Use of quinolin-4-one or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of non-alcoholic fatty liver

Abstract

The application belongs to the technical field of medicine, and particularly relates to application of a 4-(4-diethylamino-1-methylbutylamino)-7-chloroquinoline derivative or a pharmaceutically acceptable salt thereof, or a tautomer thereof, or an internal racemate thereof, or an external racemate thereof, or an enantiomer thereof, or a diastereomer thereof, or a pharmaceutical composition with the active ingredient to preparation of a medicine for preventing or treating obesity and obesity-related metabolic diseases. The 4-(4-diethylamino-1-methylbutylamino)-7-chloroquinoline derivative has a significant curative effect on obesity and related metabolic diseases. After intragastric administration of the 4-(4-diethylamino-1-methylbutylamino)-7-chloroquinoline derivative to obese mice induced by high-fat feed and high-fructose drinking water, after 6 weeks, compared with a high-fat control group, the 4-(4-diethylamino-1-methylbutylamino)-7-chloroquinoline derivative inhibits more than 70% of weight gain of the mice, and there is no significant difference in food intake and water intake, and the 4-(4-diethylamino-1-methylbutylamino)-7-chloroquinoline derivative has a good clinical application prospect.

Description

[0001] This case is a divisional application of the original application, with original patent number 202111657428X; original patent title: Application of 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative in the preparation of a drug for treating obesity; original application date: December 30, 2021, and the priority date of the original application was July 12, 2021. Technical Field

[0002] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of a 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative in the preparation of drugs for the treatment and prevention of obesity and related metabolic diseases. Background Technology

[0003] Obesity is a chronic metabolic disease caused by a combination of factors, including genetics, biological behavior, and environment. It is characterized by an abnormally high proportion of body fat due to an increase in the size and number of fat cells, leading to excessive fat deposition in certain areas. With rapid global economic development, improved living standards, and the emergence of unhealthy dietary structures and lifestyles, obesity has become a global epidemic, with its prevalence increasing and its onset occurring at increasingly younger ages. According to the World Health Organization, in 2016, 39% of adults aged 18 and over worldwide were overweight, and 13% were obese. There were also 41 million overweight preschool children globally. Based on current trends, it is projected that by 2050, the obesity rate will increase to 60% for adult men, 40% for adult women, and 25% for children.

[0004] Therefore, obesity has become a serious global medical and social problem. Obese individuals have a 1-4 times higher incidence of diseases such as cerebral embolism, heart failure, coronary heart disease, and diabetes compared to those of normal weight, and their lifespan is shortened by 4 years. Approximately 8% of all deaths worldwide each year are due to obesity. Furthermore, obesity triggers a series of related metabolic diseases, including hyperlipidemia, hypertension, non-alcoholic fatty liver disease, diabetes, and atherosclerosis. Not only do obese individuals and their families suffer, but society as a whole also incurs significant economic costs due to obesity.

[0005] Although the exact origins of obesity are not fully understood, from a pathogenesis perspective, obesity can be divided into two main categories: simple obesity and secondary obesity. Simple obesity accounts for more than 95% of all obese individuals, and it can be further divided into idiopathic obesity and acquired obesity.

[0006] Currently, there are five main strategies for treating obesity: diet, exercise, behavioral therapy, drug therapy, and rehabilitation surgery. Among these, drug therapy is the primary clinical treatment for obesity and other obesity-related metabolic diseases. The weight-loss drugs currently on the market mainly include pancreatic lipase inhibitors, such as orlistat; appetite suppressants acting on the central nervous system, such as lorcaserin; and combination weight-loss drugs (Qsymia). Orlistat inhibits pancreatic lipase activity, thereby inhibiting the breakdown and absorption of fat from food, but it can cause steatorrhea, leading to a deficiency of fat-soluble vitamins, and there are reports of it causing liver damage. Appetite suppressants are restricted in their use due to adverse reactions in the nervous system. Although the central nervous system appetite suppressants lorcaserin and Qsymia have become new weight-loss drugs approved by the FDA in recent years, their safety uncertainties regarding the central nervous system and cardiovascular system remain. Furthermore, the efficacy of these weight-loss drugs is low, with only 1-5% weight loss per year compared to a placebo. Therefore, the choices in the current weight-loss drug market are limited. If a weight-loss drug with good efficacy and low adverse reactions were to emerge, its market would expand significantly.

