Application of hyodeoxycholic acid in preparation of medicine for preventing or treating medicine-induced liver injury
The drug, prepared using porcine deoxycholic acid, solved the problem of drug-induced liver injury caused by APAP, significantly reduced liver damage indicators, restored liver function and structure, and provided better treatment results.
Patent Information
- Application Number
- CN202511763300.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-27
- Publication Date
- 2026-01-20
AI Technical Summary
Current technology lacks effective drugs for the prevention or treatment of drug-induced liver injury caused by acetaminophen (APAP), especially in the case of acute liver injury. N-acetylcysteine has limited effectiveness in the early stages of poisoning, and new treatment options are urgently needed.
Using porcine deoxycholic acid as the active ingredient, a drug was prepared to prevent or treat drug-induced liver injury. By administering porcine deoxycholic acid to mice by gavage, serum ALT, AST, γ-GT and other indicators were significantly reduced, liver function was restored, liver inflammatory factors IL-1β, IL-6 and TNF-α were reduced, GSH and SOD levels were increased, MDA accumulation was reduced, and oxidative stress was alleviated.
Porcine deoxycholic acid significantly improves APAP-induced liver injury, restores liver structure and function, and is superior to silymarin. It has good hepatoprotective activity, significant effects, and no toxic side effects.
Smart Images
Figure CN121360124A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of biological medicine, more particularly, the application of porcine oxyltic acid in the preparation of a drug for preventing or treating drug-induced liver injury. BACKGROUND
[0002] Acetaminophen (APAP) is one of the most commonly used non-steroidal anti-inflammatory drugs, and is allowed to be used in humans and other animals, but excessive use can cause liver damage, and severe cases can cause liver failure and even death. A global survey found that APAP-induced liver injury accounted for about 7% of all drug-induced liver injury, and excessive use of APAP became the main cause of acute liver failure in the United States and some regions of Europe, with a proportion of more than 50%. The latest evidence shows that even at therapeutic doses, fasting, excessive alcohol consumption and repeated use can greatly increase the probability of liver damage. In recent years, the incidence of APAP-induced liver injury has been rising in China. N-acetylcysteine, as the only FDA-approved drug for treating APAP poisoning, often has better therapeutic effect only in the early stage of poisoning, and it is urgent to find new therapeutic drugs or methods.
[0003] The mechanism of APAP-induced liver injury mainly includes the generation of toxic substances in the body, mitochondrial dysfunction, inflammatory response, oxidative stress, etc. When APAP is taken in excess, NAPQI produced by its metabolism will deplete GSH. The continuously accumulated NAPQI can form covalent bonds with intracellular and mitochondrial proteins, causing mitochondrial damage and leading to cell death. At the same time, GSH depletion and NAPQI accumulation can lead to oxidative stress and accumulation of free radicals in cells, causing mitochondrial dysfunction, loss of cell membrane integrity, etc. The release of cell contents such as DNA fragments, ATP, etc. from necrotic hepatocytes can stimulate the production and release of inflammatory factors such as TNF-α and IL-6, triggering an inflammatory response in the liver and causing secondary liver injury.
[0004] Porcine oxyltic acid is a cholanate extracted from pig bile, and its chemical name is 3a, 6a-dihydroxycholan-5b-24-oic acid (HDCA), and its molecular formula is C 24 H 40O4, English name, molecular weight of 392, CAS number is 83-49-8.Pig deoxycholic acid can stimulate bile secretion, make bile become thin and not increase solid amount, be applicable to cholangitis, cholecystitis, cholelithiasis and other non obstructive bile stasis;Also can accelerate gallbladder contrast medium discharge liver and be helpful to development.Application is suitable for type Ia or Ib hyperlipidemia, atherosclerosis.Pertussis bacillus, diphtheria bacillus, staphylococcus aureus, etc. have certain bacteriostatic effect.Can be used as anti-inflammatory drug, treats chronic bronchitis, pediatric viral upper respiratory inflammation, etc.At present, there is no research that shows that the drug has the effect of preventing or treating drug-induced liver injury.
