Application of riluzole in preparation of medicine for treating obesity and depression

Abstract

The invention relates to application of riluzole in preparation of medicines for treating obesity and depression, and particularly discloses application of riluzole or pharmaceutically acceptable salts or derivatives of riluzole, and application of riluzole or pharmaceutically acceptable salts or derivatives of riluzole in preparation of medicines or kits for preventing and / or treating obesity and depression codiseases.

Description

Technical Field

[0001] This invention relates to the field of biomedicine. Specifically, this invention relates to the use of riluzole in the preparation of medicaments for treating obesity and depression. Background Technology

[0002] Obesity is a common metabolic disease. It is reported that hundreds of millions of adult men and women worldwide are currently obese. It is estimated that by 2030, the global obese population will reach 1 billion. Normal adult male adipose tissue accounts for approximately 15%–18% of body weight, while for women it is approximately 20%–25%. The proportion of body fat increases with age.

[0003] Obesity significantly increases the incidence and mortality rates of hypertension, diabetes, and cardiovascular diseases, becoming a major global health problem and a focus of widespread social concern. The prevention and treatment of obesity has become a challenge for modern medicine. Currently, it is difficult to expect improvement in patients' obesity symptoms through treatment unless patients change their lifestyle habits and adopt appropriate physical exercise and a reasonable, healthy diet.

[0004] Depression is a prevalent and relapse-prone affective mental illness characterized by persistent low mood. Epidemiological surveys show that the lifetime prevalence of depression worldwide is approximately 4%, affecting about 300 million people, posing a significant threat to human health. As a global mental health disorder with high prevalence, high mortality, and high disability rates, depression has become a major public health issue, creating an urgent need for clinical research.

[0005] Because obesity and depression are two serious health conditions with distinctly different symptoms, they are currently treated differently. To date, there is a lack of satisfactory and effective treatments or preventative methods for both obesity and depression. Summary of the Invention

[0006] The purpose of this invention is to provide a new and effective treatment for or prevention of obesity and depression.

[0007] Another object of the present invention is to provide the use of riluzole in the preparation of medicaments for treating obesity and depression.

[0008] In a first aspect, the present invention provides the use of an active ingredient selected from riluzole, a pharmaceutically acceptable salt thereof, or an active derivative thereof, wherein the active ingredient is used to prepare a medicament or kit for the prevention and / or treatment of comorbid obesity and depression; or the active ingredient is used to prepare a formulation or composition for inhibiting palmitic acid-induced apoptosis of nerve cells.

[0009] In another preferred embodiment, the drug is administered to a WFS1-deficient object.

[0010] In another preferred embodiment, the WFS1 deficiency includes objects with WFS1 deletion, low (or significantly decreased) WFS1 expression, and / or decreased WFS1 protein activity.

[0011] In another preferred embodiment, the obesity is obesity caused by a high-fat diet.

[0012] In another preferred embodiment, the obesity is obesity that does not require activation of TREK1.

[0013] In another preferred embodiment, the obesity is selected from the group consisting of simple obesity and secondary obesity.

[0014] In another preferred embodiment, the depression is depression caused by downregulation of the WFS1 gene.

[0015] In another preferred embodiment, comorbidity refers to a situation in which the same individual simultaneously or sequentially develops more than one disease within a specific period of time or throughout their life.

[0016] In another preferred embodiment, the derivative comprises a prodrug.

[0017] In another preferred embodiment, the drug is also used for the following purposes:

[0018] (y1) promotes upregulation of WFS1 expression;

[0019] (y2) Reduce food intake, thereby slowing down obesity;

[0020] (y3) Alleviates depressive-like behavior;

[0021] (y4) Increase brain volume;

[0022] (y5) Reduces nerve cell apoptosis.

[0023] In another preferred embodiment, the drug is also used for the following purposes:

[0024] 1) Used to shorten immobility time in patients with comorbid obesity and depression;

[0025] 2) To reduce the level of despair in patients with comorbid obesity and depression;

[0026] 3) Used to reduce anxiety levels in patients with comorbid obesity and depression.

[0027] In another preferred embodiment, the dosage of the drug, based on the dosage of riluzole, is 1-6 mg / kg / day, more preferably 2-4 mg / kg / day, and even more preferably 2 mg / kg / day.

[0028] In another preferred embodiment, the drug is in the form of an oral dosage form.

[0029] In another preferred embodiment, the drug is administered to an animal selected from the group consisting of rodents or primates (humans or non-human primates).

[0030] In another preferred embodiment, the object is a person on a high-fat diet.

[0031] In another preferred embodiment, the subject is a patient with a comorbid obesity and depression.

[0032] In another preferred embodiment, the object is an object that ingests palmitic acid.

[0033] In another preferred embodiment, the object is an object whose daily palmitic acid intake significantly exceeds the recommended level.

[0034] In another preferred embodiment, the significant exceedance is defined as the ratio of the subject's daily dietary C1 to the daily dietary C0 of palmitic acid in a normal diet (ND) (i.e., C1 / C0) ≥ 1.5, preferably ≥ 2, and more preferably ≥ 3.

[0035] In another preferred embodiment, the subject is (a) a subject whose daily palmitic acid intake is significantly excessive or who is on a high-fat diet (HFD) and (b) who is obese.

[0036] In another preferred embodiment, the subject is (a) a subject whose daily palmitic acid intake is significantly excessive or who is on a high-fat diet (HFD) and (b) a subject of normal or slightly overweight weight.

[0037] In another preferred embodiment, the nerve cells include neural progenitor cells or mature nerve cells.

[0038] In another preferred embodiment, the object has one or more features selected from the group consisting of:

[0039] (y1) The WFS1 expression level of the object is low;

[0040] (y2) The subject's food intake was excessive;

[0041] (y3) The subjects exhibited elevated levels of depressive-like behaviors;

[0042] (y4) The brain volume of the subject is low;

[0043] (y5) The number of apoptotic neural progenitor cells in the subject was high.

[0044] In another preferred embodiment, "low" refers to an expression level or brain volume that is significantly lower than the baseline value; "high" refers to an intake, quantity, or level of food that is significantly higher than the baseline value.

[0045] In another preferred embodiment, the baseline values ​​are the expression level of the corresponding gene, food intake, brain volume, number of apoptosis, or level of depressive behavior in normal subjects (subjects who are not obese or depressed).

