Application of hydroxynidone in preparation of medicine for treating or preventing chronic hepatitis B with hepatic fibrosis

Abstract

The invention provides application of a compound as shown in a formula (I), a solvate, a hydrate, a prodrug or a pharmaceutically acceptable salt of the compound in preparation of drugs for treating or preventing chronic hepatitis B with hepatic fibrosis. Clinical research proves that the compound can effectively reverse hepatic fibrosis; and particularly, the traditional Chinese medicine composition has a better effect on treating fibrosis in a converging region, remarkable fibrosis, hepatic fibrosis in a progressive stage or liver cirrhosis.

Description

[0001] This application is a divisional application of patent application No. 202110531848.7 filed with the China National Intellectual Property Administration on May 14, 2021, entitled "Use of Hydroxynesone in the Preparation of Drugs for the Treatment or Prevention of Chronic Hepatitis B with Liver Fibrosis". Technical Field

[0002] This invention belongs to the field of pharmaceuticals, specifically relating to the use of hydroxynidone in the preparation of drugs for the treatment or prevention of chronic hepatitis B with liver fibrosis. Background Technology

[0003] Liver fibrosis is a common pathological basis in the progression of chronic liver disease. Various chronic injuries cause hepatocyte degeneration and necrosis, abnormal proliferation and excessive deposition of fibrous connective tissue, which encapsulates regenerating hepatocytes, forming "pseudolobules" that destroy the original tissue structure of the liver. Ultimately, this leads to nodular hardening of the liver, impaired liver function, and even complete loss of liver function, resulting in cirrhosis. Liver fibrosis is histologically reversible, while cirrhosis is more difficult to reverse, but a small percentage of cases are still reversible.

[0004] Many chronic diseases can cause liver fibrosis, such as chronic viral hepatitis, chronic alcoholism, cholestasis, congenital metabolic disorders due to enzyme deficiencies, and long-term exposure to toxins and drugs. Liver fibrosis and cirrhosis are among the main reasons affecting the quality of life and medical expenses of liver disease patients. The market demand for liver-protective drugs is increasing year by year.

[0005] Liver fibrosis is a key pathological process in the progression of chronic hepatitis B to cirrhosis. Reversing or delaying the natural progression of liver fibrosis will significantly reduce the incidence of cirrhosis and liver cancer, and lower patient mortality.

[0006] Currently, there are no specific and effective drugs for treating liver fibrosis in clinical practice. Treatment primarily focuses on alleviating liver damage and inflammation by treating the underlying diseases causing liver injury, and on preventing and treating liver fibrosis. Liver fibrosis treatment includes both etiological treatment and anti-fibrotic therapy. For chronic hepatitis B-related liver fibrosis, inhibiting and clearing HBV can reduce persistent liver damage and, to some extent, promote the repair of fibrotic liver tissue. However, for patients with hepatitis B-related liver fibrosis / cirrhosis, antiviral therapy alone cannot completely resolve the problem. The annual incidence of compensated cirrhosis progressing to decompensated cirrhosis is 3%–5%, the 5-year survival rate for decompensated cirrhosis is only 14%–35%, and the annual incidence of hepatocellular carcinoma in cirrhotic patients is 3%–6%. Therefore, there is an urgent need for effective anti-fibrotic drugs, especially those for treating or preventing progressive liver fibrosis or compensated cirrhosis caused by chronic hepatitis B, for which there are very few reports. Patients and doctors urgently need to find safe and effective treatments. Summary of the Invention

[0007] This invention provides the use of the compound of formula (I) (i.e., hydroxynidone), its solvate, hydrate, prodrug, or pharmaceutically acceptable salt thereof in the preparation of medicaments for the treatment and / or prevention of chronic hepatitis B with liver fibrosis. .

[0008] The present invention also provides compounds of formula (I), their solvates, hydrates, prodrugs or pharmaceutically acceptable salts thereof, for the treatment and / or prevention of chronic hepatitis B with liver fibrosis.

