" are used interchangeably to refer to feed with a high lipid content.

[0133] For the purposes of this invention, "high-fat diet-induced obese mice" refer to C57BL / 6J mice that have been fed a high-fat diet (60% energy from fat and 35% by weight) for approximately 20 weeks and weigh at least 45g. "High-fat diet-induced obese mice" may sometimes be referred to simply as "HFD mice."

[0134] For purposes of this invention, "healthy mice" refer to non-obese C57BL / 6J mice weighing between 20 and 35 grams. For purposes of this invention, "healthy mice fed a standard diet" refer to C57BL / 6J mice that have been fed a standard diet and not a high-fat diet, and weigh between 20 and 35 grams. For purposes of this invention, "wild-type mice fed a high-fat diet for two weeks" refer to healthy C57BL / 6J mice that have been fed a high-fat diet instead of a standard diet for two weeks to simulate the early stages of obesity.

[0135] Leptin is a polypeptide hormone encoded by the ob gene and is primarily expressed in fat cells. After eating, leptin levels in serum rise and act on the hypothalamus in the central nervous system, inhibiting feeding behavior and promoting energy metabolism.

[0136] In the present invention, the terms "leptin" and "lep" refer to endogenous leptin secreted by a subject or exogenously applied leptin protein.

[0137] The term "ob gene" refers to the obese gene, which encodes the leptin protein. This gene is located on chromosome 6 in mice. A homozygous "ob / ob" mutant of the ob gene results in a mouse inability to produce leptin, leading to severe obesity and excessive food intake. As used herein, "ob / ob" refers to a mouse strain carrying a homozygous mutation in the ob gene.

[0138] The term "db gene" refers to the diabetes gene, which encodes the leptin receptor (lepR). This gene is located on chromosome 4 in mice. Homozygous mutations of the db gene, "db / db," result in subjects being unable to receive leptin signals and exhibiting an obesity and excessive food intake phenotype. In this invention, "db / db" refers to a mouse strain carrying a homozygous mutation in the db gene.

[0139] As used herein, the term "intact leptin signaling pathway" means that the subject has no defects in any of the components of the leptin signaling pathway and is able to produce leptin protein and leptin receptors. "Intact leptin signaling pathway" is unrelated to the amount of leptin produced by the subject, serum leptin levels, and sensitivity to leptin. In mice, "intact leptin signaling pathway" means that there are no mutations in genes related to the leptin signaling pathway, such as the ob or db genes.

[0140] Similar to the role of insulin in diabetes, obesity can be divided into obesity caused by leptin signaling deficiency and obesity due to decreased leptin sensitivity, depending on whether the leptin signaling pathway is intact. Obesity caused by decreased leptin sensitivity is more common in obese people and has more complex causes.

[0141] In the present invention, the term "hyperleptinemia" refers to a state in which serum leptin levels remain elevated due to long-term fat accumulation in subjects with intact leptin signaling pathways. Improving hyperleptinemia refers to reducing serum leptin levels.

[0142] In the present invention, the term "leptin sensitivity" refers to the degree of response of a subject to endogenous or exogenous leptin. Long-term maintenance of high serum leptin levels will lead to decreased leptin sensitivity, which in turn makes it more likely to develop an obese phenotype.

[0143] In the present invention, the term "endogenous leptin" refers to leptin secreted by the subject itself. The term "exogenous leptin" refers to high-dose leptin administered during leptin sensitivity testing.

[0144] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier or excipient. The carrier includes, but is not limited to, an ion exchanger, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human albumin, buffer substances such as phosphates, glycerol, sorbic acid, potassium sorbate, a partial glyceride mixture of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, and lanolin.

[0145] The excipients include, but are not limited to, fillers and diluents, such as lactose, starch, and microcrystalline cellulose; binders, such as hydroxypropyl methylcellulose (HPMC), polyvinyl pyrrolidone (PVP), and starch slurry; disintegrants, such as sodium carboxymethyl starch (CMS-Na), low-substituted hydroxypropyl cellulose (L-HPC), and cross-linked polyvinyl pyrrolidone (PVPP); lubricants, such as magnesium stearate (MS), talc, and micronized silica; surfactants, such as sodium lauryl sulfate (SDS) and Tween 80; thickeners / gelling agents, such as carbomer and xanthan gum; preservatives, such as sodium benzoate, potassium sorbate, and parabens (parabens); and flavoring / coloring agents, such as aspartame (sweetener), menthol (flavoring), titanium dioxide (opacifier), and tartrazine (coloring).

