Application of RAB7 agonist in preparation of medicine for treating MASLD, CHB or co-disease of MASLD and CHB

Abstract

The invention belongs to the technical field of biological medicines, and discloses an application of an RAB7 agonist in preparation of a medicine for treating MASLD, CHB or co-diseases thereof, and the RAB7 agonist activates Rab7 and simultaneously promotes HBV DNA and HBsAg degradation and autophagy-lysosome functions to realize double effects of targeted improvement of virus and metabolism at the same time. The technical problem that an existing treatment scheme cannot cooperatively regulate and control virus replication and metabolic disorder is solved, and an efficient and safe treatment strategy is provided for clinical refractory common diseases.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to application of a RAB7 agonist in preparation of a medicament for treating MASLD, CHB or a comorbidity disease thereof. BACKGROUND

[0002] Chronic hepatitis B virus (CHB), often simply referred to as "hepatitis B", is caused by hepatitis B virus (HBV) infection, and the liver appears different degrees of inflammation and (or) liver fibrosis. The early symptoms of the patient mainly include repeated fatigue, dizziness, loss of appetite, etc., and as the disease worsens, it can show liver palms, spider nevi, hepatosplenomegaly, etc. Hepatitis B virus is mainly transmitted through mother-to-child, blood, broken skin and mucous membranes, and sexual contact. For the treatment of chronic hepatitis B, the primary goal is antiviral therapy, and a comprehensive and individualized treatment method is usually adopted, including adequate rest, reasonable diet and nutrition, etc. The treatment and maintenance of antiviral drug regimen should be adjusted in time according to the severity of the disease. The antiviral drugs include nucleoside (acid) antiviral drugs (such as entecavir, tenofovir, etc.) and interferon antiviral drugs (such as interferon, pegylated interferon, etc.).

[0003] Metabolic dysfunction-associated fatty liver disease (MASLD) is the most common chronic liver disease worldwide. 25% of patients with MASLD will develop metabolic dysfunction-associated steatohepatitis (MASH) and have fibrosis progression within 3 years. The occurrence of MASLD is related to many factors, including obesity, type 2 diabetes, hypertension and lipid disorders, and metabolic syndrome. MASLD can be roughly divided into two categories: metabolic dysfunction-associated fatty liver (MAFL), which is non-progressive MASLD; and metabolic dysfunction-associated steatohepatitis (MASH), which is progressive MASLD. MAFL is liver steatosis, usually occurring in the 3rd zone of liver lobule, and may or may not be accompanied by mild inflammation. The histological features of MAFL progressing to MASH are characterized by the presence of three main features: steatosis, lobular inflammation and hepatocyte ballooning, possibly with fibrosis. The process of MASH reversing back to MAFL can be highly dynamic, and may even occur in a short period of time. Abnormal lipid metabolism is an important inducer of MASLD, and the endoplasmic reticulum, as an important organelle for lipid metabolism and protein synthesis, plays a key role in the regulation of MASLD occurrence. Inflammatory response is a key factor for MASLD to progress to MASH. Endoplasmic reticulum stress promotes the release of inflammatory factors by activating signaling pathways such as NF-κB, which further exacerbates liver inflammation. At present, although the pathogenesis of MAFLD has been deeply studied at home and abroad, the exact pathogenesis and treatment of MASLD have not been fully determined. Therefore, finding high-efficiency, low-toxicity small molecule drugs has become a new strategy for clinical prevention and treatment of MASLD. The treatment drugs of metabolic dysfunction-associated fatty liver disease (MASLD, formerly known as non-alcoholic fatty liver disease / NAFLD) currently still focus on improving metabolic disorders and anti-inflammatory and anti-fibrosis. Drugs that have been approved or widely studied include Pioglitazone, Vitamin E (alpha-tocopherol), Semaglutide, Tirzepatide, SGLT2 inhibitors (such as Englitazone, Dapagliflozin), Resmetirom, Obeticholic Acid (OCA), etc.