[0007] 4-(4-Diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives, specifically including chloroquine, quinacrine dihydrochloride, and hydroxychloroquine, are a class of antimalarial drugs that can kill malaria parasites and are the first choice for controlling relapse and preventing malaria transmission in clinical practice. However, currently, these 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives are not used for the treatment and prevention of obesity and related metabolic diseases. This invention unexpectedly discovered that 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives, represented by chloroquine, quinacrine dihydrochloride, and hydroxychloroquine, have significant efficacy in the treatment or prevention of obesity and related metabolic diseases, and can be used to prepare drugs for the treatment or prevention of obesity and related metabolic diseases. Summary of the Invention

[0008] The purpose of this invention is to provide a novel use of a class of 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives for the prevention or treatment of obesity or obesity-related metabolic diseases, wherein the obesity includes simple obesity and secondary obesity; and the obesity-related metabolic diseases include non-alcoholic fatty liver disease and hyperlipidemia; specifically, it includes the following:

[0009] In a first aspect, the present invention provides the use of a 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative in the preparation of a drug for treating or preventing obesity. The use of a pharmaceutical composition comprising a 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative, or a pharmaceutically acceptable salt thereof, or a tautomer thereof, or a meso compound thereof, or a racemic mixture thereof, or an enantiomer thereof, or a diastereomer thereof, as an active ingredient, in the preparation of a drug for preventing or treating obesity, wherein the structural formula of the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative is shown in Formula (I).

[0010]

[0011] Wherein, R1 and R2 are selected from H, hydroxyl, and alkyl, respectively, or R1 and R2 can form an aromatic ring; R3 is selected from H, hydroxyl, and alkyl.

[0012] Preferably, the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative is selected from the following compounds:

[0013]

[0014] Preferably, pharmaceutically acceptable salts of the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives include quinacrine dihydrochloride, chloroquine phosphate, and hydroxychloroquine sulfate.

[0015] Preferably, the obesity includes simple obesity or pathological obesity.

[0016] Preferably, the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative, or a pharmaceutically acceptable salt thereof, or a tautomer thereof, or a racemic mixture thereof, or a racemic mixture thereof, or an enantiomer thereof, or a diastereomer thereof, or a pharmaceutical composition thereof as an active ingredient, is incorporated into a pharmaceutically acceptable carrier to form any pharmaceutically acceptable dosage form.

[0017] Preferably, the dosage form includes injections, tablets, capsules, granules, and pills.

[0018] Secondly, the present invention provides the use of a pharmaceutical composition comprising a 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative, or a pharmaceutically acceptable salt thereof, or a tautomer thereof, or a meso compound thereof, or a racemic compound thereof, or an enantiomer thereof, or a diastereomer thereof, or an active ingredient thereof, in the preparation of a medicament for the prevention or treatment of obesity-related metabolic diseases, wherein the structural formula of the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative is shown in Formula (I).

[0019]

[0020] Wherein, R1 and R2 are selected from H, hydroxyl, and alkyl, respectively, or R1 and R2 can form an aromatic ring; R3 is selected from H, hydroxyl, and alkyl.

[0021] Preferably, the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative is selected from the following compounds:

[0022]

[0023] Preferably, pharmaceutically acceptable salts of the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives include quinacrine dihydrochloride, chloroquine phosphate, and hydroxychloroquine sulfate.

[0024] Preferably, the obesity-related metabolic diseases include non-alcoholic fatty liver disease and hyperlipidemia.

[0025] Preferably, the 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivative, or a pharmaceutically acceptable salt thereof, or a tautomer thereof, or a racemic mixture thereof, or a racemic mixture thereof, or an enantiomer thereof, or a diastereomer thereof, or a pharmaceutical composition thereof as an active ingredient, is incorporated into a pharmaceutically acceptable carrier to form any pharmaceutically acceptable dosage form.