[0009] The present invention also provides a method for treating and / or preventing chronic hepatitis B with liver fibrosis, comprising the step of administering to a patient in need a therapeutic or preventative effective amount of the compound of formula (I), its solvate, hydrate, prodrug, or a pharmaceutically acceptable salt thereof.

[0010] According to some embodiments of the present invention, the chronic hepatitis B with liver fibrosis can be portal fibrosis, significant liver fibrosis, progressive liver fibrosis, or cirrhosis.

[0011] According to some embodiments of the present invention, the cirrhosis may be compensated cirrhosis or decompensated cirrhosis.

[0012] According to some embodiments of the present invention, the portal fibrosis can be characterized by the Ishak scoring system, for example, it can refer to liver fibrosis with an Ishak score ≥1. For example, liver fibrosis with an Ishak score ≥1, an Ishak score ≥2, or an Ishak score of approximately 1-3, approximately 1-2, or approximately 2-3, etc.

[0013] According to some embodiments of the present invention, the portal fibrosis can be characterized by the Scheuer scoring system, for example, it can refer to liver fibrosis with a Scheuer score of approximately ≥1, or liver fibrosis with a Scheuer score of approximately 1-2.

[0014] According to some embodiments of the present invention, the portal fibrosis can be characterized by the METAVIR scoring system, for example, it can refer to liver fibrosis with a METAVIR score of approximately ≥1, or liver fibrosis with a METAVIR score of approximately 1-2.

[0015] According to some embodiments of the present invention, the portal fibrosis can be characterized by the Knodell scoring system, for example, it can refer to liver fibrosis with a Knodell score of approximately ≥1, or liver fibrosis with a Knodell score of approximately 1-2.

[0016] According to some embodiments of the present invention, the significant liver fibrosis can be characterized by the Ishak scoring system, for example, it can refer to liver fibrosis with an Ishak score ≥3. For example, liver fibrosis with an Ishak score ≥4, or an Ishak score of approximately 3-6, approximately 3-5, approximately 3-4, approximately 4-6, approximately 4-5, or approximately 5-6, etc.

[0017] According to some embodiments of the present invention, the significant liver fibrosis can be characterized by the Scheuer scoring system, for example, it can refer to liver fibrosis with a Scheuer score of approximately ≥2 or ≥3, or liver fibrosis with a Scheuer score of approximately 2-4, 3-4 or 2-3.

[0018] According to some embodiments of the present invention, the significant liver fibrosis can be characterized by the METAVIR scoring system, for example, it can refer to liver fibrosis with a METAVIR score of approximately ≥2 or ≥3, or liver fibrosis with a METAVIR score of approximately 2-4, 3-4 or 2-3.

[0019] According to some embodiments of the present invention, the significant liver fibrosis can be characterized by the Knodell scoring system, for example, it can refer to liver fibrosis with a Knodell score of approximately ≥2 or ≥3, or liver fibrosis with a Knodell score of approximately 2-4, 3-4 or 2-3.

[0020] According to some embodiments of the present invention, the significant liver fibrosis can be characterized by liver stiffness measurement (LSM), for example, it can refer to liver fibrosis with an LSM of approximately ≥5.8 kPa, such as approximately 5.8-12.4 kPa, approximately 5.8-9.4 kPa, approximately 5.8-8 kPa, approximately 5.8-7.4 kPa, approximately 5.8-6 kPa, or approximately 9.4-12.4 kPa. The liver stiffness measurement can be measured using conventional equipment in the art, such as FibroTouch or FibroScan devices.

[0021] According to some embodiments of the present invention, the advanced liver fibrosis can be characterized by the Ishak scoring system, for example, it can refer to liver fibrosis with an Ishak score ≥4. For example, an Ishak score of approximately ≥5, such as approximately 4-6, approximately 4-5, or approximately 5-6 indicates liver fibrosis.

[0022] According to some embodiments of the present invention, the advanced liver fibrosis can be characterized by the Scheuer scoring system, for example, it can refer to liver fibrosis with a Scheuer score of approximately ≥3, or liver fibrosis with a score of approximately 3-4.