[0004] The coexistence of chronic hepatitis B (CHB) and fatty liver disease (especially metabolic associated fatty liver disease, MASLD) has become an increasingly prominent clinical challenge and research hotspot in the field of liver disease. A large number of epidemiological data show that with the global prevalence of obesity and metabolic syndrome, the prevalence of fatty liver in CHB patients has increased significantly. Studies have shown that the coexistence of fatty liver not only can accelerate the progression of liver fibrosis in CHB patients, increase the risk of cirrhosis and even hepatocellular carcinoma, but also can affect the response and effect of antiviral therapy, and have a complex impact on liver-related and all-cause mortality. However, due to the lack of existing research, the specific molecular mechanisms of their interaction have not been fully elucidated, and there is a lack of precise treatment for such patients. Therefore, in-depth exploration of the pathophysiological mechanisms and treatment strategies of CHB and MASLD co-morbidity is an urgent need and important direction of current liver disease research.

[0005] The current clinical treatment of CHB and MASLD single disease and co-morbidity faces significant limitations, and the existing scheme relies on antiviral drugs such as nucleos(t)ide analogues, peg-IFN, and simple superposition of metabolic intervention means (such as lifestyle adjustment, GLP-1 receptor agonists). This therapy is a simple superposition of single disease treatment, and the existing technology lacks a dual treatment strategy that can simultaneously target viral clearance and metabolic / inflammation regulation, resulting in insufficient synergistic intervention for co-morbidity patients and difficulty in effectively blocking disease progression.

[0006] The interaction mechanism of CHB and MASLD is complex, and this disease lacks a common core treatment target, making it difficult for a single drug to achieve synergistic intervention. Not only that, the pathological process of the two diseases influences each other, with viral activity promoting metabolic disorders, and metabolic disorders affecting antiviral immunity, further increasing the difficulty of treatment. Due to the lack of large clinical cohorts of co-morbidity patients in existing clinical practice and superficial basic research, it is difficult to find compounds that can simultaneously and efficiently and safely act on both viral and metabolic pathways.

[0007] Rab7, as a key GTPase regulating the fusion process of late endosomes / autophagosomes and lysosomes, is essential for mediating lysosomal degradation of substrates such as abnormal proteins, damaged organelles (such as mitochondria), and exogenous pathogens. Studies have found that in CHB, HBV infection can significantly impair the function of Rab7, leading to excessive consumption and imbalance of lysosomal activity, thereby weakening the ability to clear viral components and promoting disease progression. In MASLD, the autophagic flux is blocked, characterized by a significant decrease in autophagosome-lysosome fusion efficiency, which is the core pathological link leading to liver lipid metabolism disorder and inflammatory damage. Based on the above mechanism, theoretically, Rab7 agonists serve as an innovative intervention means. The agonist can specifically enhance the activity of Rab7, effectively restore the inhibited Rab7 function in the CHB model, and significantly improve the blocked autophagic flux and promote efficient autophagosome-lysosome fusion in the MASLD model. It can target the core pathological mechanisms of CHB and MASLD from multiple aspects, namely, enhancing viral clearance in CHB, promoting lipid degradation and inhibiting inflammatory response in MASLD. Rab7 agonists can provide a unified molecular theoretical basis for the treatment of these two diseases, especially their co-occurrence (comorbidity). SUMMARY

[0008] The purpose of the present application is to provide the use of RAB7 agonists in the preparation of a drug for treating MASLD, CHB or comorbid diseases, to solve the technical problem that existing treatment regimens cannot synergistically regulate viral replication and metabolic disorders, and to provide an efficient and safe treatment strategy for clinically refractory comorbidities.

[0009] 1. Simple CHB treatment cannot improve metabolic disorders, leading to continuous progression of liver damage.

[0010] 2. Simple MASLD treatment has no effect on HBV infection, and comorbid patients require multiple drug combinations with poor compliance.

[0011] 3. Comorbid treatment lacks target points for synergistically regulating viral replication and metabolic inflammation, resulting in low treatment efficiency.