[0026] Preferably, the dosage form includes injections, tablets, capsules, granules, and pills.

[0027] The beneficial effects of this invention are: ① This invention provides the application of 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives in the preparation of drugs for treating and preventing obesity and obesity-related metabolic diseases. The 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives can significantly reduce the body weight of obese mice induced by high-fat diets and high-fructose drinking water, inhibiting 70% of the weight gain in mice; simultaneously, it does not affect food intake or water intake, demonstrating significant therapeutic effects; ② Chloroquine and hydroxychloroquine are antimalarial drugs that can kill malaria parasites, providing a basis for clinical control of relapse and prevention of malaria. The invention unexpectedly discovered that chloroquine and hydroxychloroquine can significantly reduce the body weight of obese mice induced by high-fat diet and high-fructose drinking water, and can be used to treat obesity and obesity-related metabolic diseases; ③ Quinacrine is a non-selective monoamine oxidase inhibitor, but it has been reported that monoamine oxidase inhibitors (such as Moclobemide, Selegiline, and Glibenclamide) can promote lipid droplet accumulation and fat formation, which can easily lead to obesity. However, the invention unexpectedly discovered that quinacrine has the effect of inhibiting obesity, achieving unexpected results. Attached Figure Description

[0028] Figure 1 The effect of quinacrine dihydrochloride on body weight in obese mice, among which, # p<0.05, ## p<0.01, ### p<0.001 vs. ND group; * p<0.05, ** p<0.01, *** p<0.001 vs. HFD group;

[0029] Figure 2 The effect of quinacrine dihydrochloride on food intake in mice, among which, ### p<0.001 vs. ND group;

[0030] Figure 3 The effect of quinacrine dihydrochloride on water intake in mice, among which, ## p<0.01, ### p<0.001 vs. ND group;

[0031] Figure 4 The effect of quinacrine dihydrochloride on energy intake in mice, among which, ### p<0.001 vs. ND group;

[0032] Figure 5 The effect of quinacrine dihydrochloride on blood lipids in mice, among which, # p<0.05, ## p<0.01, ### p<0.001 vs. ND group; * p<0.05, *** p<0.001 vs. HFD group;

[0033] Figure 6 The effects of quinacrine dihydrochloride on liver lipids in mice, among which ## p<0.01, ### p<0.001 vs. ND group; * p<0.05, ** p<0.01 vs. HFD group;

[0034] Figure 7 The effects of chloroquine and hydroxychloroquine sulfate on mouse body weight, among which, ### p<0.001 vs. ND group; * p<0.05, ** p<0.01 vs. HFD group;

[0035] Figure 8 The effects of chloroquine and hydroxychloroquine sulfate on food intake in mice, among which, ** p<0.01 vs. HFD group;

[0036] Figure 9 The effects of chloroquine and hydroxychloroquine sulfate on water intake in mice, among which, ### p<0.001 vs. ND group; *** p<0.001 vs. HFD group;

[0037] Figure 10 The effects of chloroquine and hydroxychloroquine sulfate on energy intake in mice, among which, ### p<0.001 vs. ND group; ** p<0.01, *** p<0.001 vs. HFD group;

[0038] Figure 11 The effects of chloroquine and hydroxychloroquine sulfate on blood lipids in mice, among which, ## p<0.01, ### p<0.001 vs. ND group; * p<0.05, ** p<0.01, *** p<0.001 vs. HFD group;

[0039] Figure 12 The effects of chloroquine and hydroxychloroquine sulfate on liver lipids in mice, among which, # p<0.05, ## p<0.01 vs. ND group; * p<0.05 vs. HFD group. Detailed Implementation

[0040] The scope of protection of this invention is described in detail below with reference to specific embodiments. However, it should be noted that the scope of protection of this invention is not limited to the following embodiments, and also protects the therapeutic effects of 4-(4-diethylamino-1-methylbutanamino)-7-chloroquinoline derivatives in all obesity and related metabolic diseases, including different dosage forms, dosages, and combination therapies. Any technical solutions derived by those skilled in the art based on the concept of this invention through logical analysis, deduction, and experimentation in the prior art are all within the scope of protection claimed by this invention.