[0046] In another preferred embodiment, the value is significantly lower than the index expression value ≤ 2 / 3 of the reference value, preferably ≤ 1 / 2 of the reference value, and more preferably ≤ 1 / 3 of the reference value.

[0047] In another preferred embodiment, the value is significantly higher than the index expression value by ≥4 / 3 of the reference value, more preferably ≥3 / 2 of the reference value, and even more preferably ≥2 times the reference value.

[0048] In another preferred embodiment, the level of depressive-like behavior is selected from one or more features of the following group:

[0049] 1) Immobility time in patients with comorbid obesity and depression;

[0050] 2) The level of despair in patients with comorbid obesity and depression;

[0051] 3) Anxiety levels in patients with comorbid obesity and depression.

[0052] In another preferred embodiment, the drug is administered before the model of a patient with comorbid obesity and depression is established; and / or the drug is administered after the model of a patient with comorbid obesity and depression is established.

[0053] In another preferred embodiment, the comorbid obesity and depression patient model is performed using a method selected from the following group:

[0054] (m1) A mouse obesity model was established using a high-fat diet;

[0055] (m2) Mice with Wfs1 knockout specific in the nervous system serve as a model of neurodegenerative disease;

[0056] (m3) Human embryonic stem cells with WFS1 gene deletion were constructed using CRISPR / Cas9 technology;

[0057] (m4) Human brain organoids with WFS1 gene knockout derived from human pluripotent stem cells were used as in vitro models of lipid metabolism disorders and neuropathy.

[0058] In another preferred embodiment, the drug comprises:

[0059] (z1) The first active ingredient, riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof;

[0060] (z2) Optional, second active ingredient, other medications for the prevention and / or treatment of obesity and depression; and

[0061] (z3) Pharmaceutically acceptable carrier.

[0062] In another preferred embodiment, the component (z1) accounts for 1-99 wt% of the total weight of the drug, more preferably 10-90 wt%, and even more preferably 30-70 wt%.

[0063] In another preferred embodiment, the other drugs for the prevention and / or treatment of obesity and depression are selected from the group consisting of orlistat, smegglutide, liraglutide, semaglutide, tezepamide, metformin, bupropion, lorcaserin, phentermine, diglucose-dependent insulinotropic peptides, or combinations thereof.

[0064] In another preferred embodiment, the drug may be a single compound or a drug composition consisting of a mixture of multiple compounds.

[0065] In another preferred embodiment, the drug is used to prepare a drug or preparation for the prevention and / or treatment of obesity and depression.

[0066] In another preferred embodiment, the drug dosage form is an oral or non-oral dosage form.

[0067] In another preferred embodiment, the oral dosage form is a tablet, powder, granule or capsule, or an emulsion or syrup.

[0068] In another preferred embodiment, the non-oral dosage form is an injection or syringe.

[0069] In another preferred embodiment, the drug is selected from the group consisting of: injections, inhalers, tinctures, powders, granules, capsules, oral liquids, tablets, pills, suspensions, emulsions, lozenges, or drops.

[0070] In another preferred embodiment, the drug is administered via intravenous injection or intraperitoneal injection.

[0071] In another preferred embodiment, the drug is administered to a human or a non-human mammal.

[0072] In another preferred embodiment, the kit comprises:

[0073] (f1) A first pharmaceutical composition comprising (i) riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof, as a first active ingredient; and (ii) a pharmaceutically acceptable carrier; and

[0074] (f2) Reagents for detecting WFS1 levels.

[0075] In a second aspect of the invention, a method for preventing and / or treating obesity and depression is provided, comprising the steps of:

[0076] Administer a safe and effective amount of riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof to the recipient.

[0077] In another preferred embodiment, the administration includes oral administration.

[0078] In another preferred embodiment, the object includes a human or a non-human mammal.

[0079] In another preferred embodiment, the non-human mammals include rodents and primates, preferably mice, rats, rabbits, and monkeys.

[0080] In another preferred embodiment, the object is a person on a high-fat diet.

[0081] In another preferred embodiment, the subject is a patient with a comorbid obesity and depression.

[0082] In another preferred embodiment, the object has one or more features selected from the group consisting of:

[0083] (y1) The WFS1 expression level of the object is low;

[0084] (y2) The subject's food intake was excessive;

[0085] (y3) The subjects exhibited elevated levels of depressive-like behaviors;

[0086] (y4) The brain volume of the subject is low;

[0087] (y5) The number of apoptotic neural progenitor cells in the subject was high.

[0088] In another preferred embodiment, the level of depressive-like behavior is selected from one or more features of the following group:

[0089] 1) Immobility time in patients with comorbid obesity and depression;

[0090] 2) The level of despair in patients with comorbid obesity and depression;

[0091] 3) Anxiety levels in patients with comorbid obesity and depression.

[0092] In another preferred embodiment, "low" refers to an expression level or brain volume that is significantly lower than the baseline value; "high" refers to an intake, quantity, or level of food that is significantly higher than the baseline value.

[0093] In another preferred embodiment, the baseline values ​​are the expression level of the corresponding gene, food intake, brain volume, number of apoptosis, or level of depressive behavior in normal subjects (subjects who are not obese or depressed).

[0094] In another preferred embodiment, the riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof, is administered 1-7 days per week, more preferably 2-5 days per week, and even more preferably 2-3 days per week.

[0095] In another preferred embodiment, the riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof, is administered for 1-20 weeks, more preferably 2-12 weeks, and even more preferably 4-8 weeks.

[0096] In another preferred embodiment, the dosage of the riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof, is 1 to 10 mg / kg / day, preferably 2 to 8 mg / kg / day, for example about 2 to 6 mg / kg / day.

[0097] In a third aspect of the invention, a method for reducing neuronal apoptosis in vitro is provided, comprising the steps of:

[0098] (a) In the presence of riluzole, nerve cells or brain organoids are cultured to reduce apoptosis of the nerve cells.

[0099] In another preferred embodiment, the concentration of riluzole is 1–50 μM.

[0100] In another preferred embodiment, in step (a), the culture system also contains palmitic acid (PA).

[0101] In another preferred embodiment, the PA concentration is 1–50 mM.

[0102] In another preferred embodiment, the reduction of nerve cell apoptosis refers to the reduction of CAS3. + / SOX2 + The number or proportion of cells.