[0012] To achieve the above purpose, the following technical scheme is adopted in the present application: The present application provides the use of RAB7 agonists in the preparation of a drug for treating liver diseases, wherein the liver diseases are metabolic dysfunction-related fatty liver diseases, chronic hepatitis B or comorbid diseases.

[0013] In the above technical scheme, the RAB7 agonist is ML-098.

[0014] In the above technical scheme, in the preparation of a drug for treating metabolic dysfunction-related fatty liver diseases, the drug uses RAB7 agonists as active ingredients; or The drug contains a RAB7 agonist and a STING inhibitor as its active ingredients.

[0015] In the above technical solutions, the STING inhibitor is one of H-151, compound 18, SN-011, 4-OI, BB-Cl-amidine, and LB244.

[0016] In the above technical solutions, in the preparation of a drug for treating chronic hepatitis B, the drug uses a RAB7 agonist as the active ingredient; or The drug contains RAB7 agonist and STING agonist as active ingredients.

[0017] In the above technical solutions, the STING agonist is one of DMXAA, ADU-S100, E7766, IMSA101, MV-626, compound 31, SR-8541A, TXN10128, RBS2418, diABZI, GSK3745417, CRD3874-SI, and KL340399.

[0018] In the above technical solutions, in the preparation of a drug for treating comorbidities of chronic hepatitis B and metabolic dysfunction-related fatty liver disease, the drug uses a RAB7 agonist as the active ingredient.

[0019] In the above technical solution, the RAB7 agonist promotes HBV virus degradation and liver autophagy by activating Rab7 protein, and simultaneously reduces HBV DNA and HBsAg and improves blood lipids and blood sugar.

[0020] In the above technical solution, the RAB7 agonist reduces the levels of liver tissue inflammatory factors IL-6, IL-1β, TNF-α and serum ALT and AST, thereby alleviating liver fibrosis.

[0021] Compared with the prior art, the outstanding advantages of the present invention are reflected in the following aspects: 1. Multi-target synergistic regulation Traditional antiviral drugs (such as entecavir) only inhibit HBV polymerase and have no effect on metabolic disorders; RAB7 agonists (ML-098) activate Rab7, thereby promoting the degradation of HBV DNA and HBsAg and autophagy-lysosomal function, achieving a dual effect of simultaneously targeting and improving both the virus and metabolism.

[0022] 2. Clinical advantages of comorbid treatment The current treatment regimen for patients with comorbidities is antiviral therapy combined with MASLD treatment, however, there is no better drug for MASLD at present, and this simple superposition treatment regimen will increase the incidence of adverse reactions. The RAB7 agonist (ML-098) of the present application is administered once a week, and no adverse reactions such as diarrhea and hypoglycemia are observed in mice in the experiment, and the administration cycle is short, which provides a strong basis for clinical conversion. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 : Schematic diagram of the experimental design of CHB mice with MASLD treated by ML-098.

[0024] Figure 2 : Schematic diagram of the experimental design of MASLD mice treated by ML-098 / H-51.

[0025] Figure 3 : Schematic diagram of the experimental design of HBV-tg mice treated by ML-098 / DMXAA.

[0026] Figure 4 : ML-098 improves the lipid metabolism of CHB mice with MASLD; (A) Representative images of oil red O staining of the control group, pAAV / HBV1.2+HFD mice, and pAAV / HBV1.2+HFD+ML-098 mice, scale bar: 50 μm; (B) Expression of serum TC, TG, and NEFA; (C) Subcutaneous fat weight, epididymal fat weight, and fasting blood glucose values of mice.

[0027] Figure 5 : ML-098 improves the liver inflammation of CHB mice with MASLD; (A) Representative images of H&E and Sirius red staining of the control group, pAAV / HBV1.2+HFD mice, and pAAV / HBV1.2+HFD+ML-098 mice, scale bar: 50 μm; (B) Serum HBV DNA and HBsAg levels of mice; (C) Expression of serum ALT and AST of mice; (D) Expression levels of liver inflammation factors TNFα, IL-1β, and IL-6 of mice.

