Use of cucurbitacin C in the preparation of a product for preventing and / or improving liver injury and related metabolic disorders

Abstract

The application belongs to the field of biological medicine, and particularly relates to application of cucurbitacin C in preparation of products for preventing and / or improving liver injury and related metabolic disorders. The application provides a new application of cucurbitacin C, which can effectively reduce liver injury and related metabolic disorders caused by high-fat diet, and provides a new active material selection for preparing products with liver protection and dual functions of glycolipid metabolism regulation.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of cucurbitacin C in the preparation of products for the prevention and / or improvement of liver damage and related metabolic disorders. Background Technology

[0002] Long-term high-fat intake can lead to a series of metabolic pathological changes, including elevated fasting blood glucose, impaired glucose tolerance, insulin resistance, abnormal lipid profiles, and hepatocellular damage. Further progression can result in glucose and lipid metabolism disorders and liver damage. Currently, interventions for glucose and lipid metabolism disorders and liver damage caused by high-fat diets mainly include lifestyle modifications and the use of various hypoglycemic / lipid-lowering or hepatoprotective drugs. However, these solutions still have significant room for improvement in terms of the clarity of active ingredients, the selectivity of targets, and the safety of long-term use.

[0003] Cucurbitacins belong to the highly oxidized cucurbitane-type tetracyclic triterpenoids. Different cucurbitacin monomers exhibit significant differences in chemical structure. Specifically, all cucurbitacins belong to the cucurbitane-type tetracyclic triterpenoid parent nucleus, with distinct differences in hydroxyl groups, acetyl substitution sites, double bond positions, and stereoconfigurations, leading to substantial variations in physicochemical properties and in vivo pharmacodynamic activities. For example, cucurbitacin C differs structurally from common family members such as cucurbitacin B and cucurbitacin E in the A-ring functional group, the structure of the corresponding C-19 region, and the degree of unsaturation. These structural differences result in different physicochemical properties and biological effects among the cucurbitacin monomers. Currently, relevant technologies have not established a universal structure-activity relationship for cucurbitacins in improving metabolic disorders, nor have they provided technical insights into the role of cucurbitacin C in improving glucose and lipid metabolism disorders, liver function damage, and liver histopathological changes caused by a high-fat diet.

[0004] Therefore, there is an urgent need to explore an active substance that is safe in composition and has dual functions of efficient regulation of glucose and lipid metabolism and liver protection. Summary of the Invention

[0005] This invention aims to at least partially address at least one technical problem existing in the prior art. To this end, this invention provides the application of cucurbitacin C in the preparation of products for improving liver damage and related metabolic disorders. This invention provides a new application of cucurbitacin C, which can effectively reduce glucose and lipid metabolism disorders and liver damage caused by a high-fat diet, providing a new active ingredient option for the preparation of products with dual functions of glucose and lipid metabolism regulation and liver protection.

[0006] In a first aspect, the present invention proposes the use of cucurbitacin C in the preparation of products for the prevention and / or improvement of liver injury and related metabolic disorders.

[0007] According to embodiments of the present invention, the above application may also have at least one of the following additional technical features: According to an embodiment of the present invention, the metabolic disorder is a glucose and lipid metabolism disorder.

[0008] According to an embodiment of the present invention, the liver damage and metabolic disorder are caused by a high-fat diet.

[0009] According to embodiments of the present invention, the improvement of liver injury includes at least one of the following: (a) improvement of abnormal liver function; (b) reduction of hepatic steatosis.

[0010] According to embodiments of the present invention, the indicators of abnormal liver function include alanine aminotransferase levels and aspartate aminotransferase levels.

[0011] According to embodiments of the present invention, the improvement of metabolic disorder includes at least one of the following: (a) regulating blood glucose levels; (b) regulating blood lipid levels; and (c) improving insulin resistance.

[0012] According to an embodiment of the present invention, the indicators of blood lipid levels include one or more of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol.

[0013] According to an embodiment of the present invention, the product is a drug.

[0014] In a second aspect, the invention provides for the use of cucurbitacin C in the preparation of medicaments for the prevention and / or treatment of type 2 diabetes. The cucurbitacin C is capable of significantly improving insulin sensitivity and glucose tolerance, and of significantly reducing fasting blood glucose.

[0015] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0016] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which: Figure 1 The figures shown are the results of the effects of cucurbitacin C on liver injury and liver pathological morphology in mice in Example 1 of this invention. Figure A shows the hematoxylin-eosin (H&E) staining results of liver tissue from each group of mice, with a scale bar of 50 μm. Figure B shows the comparison results of alanine aminotransferase (ALT) activity in the plasma of each group of mice. Figure C shows the comparison results of aspartate aminotransferase (AST) activity in the plasma of each group of mice. for P <0.01, for P <0.001, for P <0.0001.