[0103] In another preferred embodiment, the nerve cells or brain organoids are WFS1-deficient or WFS1-wild-type nerve cells or brain organoids.

[0104] In a fourth aspect of the invention, a kit is provided, the kit comprising:

[0105] (a) A first pharmaceutical composition comprising (z1a) a first active ingredient, riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof; and (z1b) a pharmaceutically acceptable carrier;

[0106] (b) An optional second pharmaceutical composition comprising (z2a) a second active ingredient different from the first active ingredient, which is another medicine for the prevention and / or treatment of obesity and depression; and (z2b) a pharmaceutically acceptable carrier; and

[0107] (c) A first reagent for detecting WFS1; preferably, the first reagent is used to detect the level of WFS1 mRNA or protein.

[0108] In another preferred embodiment, the kit further includes: (d) a second reagent for detecting palmitic acid; preferably, the second reagent is used to detect the content or level of palmitic acid in food.

[0109] In another preferred embodiment, the first and second pharmaceutical compositions are each independent pharmaceutical formulations. In yet another preferred embodiment, the kit is also provided for use in the prevention and / or treatment of comorbid obesity and depression.

[0110] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0111] Figure 1 This is a schematic diagram of the construction strategy for building Wfs1 conditional knockout mice (CKO) in Example 2 of the present invention.

[0112] Figure 2 In Example 3 of this invention, the lack of neurospecific WFS1 exacerbates the vicious cycle between obesity and depression.

[0113] Figure 2 A is a schematic diagram of the vicious cycle of obesity and depression in Embodiment 3 of the present invention.

[0114] Figure 2 B represents the proportion of depression in normal and obese individuals in Embodiment 3 of the present invention.

[0115] Figure 2 C represents the proportion of obesity in the normal and depressed populations in Embodiment 3 of the present invention.

[0116] Figure 2D represents the immunostaining results of WFS1 (red), MAP2 (green), and DAPI (blue) in the cerebral cortex of mice on normal or high-fat diets in Example 3 of this invention. The scale bar is 50 μm.

[0117] Figure 2 E represents the percentage of WFS1 positive cells in the cerebral cortex of mice on a normal diet (ND) or a high-fat diet (HFD) in Example 3 of this invention (WT+ND, n=19; WT+HFD, n=18).

[0118] Figure 2 F is a representative bright-field image of the brain of WT or CKO mice on a normal or high-fat diet in Example 3 of this invention, with a scale bar of 5 mm.

[0119] Figure 2 G represents the area of ​​the cerebral cortex (mm²) of WT or CKO mice on a normal or high-fat diet in Example 3 of this invention. 2 ). (WT+ND, n=9; WT+HFD, n=9; CKO+ND, n=11; CKO+HFD, n=10).

[0120] Figure 2 H represents the weight gain of WT or CKO mice on a normal or high-fat diet from day 0 to day 180 in Example 3 of this invention.

[0121] Figure 2 I represents the movement trajectory of WT or CKO mice on a normal or high-fat diet in the open field (OFT) experiment in Example 3 of this invention.

[0122] Figure 2 JK represents the analysis of the movement distance (J) and walking speed (K) of WT or CKO mice on normal or high-fat diets in Example 3 of this invention (WT+ND, n=10; WT+HFD, n=14; CKO+ND, n=15; CKO+HFD, n=11).

[0123] Figure 2 L is a schematic diagram of the forced swimming experiment in Embodiment 3 of the present invention.

[0124] Figure 2 M represents the immobility time analysis of WT or CKO mice on normal or high-fat diets during the forced swimming experiment in Example 3 of this invention (WT+ND, n=12; WT+HFD, n=11; CKO+ND, n=13; CKO+HFD, n=12).

[0125] Data are expressed as mean ± standard deviation. Significance was determined by an unpaired two-tailed Student's t-test. Figure 2E, G, J, K, M) and two-way ANOVA ( Figure 2 H) Calculation. Comparison of WT and CKO, * p<0.05 ** p<0.01 and *** p < 0.001 was considered statistically significant; comparing a normal diet and a high-fat diet, # p<0.05 ## p<0.01 and ### p < 0.001 is considered significant.

[0126] Figure 3 In Example 4 of this invention, WFS1 deletion exacerbated apoptosis of neural progenitor cells in the human brain.

[0127] Figure 3 A is a schematic diagram of brain organoid differentiation in Embodiment 4 of the present invention.

[0128] Figure 3 B represents WT and WFS1 treated with either the control or 1 mM PA from day 50 to day 55 in Example 4 of this invention. - / - Brain organoid area (mm) 2 ) Changes (WT, n=6; WT+PA, n=10; WFS1) - / - n=6; WFS1 - / - +PA, n=7).

[0129] Figure 3 C represents WT and WFS1 treated with control or 1mM PA in Example 4 of this invention. - / - Immunostaining results of Caspase 3 (CAS3) (green), SOX2 (magenta), and DAPI (blue) in brain organoids, scale bar 50 μm.

[0130] Figure 3 D represents WT and WFS1 treated with control or 1mM PA in Example 4 of this invention. - / - CAS3 in brain organoids + Cells account for SOX2 + Percentage of total cell count (WT, n=20; WT+PA, n=21; WFS1) - / - n=19; WFS1 - / - +PA, n=22).

[0131] Data are expressed as mean ± standard deviation. Significance was calculated using an unpaired two-tailed Student's t-test. For WT and WFS1 - / - The comparison, * p<0.05 ** p<0.01 and ***p < 0.001 was considered statistically significant. Comparison between the control group and the PA group, # p<0.05 ## p<0.01 and ### p < 0.001 is considered significant.

[0132] Figure 4 Riluzole is used in Example 5 of this invention to rescue apoptosis of neural progenitor cells in the human brain organ lacking WFS1.

[0133] Figure 4 A represents the treatment of WT and WFS1 with 5μM Riluzole or as a control in Example 5 of this invention. - / - Immunostaining with CAS3 (green), SOX2 (magenta), and DAPI (blue) after brain organoid formation. Scale bar: 50 μm.

[0134] Figure 4 B represents the WT and WFS1 treated with 1mM PA in Embodiment 5 of this invention. - / - Immunostaining results of brain organoids after treatment with 5 μM iluzole or control, showing CAS3 (green), SOX2 (magenta), and DAPI (blue), scale bar at 50 μm.