[0023] According to some embodiments of the present invention, the advanced liver fibrosis can be characterized by the METAVIR scoring system, for example, it can refer to liver fibrosis with a METAVIR score of approximately ≥3, or liver fibrosis with a score of approximately 3-4.

[0024] According to some embodiments of the present invention, the advanced liver fibrosis can be characterized by the Knodell scoring system, for example, it can refer to liver fibrosis with a Knodell score of approximately ≥3, or liver fibrosis with a Knodell score of approximately 3-4.

[0025] According to some embodiments of the present invention, the advanced liver fibrosis can be characterized by liver stiffness measurements (LSM), for example, LSM approximately ≥6.0 kPa, approximately ≥7.4 kPa, approximately ≥8.0 kPa, approximately ≥9.0 kPa, approximately ≥9.5 kPa, approximately ≥10.0 kPa, approximately ≥12.0 kPa, approximately ≥12.4 kPa; for example, LSM approximately 7.4-17.0 kPa, approximately 7.4-15.0 kPa, approximately 7.4-14.6 kPa, approximately 8.0-17.0 kPa, approximately 8.0-15.0 kPa, approximately 8.0-14.6 kPa, approximately 9.0-17.0 kPa, approximately 9.0-15.0 kPa, approximately 9.0-14.6 kPa, approximately 9.5-17.0 kPa, approximately 9.5-15.0 kPa. kPa, approximately 9.5-14.6 kPa, approximately 10.0-17.0 kPa, approximately 10.0-15.0 kPa, approximately 10.0-14.6 kPa.

[0026] According to some embodiments of the present invention, the advanced liver fibrosis can be characterized by liver stiffness measurement (LSM), for example, liver fibrosis with an LSM of approximately ≥9.0 kPa or ≥10.0 kPa when bilirubin is normal and ALT (alanine aminotransferase) <5 ULN; or, liver cirrhosis with an LSM of approximately ≥6.0 kPa when bilirubin and ALT are normal, for example, liver fibrosis with an LSM of approximately ≥7.0 kPa, approximately ≥8.0 kPa, or approximately ≥9.0 kPa.

[0027] According to some embodiments of the present invention, the cirrhosis can be characterized by the Ishak scoring system, for example, it can refer to cirrhosis with an Ishak score ≥ 5, or cirrhosis with an Ishak score of approximately 5-6.

[0028] According to some embodiments of the present invention, the cirrhosis can be characterized by the Scheuer scoring system, for example, it can refer to cirrhosis with a Scheuer score of approximately ≥4.

[0029] According to some embodiments of the present invention, the cirrhosis can be characterized by the METAVIR scoring system, for example, it can refer to cirrhosis with a METAVIR score of approximately ≥4.

[0030] According to some embodiments of the present invention, the cirrhosis can be characterized by the Knodell scoring system, for example, it can refer to cirrhosis with a Knodell score of approximately ≥4.

[0031] According to some embodiments of the present invention, the cirrhosis can be characterized by liver stiffness measurement (LSM), for example, cirrhosis with an LSM of approximately ≥12.0 kPa, approximately ≥13.0 kPa, approximately ≥15.0 kPa, approximately ≥16.0 kPa, or approximately ≥17.0 kPa.

[0032] According to some embodiments of the present invention, the cirrhosis can also be classified into compensated cirrhosis and decompensated cirrhosis by one of the following three criteria: clinical staging based on major complications, staging based on hepatic venous pressure gradient (HVPG), and staging based on liver function reserve (Child-Pugh score staging).

[0033] According to some embodiments of the present invention, the clinical staging based on major complications may further include two-stage staging, four-stage staging, five-stage staging, and seven-stage staging. The two-stage staging method is based on the natural history of cirrhosis. When there are no serious complications such as ascites, esophageal variceal bleeding (GEVB), or hepatic encephalopathy (HE), it is called compensated cirrhosis. When one of the above complications occurs, it is called decompensated cirrhosis. The four-stage staging method is defined as follows: Stage 1-2 is compensated cirrhosis, and Stage 3-4 is decompensated cirrhosis. Specifically, Stage 1 is without esophageal and gastric varices and ascites; Stage 2 is with esophageal and gastric varices but without ascites; Stage 3 is with ascites with or without esophageal and gastric varices; and Stage 4 is with ruptured esophageal varices (EVB) with or without ascites. The five-stage staging method is defined as follows: stages 1-2 are compensated cirrhosis, and stages 3-5 are decompensated cirrhosis.