[0041] In the following embodiments of the present invention, a feed containing 60% fat (Beijing Keao Xieli Feed Co., Ltd., D12492 high-fat feed) and 40% fructose water (Shandong Xiwang Sugar Industry Co., Ltd., crystalline fructose) were used to model obesity in mice.

[0042] Example 1: The therapeutic effect of quinacrine dihydrochloride on C57 obese mice.

[0043] 1. Laboratory animals, materials and their sources

[0044] Experimental animals: Six-week-old SPF-grade female C57BL / 6N mice, weighing 16–19g, were purchased from the Lanzhou Veterinary Research Institute of the Chinese Academy of Agricultural Sciences without any prior medication. They were housed in the animal facility of the Medical Laboratory Building at Lanzhou University for one week of acclimatization, provided with food and free access to distilled water, and then divided into groups for experiments.

[0045] Drug: Quinacrine dihydrochloride (QD, HPLC ≥ 98%) was purchased from Aladdin Biotechnology Co., Ltd.

[0046] 2. Preparation of the C57 mouse obesity model

[0047] Eighteen 6-week-old female C57BL / 6N mice (16–19 g) were housed in the experimental animal facility of Lanzhou University. After one week of acclimatization, the mice were randomly divided into two groups: a normal control group (n=3, weighing 17.13 ± 1.13 g) fed with standard rat and mouse maintenance diet and normal drinking water, and a model group (n=17.38 ± 0.97 g) fed with a high-fat diet and 40% fructose drinking water. After 8 weeks of continuous feeding, mice whose weight was 20% higher than the average weight of the normal control group were defined as successfully modeled obese mice.

[0048] 3. Grouping and Dosing

[0049] Eight weeks later, the weight of mice in the normal control group was 19.99±0.26g, and the weight of successfully modeled obese mice was 27.11±3.03g. Ten successfully modeled obese mice were divided into two groups: the quinacrine dihydrochloride group (27.06±3.78g) and the high-fat control group (HFD group) (27.19±1.18g). The quinacrine dihydrochloride group continued to be fed a high-fat diet and 40% fructose water in subsequent experiments. The normal control group (ND group) was fed a standard rat diet and normal drinking water. Mice in the quinacrine dihydrochloride group were administered quinacrine dihydrochloride (35mg / kg) by gavage once daily, with the quinacrine dihydrochloride dissolved in physiological saline before use. Mice in the high-fat control group (HFD group) and the normal control group (ND group) were given the same amount of physiological saline daily for a total of 6 weeks. Mice were fasted for 12 hours at the same time each week. Fasting weight was recorded, and daily food and water intake was measured. Energy obtained from food and water was calculated.

[0050] 4. Data Processing

[0051] Experimental data were analyzed using SPSS 23.0 software. Data are expressed as (x±s). One-way ANOVA and LSD-t method were used for pairwise comparisons between groups. p<0.05 was considered statistically significant.

[0052] 5. Results Analysis

[0053] (1) Results of fasting body weight observation in mice:

[0054] Fasting body weight of mice as follows Figure 1 As shown, the initial average fasting body weight of mice in the normal control group was 19.99±0.26g, the initial average fasting body weight of mice in the high-fat control group was 27.19±1.18g, and the initial average fasting body weight of mice in the quinacrine dihydrochloride group was 27.06±3.78g. There was no significant difference in body weight between the high-fat control group and the quinacrine dihydrochloride group. After 6 weeks of administration, the average fasting body weight of mice in the normal control group was 21.3±0.32g, an increase of 6.55% compared with the initial average fasting body weight; the high-fat control group... The average fasting body weight of mice was 31.25±1.53g, an increase of 14.93% compared with the initial average fasting body weight; while the average fasting body weight of mice in the quinacrine dihydrochloride group was 23.84±1.00g, a decrease of 11.9% compared with the initial average fasting body weight. Compared with the high-fat control group, quinacrine dihydrochloride significantly inhibited the weight gain of mice and significantly reduced the body weight of mice (P<0.001).