[0028] Figure 6 : ML-098 and / or H-151 improve the lipid metabolism of MASLD mice; (A) Representative images of oil red O staining of the control group, MASLD mice, and MASLD (different administration groups) mice, scale bar: 50 μm; (B) Expression of serum TC, TG, and NEFA; (C) Subcutaneous fat weight, epididymal fat weight, and fasting blood glucose values of mice.

[0029] Figure 7ML-098 and / or H-151 improve liver inflammation of MASLD mice; (A) Representative images of H&E and Sirius red staining of control group, MASLD mice, MASLD (different administration groups) mice, scale bar: 50 μm; (B) Expression of ALT and AST in serum of mice; (C) Expression levels of TNFα, IL-1β and IL-6 in liver inflammation factors of mice.

[0030] Figure 8 ML-098 or DMXAA reduces the virological level of HBV mice; (A) Expression of HBV DNA in serum of mice; (B) Expression of HBsAg in serum of mice. DETAILED DESCRIPTION

[0031] In order to better illustrate the purposes, technical solutions and advantages of the present application, the present application will be further described below in combination with specific embodiments. The present application can be implemented in many different forms, and should not be understood as being limited to the embodiments set forth herein. On the contrary, these embodiments are provided so that the present disclosure will be thorough and complete, and will fully convey the idea of the present application to those skilled in the art, and the present application will be limited only by the claims.

[0032] The present application provides the use of a RAB7 agonist in the preparation of a medicament for treating a liver disease, the liver disease being a metabolic dysfunction-related fatty liver disease (MASLD), chronic hepatitis B (CHB) or a comorbid disease thereof.

[0033] The RAB7 agonist promotes the degradation of HBV virus and liver autophagy by activating Rab7 protein, and simultaneously reduces HBV DNA, HBsAg and improves blood lipids and blood glucose. The RAB7 agonist reduces the levels of liver tissue inflammation factors IL-6, IL-1β, TNF-α and serum ALT, AST, and reduces liver fibrosis.

[0034] Preferably, the RAB7 agonist is ML-098. The chemical structural formula of ML-098 is as follows, and the CAS No. is 878978-76-8.

[0035] In addition, it should be noted that in addition to ML-098, other RAB7 agonists can also be selected, which also have the same therapeutic effect.

[0036] Preferably, in the preparation of a medicament for treating a metabolic dysfunction-related fatty liver disease (MASLD), the medicament takes a RAB7 agonist as an active ingredient; or The medicament takes a RAB7 agonist and a STING inhibitor as active ingredients.

[0037] The STING inhibitor includes: (1) Inhibitors of LBD One of the drug design strategies for STING inhibitors is to use small molecules targeting the LBD to compete with ligands for the binding site. Compound 18 inhibits the activation of STING through competitive action. SN-011 is another small molecule inhibitor of STING.

[0038] (2) Inhibitors of multimerization H-151 is a nitro-furan derivative that inhibits the palmitoylation of human STING by covalently modifying the Cys91 site. Despite its short half-life, intraperitoneal injection of the inhibitor significantly reduced the level of IFN-β in Trex1- / - mice.

[0039] 4-octyl itaconate (4-OI) can directly alkylate Cys91, thereby inhibiting the activation of the cGAS / STING signaling pathway.

[0040] (3) BB-Cl-carbamimidate inhibits the palmitoylation of STING by covalently modifying Cys148, thereby weakening the multimerization of STING, the IFN-I signaling, and the autophagy process. Subsequently, through further screening of analogs of BB-Cl-carbamimidate, LB244 was found to have higher activity and better selectivity. Intraperitoneal injection of LB244 significantly attenuated the diABZI-induced cytokine secretion. However, when LB244 is taken orally, its bioavailability is low, and its clearance rate is high, which limits its drug delivery mode.

[0041] Preferably, the STING inhibitor is one of H-151, compound 18, SN-011, 4-OI, BB-Cl-carbamimidate, and LB244.

[0042] Preferably, the STING inhibitor is H-151. The chemical structure of H-151 is as follows, and its CAS No. is 941987-60-6.

