Application of isorhynchophylline in preparation of medicine for preventing and / or treating liver diseases

Abstract

The invention relates to the field of biomedicine, in particular to application of isorhynchophylline to preparation of a medicine for preventing and / or treating liver diseases. The invention discloses a natural small molecule for treating fatty liver diseases related to metabolic dysfunction. The natural small molecules are isorhynchophylline which is a component of a traditional Chinese medicinal material uncaria. The isorhynchophylline has the effects of regulating and controlling lipid metabolism, reducing lipid accumulation and improving liver functions, and shows the potential of preventing and treating the fatty liver diseases related to metabolic dysfunction. Therefore, the compound can be used for preparing medicines for resisting fatty liver diseases related to metabolic dysfunction.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biomedicine, and particularly to the use of isorhynchophylline in the preparation of a drug for preventing and / or treating liver disease. BACKGROUND

[0002] Metabolic dysfunction-associated fatty liver disease (MAFLD) is a progressive and prevalent liver disease closely related to metabolic dysfunction. The main feature of MAFLD is hepatocyte steatosis, and with the progression of the disease, it can develop into metabolic dysfunction-associated steatohepatitis (MASH), and the degree of inflammation and liver fibrosis gradually increases. The course of the disease has high heterogeneity, and in the late stage, it can progress to cirrhosis, liver failure and even liver cancer. The global prevalence of obesity, diabetes and metabolic syndrome has led to a continuous rise in the prevalence of MAFLD. Today, MAFLD has surpassed viral hepatitis and become the leading cause of chronic liver disease. Its pathogenesis involves multiple factors interacting with each other, mainly including metabolic disorders, genetic susceptibility, lifestyle and environmental factors, etc. Based on the complex pathogenesis of MAFLD, the number of potential therapeutic targets is not mature and the number of validated targets is very limited, and the development of related targeted therapeutic drugs has stagnated. At present, only one drug for MAFLD has been approved by the FDA for marketing, and there is a lack of effective targeted treatment options in clinical practice. Therefore, it is of great clinical significance to develop and screen MAFLD prevention or treatment drugs with high efficiency and low toxicity.

[0003] Traditional Chinese medicine material Uncaria rhynchophylla, Uncaria macrophylla Wall., Uncaria hirsuta Havil. and the like. It is recorded in Bencao Huiyan that Uncaria rhynchophylla “can be easily removed by long-term boiling, and then adding it into the decoction of other drugs can restore its activity”. Modern clinical practice believes that Uncaria rhynchophylla has the effects of clearing heat and calming the liver, and calming wind and stopping convulsion. Isorhynchophylline belongs to indole alkaloids, and as the main active ingredient in Uncaria rhynchophylla, it has the pharmacological effects of lowering blood pressure, dilating blood vessels and protecting nerves. However, the pharmacological effects of isorhynchophylline on MAFLD have not been reported. SUMMARY

[0004] The present application aims to provide the use of isorhynchophylline in the preparation of a drug for preventing and / or treating liver disease.

[0005] The use of isorhizine in the preparation of drugs for the prevention and / or treatment of liver diseases, including fatty liver disease.

[0006] Preferably, the fatty liver disease includes non-alcoholic fatty liver disease. Non-alcoholic fatty liver disease is a condition characterized by impaired liver metabolic function.

[0007] Preferably, the non-alcoholic fatty liver disease is a fatty liver disease related to metabolic dysfunction.

[0008] Preferably, the drug further comprises one or more pharmaceutically acceptable excipients.

[0009] Preferably, the pharmaceutically acceptable excipients include one or more of the following: carriers, excipients, diluents, lubricants, wetting agents, emulsifiers, suspension stabilizers, preservatives, sweeteners, and flavorings.

[0010] Preferably, the dosage form of the drug includes one or more of granules, capsules, tablets, powders, oral liquids, injections, syrups, lozenges, pills, injections, and transdermal absorption preparations.

[0011] Preferably, the single application dose of isorhizine is 50-60 mg / kg.

[0012] The non-therapeutic methods in this application include, but are not limited to, health care methods and scientific research methods, and can be applied to humans or other mammals such as rats, rabbits, dogs, etc.

[0013] The symptoms of MAFLD in this application include, but are not limited to, elevated liver coefficient, liver fat accumulation, elevated serum transaminase and total cholesterol levels.

[0014] The uses include reducing liver organ coefficient, serum transaminase, and serum total cholesterol levels in MAFLD, as well as reducing liver fat accumulation and histopathological damage, thus having a protective effect against MAFLD and reducing weight and body fat percentage.