[0017] Figure 2 The figures shown in Example 1 of this invention illustrate the effects of cucurbitacin C on glucose metabolism-related indicators in mice. A represents a comparison of fasting blood glucose levels in each group of mice; B represents the blood glucose curves from the oral glucose tolerance test (OGTT) in each group of mice; C represents the statistical results of the area under the curve (AUC) of blood glucose levels from the OGTT in each group of mice; D represents the blood glucose curves from the insulin tolerance test (ITT) in each group of mice; and E represents the statistical results of the area under the curve (AUC) of blood glucose levels from the insulin tolerance test in each group of mice. for P <0.05, for P <0.01, for P <0.0001; Figure 3 The graph shows the effect of cucurbitacin C on blood lipid levels in mice in Example 1 of this invention. In the graph, A is a comparison of triglyceride levels in each group of mice, B is a comparison of total cholesterol levels in each group of mice, C is a comparison of low-density lipoprotein cholesterol levels in each group of mice, and D is a comparison of high-density lipoprotein cholesterol levels in each group of mice. for P <0.05, for P <0.01, for P <0.001, for P <0.0001; Detailed Implementation The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0018] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0019] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0020] In this document, the terms “comprising” or “including” are open-ended expressions, meaning that they include the contents specified in this invention, but do not exclude other aspects.

[0021] In this document, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition that may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0022] application This invention proposes the application of cucurbitacin C in the preparation of products for the prevention and / or improvement of liver damage and related metabolic disorders.

[0023] Cucurbitacin C intervention can simultaneously improve indicators related to glucose and lipid metabolism and reduce hepatocellular damage. Specifically, cucurbitacin C can reduce fasting blood glucose elevation induced by a high-fat diet, improve oral glucose tolerance and insulin resistance, and reduce plasma total cholesterol, triglycerides and low-density lipoprotein cholesterol levels while increasing high-density lipoprotein cholesterol levels. At the same time, it reduces alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, and alleviates hepatic steatosis and pathological damage to liver tissue. Thus, cucurbitacin C can exert a comprehensive effect from three levels: glucose metabolism regulation, lipid metabolism regulation and hepatocellular protection, providing a new active substance option for its application in the preparation of products with dual functions of glucose and lipid metabolism regulation and hepatoprotection.

[0024] According to an embodiment of the present invention, the metabolic disorder is a glucose and lipid metabolism disorder.

[0025] This invention is the first to discover that administering cucurbitacin C concurrently with the start of a high-fat diet and continuing this intervention for 12 weeks can significantly improve liver histopathological changes, liver function damage, and glucose and lipid metabolism disorders under continuous high-fat diet exposure, and alleviate abnormalities in blood glucose and lipid metabolism. These results indicate that cucurbitacin C can exert a sustained protective effect during the development and progression of high-fat diet-induced liver damage and related glucose and lipid metabolism disorders. In particular, this invention observed the aforementioned significant protective effect at an extremely low dose (0.05 mg / kg), demonstrating excellent pharmacological activity.

[0026] According to an embodiment of the present invention, the liver damage and metabolic disorder are caused by a high-fat diet.

[0027] According to embodiments of the present invention, the improvement of liver injury includes at least one of the following: (a) improvement of abnormal liver function; (b) reduction of hepatic steatosis.

[0028] According to embodiments of the present invention, the indicators of abnormal liver function include alanine aminotransferase and aspartate aminotransferase.

[0029] According to embodiments of the present invention, the improvement of glucose and lipid metabolism disorder includes at least one of the following: (a) regulating blood glucose levels; (b) regulating blood lipid levels; and (c) improving insulin resistance.

[0030] According to an embodiment of the present invention, the indicators of blood lipid levels include one or more of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol.

[0031] According to an embodiment of the present invention, the product is a drug.

[0032] In some embodiments of the present invention, the use of cucurbitacin C in the preparation of medicaments for the prevention and / or treatment of type 2 diabetes is proposed. The cucurbitacin C can significantly improve insulin sensitivity and glucose tolerance, and can significantly reduce fasting blood glucose.

[0033] According to embodiments of the present invention, the medicament further comprises pharmaceutically acceptable excipients.

[0034] According to embodiments of the present invention, the form of the drug includes liquid, semi-solid, or solid.