[0043] Preferably, in the preparation of a drug for treating chronic hepatitis B (CHB), the drug has a RAB7 agonist as an active ingredient; or The drug has a RAB7 agonist and a STING agonist as active ingredients.

[0044] Agonists of STING are mainly divided into cyclic dinucleotide (CDN) agonists, ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) inhibitors, and non-CDN agonists.

[0045] (1) CDN agonists ADU-S100, clinical trial number: NCT02675439.

[0046] E7766, its phase I clinical trial (NCT04144140) was terminated in advance. As of December 2024, IMSA101 (NCT06601296 and CTR20211689) is undergoing clinical research to comprehensively evaluate its safety and effectiveness.

[0047] (2) ENPP1 inhibitors: ENPP1 can degrade extracellular 2', 3'-cGAMP. By inhibiting ENPP1, the concentration of 2', 3'-cGAMP in the blood can be increased, thereby activating STING.

[0048] MV-626: is a selective ENPP1 inhibitor. In mouse pancreatic cancer model experiments, injecting MV-626 alone or in combination with radiotherapy enhances the body's anti-tumor immune response and improves the overall survival rate of mice. See literature: He X, Wedn A, Wang J, et al . IUPHAR ECR review: the cGAS-STING pathway: novel functions beyond innate immune and emerging therapeutic opportunities[J / OL]. Pharmacol Res , 2024, 201: 107063 [2024-12-30]. https: / / pubmed.ncbi.nlm.nih.gov / 38216006 / . DOI: 10.1016 / j.phrs.2024.107063. Compound 31: is another inhibitor targeting the catalytic pocket of ENPP1. In a mouse model of breast cancer, this compound significantly reduced the number of lung metastatic nodules and intraperitoneal nodules in mice, while the body weight of mice did not change significantly, indicating that this inhibitor has good anti-tumor activity and safety. See literature: Sun Y, Chen M, Han Y, et al.Discovery of pyrido[2,3- d ]pyrimidin- 7-one derivatives as highly potent andefficacious ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1)inhibitors for cancer treatment[J]. JMed Chem, 2024, 67(5): 3986-4006. Currently, ENPP1 inhibitors that have entered phase I clinical trials include SR-8541A (NCT06063681), TXN10128 (NCT05978492), and RBS2418 (NCT05270213).

[0049] (3) Non-CDN agonists Linked amidobenzimidazole (diABZI): triggers downstream TBK1 kinase activation by binding to the STING protein, inducing type I interferon and nuclear factor kappa B pathways. GSK3745417 is currently in phase I clinical trials (NCT03843359).

[0050] CRD3874-SI has entered phase I clinical trials for evaluating its therapeutic effect in patients with acute myeloid leukemia (NCT06626633) or advanced / metastatic malignant solid tumors (NCT06021626).

[0051] In addition, non-CDN STING agonist KL340399 has also entered clinical trials (NCT05549804 and NCT05387928).

[0052] Preferably, the STING agonist is one of DMXAA, ADU-S100, E7766, IMSA101, MV-626, Compound 31, SR-8541A, TXN10128, RBS2418, diABZI, GSK3745417, CRD3874-SI, KL340399.

[0053] Preferably, the STING agonist is DMXAA. The chemical structure of DMXAA is as follows, and the CAS No. is 117570-53-3.

[0054] Preferably, in the preparation of a medicament for treating the co-morbid disease of chronic hepatitis B and metabolic dysfunction-related fatty liver disease, the medicament takes RAB7 agonist as an active ingredient.