[0055] (2) Results of observation on food intake in mice:

[0056] The results of mouse food intake are as follows Figure 2 As shown, the food intake of mice in the normal control group was 2.64±0.05 g / mouse / day, the food intake of mice in the high-fat control group was 1.72±0.14 g / mouse / day, and the food intake of mice in the quinacrine dihydrochloride group was 1.76±0.13 g / mouse / day. There was no significant difference in food intake between the high-fat control group and the quinacrine dihydrochloride group (P>0.05), but the food intake of both groups was significantly lower than that of the normal control group (P<0.001). This is because the high-fat diet provides much more energy than the normal diet, and mice only need to consume a smaller amount of food to obtain the same amount of energy. The results indicate that quinacrine dihydrochloride has no effect on the food intake of mice.

[0057] (3) Results of observation on water intake in mice

[0058] The results of water intake in mice are as follows Figure 3As shown, the water intake of mice in the normal control group was 5.26±0.25 mL / mouse / day, the water intake of mice in the high-fat control group was 3.96±0.60 mL / mouse / day, and the water intake of mice in the quinacrine dihydrochloride group was 3.57±0.45 mL / mouse / day. Pairwise comparisons showed that the water intake of mice in the high-fat control group and the quinacrine dihydrochloride group was significantly lower than that of the normal control group (P<0.01 and P<0.001, respectively). This is because the 40% fructose water provides more energy than ordinary water, so mice only need to consume less water to survive. Furthermore, the water intake of mice in the quinacrine dihydrochloride group showed a slightly lower trend than that of the high-fat control group, indicating a reduction in energy intake from the 40% fructose water. However, there was no significant difference in water intake between the high-fat control group and the quinacrine dihydrochloride group (P>0.05). The results showed that quinacrine dihydrochloride had no significant effect on water intake in mice.

[0059] (4) Results of observation of energy intake in mice

[0060] The results of energy intake in mice are as follows: Figure 4 As shown, the energy intake of mice in the normal control group was 8.98±0.18 kcal / mouse / day, the energy intake of mice in the high-fat control group was 15.34±1.40 kcal / mouse / day, and the energy intake of mice in the quinacrine dihydrochloride group was 14.92±1.22 kcal / mouse / day. Pairwise comparisons showed that the energy intake of the high-fat control group and the quinacrine dihydrochloride group was significantly higher than that of the normal control group (P<0.001). This is because the high-fat diet and 40% fructose water provided significantly more energy than the normal diet and drinking water. There was no significant difference in energy intake between the high-fat control group and the quinacrine dihydrochloride group (P>0.05). The results indicate that quinacrine dihydrochloride has no effect on the energy intake of mice.

[0061] In conclusion, quinacrine dihydrochloride can significantly reduce the weight of obese mice, and achieves a significant weight loss effect without reducing energy intake or food intake, showing good prospects for clinical application.

[0062] (5) Detection of lipid-related biochemical indicators in mice

[0063] After the experiment, blood was collected via cardiac puncture and left at room temperature for 1 hour. The blood was then centrifuged at 860g and 4℃ for 15 minutes, and the supernatant serum was collected. Serum triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels were measured using kits (Nanjing Jiancheng Bioengineering Institute, catalog numbers A110-1, A111-1, A112-1, and A113-1).

[0064] The test results for triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) are as follows: Figure 5 As shown in the AD diagram, the serum TG, TC, HDL-C, and LDL-C levels in the ND group mice were 0.86±0.20 mmol / L, 1.63±0.05 mmol / L, 1.72±0.21 mmol / L, and 0.19±0.02 mmol / L, respectively. In the HFD group mice, the serum TG, TC, HDL-C, and LDL-C levels were 1.60±0.69 mmol / L, 3.52±0.37 mmol / L, 3.07±0.12 mmol / L, and 0.31±0.08 mmol / L, respectively. In the Quinacrine group mice, the serum TG, TC, HDL-C, and LDL-C levels were 1.00±0.30 mmol / L, 3.22±0.20 mmol / L, 2.81±0.18 mmol / L, and 0.06±0.02 mmol / L, respectively. Compared with mice in the normal diet group, mice fed a high-fat diet showed significantly increased serum TG, TC, HDL-C, and LDL-C levels, forming a significant difference. However, mice treated with quinacrine dihydrochloride showed improved serum TG, TC, HDL-C, and LDL-C levels, indicating that quinacrine dihydrochloride can regulate the lipid content in the serum of obese mice to a certain extent and prevent the occurrence of hyperlipidemia.