[0015] Available drug dosage forms for this application include various injectable and oral dosage forms, including but not limited to tablets, capsules, oral solutions, injections, powder injections, and transdermal drug delivery formulations, with oral dosage forms being particularly preferred.

[0016] In addition to isorhamnetin, the drugs described in this application may also include various pharmaceutically acceptable excipients / components, including but not limited to coating materials, solvents, solubilizers, binders, stabilizers, antioxidants, pH adjusters, flavoring agents, etc., with particular preference for various excipients / components that can be used in oral dosage forms.

[0017] The medicament of this invention may contain other traditional Chinese medicine or health products for treating MAFLD, or the medicament of this invention may be used in combination with these traditional Chinese medicine or health products or surgical treatments. The traditional Chinese medicine or health products include, but are not limited to, anti-infective drugs, antioxidants, and agents that improve energy metabolism and blood circulation.

[0018] Compared with the prior art, the beneficial effects achieved by the present invention are: This invention aims to investigate the pharmacological effects of isorhizine and discover that it can significantly reduce liver lipid accumulation, serum transaminase, triglyceride and total cholesterol levels in MAFLD mice, thereby proposing the potential application of isorhizine in the preparation of drugs for the treatment of MAFLD.

[0019] This invention provides the use of isorhizine in the preparation of drugs for treating MAFLD, wherein the structural formula of isorhizine is shown in Formula I.

[0020] Formula I Isorhizine significantly reduced lipid droplet accumulation in MAFLD model cells and exhibited a significant protective effect on HHL-5 hepatocytes. It significantly reduced serum transaminase (ALT / AST), triglyceride (TG), and total cholesterol (TC) levels in MAFLD model mice, and decreased liver fat accumulation and histopathological damage, demonstrating its significant protective effect against MAFLD in mice. Attached Figure Description

[0021] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings: Figure 1 This is an image showing the accumulation of lipid droplets in HHL-5 cells stained with Oil Red O in a MAFLD model. Figure 2 This is a graph showing the area statistics of the HHL-5 cell MAFLD model; Figure 3 This is a schematic diagram of the structure used in the toxicological experiment to verify the HHL-5 cell MAFLD model. Figure 4 This is a schematic diagram of Oil Red O staining results of mouse tissues from mice fed a high-fat diet and administered with a carrier / isohedrin for 6 weeks. Figure 5 This is a statistical diagram of lipid droplet area after Oil Red O staining of tissues from mice fed a high-fat diet and administered with a carrier / isohedrin for 6 weeks. Figure 6 This is a schematic diagram of TG / TC levels in liver tissue of mice fed a high-fat diet and administered with a carrier / isohedrin for 6 weeks. Figure 7 This is a schematic diagram of serum TG / TC levels in mice fed a high-fat diet and administered with a carrier / isohedrin for 6 weeks. Figure 8 The serum ALT / AST levels in the liver of mice fed a high-fat diet and administered a carrier / isohedrin for 6 weeks were measured. Detailed Implementation

[0022] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] 1. The effect of isorhamnetin in vitro treatment of MAFLD Materials and reagents Human hepatocyte cell line HHL-5 (purchased from Shanghai Kuisai Biotechnology Co., Ltd.); DMEM medium for cell culture, penicillin-streptomycin antibiotic solution and fetal bovine serum (FBS) (purchased from GIBCO Invitrogen); sodium oleate (purchased from Sigma, O-7501), sodium palmitate (purchased from Sigma, P-9767); isorhamnetin (purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity ≥98%), Oil Red O reagent kit (purchased from Beyotime, Cat# C0158S), Mayer hematoxylin staining solution (purchased from Yuanye Biotechnology, Cat# R20568-500mL). The experimental steps are as follows: 1) Establishment of a cellular MAFLD model: HHL-5 cells were cultured in complete DMEM medium (containing 10% FBS, 1% 100U / mL penicillin and 100μg / mL streptomycin solution) and placed in a 37℃ incubator with 5% CO2. Cells were divided into a control group (WT), a model group (FFA), and an isorhizine-treated group (IRN-FFA). When the cells adhered and grew to 50%, the control group was replaced with complete DMEM medium, while the model group and the isorhizine-treated group were replaced with complete DMEM medium containing 500μM fatty acids (sodium oleate:sodium palmitate = 2:1) to induce the MAFLD model. 2) After 24 hours of cell culture, the control group and the model group were replaced with DMEM complete culture medium, while the isorhamnetin administration group was replaced with complete DMEM culture medium containing 20 μM, 40 μM, and 60 μM isorhamnetin. 3) After culturing for another 24 hours, the cells were treated and Oil Red O staining was used to assess the degree of lipid droplet accumulation in the cells.