[0135] Figure 4 C represents the treatment of WT and WFS1 with 5 μM Riluzole or as a control in Example 5 of this invention. - / - CAS3 after brain organoid + Cells account for SOX2 + Percentage of total cell count (WT + control group, n=16; WT + Riluzole, n=15; WFS1) - / - +Control group, n=14; WFS1 - / - +Riluzole, n=13).

[0136] Figure 4 D represents the WT and WFS1 of the 1mM PA treatment in Embodiment 5 of this invention. - / - Brain organoids treated with 5 μM MRiluzole or a control group showed CAS3 + Cells account for SOX2 + Percentage of total cell count (WT+PA+control group, n=11; WT+PA+Riluzole, n=12; WFS1) - / - +PA+control group, n=13; WFS1 - / - +PA+Riluzole, n=13).

[0137] Figure 4E represents WT and WFS1 treated with 5 μM Riluzole or the control from day 50 to day 55 in Example 5 of this invention. - / - Individual area of ​​brain organoids (mm²) 2 Changes in (WT+PA+control group, n=10; WT+PA+Riluzole, n=6; WFS1) - / - +PA+control group, n=8; WFS1 - / - +PA+Riluzole, n=6).

[0138] Data are expressed as mean ± standard deviation. Significance was calculated using an unpaired two-tailed Student's t-test. WT and WFS1 - / - The comparison, * p<0.05 ** p<0.01 and *** p < 0.001 was considered statistically significant; compared with the control group and the Riluzole group, # p<0.05 ## p<0.01 and ### p < 0.001 is considered significant.

[0139] Figure 5 In Example 6 of this invention, Riluzole alleviates depressive-like behavior caused by a high-fat diet exacerbated by WFS1 deficiency.

[0140] Figure 5 A is a schematic diagram of WT and CKO mice in Example 6 of the present invention, after being fed a high-fat diet and then treated with Riluzole or a control.

[0141] Figure 5 B shows the immunostaining results of CAS3 (red), Nestin (green), and DAPI (blue) in the hippocampus of WT and CKO mice on normal or high-fat diets using Riluzole or a control in Example 6 of this invention. Scale bar is 50 μm.

[0142] Figure 5 C represents the percentage of CAS3-positive cells in the hippocampus of WT and CKO mice on normal or high-fat diets treated with Ri luzole or control in Example 6 of this invention, relative to the total number of Nestin-positive cells (WT+ND+control group, n=27; WT+HFD+control group, n=18; WT+HFD+Riluzole, n=23; CKO+ND+control group, n=32; CKO+HFD+control group, n=20; CKO+HFD+Riluzole, n=22).

[0143] Figure 5D is a representative bright-field image of the brain of WT and CKO mice on normal or high-fat diets in Example 6 of this invention after being treated with Ri luzole or a control. The scale bar is 5 mm.

[0144] Figure 5 E represents the cerebral cortex area (mm²) of WT and CKO mice subjected to Riluzole or control treatments (normal or high-fat diets) in Example 6 of this invention. 2 (WT+ND, n=16; WT+HFD+control group, n=12; WT+HFD+Riluzole, n=12; CKO+ND, n=19; CKO+HFD+control group, n=9; CKO+HFD+Riluzole, n=12).

[0145] Figure 5 F represents the representative movement trajectory of WT or CKO mice treated with Ri luzole or control on a normal or high-fat diet in a mining experiment in Example 6 of this invention.

[0146] Figure 5 GH represents the analysis of walking speed (G) and distance (H) of WT or CKO mice on normal or high-fat diets treated with Ri luzole or as controls in a mining experiment of Example 6 of this invention (WT+ND, n=11; WT+HFD+control group, n=10; WT+HFD+Ri luzole, n=12; CKO+ND, n=16; CKO+HFD+control group, n=9; CKO+HFD+Riluzole, n=8).

[0147] Figure 5 I is a schematic diagram of the forced swimming experiment in Embodiment 6 of the present invention.

[0148] Figure 5 J represents the immobility time analysis of WT or CKO mice on normal or high-fat diets in Example 6 of this invention after receiving Ri luzole or control treatment (WT+ND, n=19; WT+HFD+control group, n=12; WT+HFD+Ri luzole, n=14; CKO+ND, n=17; CKO+HFD+control group, n=10; CKO+HFD+Ri luzole, n=14).

[0149] Data are expressed as mean ± standard deviation. Significance was calculated using an unpaired two-tailed Student's t-test. WT and CKO were compared. * p<0.05 ** p<0.01 and *** p<0.001 was considered significant; comparing a normal diet and a high-fat diet, #p<0.05 ## p<0.01 and ### p < 0.001 was considered significant; compared with the control group and the Riluzole group, and Significant.

[0150] Figure 6 In Example 7 of this invention, Riluzole is used to alleviate obesity induced by a high-fat diet exacerbated by WFS1 deficiency.

[0151] Figure 6 A represents the cumulative food intake of WT and CKO mice on a high-fat diet or control diet in Example 7 of this invention (WT+HFD+control group, n=6; WT+HFD+Riluzole, n=5; CKO+HFD+control group, n=5; CKO+HFD+Riluzole, n=5).

[0152] Figure 6 B is a representative bright-field image of WT and CKO mice subjected to a high-fat diet or control treatment in Example 7 of this invention, with a scale bar of 7 cm.

[0153] Figure 6 CD represents the weight gain (C) and weight change (D) of WT and CKO mice treated with a high-fat diet using Riluzole or a control in Example 7 of this invention (WT+HFD+control group, n=12; WT+HFD+Riluzole, n=13; CKO+HFD+control group, n=10; CKO+HFD+Riluzole, n=15).

[0154] Figure 6 EF represents the oxygen consumption (VO2, mL / kg / h) (E) and carbon dioxide production (VCO2, mL / kg / h) (F) of WT and CKO mice on the HFD diet in Example 7 of this invention after 24 hours of treatment with Riluzole or control in a metabolic cage.

[0155] Figure 6 GH represents the area under the curve (AUC) of IPGTT (G) and IPGTT (H) of WT and CKO mice (WT+HFD+control group, n=11; WT+HFD+Riluzole, n=11; CKO+HFD+control group, n=7; CKO+HFD+Riluzole, n=8) treated with a high-fat diet by Riluzole or control in Example 7 of this invention.