[0034] The seven-stage staging method is as follows: stages 0-2 are compensated cirrhosis, and stages 3-6 are decompensated cirrhosis. Specifically, stage 0 is characterized by the absence of esophageal and gastric varices, with HVPG between 5-10 mmHg; stage 1 is characterized by the absence of esophageal and gastric varices, but HVPG ≥ 10 mmHg; stage 2 is characterized by the presence of esophageal and gastric varices; stage 3 is characterized by the presence of endoscopic variceal bleeding (EVB); stage 4 is characterized by the first decompensated event other than bleeding; stage 5 is characterized by a recurrence of a decompensated event; and stage 6, also the late stage of decompensation, is characterized by refractory ascites, hemorrhage (HE), severe infection, renal failure, and acute-on-chronic liver failure.

[0035] According to some embodiments of the present invention, HVPG ≤ 12 mmHg indicates compensated cirrhosis, and HVPG > 12 mmHg indicates decompensated cirrhosis; wherein HVPG < 10 mmHg indicates the absence of varicose veins, and HVPG between 10-12 mmHg indicates clinically significant portal hypertension.

[0036] According to some embodiments of the present invention, Child-Pugh classification A indicates compensated cirrhosis, and Child-Pugh classification B or C indicates decompensated cirrhosis, wherein grade B is the early decompensated stage and grade C is the late decompensated stage.

[0037] According to some embodiments of the present invention, the chronic hepatitis B with liver fibrosis does not progress to liver cancer.

[0038] The Ishak scoring system is a modified version of the Knodell scoring system, classifying liver fibrosis assessment into stages 0-6. It is currently the most sensitive and commonly used method internationally for assessing liver fibrosis in before-and-after case-control studies. According to the "Consensus on Diagnosis and Treatment of Liver Fibrosis (2019)" jointly published by the Chinese Society of Hepatology, the Chinese Society of Gastroenterology, and the Chinese Society of Infectious Diseases (Practical Hepatology Journal, November 2019, Vol. 22, No. 6, pp. 793-803), in recent years, domestic and foreign scholars have quantified the "minority" and "majority" in the Ishak score to reduce human error, and have staged it according to the number of fibrous septa and pseudolobules. "Minority" refers to 1-3 fibrous septa, and more than 3 is "majority". Therefore, the Ishak scoring system describes the staging of liver fibrosis (i.e., Ishak score) as follows: Stage I (Ishak score 1): No fibrous septa; Stage II (Ishak score 2): 1 fibrous septum; Stage III (Ishak score 3): 2-3 fibrous septa; Stage IV (Ishak score 4): 4 fibrous septa; Stage V (Ishak score 5): 4 or more fibrous septa + 1-3 clearly defined pseudolobules; Stage VI (Ishak score 6): More than 3 pseudolobules.

[0039] According to some embodiments of the invention, an Ishak score of 0 may indicate the absence of fibrosis. In some embodiments, an Ishak score of 1 may indicate some portal fibrosis (PF), with or without short fibrous septa. In some embodiments, an Ishak score of 2 may indicate majority portal fibrosis, with or without one fibrous septum. In some embodiments, an Ishak score of 3 may indicate majority portal fibrosis, with 2-3 fibrous septa. In some embodiments, an Ishak score of 4 may indicate portal fibrosis with significant portal-portal bridging fibrosis and portal-central bridging fibrosis, with 4 or more fibrous septa. In some embodiments, an Ishak score of 5 may indicate significant portal-portal bridging fibrosis and / or portal-central bridging fibrosis, with 1-3 pseudolobules. In some embodiments, an Ishak score of 6 may indicate more than 3 pseudolobules, indicating cirrhosis.