[0065] (6) Detection of lipid levels in mouse liver

[0066] Prepare a 10% homogenate of liver tissue (liver tissue: phosphate buffer = 100 mg: 0.9 mL) according to the kit instructions, and determine the triglyceride and total cholesterol content in the liver.

[0067] The results of the triglyceride and total cholesterol tests are as follows: Figure 6As shown in Figures AB, the liver TG and TC levels in the ND group mice were 1.48±0.63 mmol / L and 1.87±1.02 mmol / L, respectively; in the HFD group mice, they were 3.85±1.23 mmol / L and 4.05±1.08 mmol / L, respectively; and in the Quinacrine group mice, they were 1.95±0.65 mmol / L and 2.56±0.47 mmol / L, respectively. Compared with mice on a normal diet, the liver triglyceride and total cholesterol levels in mice fed a high-fat, high-sugar diet were significantly increased, while the levels of triglycerides and total cholesterol in the liver of mice treated with quinacrine dihydrochloride were significantly reduced. Therefore, the experimental results indicate that quinacrine dihydrochloride can effectively regulate excessive lipid accumulation in the liver caused by obesity, bringing lipid levels in liver tissue back to normal and preventing fatty liver disease.

[0068] Example 2: The therapeutic effects of chloroquine and hydroxychloroquine sulfate on C57 obese mice.

[0069] 1. Laboratory animals, materials and their sources

[0070] Experimental animals: Six-week-old SPF-grade female C57BL / 6N mice, weighing 16–19g, were purchased from the Lanzhou Veterinary Research Institute of the Chinese Academy of Agricultural Sciences without any prior medication. They were housed in the animal facility of the Medical Laboratory Building at Lanzhou University for one week of acclimatization, provided with food and free access to distilled water, and then divided into groups for experiments.

[0071] Drugs: Chloroquine (HPLC ≥ 97%) was purchased from McLlin Biochemical Technology Co., Ltd., and hydroxychloroquine sulfate (HCQ, HPLC ≥ 98%) was purchased from Aladdin Biotechnology Co., Ltd.

[0072] 2. Preparation of the C57 mouse obesity model

[0073] Thirty-five 6-week-old female C57BL / 6N mice (16–19 g) were housed in the experimental animal facility of Lanzhou University. After one week of acclimatization, the mice were randomly divided into two groups: a normal control group (n=6, weighing 18.96 ± 1.03 g) fed with standard rat and mouse diet and normal drinking water, and a model group (n=19.20 ± 0.85 g) fed with a high-fat diet and 40% fructose drinking water to induce obesity. After 11 weeks, mice with a weight 20% higher than the normal control group were selected as successfully modeled obese mice.

[0074] 3. Grouping and Dosing

[0075] After 11 weeks, the weight of mice in the normal control group was 21.50±0.80g, and the weight of mice that successfully developed the obese model was 26.11±2.30g. The 18 successfully modeled obese mice were divided into three groups: the chloroquine group (25.71±1.88g), the hydroxychloroquine sulfate group (HCQ group) (26.07±2.19g), and the high-fat control group (HFD group) (26.56±2.67g). In subsequent experiments, the mice were still fed with high-fat diet and 40% fructose water. The normal control group (ND group) mice were fed with normal rat and mouse maintenance diet and normal drinking water. Mice in the chloroquine group were administered chloroquine (15 mg / kg) by gavage once daily. The chloroquine was prepared as a suspension in physiological saline before use. Mice in the hydroxychloroquine sulfate group (HCQ group) were administered hydroxychloroquine sulfate (20 mg / kg) by gavage once daily. The hydroxychloroquine sulfate was dissolved in physiological saline before use. Mice in the high-fat control group (HFD group) and the normal control group (ND group) were given the same amount of physiological saline daily for a total of 11 weeks. Mouse body weight was recorded at the same time each week, and daily food intake and water intake were measured to calculate the energy obtained from food and water.

