Use of copper-containing compounds for the preparation of a medicament for the prevention and / or treatment of obesity and its associated metabolic disorders

Abstract

The application discloses application of a copper-containing compound in preparation of a medicine for preventing and / or treating obesity and related metabolic diseases. The application breaks through the traditional cognition on safety of high-dose copper, and proves that oral administration of the copper-containing compound (such as copper chloride) in a specific dose range (calculated according to copper element, such as 0.07-2.84 mg / kg body weight per day) can effectively inhibit weight growth of animals induced by a high-fat diet, and simultaneously has an effect on liver steatosis and body composition, without risk of liver toxicity. Compared with existing weight loss methods, the application scheme has the advantages of low raw material price, convenient oral use, wide safety window, remarkable comprehensive effect and the like, and provides a clear direction and a solid experimental basis for development of a new, economical and safe medicine or health care food for preventing and treating obesity.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more specifically, to the use of copper-containing compounds in the preparation of medicaments for the prevention and / or treatment of obesity and related metabolic diseases. Background Technology

[0002] Obesity is a chronic metabolic disease caused by multiple factors, characterized by excessive accumulation and / or abnormal distribution of body fat. Its prevalence is mainly attributed to changes in dietary structure (increased intake of high-fat and high-sugar foods) and reduced physical activity. Obesity is also a significant risk factor for type 2 diabetes, hypertension, dyslipidemia, non-alcoholic fatty liver disease, cardiovascular disease, and certain cancers. Therefore, developing effective methods for the prevention and treatment of obesity has significant clinical and social value.

[0003] Currently, obesity prevention and treatment mainly include lifestyle interventions (dietary control and exercise), drug therapy, and metabolic surgery. However, these methods all have significant limitations: lifestyle interventions rely on long-term individual adherence, resulting in poor compliance, low success rates, and a high risk of rebound. Existing drugs (such as the GLP-1 receptor agonist semaglutide) are effective, but they are generally expensive, require injection, and may cause gastrointestinal discomfort and other side effects. Furthermore, their long-term safety requires more data to support its widespread application. Metabolic surgery, while highly effective, is invasive, carries surgical risks, and carries potential complications such as malabsorption, making it only suitable for severely obese patients. Therefore, developing a novel weight management strategy that is inexpensive, convenient to take orally, and safe and effective has enormous market demand and clinical significance.

[0004] Copper is an essential trace element for the human body. As a cofactor for many key enzymes (such as cytochrome C oxidase, superoxide dismutase, and lysyl oxidase), it participates in core physiological processes such as energy metabolism, antioxidant defense, connective tissue formation, and iron metabolism. The Chinese Nutrition Society has established a Recommended Nutrient Intake (RNI) of 0.8 mg per day for adults and a Tolerable Upper Intake Level (UL) of 8.0 mg per day. This dosage range is primarily intended to meet basic physiological needs and prevent deficiency. However, the relationship between copper and obesity remains highly uncertain with current technology. Population-based observational studies have not established a positive association between copper supplementation and weight loss. Some studies have found that serum, visceral fat, and liver copper levels in obese patients are significantly higher than in people of normal weight. Data from the Chinese Nutrition Survey show that dietary copper content and obesity risk exhibit a U-shaped curve, with both excessively low and high levels increasing the risk (Front Nutr 2022;9:1009721). These observational data even suggest that elevated copper levels coexist with obesity, making it difficult for those skilled in the art to be motivated to believe that "copper supplementation can treat obesity."

[0005] Furthermore, existing technologies contain clear safety warnings and technological biases. The conventional wisdom holds that excessive copper (especially inorganic copper) may lead to liver accumulation, oxidative stress, and organ damage; these safety concerns have severely hampered its exploration in high-dose treatment of obesity.

[0006] In addition, existing sporadic studies have mostly focused on copper deficiency supplementation or low-dose effects, failing to identify a dose window that combines significant therapeutic efficacy with a clear safety margin.

[0007] Therefore, there is an urgent need in this field to solve the technical problem of how to safely utilize the biological activity of copper to intervene in obesity. Summary of the Invention

[0008] The purpose of this invention is to overcome the shortcomings of existing obesity treatment methods and provide a novel, safe and effective weight loss solution based on copper-containing compounds.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] In a first aspect, the present invention discloses the use of copper-containing compounds in the preparation of medicaments for the prevention and / or treatment of obesity and related metabolic diseases, wherein the dosage of the copper-containing compound, calculated in terms of elemental copper, is 0.07-2.84 mg / kg body weight per day; and the obesity and related metabolic diseases are induced by a high-fat diet.