[0040] According to some embodiments of the present invention, the Ishak score can be converted into another scoring system. The scoring system could be, for example, the Scheuer score, the Metavir score, or the Knodell score.

[0041] The Scheuer scoring system classifies liver fibrosis into stages 0-4. According to some embodiments of the invention, a Scheuer score of 0 may indicate no fibrosis. In some embodiments, a score of 1 may indicate portal enlargement. In some embodiments, a score of 2 may indicate portal fibrosis (PF) with fibrous septa formation. In some embodiments, a score of 3 may indicate fibrous septa with lobular disorganization. In some embodiments, a score of 4 may indicate cirrhosis. In some embodiments, the Scheuer score may be ≥1, ≥2, or ≥3, or it may be 0-4, 1-4, 2-4, 3-4, 1-3, or 2-3.

[0042] The METAVIR scoring system classifies liver fibrosis into stages 0-4. According to some embodiments of the invention, a METAVIR score of F0 may indicate no fibrosis. In some embodiments, a METAVIR score of 1 may indicate portal fibrosis without fibrotic septa. In some embodiments, a METAVIR score of 2 may indicate portal fibrosis with a small number of fibrotic septa. In some embodiments, a METAVIR score of 3 may indicate septal fibrosis and structural disorder. In some embodiments, a METAVIR score of 4 may indicate cirrhosis. In some embodiments, a METAVIR score may be ≥1, ≥2, or ≥3, or may be 0-4, 1-4, 2-4, 3-4, 1-3, 1-2, or 2-3.

[0043] The Knodell scoring system classifies liver fibrosis into stages 0-4. According to some embodiments of the invention, a Knodell score of 0 may indicate no fibrosis. In some embodiments, a Knodell score of 1 may indicate mild fibrosis with portal enlargement. In some embodiments, a Knodell score of 2 may indicate moderate fibrosis. In some embodiments, a Knodell score of 3 may indicate severe fibrosis, with portal-portal bridging fibrosis and portal-central bridging fibrosis. In some embodiments, a Knodell score of 4 may indicate cirrhosis. In some embodiments, a Knodell score may be ≥1, ≥2, or ≥3, or it may be 1-4, 1-3, 2-4, or 2-3.

[0044] The Child-Pugh classification is a clinically used grading standard for quantitatively assessing liver reserve function in patients with cirrhosis. It categorizes five indicators (general condition, ascites, serum bilirubin, serum albumin concentration, and prothrombin time; the general condition can also be represented by the presence and severity of hepatic encephalopathy) into three levels, scored as 1, 2, and 3 points respectively. The scores for the five indicators are summed, with a minimum total score of 5 and a maximum of 15. Based on this total score, liver reserve function is classified into three grades: A (5-6 points), B (7-9 points), and C (10-15 points). A higher score indicates poorer liver reserve function.

[0045] According to an embodiment of the present invention, the dosage of the compound represented by formula (I) can be 50-500 mg / day, for example 80-450 mg / day, 100-400 mg / day, 120-360 mg / day, and exemplary 180 mg / day, 270 mg / day, and 360 mg / day.

[0046] According to an embodiment of the present invention, the drug can be administered 1-5 times / day, for example 3 times / day.

[0047] According to embodiments of the present invention, the pharmaceutically acceptable salt comprises a salt formed by the compound of formula (I) with an organic acid selected from propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, and citric acid, or an acidic amino acid selected from aspartic acid and glutamic acid, and then with an inorganic base, including sodium, potassium, calcium, aluminum, and ammonium salts; or a salt formed with an organic base, including methylamine salts, ethylamine salts, and ethanolamine salts; or a salt formed with a basic amino acid selected from lysine, arginine, and ornithine, and then with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; or a salt formed with an organic acid selected from formic acid, acetic acid, picric acid, methanesulfonic acid, and ethanesulfonic acid.

[0048] According to embodiments of the present invention, the drug further includes one or more pharmaceutically acceptable carriers or excipients.