[0055] Drug preparation procedure in mice: For example, taking the preparation of 1 ml of ML-098, DMXAA or H-151 solution with a concentration of 5 mg / ml, the specific operation is as follows: first, accurately weigh 5 mg of the target drug (ML-098, DMXAA or H-151) and place it in a sterile centrifuge tube; then, add 100 μl of DMSO (10% of the final volume) and mix well with a vortex mixer until the drug is completely dissolved, at which time the solution is clear; then, slowly add 400 μl of PEG 300 (40% of the final volume) while gently inverting the centrifuge tube to mix, avoiding the formation of bubbles and ensuring complete mixing with the previous solution; then, add 50 μl of Tween-80 (5% of the final volume) and continue to vortex for 30 seconds to ensure uniform dispersion of the surfactant and prevent drug precipitation; finally, add 450 μl of physiological saline (45% of the final volume) and vortex again for 1 minute to form a uniform solution with a total volume of 1 ml, in which the proportions of the components are strictly 10% DMSO, 40% PEG 300, 5% Tween-80 and 45% physiological saline, and the drug concentration is accurately maintained at 5 mg / ml. The prepared solution can be used immediately for experimental administration or stored briefly at 4°C in the dark (not more than 24 hours) to ensure stability.

[0056] Intraperitoneal injection of drugs in mice: The mice are administered at a dose of 10 mg / kg, first gently fixed with the abdomen exposed for injection, and the skin of the abdomen is disinfected with a 75% alcohol cotton ball; then, a 1 ml syringe is held by hand, and the drug solution prepared in proportion is drawn, and the needle is inserted into the skin at a 30°-45° angle to the abdominal wall on the left or right side of the mouse (avoiding the bladder and important organ areas), and the drug is slowly pushed in. After injection, the needle is quickly withdrawn, and a dry cotton ball is pressed against the injection site for a few moments to prevent leakage of the liquid.

[0057] Frequency of administration: 1. ML-098 single drug intraperitoneal injection, 10 mg / kg, 5 times a week (continuous injection for 5 days), a total of 2 weeks.

[0058] 2. H-151 single drug intraperitoneal injection, dose 10 mg / kg, once a week, a total of 4 weeks.

[0059] 3. Combination regimen: ML-098 (10 mg / kg) + STING inhibitor H-151 (10 mg / kg) intraperitoneal co-injection, with the same frequency and cycle as the single drug regimen.

[0060] 4. DMXAA alone, i.p. at a dose of 10 mg / kg, once every 3 days for a total of 2 weeks.

[0061] 5. Combination regimen: ML-098 (10 mg / kg) + STING agonist DMXAA (10 mg / kg) i.p. at the same frequency and cycle as the monotherapy regimen.

[0062] Animal tissue sampling: 1. After the mice in each group reached the end of modeling, they were fasted overnight without water restriction; 2. After recording the body weight of the mice at the end of modeling using an electronic balance, the mice were anesthetized with ether, and blood was collected from the eyeball into a sterile eppendorf tube using the eyeball blood collection method. The blood was allowed to clot at room temperature for 2 hours, and then centrifuged at 3500 rpm for 10 minutes to collect the mouse serum, which was stored in a -80°C freezer.

[0063] 3. The mice were sacrificed by cervical dislocation, and the mice were disinfected by soaking in 75% alcohol. The limbs of the mice were fixed on the mouse killing table. The mouse abdominal skin and subcutaneous fat were separated using ophthalmic scissors and ophthalmic forceps, and the liver was fully exposed. The liver ligament was carefully separated, the gallbladder was removed, and the removed liver was washed in PBS and then dried with filter paper.

[0064] 4. Two pieces of green bean-sized tissue were fixed in 4% paraformaldehyde and OCT embedding agent, respectively, for subsequent experiments. The remaining liver tissue was cut into small pieces and placed in sterile cryotubes, frozen in liquid nitrogen, and finally transferred to a -80°C freezer for storage.

[0065] 5. For mouse intraocular venous blood collection: After the mouse was anesthetized with ether, the left thumb and index finger were lightly pressed against the skin of the head and neck to make the eyeball slightly convex. The right hand held a capillary tube and rotated it at a 30° angle from the conjunctival fornix between the inner canthus and the eyeball to the posterior orbit. When a significant emptying sensation was felt, the capillary tube was gently lifted to collect blood using negative pressure siphoning. Immediately after withdrawing the tube, a sterile cotton ball was used to press and stop bleeding for 10 seconds.

[0066] Mouse tissues and sera were used for subsequent metabolic, inflammatory, and virological tests.