[0146] The pharmaceutical composition of the present invention can be prepared into various forms according to different administration routes.

[0147] According to the present invention, the pharmaceutical composition can be administered in any of the following ways: orally, by spray inhalation, rectally, nasally, buccally, vaginally, topically, parenterally, such as by subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, and intracranial injection or infusion, or via an externally implanted reservoir. Among these, oral, intraperitoneal, or intravenous administration is preferred.

[0148] The following describes embodiments of the present invention in detail. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention. All other embodiments obtained by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort shall fall within the scope of protection of the present invention.

[0149] Example 1. Establishment of high-fat diet-induced obese mice

[0150] In animal models, high-fat diet-induced obesity can mimic the obesity caused by decreased leptin sensitivity, which is prevalent in humans. High-fat diet-induced obese animals are commonly used models for metabolic diseases or conditions such as obesity, decreased leptin sensitivity, and non-alcoholic fatty liver disease.

[0151] Select 6-8 weeks old (about 20g) male mice of C57BL / 6J strain to construct obese mouse model. Feed male mice with high fat diet (fat energy supply accounts for 60%, fat mass accounts for 35%) for about 20 weeks, and the obesity model can be successfully established. The established model is completed with the corresponding experimental index detection according to the following technical route, such as Figure 1 shown.

[0152] Obese mice with established obesity models were assigned to either an experimental group or a control group, which received a solvent. Basimglurant was administered during the administration period, and the administration schedule was fixed. Body weights of the mice in both groups were recorded before each administration.

[0153] Basimglurant was dissolved in DMSO to prepare a 25 g / L stock solution. Based on mouse body weight, an appropriate volume of the stock solution was added to a 0.5% sodium carboxymethylcellulose (CMC) solution at a dose of 3 mg / kg (mouse body weight) to form a uniform suspension for intraperitoneal injection. Ultrasonic dissolution was used for dissolution. A control group received the same volume of DMSO as the stock solution in the experimental group, added to the 0.5% CMC solution. Basimglurant was administered intraperitoneally at a dose of 3 mg / kg every two days, while control mice received blank solvent every two days. Administration was performed on days 0, 2, 4, 6, 8, 10, and 12. On day 14, the obese mice were weighed for the final time, and tissue samples were collected, primarily serum, white adipose tissue, brown adipose tissue, and liver tissue. The tissue samples were then sectioned and stained accordingly to characterize changes in fat content in white adipose tissue (WAT), brown adipose tissue (BAT), and liver tissue after two weeks of basimglurant administration.

[0154] The histological examinations used in the present invention include H&E (hematoxylin and eosin) staining and Oil Red O staining, both of which were performed using methods known in the art. Oil Red O staining used a modified Oil Red O staining kit (purchased from Beyotime, C0158S) and hematoxylin staining solution (purchased from Beyotime, C0107).

[0155] Example 2. Administration of Basimglurant has a weight-reducing effect on high-fat diet-induced obese mice

[0156] Obese mice induced by high-fat diet were intraperitoneally administered with solvent or basimglurant (3 mg / kg / 2 days) for 2 weeks. Figure 2 A). Basimglurant administration significantly reduced the body weight of obese mice induced by a high-fat diet. After two weeks of administration, the body weight of the Basimglurant administration group decreased by 9.62 g ( Figure 2 B), the body weight decreased by about 20% compared with the initial body weight. The body weight of the control mice administered with the solvent did not change significantly ( Figure 2 B).

[0157] At the same time, the food intake of high-fat diet-induced obese mice administered with basimglurant was significantly reduced compared with the vehicle-administered group ( Figure 2 C).