[0011] Furthermore, the copper-containing compound is selected from one or more of soluble inorganic copper salts and copper gluconate.

[0012] Furthermore, the copper-containing compound is selected from copper chloride (CuCl2) or copper sulfate (CuSO4), more preferably copper chloride (CuCl2).

[0013] Furthermore, the copper-containing compound reduces body weight or inhibits abnormal weight gain, lowers serum triglycerides (TG) and / or total cholesterol (TC), increases high-density lipoprotein cholesterol (HDL-C), alleviates hepatic steatosis, reduces hepatic transaminase (ALT and / or AST) activity, reduces body fat percentage, and increases lean body mass ratio.

[0014] Furthermore, the dosage of the copper-containing compound, calculated in terms of elemental copper, is 2.84 mg / kg body weight per day.

[0015] Furthermore, the administration method is oral.

[0016] In a second aspect, the present invention provides the use of copper-containing compounds in the preparation of medicaments for alleviating hepatic steatosis or improving non-alcoholic fatty liver disease (NAFLD), wherein the copper-containing compounds are selected from one or more of soluble inorganic copper salts and copper gluconate.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] This invention reveals that copper supplementation in a high-fat diet-induced obesity animal model not only sustainably and significantly inhibits weight gain (with a maximum weight loss exceeding 20% ​​in the later stages of intervention), but also systematically corrects multiple metabolic abnormalities, significantly reducing the degree of hepatic steatosis and decreasing hepatic transaminase (ALT / AST) activity. This provides an important theoretical basis and experimental evidence for the subsequent development of novel weight loss strategies based on copper homeostasis regulation.

[0019] Furthermore, this invention successfully overcomes the long-standing technical prejudice that "high-dose copper is hepatotoxic," establishing for the first time a safe and broad therapeutic window. Existing technologies generally believe that inorganic copper supplementation easily leads to copper accumulation in the liver and induces oxidative stress damage. However, this invention, through rigorous animal experiments, demonstrates that even with relatively high doses (e.g., 6 mg / kg / day of copper chloride, equivalent to 2.84 mg / kg / day of elemental copper) administered orally for six consecutive weeks, the liver copper content and liver function indicators (ALT, AST) in the experimental animals did not show abnormal increases, with no significant difference compared to the control group. This finding strongly demonstrates that within this dose range, the body is fully capable of effectively metabolizing and excreting ingested copper without causing liver damage, thus removing the most critical safety obstacle for the clinical application of copper-containing compounds. Attached Figure Description

[0020] Figure 1 The effect of different doses of copper chloride on the body weight of mice fed a standard diet.

[0021] Figure 2 The results of liver copper content and liver function indicators (ALT, AST) in mice fed with standard diet were obtained after 6 weeks of gavage administration of high dose (6 mg / kg / day) of copper chloride.

[0022] Figure 3 The results show the effect of high-dose (6 mg / kg / day) copper chloride gavage on the body weight of mice fed a standard diet after 6 weeks.

[0023] Figure 4 Images and weight changes of mice fed a high-fat diet after copper chloride intervention are shown. A is an image of male mice, B is a weight gain curve of male mice, C is an image of female mice, and D is a weight gain curve of female mice.

[0024] Figure 5 The images show histopathological sections of liver tissue from mice on a high-fat diet after copper chloride intervention. The scale bar is 250 μm.

[0025] Figure 6The effect of copper chloride intervention on liver function in mice fed a high-fat diet is shown in Figure 1. A represents serum ALT activity in male mice, B represents serum AST activity in male mice, C represents serum ALT activity in female mice, and D represents serum AST activity in female mice.

[0026] Figure 7 The effect of copper chloride intervention on body composition (fat-to-body weight ratio and lean-to-body weight ratio) in mice fed a high-fat diet is shown in Figure 1. A represents the fat-to-body weight ratio of male mice, B represents the lean-to-body weight ratio of male mice, C represents the fat-to-body weight ratio of female mice, and D represents the lean-to-body weight ratio of female mice.

[0027] Figure 8 The effect of copper chloride intervention on the diameter of adipocytes in mice on a high-fat diet is shown. The scale bar is 250 μm. Detailed Implementation

[0028] The present invention is further illustrated below through specific embodiments, but the scope of protection of the present invention is not limited thereto.