[0076] 4. Data Processing

[0077] Experimental data were analyzed using SPSS 23.0 software. Data are expressed as (x±s). One-way ANOVA and LSD-t method were used for pairwise comparisons between groups. p<0.05 was considered statistically significant.

[0078] 5. Results Analysis

[0079] (1) Results of mouse body weight observation:

[0080] The results are as follows Figure 7As shown, the initial average weight of mice in the ND group was 21.50 g, the initial average weight of mice in the HFD group was 26.56 g, the initial average weight of mice in the Chloroquine group was 25.71 g, and the initial average weight of mice in the HCQ group was 26.07 g. There was no significant difference in weight among the HFD, Chloroquine, and HCQ groups. After 11 weeks of administration, the weight of mice in the ND group was 23.16 ± 1.11 g, an increase of 7.77% compared to the initial weight; the weight of mice in the HFD group was 30.93 ± 3.55 g, an increase of 16.45% compared to the initial weight; while the average weight of mice in the Chloroquine group was 25.93 ± 2.80 g, an increase of only 0.86% compared to the initial weight; and the average weight of mice in the HCQ group was 25.90 ± 2.13 g, a decrease of 0.65% compared to the initial weight. Compared with the high-fat control group, chloroquine and hydroxychloroquine sulfate significantly inhibited the weight gain of mice. * P<0.05, * P<0.01).

[0081] (2) Results of observation on food intake in mice:

[0082] like Figure 8 As shown, the food intake of mice in the ND group was 2.93±0.18 g / mouse / day, the food intake of mice in the HFD group was 3.13±0.92 g / mouse / day, the food intake of mice in the Chloroquine group was 2.19±0.53 g / mouse / day, and the food intake of mice in the HCQ group was 1.96±1.02 g / mouse / day. There was no significant difference in food intake between the HFD and ND groups (P>0.05), but the food intake of mice in the HFD group was slightly higher than that in the ND group, while the food intake of mice in the Chloroquine and HCQ groups was significantly lower than that in the HFD group (P<0.01). These results indicate that chloroquine and hydroxychloroquine sulfate have an appetite-suppressing effect, which is one of the reasons why the weight gain of mice treated with chloroquine and hydroxychloroquine sulfate was inhibited.

[0083] (3) Results of observation on water intake in mice

[0084] The results are as follows Figure 9As shown, the water intake of mice in the ND group was 4.43±0.21 mL / mouse / day, the HFD group was 3.39±0.34 mL / mouse / day, the Chloroquine group was 2.67±0.26 mL / mouse / day, and the HCQ group was 2.53±0.18 mL / mouse / day. Compared pairwise, the water intake of mice in the HFD group was significantly lower than that in the ND group (P<0.001). This is because the 40% fructose water provides more energy than ordinary water, so mice only need to consume less water to survive. Conversely, the water intake of mice in the Chloroquine and HCQ groups was significantly less than that in the HFD group (P<0.001), indicating reduced energy intake from the 40% fructose water and inhibited weight gain.

[0085] (4) Results of observation of energy intake in mice

[0086] The results are as follows Figure 10 As shown, the energy intake of mice in the ND group was 10.00±0.65 kcal / mouse / day, the energy intake of mice in the HFD group was 21.67±4.51 kcal / mouse / day, the energy intake of mice in the Chloroquine group was 15.56±2.60 kcal / mouse / day, and the energy intake of mice in the HCQ group was 14.10±5.45 kcal / mouse / day. Compared pairwise, the energy intake of the HFD group was significantly higher than that of the normal control group (P<0.001). This is because the high-fat diet and 40% fructose water provided far more energy than the normal diet and drinking water, which explains why the mice in the HFD group remained obese and continued to gain weight. Conversely, the energy intake of mice in the Chloroquine and HCQ groups was significantly lower than that of the HFD group (P<0.01, P<0.001). These results indicate that chloroquine and hydroxychloroquine sulfate can reduce energy intake in mice, thereby combating obesity.