[0158] Example 3. Administration of Basimglurant has an anti-obese effect on high-fat diet-induced obese mice

[0159] Histological examination revealed that the lipid droplets in the white adipocytes of mice in the Basimglurant-treated group were smaller than those in the control group ( Figure 3 A, B). In mice that received basimglurant, brown adipose tissue also had smaller lipid droplets ( Figure 3 A, C)

[0160] At the whole tissue level, long-term basimglurant administration (2 weeks) significantly reduced the mass of subcutaneous white adipose tissue (sWAT) and the subcutaneous white adipose tissue-to-body weight ratio (sWAT / Weight) in high-fat diet-induced obese mice ( Figure 3 D, F). Long-term basimglurant administration also slightly reduced the mass of gonadal white adipose tissue (gWAT) in high-fat diet-induced obese mice, but the statistical results were not significant ( Figure 3 E). Basimglurant administration had little effect on the ratio of gonadal white adipose tissue to body weight (gWAT / Weight), which may be due to the significant effect of basimglurant on reducing body weight, resulting in a minor decrease in this ratio ( Figure 3 G).

[0161] In short, basimglurant reduced fat storage in high-fat diet-induced obese mice, that is, basimglurant administration achieved a weight loss effect.

[0162] Example 4. Administration of Basimglurant Improves Hepatic Steatosis in High-Fat Diet-Induced Obese Mice

[0163] Obese mice induced by a high-fat diet are also a commonly used model for non-alcoholic fatty liver disease. Long-term high-fat diet induces severe hepatic steatosis in liver tissue ( Figure 4 A) Two weeks of basimglurant administration improved hepatic steatosis induced by a high-fat diet. H&E staining results showed that the lipid droplets in the hepatocytes of mice in the basimglurant-treated group were smaller than those in the control group ( Figure 4 A and B). Oil Red O is a fat-soluble dye that dissolves in triglycerides in tissues, making them appear red. Oil Red O staining results showed that Basimglurant administration can alleviate the ectopic accumulation of fat in liver cells induced by a high-fat diet and significantly reduce the size of lipid droplets in liver cells ( Figure 4 A and C). There were no significant changes in liver mass and liver-to-body weight ratio ( Figure 4D and E). The slight increase in liver-to-body weight ratio is due to body weight loss.

[0164] In addition, biochemical results showed that basimglurant administration significantly reduced the triglyceride content in the liver ( Figure 4 F), and had no significant effect on the total cholesterol content in the liver ( Figure 4 G). In brief, basimglurant ameliorates ectopic fat accumulation in the liver of mice with high-fat diet-induced obesity.

[0165] In conclusion, 2 weeks of basimglurant administration had an anti-obesity effect in high-fat diet-induced obese mice: it reduced body weight, food intake, and lipid accumulation in white and brown fat; in addition, 2 weeks of basimglurant administration had an ameliorative effect on non-alcoholic fatty liver disease: it reduced hepatic steatosis.

[0166] Example 5. Administration of basimglurant had no significant effect on body weight or food intake in healthy mice fed a normal diet

[0167] After confirming that Basimglurant can reduce the body weight of mice with high-fat diet-induced obesity, the inventors also tested the effect of Basimglurant on healthy C57BL / 6J mice fed a normal diet. The experimental results showed that two weeks of Basimglurant administration had no significant effect on the body weight and food intake of healthy mice ( Figure 5 ).

[0168] Example 6. Administration of Basimglurant has no significant effect on lipid accumulation in healthy mice fed a normal diet

[0169] Compared to the obese mice induced by a high-fat diet mentioned above, the lipid droplets of healthy mice fed a normal diet were smaller in both white adipose tissue and brown adipose tissue. Two weeks of basimglurant administration did not significantly affect the lipid droplet size of adipocytes in white adipose tissue and brown adipose tissue ( Figure 6 AC). In addition, basimglurant administration did not significantly affect the mass of sWAT and gWAT and the white fat-to-body weight ratio in healthy mice fed a normal diet ( Figure 6 DG).

[0170] In conclusion, basimglurant did not significantly affect lipid accumulation in adipose tissue of healthy mice fed a normal diet.

[0171] Example 7. Administration of Basimglurant has no significant effect on hepatic steatosis in healthy mice fed a normal diet

[0172] Healthy C57BL / 6J mice fed a normal diet had healthy liver tissue, and no ectopic fat accumulation in the liver was detected by H&E staining. Basimglurant administration did not change the morphology of liver cells ( Figure 7 A), and had no significant effect on liver mass, liver weight ratio, liver triglyceride and total cholesterol content ( Figure 7 BE). In brief, basimglurant had no significant effect on ectopic fat accumulation in the liver of healthy mice fed a normal chow diet.