[0029] Example 1: Safety verification of copper-containing compounds (taking CuCl2 as an example)

[0030] This embodiment verifies whether oral administration of copper-containing compounds, such as copper chloride, under the preferred dosage conditions of the present invention will lead to copper accumulation in the liver and liver function damage, thereby evaluating its basic safety for oral weight loss.

[0031] Wild-type male C57BL / 6 mice were selected as experimental animals. The mice were 6-8 weeks old, with an initial weight of 18-25g, and were purchased from Hangzhou Ziyuan Experimental Animal Technology Co., Ltd.

[0032] Mice were acclimatized for 7 days in an SPF-grade environment with the following conditions: temperature 20-26℃, humidity 30-70%, 12 / 12-hour light cycle, and free access to food and water, fed a standard diet. The experimental animals were randomly divided into two groups of 10 each. The treatment group received 6 mg / kg / day of copper chloride (CuCl2) via gavage (corresponding to a copper element dose of 2.84 mg / kg / day), once daily for 6 weeks. The control group received an equal volume of water via gavage, with all other conditions the same.

[0033] After drug administration, the mice were euthanized, and liver tissue was collected for the detection of liver copper content and related liver function indicators.

[0034] (1) Detection of copper content in the liver

[0035] Approximately 50 mg of liver tissue was collected, and the copper content in the tissue was detected using ICP-MS (inductively coupled plasma mass spectrometry).

[0036] (2) Liver function index testing

[0037] A portion of liver tissue was taken and added to physiological saline to prepare a 10% tissue homogenate. After centrifugation, the supernatant was collected, and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using a fully automated biochemical analyzer (HITACHI 3100).

[0038] All data are expressed as mean ± standard deviation. Statistical analysis was performed using SPSS software, and independent samples t-tests were used for comparisons between two groups. A p-value < 0.05 was considered statistically significant.

[0039] The above results are as follows Figure 2 As shown in the figure; ICP-MS detection results showed that there was no significant difference in copper content in the liver tissue of mice in the CuCl2 treatment group and the control group (P > 0.05); the detection results of ALT and AST in liver tissue homogenate showed that there was no significant difference in ALT and AST levels between the CuCl2 treatment group and the control group (P > 0.05).

[0040] The above results indicate that, under standard feed conditions, oral administration of CuCl2 at a dose of 6 mg / kg / day for 6 consecutive weeks did not lead to copper accumulation in the liver, suggesting that this dose did not exceed the body's copper excretion capacity and did not impair liver function, demonstrating good basic safety.

[0041] At the same time, such as Figure 3 As shown, under standard feed conditions, compared with the control group, continuous gavage administration of CuCl2 6 mg / kg / day for 6 weeks reduced the body weight of mice.

[0042] Example 2: Intervention effect of copper-containing compounds on obesity induced by a high-fat diet

[0043] This embodiment was used to investigate the effect of copper-containing compounds (taking copper chloride as an example) on weight loss in high-fat diet (HFD) induced obese mice and its effects on hepatic steatosis, liver function and body composition.

[0044] C57BL / 6 mice of the same age and strain as those in Example 1, including both males and females, were selected. They were fed a high-fat diet (HFD, energy ratio: 60% fat) for 9 consecutive weeks to establish an obesity model. A group of age-matched mice was fed a normal diet as a baseline control (Chow group).

[0045] After successful modeling, the HFD-fed mice were randomly divided into two groups of 10 each, and continued to be fed the HFD diet.

[0046] Model control group (HFD): Administered an equal volume of purified drinking water by gavage once daily;

[0047] The treatment group (HFD+CuCl2): 6 mg / kg / day CuCl2 was administered by gavage once daily.

[0048] The intervention lasted for 10 weeks. The following indicators were measured during or after the intervention:

[0049] (1) Body weight: During the entire intervention period, the mice were weighed at a fixed time every week, and the relative body weight change was calculated based on the body weight at the beginning of the intervention (recorded as 100%), and a body weight growth curve was plotted.

[0050] like Figure 4 As shown, the HFD-fed mice gained weight significantly faster than the Chow group. CuCl2 intervention significantly inhibited HFD-induced weight gain, and was effective in both sexes, with particularly significant effects on male mice. At the end of the intervention (day 133), the body weight of male mice in the HFD+CuCl2 group was 18.58±7.06% less than that of the HFD model control group; the body weight of female mice was 18.66±12.31% less.

[0051] (2) Liver histopathology: After the intervention, liver tissues of mice in each group were taken, fixed with 4% paraformaldehyde, embedded in paraffin, sectioned, and stained with hematoxylin-eosin (HE); another portion of liver tissue was taken, embedded in OCT, frozen sectioned, and stained with Oil Red O. The histopathological changes and lipid deposition of liver tissues were observed under a light microscope.