[0005] Silymarin can be used for the treatment of toxic liver damage and the supportive treatment of chronic hepatitis and cirrhosis, and the Chinese Drug-induced Liver Injury Basic Diagnosis and Treatment and Management Guidelines (2024) points out that for mild to moderate hepatocyte injury type and mixed type DILI patients without jaundice, silymarin can be reasonably used, so silymarin is selected as the control drug. SUMMARY
[0006] The technical problem to be solved by the present application is to provide an application of pig deoxycholic acid in preparing a drug for preventing or treating drug-induced liver injury, especially having a good relieving effect on APAP-induced acute liver injury, having strong liver-protecting activity, and showing a good development and utilization prospect in preparing a drug for preventing and treating acute liver injury.
[0007] To solve the above technical problems, the present application adopts the following technical solutions:
[0008] The application of pig deoxycholic acid in preparing a drug for preventing or treating drug-induced liver injury. The structural formula of pig deoxycholic acid is as follows:
[0009]
[0010] Further, the type of drug-induced liver injury is caused by acetaminophen.
[0011] Further, the purity of acetaminophen is ≥98%.
[0012] The present application also discloses a drug for preventing or treating drug-induced liver injury, wherein the drug takes pig deoxycholic acid as an active ingredient.
[0013] Further, the drug takes pig deoxycholic acid as a single active ingredient.
[0014] Further, the purity of pig deoxycholic acid is ≥98%.
[0015] Further, the drug takes pig deoxycholic acid and other drugs for preventing and treating drug-induced liver injury as active ingredients.
[0016] The application further discloses a medicine preparation comprising the medicine.
[0017] Further, the dosage form of the medicine preparation comprises one or more of a capsule, a tablet, an injection, a granule, an emulsion, a paste, a patch, a pill and a syrup.
[0018] Further, the medicine preparation comprises a pharmaceutically acceptable carrier or excipient.
[0019] Further, the pharmaceutically acceptable carrier or excipient comprises one or more of a diluent, a wetting agent, a binding agent, a disintegrant, a lubricant, a flavoring agent, a solvent, a solubilizer, a cosolvent, an emulsifier, an antioxidant, a metal complexing agent, a preservative, a pH regulator, a surfactant, an excipient, a filler and a synergist.
[0020] Compared with the prior art, the application has the following beneficial effects:
[0021] Through research, the application first finds a new use of hyodeoxycholic acid, i.e. application in preparation of a medicine for preventing or treating drug-induced liver injury. By establishing a liver injury model of a mouse by using acetylaminophenol, the ALT, AST and γ-GT in serum of the mouse given hyodeoxycholic acid are significantly reduced, and liver function is basically recovered; histopathology shows that liver morphology and structure tend to be normal; the levels of inflammatory factors IL-1β, IL-6 and TNF-α in the liver are significantly reduced, and liver inflammation is reduced; meanwhile, the levels of GSH and SOD in the liver tissue are increased, and the accumulation of MDA is reduced, and oxidative stress of the liver is reduced. Therefore, hyodeoxycholic acid can effectively relieve drug-induced liver injury induced by APAP, and the effect is better than that of silymarin, and the hyodeoxycholic acid has great potential in development of a liver-protecting medicine. BRIEF DESCRIPTION OF DRAWINGS
[0022] Figure 1 Fig. 1 is a diagram showing the effect of hyodeoxycholic acid on liver tissue morphology of a mouse.
[0023] Figure 2 Fig. 2 is a diagram showing the effect of hyodeoxycholic acid on liver tissue structure of a mouse.
[0024] Figure 3 Fig. 3 is a diagram showing the effect of hyodeoxycholic acid on serum biochemical indexes of a mouse. In the diagram, A represents alanine aminotransferase (ALT), B represents aspartate aminotransferase (AST), and C represents γ-glutamyl transpeptidase (γ-GT).
[0025] Figure 4 Fig. 4 is a diagram showing the effect of hyodeoxycholic acid on oxidative indexes of a mouse. In the diagram, A represents malondialdehyde (MDA), B represents total superoxide dismutase (SOD), and C represents reduced glutathione (GSH).