[0035] According to embodiments of the present invention, the medicament further comprises a pharmaceutically acceptable carrier, including any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and delayed absorption agents, etc. Specific examples may be one or more of water, saline, phosphate-buffered saline, glucose, glycerol, ethanol, etc., and combinations thereof. In many cases, the pharmaceutical composition includes isotonic agents, such as sugars, polyols (e.g., mannitol, sorbitol), or sodium chloride. Of course, a pharmaceutically acceptable carrier may also include trace amounts of excipients, such as wetting agents or emulsifiers, preservatives, or buffers, to extend the shelf life or potency of the cucurbitacin C. The use of the carrier in therapeutic or pharmaceutical compositions is covered, except in cases where any conventional carrier is incompatible with the active ingredient.

[0036] For pharmaceuticals or pharmacologically active agents, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the dosage forms of this invention, the "effective amount" of one active substance in the composition refers to the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0037] According to embodiments of the present invention, the dosage form of the drug is at least one of an injectable formulation, an oral formulation, or an inhaled formulation.

[0038] According to an embodiment of the present invention, the injectable formulation is an injection solution or a lyophilized powder for injection.

[0039] According to embodiments of the present invention, the oral preparation is a tablet, capsule, granule, oral liquid / solution, or droplet.

[0040] According to embodiments of the present invention, the inhaled formulation is a solution, powder, or suspension thereof.

[0041] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0042] Example 1: The intervention effect of cucurbitacin C on liver damage and glucose / lipid metabolism disorders 1. Animal experiments Animal strain: Six-week-old SPF-grade male C57BL / 6J mice (purchased from Beijing Yaokang Biotechnology Co., Ltd.), weighing 18-22 g, were selected.

[0043] Basic feeding conditions: temperature 22±2℃, relative humidity 50~60%, 12 h / 12 ​​h light and dark cycle, free access to water.

[0044] Animal grouping: Fifteen mice were randomly divided into three groups of five each, with the following treatments for each group (intervention for 12 weeks): (1) Normal diet control group (Ctrl group): fed with ordinary feed (purchased from Beijing SPF Biotechnology Co., Ltd., product number SPF-F02-003) and administered 0.2 mL of 0.5% carboxymethyl cellulose (CMC) solution by gavage daily.

[0045] (2) High-fat diet model group (HFD group): High-fat diet (purchased from Research Diets, Inc. (USA), catalog number D12492) was administered, and 0.2 mL of 0.5% CMC solution was administered by gavage daily.

[0046] (3) Cucurbitacin C intervention group (CuC group): given a high-fat diet and 0.2 mL of cucurbitacin C (CuC) solution (dissolved in 0.5% CMC, dose of 0.05 mg / kg) by gavage daily.

[0047] 2. Detection indicators (1) Fasting blood glucose test Before the end of the intervention in week 12, mice in each group were fasted for 14 hours, blood was collected from the tail tip, and fasting blood glucose (FBG) was measured using a blood glucose meter.

[0048] (2) Oral glucose tolerance test At 11 weeks, after fasting for 14 hours, mice in each group were given glucose solution by gavage at a dose of 2 g / kg body weight. Blood glucose was measured by collecting blood from the tail tip at 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min after glucose administration, and the area under the blood glucose curve (AUC) was calculated.

[0049] (3) Insulin tolerance test At 12 weeks, after fasting for 6 hours, insulin was injected intraperitoneally at a dose of 0.75 U / kg body weight. Blood glucose was measured at 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min after injection, and the AUC and the decrease in blood glucose were calculated.

[0050] (4) Blood lipid level testing After 12 weeks of intervention, mice in each group were sacrificed, blood was collected, and the levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) in plasma were measured.

[0051] (5) Detection of plasma biochemical indicators for liver function After 12 weeks of intervention, mice in each group were sacrificed, plasma was collected, and the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured.

[0052] (6) Liver histopathological examination After 12 weeks of intervention, mice in each group were sacrificed, liver tissue was collected, fixed with 4% paraformaldehyde, routinely dehydrated, embedded in paraffin, sectioned, stained with hematoxylin and eosin (H&E), and observed under an optical microscope for hepatocyte morphology, lipid vacuoles and tissue structure.

[0053] 3. Experimental Results The effects of cucurbitacin C on liver injury and liver pathological morphology in mice are shown in the following results. Figure 1 .