[0067] Example 1 This example verifies the therapeutic effect of ML-098 through in vivo animal experiments, and the specific technical solutions are as follows: (I) Establishment of experimental model 1. CHB combined with MASLD co-disease model: 10 μg of pAAV / HBV1.2 plasmid was injected into the mouse tail vein by high-pressure hydrodynamic injection, and then the successfully modeled mice with HBsAg > 500 mIU / ml were selected, and randomly given HFD for 20 weeks.

[0068] 2, Simple MASLD model: 6-8 weeks old male C57BL / 6 mice were selected, and high-fat diet (HFD, 60% fat calories) was fed for 20 weeks.

[0069] 3, Simple HBV infection model: 6-8 weeks old male HBV-tg transgenic mice (C57BL / 6 background, stably expressing HBV (type A) full genome) were selected, and the serum HBV DNA baseline level was 1×10 8 -2×10 8 IU / mL, and the HBsAg baseline was 15000-25000 IU / mL.

[0070] (II) Design of drug administration scheme: Drug dissolution scheme: for drug dissolution of ML-098, DMXAA and H-151, 10% DMSO, 40% PEG 300, 5% Tween-80 were added in turn, and finally 45% physiological saline was prepared.

[0071] 1, CHB combined with MASLD treatment: Single drug regimen: ML-098 single drug intraperitoneal injection, 10 mg / kg, 5 times a week (continuous injection for 5 days), a total of 2 weeks. Figure 1 ) 2, Simple MASLD treatment: Single drug regimen: ML-098 intraperitoneal injection, dose 10 mg / kg, 5 times a week (continuous injection for 5 days), a total of 2 weeks. H-151 intraperitoneal injection, dose 10 mg / kg, once a week, a total of 4 weeks.

[0072] Combined regimen: ML-098 (10 mg / kg) + STING inhibitor H-151 (10 mg / kg) intraperitoneal injection, the frequency and cycle of administration are the same as those of the single drug regimen. Figure 2 ) 3, Simple CHB treatment: Single drug regimen: ML-098 intraperitoneal injection, dose 10 mg / kg, 5 times a week (continuous injection for 5 days), a total of 2 weeks. DMXAA intraperitoneal injection, dose 10 mg / kg, once every 3 days, a total of 2 weeks.

[0073] Combined regimen: ML-098 (10 mg / kg) + STING agonist DMXAA (10 mg / kg) intraperitoneal injection, the frequency and cycle of administration are the same as those of the single drug regimen. Figure 3 ) (III) Detection index and method: 1, Antiviral effect: Serum HBV DNA and serum HBsAg.

[0074] 2. Metabolic indicators: Liver steatosis: H&E staining of paraffin sections of liver tissue.

[0075] Blood lipids (TC, TG, NEFA) were detected.

[0076] Fasting blood glucose (FBG): tail vein blood sampling, blood glucose meter detection.

[0077] Fat weight: epididymal fat (eWAT) and subcutaneous fat (iWAT) were separated by dissection and weighed on an electronic balance.

[0078] 3. Liver injury and inflammation: Liver enzymes (ALT, AST) were detected by biochemical analyzer.

[0079] Inflammatory factors (IL-6, IL-1β, TNF-α) were detected by ELISA kit.

[0080] Liver fibrosis: Sirius red trichrome staining was used to observe the area of collagen deposition.

[0081] (IV) Experimental results: 1. Effect of comorbidity treatment 1) Improvement of glucose and lipid metabolism disorder: ML-098 reduced lipid deposition in the liver (A); decreased triglycerides (TG), cholesterol (TC), and free fatty acids (NEFA) (B); reduced subcutaneous fat weight, epididymal fat weight, and fasting blood glucose level (C). Figure 4 A); decreased triglycerides (TG), cholesterol (TC), and free fatty acids (NEFA) (B), subcutaneous fat weight, epididymal fat weight, and fasting blood glucose level (C). Figure 4 B). Figure 4 C).

[0082] 2) Improvement of liver inflammation: reduced inflammatory cell infiltration (A); decreased liver function ALT and AST levels (C); decreased inflammatory factors TNF-α, IL-1β, and IL-6 (D). Figure 5 C). Figure 5 D). Figure 5 B).