[0173] In summary, 2 weeks of basimglurant administration had no significant effect on healthy mice fed a normal diet: no significant changes were observed in body weight, food intake, adipose tissue, or liver tissue. Basimglurant only had a weight loss effect on wild-type obese mice induced by a high-fat diet, but had no significant weight loss effect or improvement in non-alcoholic fatty liver disease in healthy mice fed a normal diet.

[0174] Example 8. Basimglurant prevents high-fat diet-induced weight gain in healthy mice

[0175] We previously found that Basimglurant has the effect of treating obesity in high-fat diet-induced obese mice. In order to further explore whether Basimglurant has the effect of preventing obesity, Figure 8 A, In this study, wild-type healthy mice with intact leptin pathways were fed a high-fat diet for 2 weeks to simulate the initial stage of obesity, during which time they were administered with either a vehicle or basimglurant.

[0176] A 2-week high-fat diet significantly induced weight gain in mice, whereas basimglurant administration during this period prevented the high-fat diet-induced weight gain ( Figure 8 B and C). Basimglurant administration slightly reduced the intake of high-fat feed in healthy mice compared to the high-fat diet-vehicle administration group ( Figure 8 D).

[0177] These data indicate that basimglurant administration has the effect of preventing high-fat diet-induced obesity and suppressing the intake of high-fat diet in healthy mice at the early stage of obesity, thereby preventing weight gain.

[0178] Example 9. Basimglurant administration to healthy mice prevents high-fat diet-induced hepatic steatosis and lipid accumulation

[0179] A 2-week high-fat diet induced ectopic accumulation of lipid droplets in hepatocytes, and basimglurant administration during this period prevented high-fat diet-induced hepatic steatosis ( Figure 9 A). However, a high-fat diet for only 2 weeks was not sufficient to induce changes in the liver-to-body weight ratio, and basimglurant administration did not cause changes in the liver-to-body weight ratio ( Figure 9 B).

[0180] In addition, a 2-week high-fat diet induced an increase in lipid droplets in white adipose tissue and brown adipose tissue. Basimglurant administration during the high-fat diet induction period partially reversed the high-fat diet-induced increase in lipid droplets ( Figure 9 A) Compared with the high-fat diet-vehicle administration group, the lipid droplets in white adipose tissue and brown adipose tissue of mice in the high-fat diet-basimglurant administration group were smaller, but not reaching the levels of healthy mice without high-fat diet induction.

[0181] Regarding the mass of white adipose tissue, a 2-week high-fat diet significantly increased the mass of sWAT and gWAT, whereas basimglurant administration during this period reduced the mass of sWAT and gWAT, with basimglurant administration having a significant effect on the reduction of sWAT mass ( Figure 9 C and D).

[0182] These data suggest that basimglurant administration has a protective effect against high-fat diet-induced obesity and prevents lipid accumulation in hepatocytes and adipose tissue at the early stages of obesity development.

[0183] Example 10. Administration of Basimglurant has no significant effect on body weight or food intake in ob / ob or db / db mutant mice

[0184] Leptin is one of the key hormones associated with obesity. Secreted by adipose tissue, leptin acts on the hypothalamus, suppressing appetite and promoting energy expenditure. The ob / ob mutant mouse has a mutation in the leptin (obese) gene on chromosome 6, preventing it from producing leptin protein. Its phenotype is characterized by severe obesity and excessive food intake. The db / db mouse has a mutation in the gene encoding the leptin receptor on chromosome 4, preventing it from responding to leptin signals. The db / db mouse exhibits a degree of obesity and excessive food intake. The ob / ob and db / db mutant mice are two commonly used models of obesity.

[0185] To investigate whether basimglurant-mediated weight loss is dependent on leptin signaling, we administered basimglurant to ob / ob and db / db mice, two leptin signaling-deficient mice. We found that even at a higher dose than that of high-fat diet-induced obese wild-type mice (vehicle or basimglurant (3 mg / kg / day) intraperitoneal administration for 3 weeks), basimglurant failed to significantly change the body weight of ob / ob or db / db mice. Figure 10 B). Meanwhile, basimglurant did not reduce food intake in ob / ob or db / db mice ( Figure 10 C).

[0186] In brief, basimglurant had no significant effect on body weight and food intake in ob / ob and db / db mice.