[0054] The results showed that the liver tissue of mice in the control group (Ctrl group) was intact, with hepatocytes arranged neatly and no obvious fat vacuoles. In contrast, the hepatocytes of mice in the high-fat diet group (HFD group) showed numerous fat vacuoles of varying sizes and disordered hepatocyte arrangement, indicating significant hepatic steatosis. Compared with the high-fat diet group (HFD group), the cucurbitacin C intervention group (CuC group) showed a significant reduction in fat vacuoles in the liver tissue, and improved hepatocyte arrangement and tissue structure, indicating that CuC can alleviate high-fat diet-induced hepatic lipid deposition and pathological damage (see details). Figure 1 (A) Meanwhile, compared with the control group (Ctrl group), the plasma ALT and AST activities of mice in the high-fat diet group (HFD group) were significantly increased, suggesting that high-fat diet induced significant liver function damage. Compared with the high-fat diet group (HFD group), the plasma ALT and AST activities of mice in the cucurbitacin C intervention group (CuC group) were significantly decreased, suggesting that CuC can effectively alleviate high-fat diet-induced liver function damage (see A for details). Figure 1 (B and C in the middle).

[0055] The above results indicate that CuC can effectively improve liver damage induced by a high-fat diet.

[0056] The effects of cucurbitacin C on glucose metabolism-related indicators in mice are shown in the following results. Figure 2 .

[0057] The results showed that compared with the control group (Ctrl group), the fasting blood glucose of mice in the high-fat diet group (HFD group) was significantly increased, while CuC intervention significantly reduced the fasting blood glucose of mice in the high-fat diet group (corresponding to the CuC group) (see details). Figure 2 (A) Meanwhile, compared with the control group (Ctrl group), the high-fat diet group mice (HFD group) showed significantly higher blood glucose levels after glucose loading and a significantly increased area under the OGTT curve, suggesting that a high-fat diet leads to impaired glucose tolerance in mice. Conversely, compared with the high-fat diet group mice (HFD group), the cucurbitacin C intervention group mice (CuC group) showed lower blood glucose levels and a significantly reduced area under the OGTT curve, indicating that CuC can improve high-fat diet-induced impaired glucose tolerance (see details in A). Figure 2 (See B and C in the original text). Furthermore, mice in the high-fat diet group (HFD group) showed insufficient glucose reduction after insulin injection and had a higher area under the curve (AUC) during the ITT, indicating that a high-fat diet leads to decreased insulin sensitivity in mice. CuC intervention (corresponding to the CuC group) reduced glucose levels and AUC during the ITT, suggesting that CuC can improve high-fat diet-induced insulin resistance (see details in the original text). Figure 2 (D and E in the text).

[0058] The above results indicate that CuC intervention can effectively alleviate glucose metabolism disorders induced by a high-fat diet.

[0059] The effect of cucurbitacin C on blood lipid levels in mice is shown in the following results. Figure 3 .

[0060] The results showed that the plasma TG, TC, and LDL-C levels in mice in the high-fat diet group (HFD group) were significantly increased, while the HDL-C level was significantly decreased. However, after CuC intervention (corresponding to the CuC group), the plasma TG, TC, and LDL-C levels were effectively reduced, while the plasma HDL-C level was increased.

[0061] The above results indicate that CuC can improve lipid metabolism disorders induced by a high-fat diet in mice.

[0062] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0063] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. Application of cucurbitacin C in the preparation of products for the prevention and / or improvement of liver damage and related metabolic disorders.

2. The application according to claim 1, characterized in that, The metabolic disorder is a glucose and lipid metabolism disorder; Optionally, the liver damage and metabolic disorder are caused by a high-fat diet.

3. The application according to claim 1, characterized in that, The improvement of liver injury includes at least one of the following: (a) Improves abnormal liver function; (b) Reduces hepatic steatosis.

4. The application according to claim 3, characterized in that, The indicators of abnormal liver function include alanine aminotransferase levels and / or aspartate aminotransferase levels.

5. The application according to claim 1, characterized in that, The improvement of the metabolic disorder includes at least one of the following: (a) Regulates blood glucose levels; (b) Regulate blood lipid levels; (c) Improve insulin resistance.

6. The application according to claim 5, characterized in that, The indicators of blood lipid levels include one or more of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol.

7. The application according to claim 1, characterized in that, The product is a drug.

8. The application according to claim 7, characterized in that, The drug further includes pharmaceutically acceptable excipients.

9. The application according to any one of claims 7 or 8, characterized in that, The drug dosage form is at least one of an injectable formulation, an oral formulation, or an inhaled formulation.

10. Use of cucurbitacin C in the preparation of medicines for the prevention and / or treatment of type 2 diabetes.

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