[0156] Data are expressed as mean ± standard deviation. Significance was determined using unpaired two-tailed Student's t-test (A, H) and two-way ANOVA (CG). Comparison of WT and CKO, * p<0.05 ** p<0.01 and *** p<0.001 was considered statistically significant; comparison between the WT+HFD group, the control group, and Riluzole, # p<0.05 ## p<0.01 and ### p < 0.001 was considered statistically significant. Comparison of the CKO+HFD group, control group, and Riluzole group... It is considered significant. Detailed Implementation

[0157] Through extensive and in-depth research and screening, the inventors unexpectedly discovered for the first time that riluzole, a known drug, can effectively prevent and / or treat the comorbidity of obesity and depression in specific populations. Based on this discovery, the inventors completed this invention.

[0158] the term

[0159] As used herein, the terms "pharmaceutical composition of the present invention" and "pharmaceutical of the present invention" are used interchangeably and both refer to a preparation or pharmaceutical composition containing riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof as an active ingredient.

[0160] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0161] As used herein, the terms “subject” and “required subject” refer to any mammal or non-mammal. Mammals include, but are not limited to, humans, vertebrates such as rodents, non-human primates, cattle, horses, dogs, cats, pigs, sheep, and goats.

[0162] Comorbidity of obesity and depression

[0163] As used in this article, the terms "obesity" and "obesity disorder" are used interchangeably, both referring to a chronic metabolic disease caused by the interaction of multiple factors, including genetic and environmental factors, where the amount of fat stored in the body exceeds 20% of ideal body weight. Its pathogenesis involves energy intake exceeding energy expenditure, with a high-fat diet leading to excessive fat accumulation and excessive weight. Long-term obesity can trigger various diseases, including but not limited to: being overweight and having high cholesterol. Furthermore, obesity increases the burden on joints, exacerbating joint degeneration and leading to osteoarthritis; it restricts chest wall movement, affecting respiratory function; it causes sleep apnea syndrome; and it leads to chronic hypoxic damage to the brain and body.

[0164] Obesity and depression are two major global health risks. Studies have shown a vicious cycle between depression and obesity: people with depression are at higher risk of obesity, and obese people are at higher risk of depression. The prevalence of depression among obese individuals reaches 23%. Compared to the risk of individual conditions, men with both depression and obesity have a 68% higher risk of developing diabetes and a 57% higher risk of developing hypertension. This vicious cycle often leads to the simultaneous occurrence or exacerbation of depression and obesity.

[0165] As used in this article, the term "comorbidity" refers to the situation in which the same individual has more than one disease occurring simultaneously or sequentially within a specific period of time or throughout their life.

[0166] In this application, comorbidity refers to the simultaneous or sequential occurrence of obesity and depression in the same individual during a specific period of time or throughout their life, especially the simultaneous occurrence or suffering from obesity and depression.

[0167] WFS1 gene

[0168] The WFS1 gene is located on chromosome 4p16.1 and consists of 8 exons (33.4kb). It encodes Wolframin, a transmembrane protein on the endoplasmic reticulum composed of 890 amino acids. It is highly expressed in pancreatic β cells and brain cells and is mainly involved in membrane transport, protein processing, and cell inhibition of apoptosis.

[0169] Currently, direct evidence linking WFS1 to obesity is lacking. Studies have shown that knocking out Wfs1 leads to age-dependent behavioral deficits and neurodegeneration in fruit flies. Furthermore, the brainstem of Wfs1 knockout mice and rats is significantly smaller, revealing a conserved and important role for WFS1 in brain development and neurodegeneration.

[0170] Riluzole

[0171] Riluzole (CAS No.: 1744-22-5, Molecular Formula: C8H5F3N2OS, 2-amino-6-trifluoromethoxybenzothiazole), its chemical structural formula is shown below:

[0172]

[0173] Riluzole is a drug approved by the U.S. Food and Drug Administration (FDA) for the treatment of ALS. The main neuroprotective mechanism of riluzole in ALS patients is the inhibition of glutamate release at nerve endings.

[0174] Palmitic acid (PA)

[0175] Palmitic acid (PA) (CAS No.: 3343-33-7, Molecular formula: C 16 H 30 O2T2), the chemical structural formula of which is shown below:

[0176]

[0177] Palmitic acid is the most common saturated fatty acid that induces obesity.

[0178] Pharmaceutical composition and administration

[0179] The pharmaceutical composition provided by the present invention preferably contains 0.1-99 wt% of a first active ingredient, with the remainder being a second active ingredient, a pharmaceutically acceptable carrier, a diluent or solution or a salt solution.

[0180] The first active ingredient of this invention is riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof. Furthermore, it can be used in combination with other therapeutic agents, i.e., a second active ingredient.

[0181] The second active ingredient can be used to prevent and / or treat conditions such as obesity and depression.

[0182] When necessary, one or more pharmaceutically acceptable carriers may be added to the drug of this invention. These carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, etc., which are conventional in the pharmaceutical field.

[0183] The compounds and pharmaceutical compositions provided by this invention can be in various forms, such as tablets, injections, capsules, powders, syrups, solutions, suspensions, and aerosols, and can be present in suitable solid or liquid carriers or diluents and in suitable sterilization devices for injection or infusion.

[0184] Various dosage forms of the pharmaceutical compositions of the present invention can be prepared according to conventional pharmaceutical preparation methods. The dosage form typically contains 0.05-1000 mg of the active compound of the present invention per unit volume, preferably 1-500 mg per unit volume.

[0185] The pharmaceutical compositions of the present invention can be used clinically in mammals, including humans and animals, via routes of administration such as mouth, nose, skin, lungs, or gastrointestinal tract. Oral administration is most preferred. The most preferred daily dose is 0.01-400 mg / kg body weight, taken as a single dose, or 0.01-200 mg / kg body weight, taken in divided doses. Regardless of the method of administration, the optimal dose for an individual should be determined based on the specific treatment. Generally, a low dose is started, and the dose is gradually increased until the most suitable dose is found.

[0186] The drugs or inhibitors of the present invention can be administered in various ways, such as by injection, spray, nasal drops, eye drops, penetration, absorption, physical or chemical mediated methods, into the body such as muscles, intradermal, subcutaneous, veins, and mucous membranes; or by being mixed with or encapsulated by other substances and introduced into the body.