[0083] 3) Reduction of HBV DNA and HBsAg levels (B). Figure 5 B).

[0084] 4) Improvement of liver fibrosis: reduced collagen deposition area (A). Figure 5 A).

[0085] 2. Effect of simple MASLD treatment 1) Improvement of glucose and lipid metabolism disorder: single drug ML-098 group, single drug H-151 group, and combined drug ML-098+H-151 group all showed reduced lipid deposition in the liver (A), decreased TG, TC, and NEFA (B). Figure 6 B). Figure 6B) and epididymal fat weight loss Figure 6 C). Only the single drug ML-098 group and the combination drug ML-098+H-151 group can improve the fasting blood glucose level, and the single drug H-151 group cannot improve the blood glucose of MASLD mice Figure 6 C).

[0086] 2) Inflammation and liver damage: the single drug ML-098 group, the single drug H-151 group, and the combination drug ML-098+H-151 group can all see a reduction in liver inflammatory cell infiltration Figure 7 A), and only the single drug ML-098 group improves the ALT and AST of MASLD mice Figure 7 B). The single drug ML-098 group, the single drug H-151 group, and the combination drug ML-098+H-151 group can all reduce the levels of inflammatory factors TNF-α, IL-1β, and IL-6 in MASLD mice Figure 7 C).

[0087] 3, the treatment effect of simple CHB 1) The single drug ML-098 group and the single drug DMXAA group can both reduce the HBV DNA level of HBV-tg mice, while the combination drug ML-098+DMXAA group is less effective than the single drug group in reducing HBV DNA Figure 8 A).

[0088] 2) The single drug DMXAA group and the combination drug ML-098+DMXAA group are less effective in reducing the level of HBsAg in mice than the single drug ML-098 group Figure 8 B).

[0089] The RAB7 agonist single drug or combination regimen of the present application significantly reduces HBV replication indicators (HBV DNA, HBsAg); improves liver steatosis, dyslipidemia, and glucose disorders in MASLD; simultaneously inhibits liver inflammation and fibrosis, reduces liver damage markers; provides an integrated treatment regimen for CHB combined with MASLD comorbidity, and reduces adverse reactions caused by drug combination.

[0090] Obviously, the above examples are merely examples for clarity and do not limit the embodiments. For those of ordinary skill in the art, other different forms of changes or variations can be made on the basis of the above description. Here, it is not necessary and impossible to exhaust all embodiments. The obvious changes or variations derived therefrom are still within the protection scope of the present application.

Claims

1. Use of a RAB7 agonist in the manufacture of a medicament for treating a liver disease, which is chronic hepatitis B or a comorbid disease of chronic hepatitis B and metabolic dysfunction-related fatty liver disease.

2. Use according to claim 1, characterized in that: The RAB7 agonist is ML-098.

3. Use according to claim 1, characterized in that: In the manufacture of a medicament for treating chronic hepatitis B, the medicament takes the RAB7 agonist as an active ingredient; or The medicament takes the RAB7 agonist and a STING agonist as active ingredients.

4. Use according to claim 3, characterized in that: The STING agonist is one of DMXAA, ADU-S100, E7766, IMSA101, MV-626, Compound 31, SR-8541A, TXN10128, RBS2418, diABZI, GSK3745417, CRD3874-SI, and KL340399.

5. The use according to claim 1, characterized in that: In the manufacture of a medicament for treating a comorbid disease of chronic hepatitis B and metabolic dysfunction-related fatty liver disease, the medicament takes the RAB7 agonist as an active ingredient.

6. The use according to claim 1, characterized in that: The RAB7 agonist promotes HBV degradation and liver autophagy by activating Rab7 protein, and synchronously reduces HBV DNA, HBsAg and improves blood lipids and blood glucose.

7. Use according to claim 1, characterized in that: The RAB7 agonist reduces liver tissue inflammatory factors IL-6, IL-1β, TNF-α and serum ALT, AST levels, and reduces liver fibrosis.

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