[0024] The results are as follows Figures 1-3 As shown: Figure 1 Oil Red O staining showed that the lipid droplet accumulation in the isorhamnetin-treated group (IRN-FFA) was significantly reduced compared to the model group (FFA), and the effect of inhibiting lipid droplet accumulation was concentration-dependent. Figure 2 After Oil Red O staining, the lipid droplet area in the IRN-FFA group was significantly lower than that in the FFA group. Figure 3 In China: Toxicological experiments verified cell viability after administration of isorhizine, showing that isorhizine did not exhibit significant toxicity with increasing concentration.

[0025] 2. The effect of isorhamnetin in vivo treatment of MAFLD The main drugs and reagents used in this experiment included: high-fat diet (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.); physiological saline (purchased from Sewell Biotechnology Co., Ltd.); universal tissue fixative (Sangon Biotech, A500684-0500); and ALT, AST, TG, and TC reagent kits (purchased from Shanghai Enzyme-Linked Immunosorbent Assay Biotechnology Co., Ltd.). The experimental procedures are as follows: A high-fat diet model was established in mice: Six-week-old C57 mice were acclimatized for 7 days and randomly divided into a high-fat diet group (HFD) and a normal diet group (WT). The WT group was fed a normal maintenance roughage diet, while the HFD group was fed a high-fat diet (60% fat-based purified high-fat diet: 20% protein, 20% carbohydrates, and 60% fat). Feeding continued for 6 weeks, and body weight changes were recorded weekly.

[0026] Six weeks after modeling the high-fat diet in mice, the HFD group was randomly divided into an HFD-treatment group and an HFD-control group, with six mice in each group. The HFD-treatment group was administered 60 mg / kg of isorhamnetin (the drug was weighed and dissolved in 0.5% sodium carboxymethyl cellulose solution) by gavage daily, while the HFD-control group was administered 200 μL of 0.5% sodium carboxymethyl cellulose solution by gavage daily. The body weight of the mice in each group was recorded daily. After 6 weeks of continuous administration, blood was collected from the heart of anesthetized mice, and liver tissue was collected for biochemical index detection. Serum TG, TC, ALT, and AST levels and tissue TG and TC levels were detected using kits, and the therapeutic effect of isorhamnetin on MAFLD was evaluated by Oil Red O staining.

[0027] The results are as follows Figures 4-8 As shown: Figure 4 In a study of mice fed a high-fat diet and administered a carrier / isorphine for 6 weeks, Oil Red O staining of tissues showed that lipid droplet accumulation was significantly reduced in the isorphine-treated group (IRN) compared to the model group (HFD).

[0028] Figure 5In the study of lipid droplet area after Oil Red O staining, the lipid droplet area in the IRN group was significantly reduced compared to that in the HFD group (WT: n = 6, HFD: n = 6, IRN: n = 6).

[0029] Figure 6 In mice fed a high-fat diet and administered a carrier / isohedrin for 6 weeks, the TG / TC levels in liver tissue of the IRN group were lower than those in the HFD group.

[0030] Figure 7 In mice fed a high-fat diet and administered a carrier / isohedrin for 6 weeks, serum TG / TC levels in the liver tissue of the IRN group were lower than those in the HFD group.

[0031] Figure 8 In a study of mice fed a high-fat diet and administered a carrier / isohedrin for 6 weeks, serum ALT / AST levels in the liver were measured, and IRN was found to improve liver damage induced by HFD feeding.

[0032] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0033] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The use of isorhamnetin in the preparation of drugs for the prevention and / or treatment of liver diseases, characterized in that, The liver diseases mentioned include fatty liver disease.

2. The application according to claim 1, characterized in that, The fatty liver disease mentioned includes non-alcoholic fatty liver disease.

3. The application according to claim 2, characterized in that, Nonalcoholic fatty liver disease (NAFLD) is a fatty liver disease associated with metabolic dysfunction.

4. The application according to any one of claims 1 to 3, characterized in that, The drug also contains one or more pharmaceutically acceptable excipients.

5. The application according to claim 4, characterized in that, The pharmaceutically acceptable excipients include one or more of the following: carriers, excipients, diluents, lubricants, wetting agents, emulsifiers, suspension stabilizers, preservatives, sweeteners, and flavorings.

6. The application according to any one of claims 1 to 3, characterized in that, The dosage form of the drug includes one or more of the following: granules, capsules, tablets, powders, oral liquids, injections, syrups, lozenges, pills, injections, and transdermal absorption preparations.

7. The application according to any one of claims 1 to 3, characterized in that, The single-dose dose of the aforementioned isorhamnetin is 50-60 mg / kg.

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