[0026] Figure 5 Figure of the effect of poractant alfa on inflammatory indicators in the liver of mice. In the figure: A interleukin-1 beta (IL-1 beta); B interleukin-6 (IL-6); C tumor necrosis factor alpha (TNF-alpha). DETAILED DESCRIPTION
[0027] 1 MATERIALS
[0028] 1.1 Experimental animals
[0029] 4-week-old (body weight 20 ± 2 g) SPF male KM mice were purchased from Changsha Tianqin Biotechnology Co., Ltd., license number SCXK (Xiang) 2024-0021.
[0030] 1.2 Main reagents
[0031] Poractant alfa (≥98%), acetaminophen (AR, ≥98%), sodium carboxymethylcellulose (600-1000 mpa.s, USP grade), silymarin (≥80%) were all purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; hematoxylin-eosin staining solution was purchased from Wuhan Savel Biological Technology Co., Ltd.; ALT, AST, gamma-GT, SOD, MDA and GSH assay kits were purchased from Nanjing Jiancheng Biological Engineering Institute; IL-1 beta, IL-6 and TNF-alpha mouse ELISA detection kits were purchased from Jiangsu Zimei Industry Co., Ltd.
[0032] 2 METHODS
[0033] 2.1 Grouping and administration of experimental animals
[0034] 40 male KM mice were randomly divided into normal group, model group, poractant alfa group (150 mg / kg), silymarin group (150 mg / kg), and each group of mice was caged and fed for 14 d. Poractant alfa and silymarin were dissolved in 1% sodium carboxymethylcellulose, and the intragastric volume was 0.1 mL / 10 g. The normal group and the model group were intragastrically administered with the same volume of 1% sodium carboxymethylcellulose; after the last administration, the mice were fasted for 16 h, and except for the normal group, the mice were intragastrically administered with APAP (200 mg / kg) to establish a liver injury model. After 12 h, the mice were weighed, and the orbital blood was collected immediately after dissection. The liver was weighed and recorded, and part of the left lobe was fixed in 4% paraformaldehyde. The remaining liver was quickly frozen in liquid nitrogen and transferred to a -80 °C refrigerator for subsequent detection.
[0035] 2.2 Determination of blood biochemical indicators
[0036] After the mouse blood was placed at room temperature for 2 h, it was centrifuged at 3500 rpm for 15 min to separate the serum, and the ALT, AST and gamma-GT activities were detected according to the kit instructions.
[0037] 2.3 Liver histopathological examination
[0038] The fixed left lobe of the liver was sequentially dehydrated, transparent, immersed in wax, embedded, and other processes to make sections and perform H&E staining. Observations were made under a microscope and photographs were taken.
[0039] 2.4 Determination of antioxidant capacity of the liver
[0040] Prepare liver homogenate according to the kit instructions, and detect liver MDA, SOD and GSH.
[0041] 2.5 Detection of inflammatory factors in the liver
[0042] According to the mouse ELISA kit instructions, detect IL-1β, IL-6 and TNF-α in the liver.
[0043] 3 Results and analysis
[0044] 3.1 Protective effect of porcine ox bile acid on liver injury mice
[0045] The morphological changes of the liver can reflect the degree of liver damage. The liver of the normal group of mice was reddish brown in color and soft in texture. The liver of the model group of mice was not uniform in appearance, with obvious bleeding and congestion. The liver of the porcine ox bile acid group showed varying degrees of recovery, with the liver of the porcine ox bile acid group basically showing normal morphology. Figure 1 ).
[0046] 3.2 Effect of porcine ox bile acid on liver structure in liver injury mice
[0047] Complete tissue structure is the basis for ensuring normal organ function. The structure of the liver lobule of the normal group of mice was complete, the liver cords were arranged in order, and the liver cells were round and full. The liver lobule structure of the model group of mice was severely damaged, with obvious bleeding and inflammatory infiltration. The liver cells around the central vein were necrotic, and the liver cell nuclei were dissolved and disappeared. The liver tissue structure of the porcine ox bile acid group was basically normal, and the effect was better than that of the silymarin group. Figure 2 ).