[0187] Typically, the active ingredient of the present invention or a pharmaceutical composition containing it can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0188] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including O / W, W / O, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0189] The active ingredients of this invention can be formulated into ordinary formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0190] In order to formulate the active ingredient of the present invention into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0191] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0192] To formulate the drug delivery unit into capsules, the active ingredient of this invention can be mixed with a diluent and a flow aid, and the mixture can be placed directly into hard or soft capsules. Alternatively, the active ingredient can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. The diluents, binders, wetting agents, disintegrants, and flow aids used to prepare the tablets of this invention can also be used to prepare the capsules of this invention.

[0193] To prepare the active ingredient of this invention into an injection, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure regulators can be added. Solubilizers or co-solvents can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters can be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure regulators can be sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing a lyophilized powder injection, mannitol, glucose, etc., can also be added as a support agent.

[0194] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0195] The active ingredients or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs.

[0196] When the active ingredient of this invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.

[0197] The main advantages of this invention include:

[0198] (1) This invention provides the application of riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof, in the treatment of obesity and depression. This invention utilizes a mouse neurodegenerative disease model with WFS1 knockout in the nervous system and an in vitro human brain organoid model with WFS1 gene knockout in a high-fat pathological microenvironment to verify that riluzole can effectively alleviate obesity-like and depression-like behaviors induced by a high-fat diet.

[0199] (2) This invention utilizes a mouse neurodegenerative disease model with WFS1 knockout in the nervous system and an in vitro model of human brain organoids with WFS1 gene knockout in a high-fat pathological microenvironment to verify that riluzole can effectively reduce apoptosis of neural progenitor cells caused by high fat.

[0200] (3) Through the three-dimensional human brain organoids derived from pluripotent stem cells and the mouse model of WFS1 conditional knockout of the nervous system, the inventors studied obesity and neurological dysfunction caused by lipid metabolism disorders and confirmed that a high-fat diet (HFD) induces apoptosis of neural stem cells, thereby reducing the brain volume of human brain organoids and mice.

[0201] (4) In the present invention, WFS1 deficiency exacerbates obesity and depressive-like behavior in mice induced by high-fat diet in a conditional knockout mouse model of the nervous system. Riluzole can not only reduce apoptosis of neural progenitor cells in human brain organoids with WFS1 gene knockout, but also more effectively alleviate obesity and depressive-like behavior induced by high-fat diet in mice.

[0202] (5) Riluzole, as a marketed drug, has high safety and very low side effects, making it suitable for long-term use.

[0203] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0204] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available products.

[0205] Example 1: Construction of a WFS1 gene knockout human embryonic stem cell line

[0206] 1. Plasmid construction

[0207] 1.1 Plasmid Design and sgRNA Screening

[0208] Multiple sgRNAs were designed based on the WFS1 genome sequence, and two sgRNAs were selected. The modified sgRNA fragments were then inserted into a vector (P2u6) carrying the Cas9 protein gene and the puromycin resistance gene. Based on a combined score of sgRNA targeting and off-target rates, the following sgRNA sequences were selected:

[0209] sgRNA1: attgagtcgggaacgcgcc (SEQ ID NO.1)

[0210] sgRNA2: gcggagcagtgttggagtc (SEQ ID NO.2)

[0211] 1.2 Plasmid transformation and sequencing

[0212] Thaw DH5α chemocompetent cells on ice for 5 min. Add the ligation product to 100 μL of competent cells, gently tap the tube wall to mix (avoid pipetting), and incubate on ice for 25 min. Transfer to a 42°C water bath for heat shock for 45 s, then cool on ice for 2 min. Add 900 μL of LB broth (antibiotic-free) to a clean bench, incubate at 37°C for 1 h (225 rpm), then centrifuge at 5000 rpm for 1 min. Discard 850 μL of supernatant, resuspend approximately 150 μL on a plate, spread with spreader beads, and incubate upside down at 37°C for 12–16 h. Pick single colonies and send them to a sequencing company for sequencing. Screen for successfully sequenced plasmids and perform large-scale plasmid extraction.

[0213] 2. Embryonic stem cell culture conditions:

[0214] (1) Dilute Matrigel at 1:80 and use it to coat a 3.5cm cell culture dish. Let it stand for 1 hour to allow the gel to solidify. Wash the cells twice with DMEM / F12 medium before seeding.

[0215] (2) Human embryonic stem cells (H1) were revived and seeded into Matrigel-coated culture dishes and cultured in mTeSR medium (Stem Cell, #85850). Y-27632 (1000X) was added on the first day of culture.

[0216] (3) After inoculation, the cells were placed in a 37°C incubator containing 5% CO2 for cell culture. The medium was completely changed on the second day. After removing Rock Inhibitor (Y-27632) from the medium, the medium was changed to fresh medium every day for cell passage.

[0217] 3. Plasmid electroporation and cell line selection

[0218] (1) After the embryonic stem cells are cultured to a density of about 80%, they are digested using TrypLE and filtered using a 40μm cell filter to prepare for electroporation by dispersing them into single cells as much as possible.

[0219] (2) After cell counting, take 5 × 10⁻⁶ cells. 5 Electrode solutions were prepared in the cells on both sides, and recombinant plasmids carrying sgRNA and Cas9 protein genes were introduced into the cells via electrotransfection.

[0220] (3) Resuspend the electroporated embryonic stem cells in fresh mTeSR medium and seed them into a 3.5cm cell culture dish. Observe the cell status using EVOS.

[0221] (4) Change the medium every other day. After two consecutive days of culture, add 500 ng / mL of puromycin for cell selection. After two consecutive days of selection, replace the medium with one that does not contain puromycin and continue culturing until the clones reach a size of 30-50 μM.

[0222] (5) Select single clones of cells, transfer them to 96-well plates for expansion culture, scrape off some cells for genotyping, and verify that the WFS1 gene has been knocked out by sequencing.

[0223] 4. Inducing embryonic stem cells to differentiate into human brain organoids in vitro.

[0224] WFS1 knockout human embryonic stem cells were naturally sedimented and aggregated into embryoid bodies (EBs) using low-adsorption 96-well plates. On day 5 of continuous culture, the medium was switched to induce neural differentiation, and the medium was changed every other day. On day 11, the EBs were embedded in Matrigel and transferred to 10 cm plates for further culture. On day 16, a new differentiation medium (without vitamin A) was used, and the cells were placed in a shaker (57 rpm) for suspension culture. On day 21, a new differentiation medium (with vitamin A) was used, and the cells were placed in a shaker (57 rpm) for continued suspension culture until day 50, at which point experiments could begin.