[0187] Example 11. Administration of Basimglurant has no significant effect on lipid accumulation in ob / ob or db / db mutant mice

[0188] Both ob / ob and db / db have severe obesity, with larger lipid droplets in both white and brown fat. Basimglurant administration did not reduce lipid droplet size in adipocytes in WAT or BAT of ob / ob or db / db mutants. Figure 11 AC, Figure 12 AC). In addition, basimglurant administration did not significantly affect the sWAT-to-body weight ratio (sWAT / Weight) and gWAT-to-body weight ratio (gWAT / Weight) of the two mutant mice ( Figure 11 D and E, Figure 12 D and E).

[0189] In conclusion, basimglurant had no effect on lipid accumulation in adipose tissue of ob / ob and db / db mice.

[0190] Example 12. Administration of Basimglurant has no significant effect on hepatic steatosis in ob / ob or db / db mutant mice

[0191] In addition to the obesity phenotype, both ob / ob and db / db mutants have severe hepatic steatosis. Many large lipid droplets accumulate in the liver cells of ob / ob mice, and long-term administration of basimglurant does not improve the ectopic accumulation of fat in the liver ( Figure 13 A and B), and had no significant effect on liver weight ratio ( Figure 13 C). The liver cells of db / db mice have many dense accumulations of small lipid droplets. Long-term administration of Basimglurant cannot improve the ectopic accumulation of fat in the liver ( Figure 14 A and 14B), and had no significant effect on liver weight ratio ( Figure 14 C). In summary, basimglurant administration had no significant effect on ameliorating ectopic fat accumulation in the liver of ob / ob or db / db mutant mice.

[0192] These data suggest that long-term basimglurant administration has no weight-loss effect in ob / ob or db / db mutant mice: it does not reduce body weight, food intake, fat accumulation in adipose tissue, or hepatic steatosis. Given that both ob / ob and db / db mutants lack leptin signaling, the weight-loss effect of basimglurant is mediated by leptin signaling.

[0193] Example 13. Administration of Basimglurant Improves Hyperleptinemia

[0194] Obese mice induced by a high-fat diet also suffer from hyperleptinemia, which is manifested by serum leptin levels much higher than those of mice with normal weight. Long-term hyperleptinemia can lead to a decrease in the sensitivity of the mouse central nervous system to leptin. Figure 15 As shown, the inventors tested the effect of basimglurant on serum leptin levels in normal-weight mice, obese mice, and wild-type mice fed a high-fat diet for 2 weeks (at the early stage of obesity).

[0195] In normal-weight mice, serum leptin levels were maintained at low levels, and 2-week administration of basimglurant had no significant effect on serum leptin levels in normal-weight mice ( Figure 15 A)

[0196] Basimglurant administration for 2 weeks significantly reduced serum leptin levels in obese mice induced by long-term high-fat diet. Figure 15 B).

[0197] Shortening the administration time of Basimglurant to 4 days in high-fat diet-induced obese mice and detecting serum leptin levels before Basimglurant exerted its weight-loss effect revealed that short-term Basimglurant administration could significantly reduce serum leptin levels ( Figure 15 C).

[0198] The diet of healthy mice was changed from ordinary diet to high-fat diet for 2 weeks to simulate the early stage of obesity. During this period, solvent or Basimglurant (3 mg / kg / 2 days) was intraperitoneally administered. Another group continued to be fed with ordinary diet as a negative control. In the early stage of obesity development, 2 weeks of high-fat diet feeding can significantly increase serum leptin levels. While feeding with high-fat diet, long-term administration of Basimglurant (3 mg / kg, administered once every 2 days for 2 weeks) can improve the hyperleptinemia induced by 2 weeks of high-fat diet ( Figure 15 D).

[0199] The above results indicate that basimglurant can improve hyperleptinemia and has no effect on leptin levels in normal weight mice.

[0200] Example 14. Administration of Basimglurant Improves Leptin Sensitivity in High-Fat Diet-Induced Obese Mice

[0201] The JAK-STAT pathway is currently considered the primary pathway for leptin signaling. In obese mice, hyperleptinemia leads to decreased leptin sensitivity, preventing leptin from functioning normally and thus failing to achieve the weight-loss effects of suppressing appetite and promoting metabolism.