[0048] 3.3 Effect of porcine ox bile acid on liver blood biochemistry in liver injury mice
[0049] The most prominent characteristic of liver injury induced by APAP is hepatocyte necrosis. Following hepatocyte necrosis, the cellular contents are released into the bloodstream, causing changes in relevant blood biochemical indicators. Serum ALT, AST, and γ-GT levels directly reflect liver function. Compared to the normal group, the model group showed significantly elevated serum ALT, AST, and γ-GT levels, indicating successful liver injury modeling. Simultaneously, elevated γ-GT indicated a certain degree of damage to the hepatobiliary system. Compared to the model group, the porcine deoxycholic acid group showed significantly decreased serum AST, ALT, and γ-GT levels, achieving or even exceeding the efficacy of silymarin. Figure 3 ).
[0050] 3.4 Effects of porcine deoxycholic acid on liver oxidative parameters in mice
[0051] Dysregulation of the body's oxidative and antioxidant systems is a key characteristic of acute liver injury caused by acute oxidative stress (APAP). During oxidative stress, changes in the levels of superoxide dismutase (SOD), reduced glutathione (GSH), and malondialdehyde (MDA) are most common. Compared to the normal group, the model group showed a significantly increased MDA level, along with a significant decrease in various antioxidant substances (such as SOD and GSH) in both the major enzyme defense system and non-enzyme defense systems, indicating severe oxidative stress in the liver. Compared to the model group, the porcine deoxycholic acid group showed a significantly decreased MDA level, and the restored GSH and SOD levels were close to those of the normal group. Figure 4 This indicates that porcine deoxycholic acid has excellent antioxidant capacity, and its effect can reach or even exceed that of silymarin.
[0052] 3.5 Effects of porcine deoxycholic acid on liver inflammatory markers in liver-injured mice
[0053] IL-1β, IL-6, and TNF-α play important roles in the occurrence and development of APAP-induced liver injury, leading to accelerated hepatocyte damage and even death. Compared with the normal group, the levels of IL-1β, IL-6, and TNF-α in the liver of the model group were significantly increased; compared with the model group, the levels of IL-1β, IL-6, and TNF-α in the liver of the porcine deoxycholic acid group were significantly decreased, with effects similar to those of silymarin. Figure 5 The above results indicate that porcine deoxycholic acid has a certain anti-inflammatory effect.
[0054] The pharmacodynamic studies described above have demonstrated that porcine deoxycholic acid has a significant protective effect against liver injury. It can reverse serum biochemical indicators, liver tissue oxidative stress indicators, and inflammatory factor indicators in APAP-induced drug-induced liver injury in mice, and improve APAP-induced liver congestion, edema, and liver tissue lesions. Furthermore, the effective dose is low and there are no toxic side effects. Therefore, porcine deoxycholic acid can be considered a candidate drug for the prevention and treatment of drug-induced liver injury.
Claims
1. Use of hyodeoxycholic acid in the preparation of a drug for preventing or treating drug-induced liver injury.
2. Use according to claim 1, wherein The type of drug-induced liver injury is caused by acetaminophen.
3. Use according to claim 2, wherein the compound is ###0002### The purity of the acetaminophen is ≥98%.
4. The use according to claim 1, wherein The drug has hyodeoxycholic acid as a single active ingredient.
5. The use according to claim 4, wherein the compound is ###0002### The purity of the hyodeoxycholic acid is ≥98%.
6. The use according to claim 1, wherein The drug has hyodeoxycholic acid and other drugs for preventing and treating drug-induced liver injury as active ingredients.
7. A pharmaceutical preparation, characterized by, The drug comprises the drug of claim 4.
8. The pharmaceutical preparation according to claim 7, characterized in that The administration form of the drug preparation comprises one or more of capsules, tablets, injections, granules, emulsions, ointments, patches, pills, and syrups.
9. The pharmaceutical preparation according to claim 7 or 8, characterized in that The drug preparation contains a pharmaceutically acceptable carrier or excipient.
10. The pharmaceutical preparation according to claim 9, characterized in that The pharmaceutically acceptable carrier or excipient comprises one or more of diluents, wetting agents, binding agents, disintegrating agents, lubricating agents, flavoring agents, solvents, solubilizing agents, co-solvents, emulsifying agents, antioxidants, metal complexing agents, preservatives, pH adjusting agents, surfactants, excipients, fillers, and synergists.