[0052] like Figure 5 As shown, the livers of mice in the HFD model control group exhibited significant hepatocyte steatosis, with large and numerous lipid droplet vacuoles and large, dark-colored Oil Red O staining positive areas. In contrast, the HFD+CuCl2 group showed a significant reduction in hepatocyte steatosis, with smaller and fewer lipid droplets and a marked decrease in Oil Red O staining positive areas. This indicates that CuCl2 intervention can effectively alleviate HFD-induced hepatic steatosis.

[0053] (3) Liver function indicators: After the intervention, liver tissue was taken to prepare homogenate and ALT and AST levels were detected, using the same method as in Example 1.

[0054] like Figure 6 As shown, in male mice, HFD feeding significantly increased serum AST activity, and ALT activity also showed an increasing trend. After CuCl2 intervention, both AST and ALT activities significantly decreased, recovering to levels close to those of the Chow group. There were no significant differences in transaminase levels among the groups in female mice.

[0055] (4) Body composition analysis: After the intervention, the fat-to-body weight ratio and lean-to-body weight ratio of live mice were detected using a small animal nuclear magnetic resonance body composition analyzer (NIUMAGQMR06-EDU).

[0056] like Figure 7 As shown, HFD feeding significantly increased the fat-to-body-weight ratio and decreased the lean-to-body-weight ratio in mice. CuCl2 intervention significantly reduced the fat-to-body-weight ratio in male mice and correspondingly increased the lean-to-body-weight ratio, indicating its effect on improving body composition. The effects on female mice showed a consistent trend but did not reach statistical significance.

[0057] (5) Adipocyte morphology: After the intervention, the epididymal fat pad (male) or inguinal fat (female) of mice was taken, fixed, embedded, sectioned and stained with HE, and the adipocyte diameter was observed and measured under a microscope.

[0058] like Figure 8 As shown, the adipocyte diameter in the HFD model control group mice was significantly larger than that in the Chow group. After CuCl2 intervention, the adipocyte diameter decreased significantly, approaching the normal level, indicating that it can effectively inhibit HFD-induced adipocyte hypertrophy.

[0059] Data are presented as mean ± standard deviation. One-way ANOVA was used for comparisons among multiple groups, followed by Tukey's multiple comparison test. A p-value < 0.05 was considered statistically significant.

[0060] In summary, CuCl2 intervention can effectively combat HFD-induced obesity and related metabolic disorders, manifested in inhibiting weight gain, reducing hepatic steatosis, improving liver function, optimizing body composition, and shrinking fat cells.

[0061] Based on a renewed understanding of the biological effects of copper and successful exploration of safe dosage, this invention proposes for the first time a novel use of copper-containing compounds (especially soluble inorganic copper salts) in the preparation of safe and effective oral weight-loss products. This approach, with its comprehensive advantages of low cost, high efficacy, convenient oral administration, and well-established safety profile, provides a promising new strategy for the prevention and treatment of obesity and its complications.

[0062] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of copper-containing compounds in the preparation of medicaments for the prevention and / or treatment of obesity and related metabolic diseases, characterized in that, The dosage of the copper-containing compound, calculated as elemental copper, is 0.07-2.84 mg / kg body weight per day; the obesity and related metabolic diseases are induced by a high-fat diet, and the administration route is oral.

2. The application according to claim 1, characterized in that, The copper-containing compound is selected from one or more of soluble inorganic copper salts and copper gluconate.

3. The application according to claim 2, characterized in that, The copper-containing compound is selected from copper chloride or copper sulfate.

4. The application according to claim 3, characterized in that, The copper-containing compound is copper chloride.

5. The application according to claim 1, characterized in that, The copper-containing compounds reduce weight or inhibit abnormal weight gain, lower serum triglycerides and / or total cholesterol, increase high-density lipoprotein cholesterol, alleviate hepatic steatosis, reduce hepatic transaminase activity, reduce body fat percentage, and increase lean body mass ratio.

6. The application according to claim 1, characterized in that, The dosage of the copper-containing compound, calculated in terms of elemental copper, is 2.84 mg / kg body weight per day.

7. The use of copper-containing compounds in the preparation of medicaments for alleviating hepatic steatosis or improving non-alcoholic fatty liver disease, characterized in that, The copper-containing compound is selected from one or more of soluble inorganic copper salts and copper gluconate.

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