[0087] (5) Detection of lipid-related biochemical indicators in mice

[0088] After the experiment, blood was collected via cardiac puncture. After being left at room temperature for 1 hour, the blood was centrifuged at 860g and 4℃ for 15 minutes to obtain the supernatant serum. Serum levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured using kits (Nanjing Jiancheng Bioengineering Institute, catalog numbers A110-1, A111-1, A112-1, and A113-1). The results of triglyceride (TG), total cholesterol (TC), HDL-C, and LDL-C measurements are shown below. Figure 11 As shown in the AD diagram, the serum TG, TC, HDL-C, and LDL-C levels in the ND group mice were 1.17±0.20 mmol / L, 0.72±0.13 mmol / L, 3.21±0.48 mmol / L, and 0.40±0.13 mmol / L, respectively. In the HFD group mice, the serum TG, TC, HDL-C, and LDL-C levels were 1.55±0.04 mmol / L, 1.89±0.30 mmol / L, 4.95±0.43 mmol / L, and 0.99±0.16 mmol / L, respectively. The serum TG, TC, HDL-C, and LDL-C levels in the hloroquine group mice were 1.23±0.19 mmol / L, 1.33±0.21 mmol / L, 3.96±0.97 mmol / L, and 0.56±0.14 mmol / L, respectively. The serum TG, TC, HDL-C, and LDL-C levels in the HCQ group mice were 1.49±0.06 mmol / L, 1.76±0.17 mmol / L, 4.44±0.17 mmol / L, and 0.81±0.15 mmol / L, respectively.

[0089] Compared to mice on a normal diet, mice fed a high-fat diet showed significantly increased serum TG, TC, HDL-C, and LDL-C levels, indicating a substantial difference. In contrast, mice treated with chloroquine showed improved serum TG, TC, HDL-C, and LDL-C levels, significantly lower than the HFD group. This suggests that chloroquine can regulate serum lipid levels in obese mice to some extent, preventing hyperlipidemia. Mice treated with hydroxychloroquine sulfate also showed varying degrees of reduction in serum TG, TC, HDL-C, and LDL-C levels, indicating that hydroxychloroquine sulfate also exhibits a lipid-regulating effect.

[0090] (6) Detection of lipid levels in mouse liver

[0091] Prepare a 10% liver tissue homogenate medium (liver tissue: phosphate buffer = 100 mg: 0.9 mL) according to the kit instructions, and determine the triglyceride and total cholesterol content in the liver. The results of the triglyceride and total cholesterol content detection are as follows: Figure 12As shown in Figures AB, the liver TG and TC levels in the ND group mice were 0.46±0.18 mmol / L and 1.69±0.30 mmol / L, respectively; in the HFD group mice, they were 1.07±0.18 mmol / L and 2.64±0.86 mmol / L, respectively; in the Chloroquine group mice, they were 0.98±0.34 mmol / L and 1.82±0.50 mmol / L, respectively; and in the HCQ group mice, they were 1.25±0.27 mmol / L and 1.80±0.29 mmol / L, respectively. Compared with mice on a normal diet, the liver triglyceride and total cholesterol levels in mice fed a high-fat, high-sugar diet were significantly increased. After chloroquine treatment, there was no significant difference in liver triglyceride levels compared to the HFD group, but the total cholesterol content in the liver was significantly decreased. The total cholesterol content in the liver of mice treated with hydroxychloroquine sulfate was also significantly decreased. Experimental results show that chloroquine and hydroxychloroquine sulfate can, to some extent, inhibit excessive accumulation of lipids in the liver caused by obesity, especially cholesterol accumulation, bringing cholesterol levels in liver tissue closer to normal and reducing the risk of fatty liver disease.

Claims

1. Use of quinacrine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of non-alcoholic fatty liver, characterized in that, The structural formula of the quinolinic acid is shown as formula (I), 2. Use according to claim 1, wherein The pharmaceutically acceptable salt of the quinolinic acid is quinolinic acid dihydrochloride.

3. Use according to claim 1 or 2, characterized in that, The quinolinic acid or its pharmaceutically acceptable salt is added into a pharmaceutically acceptable carrier to form a pharmaceutically acceptable dosage form.

4. Use according to claim 3, characterized in that, The dosage form includes injection, tablet, capsule, granule and pill.

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