[0225] Example 2: Construction of Wfs1 conditional knockout mice

[0226] To construct Wfs1 conditional knockout mice for the nervous system, Nestin-Cre tool mice and Wfs1 mice were used. flox / floxMice were purchased from Shanghai Southern Model Biotechnology Co., Ltd. The construction strategy involved inserting loxp sequences flanking exon 3 of the mouse Wfs1 gene using CRISPR-Cas9 technology. Figure 1 The Wfs1 gene targets the transcript Wfs1-201 (ENSMUST00000043964.13). Through Wfs1... flox / flox Mice were mated with Nestin-Cre mice, and the offspring were further mated to obtain Wfs1 conditional knockout mice.

[0227] Example 3: Nervous system-specific knockout of Wfs1 exacerbates the vicious cycle of obesity and depression.

[0228] Experiments showed that WFS1-positive cells were significantly reduced in the cortex of mice fed a high-fat diet (HFD). Figure 2 DE).

[0229] To investigate the effects of WFS1 deficiency in the nervous system on obesity, Wfs1-flox mice (Wfs1 f / f Mating WFS1 knockout mice with Nestin-Cre transgenic mice produced conditional WFS1 knockout (CKO) mice. Unexpectedly, the brain volume of WFS1 knockout mice was reduced, and a high-fat diet significantly exacerbated this reduction. Figure 2 FG).

[0230] The results are shown in Table 1:

[0231] Table 1

[0232]

[0233] To illustrate the vicious cycle between obesity and depression, an obesity model was established by feeding mice a high-fat diet, and depressive-like behavioral experiments were conducted, including the open field test and forced swimming test. From 6 to 30 weeks, the weight gain of CKO mice on a normal diet was much lower than that of WT mice, but the weight gain of CKO mice on a high-fat diet was higher than that of WT mice on a high-fat diet. This indicates that Wfs1 deficiency increases susceptibility to obesity induced by a high-fat diet. Figure 2 H).

[0234] Previous studies have found that the absence of Wfs1 in the nervous system leads to depressive-like behavior in ND mice.

[0235] In the open field experiment, as shown in Table 1, the movement distance and walking speed of WT mice on a high-fat diet were significantly lower than those on a normal diet. This phenotype was even more pronounced in CKO mice. Figure 2 IK).

[0236] In the forced swimming experiment, compared with mice on a normal diet, WT mice on a high-fat diet had a longer immobile time (i.e., immobile state time in Table 1) and exhibited severe depressive-like behavior. CKO mice on a high-fat diet showed even more severe depressive-like behavior, indicating that the depressive-like behavior in mice was induced by HFD and exacerbated by Wfs1 deficiency. Figure 2 In summary, these observations suggest that Wfs1 deficiency exacerbates HFD-induced obesity and depressive-like behaviors.

[0237] Example 4: WFS1 deficiency exacerbates apoptosis of neural progenitor cells in the human brain.

[0238] To reproduce the key role of WFS1 in human obesity and depression, human brain organoids were cultured in vitro using a three-dimensional culture system. Brain organoids generated from WT and WFS1-deficient H1 hESCs were used to visually evaluate the effects of WFS1 on the in vitro nervous system. Figure 3 A). During the growth of brain organoids, the volume of WFS1-deficient brain organoids was consistently smaller than WT, and the difference was significant at day 50.

[0239] Palmitic acid (PA) is the most common saturated fatty acid that induces obesity. PA-induced lipotoxicity has been reported to lead to type 2 diabetic neuropathy and neurodegenerative diseases. To more accurately simulate the pathological environment of human obesity and explore the regulatory role of WFS1 in obesity, 1 mM PA was added to brain organoids at 50 days, and the size of the brain organoids was measured 5 days later.

[0240] The results showed that WFS1 deficiency led to a slower growth rate of brain organoids, and the growth rate was even slower after the addition of PA. Figure 3 B). This indicates that the presence of palmitic acid (PA) leads to apoptosis of hypothalamic neurons.

[0241] The results of immunofluorescence staining are shown in Table 2:

[0242] Table 2

[0243] Group <![CDATA[CAS3 + / SOX2 + Cell percentage (%) WT+ Reference 5.94 WT+PA 12.30 <![CDATA[WFS1 - / - +Comparison]]> 12.53 <![CDATA[WFS1 - / - +PA]]> 19.87

[0244] The above results further show that certain or high concentrations of palmitic acid (PA) (e.g., 1 mM PA) can induce neuronal apoptosis, and the lack of WFS1 further exacerbates apoptosis. Figure 3 CD).

[0245] Example 5: Riluzole rescues WFS1 deficiency-induced lipotoxicity-induced apoptosis of neural progenitor cells in human brain organs.

[0246] Riluzole is a glutamate modulator that has been approved for the treatment of amyotrophic lateral sclerosis (ALS). To verify the effect of riluzole on WFS1 deficiency-induced apoptosis in brain organoids, human brain organoids were treated with 5 μM riluzole for 5 days.

[0247] Immunofluorescence staining results showed that treatment of WFS1-deficient brain organoids with riluzole significantly reduced the proportion of CAS3-positive cells in SOX2-positive cells, indicating that riluzole can rescue WFS1 deficiency-induced apoptosis. Figure 4 A, C).

[0248] Given that lipotoxicity exacerbated apoptosis and led to a reduction in size in WFS1 knockout brain organoids in previous experiments, this study further investigated whether riluzole could alleviate the effects of lipotoxicity.

[0249] The results showed that riluzole treatment could alleviate lipotoxic-induced brain organoid shrinkage and NPC apoptosis in WFS1-deficient human brain organoids. Figure 4 B,DE).

[0250] Example 6: Riluzole alleviates WFS1 deficiency-induced high-fat diet-induced depressive-like behavior in mice.

[0251] PA is an important fatty acid component of a high-fat diet, therefore, the next step is to investigate the effects of riluzole on the nervous system of obese and depressed mice on a high-fat diet. Figure 5 A). Studies have shown that riluzole can reduce the reduction in brain organoid volume and NPC apoptosis. Therefore, in this embodiment, we further investigated whether riluzole could produce the same effect in mice.