[0202] The inventors investigated whether leptin sensitivity is restored after Basimglurant improves hyperleptinemia. STAT3 is a downstream of the leptin receptor, and the phosphorylation level of STAT3 (Tyr705) is an important marker for testing hypothalamic leptin sensitivity. Figure 16 As shown in Figure A, the inventors used solvent or basimglurant (3 mg / kg) for short-term intraperitoneal administration to high-fat diet-induced obese mice for 4 days. 16 hours after the last administration, normal saline or leptin (1 mg / kg) was intraperitoneally injected. 40 minutes later, the hypothalamus of the mice was obtained and the total STAT3 and STAT3 (Tyr705) phosphorylation levels in the hypothalamus were analyzed by immunoblotting.

[0203] Immunoblotting was performed according to methods known in the art. The anti-STAT3 antibody was a mouse antibody (Cat. No. CST#9139) and was used at a dilution of 1:2000. The anti-phospho-STAT3 (Tyr705) antibody was a rabbit antibody (Cat. No. CST#9145) and was used at a dilution of 1:1000. The anti-β-actin antibody was a rabbit antibody purchased from ABclonal (Cat. No. AC026) and was used at a dilution of 1:500.

[0204] In the solvent-pretreated mice, injection of exogenous leptin did not increase the phosphorylation level of STAT3 (Tyr705). Figure 16B, Lanes 1-3 vs. Lanes 4-6). However, in high-fat diet-induced obese mice pretreated with basimglurant, exogenous leptin injection significantly increased the phosphorylation level of STAT3 (Tyr705) in the hypothalamus ( Figure 16 B, Lanes 7-9 vs. Lanes 10-12). Immunoblotting results showed that basimglurant enhanced leptin sensitivity in high-fat diet-induced obese mice.

[0205] In summary, the inventors found that Basimglurant can effectively reduce the body weight of obese subjects induced by a high-fat diet, and its mechanism of action is to reduce the serum leptin level of the subjects, improve hyperleptinemia, and increase leptin sensitivity, thereby inhibiting food intake to achieve a weight loss effect.

[0206] The above describes exemplary embodiments of the present invention. However, the scope of protection of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

[0207] References

[0208] 1.GBD 2015Obesity Collaborators,Afshin A,Forouzanfar MH,et al.HealthEffects of Overweight and Obesity in 195Countries over25Years.N Engl JMed.2017;377(1):13-27

[0209] 2. Wilding JPH, Jacob S. Cardiovascular outcome trials in obesity: Areview. Obes Rev. 2021; 22(1):e13112.

[0210] 3.Gregory KJ, Noetzel MJ, Niswender CM. Pharmacology of metabotropicglutamate receptor allosteric modulators: structural basis and therapeutic potential for CNS disorders. Prog MolBiol Transl Sci. 2013; 115:61-121.

[0211] 4.Lindemann L,Porter RH,Scharf SH,et al.Pharmacology of basimglurant(RO4917523,RG7090),a unique metabotropic glutamate receptor 5negativeallosteric modulator in clinical development for depression.J Pharmacol ExpTher.2015;353(1):213-233。

Claims

1. Use of an mGlu5 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing the following conditions in a subject: Nonalcoholic fatty liver disease, obesity, and / or hyperleptinemia.

2. Use of an mGlu5 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of a medicament for improving leptin sensitivity in a subject.

3. The use according to claim 1 or 2, wherein the subject has an intact leptin signaling pathway.

4. The use according to any one of claims 1 to 3, wherein the mGlu5 inhibitor comprises one or more of the group consisting of: Basimglurant, CTEP, AZD 2066: The use according to claim 4 , wherein the mGlu5 inhibitor is Basimglurant.

6. The use according to any one of claims 1 to 5, wherein the subject is selected from rodents and primates.

7. Use of a pharmaceutical composition in the preparation of a medicament for treating or preventing the following conditions in a subject: non-alcoholic fatty liver disease, obesity and / or hyperleptinemia; The pharmaceutical composition comprises one or more mGlu5 inhibitors selected from the group consisting of: Basimglurant, CTEP, AZD 2066, and a pharmaceutically acceptable carrier or excipient. Preferably, the mGlu5 inhibitor is Basimglurant.

8. Use of a pharmaceutical composition in the preparation of a medicament for improving leptin sensitivity in a subject; The pharmaceutical composition comprises one or more mGlu5 inhibitors selected from the group consisting of: Basimglurant, CTEP, AZD 2066, and a pharmaceutically acceptable carrier or excipient. Preferably, the mGlu5 inhibitor is Basimglurant.

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