[0252] The results are as follows Figure 5 As shown in Table 3:

[0253] Table 3

[0254]

[0255] The results showed that riluzole reduced apoptosis of NPCs in the hippocampus of mice and rescued the brain volume reduction in WT and CKO mice under a high-fat diet. Figure 5 BE).

[0256] In addition, to investigate whether riluzole alleviated the depressive-like phenotype in high-fat diet-induced obese mice, depressive behavioral assessments were performed, including the open field test and the forced swimming test. The methods are as follows:

[0257] WT and CKO mice were fed a high-fat diet for 6 consecutive months (obesity model), and then treated with 50 mg / kg riluzole daily for 2 months.

[0258] The results are as follows Figure 5As shown. It is noteworthy that the recovery of walking speed and motor ability in mice during the open field experiment (…) Figure 5 FH) and the reduction in immobility time in the forced swimming test ( Figure 5 The results (IJ) indicate that riluzole can effectively alleviate depressive-like behavior in obese mice induced by a high-fat diet.

[0259] These results indicate that, unexpectedly, riluzole can reverse high-fat diet-induced depressive-like behaviors, thereby simultaneously treating the comorbidity of obesity and depression.

[0260] Example 7: Riluzole alleviated obesity induced by a high-fat diet exacerbated by WFS1 deficiency.

[0261] To investigate the effect of riluzole on obesity induced by a high-fat diet, obese mice were treated with riluzole for 2 months and then evaluated.

[0262] The results are as follows Figure 6 As shown, a high-fat diet prevented weight loss in Wfs1 mutant mice, suggesting that the Wfs1 mutation increases susceptibility to high-fat diet-induced diabetes or metabolic syndrome. However, the mechanism of Wfs1-mediated food intake inhibition requires further investigation.

[0263] In addition, WT and CKO mice on a high-fat diet showed decreased appetite for high-fat foods. Figure 6 A) Significant weight loss ( Figure 6 BD), O2 consumption (VO2) and CO2 production (VCO2) increased ( Figure 6 EF).

[0264] Surprisingly, riluzole was able to effectively reduce obesity induced by a high-fat diet (HFD).

[0265] Intraperitoneal glucose tolerance test (IPGTT) results showed that riluzole treatment improved glucose tolerance in mice, indicating that riluzole can reduce the risk of diabetes by inhibiting high-fat diet-induced obesity. Figure 6 GH).

[0266] In summary, this invention effectively overcomes the various shortcomings of the prior art and has high industrial application value.

[0267] discuss

[0268] Studies have shown that obese individuals have a 1.48 times higher risk of developing depression (16.7%) than the general population (11.3%). Similarly, individuals with depression are 1.4 times more likely to be obese (29.6%) than non-depressed individuals (the normal group) (21.1%). Figure 2These results indicate that this bidirectional relationship often leads to the co-occurrence of depression and obesity, creating a vicious cycle. The high incidence of depression among obese individuals and the high incidence of obesity among depressed individuals both suggest that the comorbidity of obesity and depression poses a greater threat to human health, implying that research and solutions to the challenges of obesity and depression complications are urgently needed.

[0269] For cognitive decline, some key symptoms manifest as abnormalities in brain structure, including reduced brain volume, brain atrophy, and white matter hyperintensity. A high-fat diet (HFD) is a major risk factor for obesity and type 2 diabetes (T2D).

[0270] This invention is the first to discover and confirm a correlation between obesity, cognitive function, and depression in a specific population (Wfs1-deficient type), although the underlying mechanism is not yet clear. Furthermore, this invention is the first to demonstrate that riluzole, a known drug, can effectively prevent and / or treat the comorbidity of obesity and depression in a specific population, thus providing a more effective and safer approach to preventing and / or treating the comorbidity of obesity and depression.

[0271] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. Use of an active ingredient, characterized in that The active ingredient is selected from the group consisting of riluzole, or a pharmaceutically acceptable salt thereof, or an active derivative thereof, and the active ingredient is used for the manufacture of a medicament or a kit for preventing and / or treating obesity and depression comorbidity; or the active ingredient is used for the manufacture of a preparation or a composition for inhibiting apoptosis of neural cells caused by palmitate.

2. Use according to claim 1, characterized in that, The medicament is administered to a subject with WFS1 deficiency.

3. Use according to claim 1, characterized in that, The obesity is obesity caused by high fat diet (HFD).

4. The use according to claim 1, characterized in that, The medicament is also used for the following uses: (y1) promoting up-regulation of WFS1 expression; (y2) reducing food intake, thereby slowing down obesity; (y3) alleviating depression-like behavior; (y4) expanding brain volume; (y5) reducing apoptosis of neural cells.

5. The use according to claim 1, characterized in that, The medicament is also used for the following uses: 1) for shortening immobility time of a patient with obesity and depression comorbidity; 2) for reducing hopelessness of a patient with obesity and depression comorbidity; 3) for reducing anxiety of a patient with obesity and depression comorbidity.

6. The use according to claim 1, characterized in that, The amount of the medicament used is 1-6 mg / kg / day of riluzole, preferably 2-4 mg / kg / day, more preferably 2 mg / kg / day.

7. The use according to claim 1, characterized in that, The medicament comprises: (z1) a first active ingredient, riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof; (z2) optionally, a second active ingredient, other medicaments for preventing and / or treating obesity and depression; and (z3) a pharmaceutically acceptable carrier.

8. A method of reducing apoptosis in neural cells in vitro, comprising contacting the neural cells with a compound of claim 1. 5 The method comprises the step of: (a) culturing neural cells or brain organoids in the presence of riluzole, thereby reducing apoptosis of the neural cells.

9. The method of claim 8, wherein, In step (a), the culturing system also contains palmitate (PA).

10. A kit comprising: (a) a first pharmaceutical composition containing (z1a) a first active ingredient, riluzole, or a pharmaceutically acceptable salt thereof, or a derivative thereof; and (z1b) a pharmaceutically acceptable carrier; (b) optionally, a second pharmaceutical composition containing (z2a) a second active ingredient different from the first active ingredient, which is other medicaments for preventing and / or treating obesity and depression; and (z2b) a pharmaceutically acceptable carrier; and (c) a first reagent for detecting WFS1; preferably, the first reagent is for detecting the level of mRNA or protein of WFS1. ​ ​

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