[0049] According to embodiments of the present invention, the drug may be an oral preparation or a parenteral preparation. Further, the oral preparation may be a tablet, capsule, pill, dispersible powder, granule, oral liquid, syrup, elixir, etc.; the parenteral preparation may be an injection, powder for injection, etc.

[0050] Some examples of suitable excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. Formulations may also contain: lubricants such as talc, magnesium stearate, and mineral oil; humectants; emulsifiers and suspending agents; preservatives such as methyl benzoate and hydroxypropyl benzoate; sweeteners and flavoring agents. The medicaments of the present invention can be formulated using methods known in the art to provide immediate, sustained, or delayed release of the active ingredient upon administration to a patient.

[0051] The tablets or pills of the present invention can be coated or compounded to obtain dosage forms that provide the advantage of long-lasting action. For example, the tablets or pills contain an internal dose and an external dose component, the latter being a coated form of the former. The two components can be isolated by an enteric coating layer, which is used to prevent disintegration in the stomach, allowing the internal component to pass through the duodenum intact or to delay release. A variety of substances can be used for such enteric coatings or coatings, including a variety of high molecular weight acids and mixtures of high molecular weight acids with such substances such as shellac, cetyl alcohol, and cellulose acetate. The compounds of the present invention can be incorporated into liquid forms for oral or injectable administration, including aqueous solutions, appropriately flavored syrups, water or oil suspensions; and emulsions prepared using edible oils such as cottonseed oil, sesame oil, medium-chain triglycerides, coconut oil, or peanut oil; as well as elixirs and similar pharmaceutical solvents.

[0052] Beneficial effects The present invention provides the use of the compound of formula (I), its solvate, hydrate, pharmaceutical prodrug, or pharmaceutically acceptable salt thereof, in the preparation of a medicament for the treatment or prevention of chronic hepatitis B with liver fibrosis, which can effectively reverse liver fibrosis, particularly in the treatment of significant fibrosis, advanced liver fibrosis, or cirrhosis. Attached Figure Description

[0053] Figure 1 The distribution of Ishak scores (FAS) for liver fibrosis before and after treatment.

[0054] Figure 2 The distribution of Ishak scores (PPS) for liver fibrosis before and after treatment.

[0055] Figure 3A graph showing the intergroup difference (FAS) for a decrease of ≥1 point in Ishak score from baseline after 52 weeks of treatment for liver fibrosis.

[0056] Figure 4 Plot of intergroup differences (PPS) for Ishak score decrease of ≥1 point from baseline after 52 weeks of treatment for liver fibrosis. Detailed Implementation

[0057] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0058] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0059] Preparation example: Preparation of hydroxynidone capsules Prescription: The prescription composition of each unit dose product is detailed in the table below:

[0060] Preparation method: The detailed production process is described below, using the process validation batch (batch size: 900,000 units) as an example: 1) Preparation of raw and auxiliary materials: - Collect raw and auxiliary materials and carefully check the product name, batch number, weight, manufacturer, etc., and check that the packaging is intact.

[0061] - Pass lactose monohydrate, magnesium stearate, silicon dioxide, and hydroxynidone through an 80-mesh sieve for later use.

[0062] 2) Weighing and batching: Weigh each raw and auxiliary material accurately according to the batch instructions and verify the weighing.

[0063] 3) Mixing: Add all materials to the three-dimensional motion mixer, turn on the machine, slowly increase the speed to 800 r / min, mix for 15 minutes to obtain the intermediate product of mixed powder, check the material balance, which should be between 98.00% and 100.00%.

[0064] 4) Capsule filling: Using a No. 4 automatic hard capsule filling machine, the filling amount is adjusted according to the content of intermediate products to ensure that the weight difference is within the specified range before filling.

[0065] 5) Polishing and Capsule Removal: Polish the filled capsules. Collect the polished capsules and place them in a tray. Sift out the fine powder adhering to the capsule surface and remove defective products such as those with lenticular ends, wrinkles, gaps, air bubbles, breaks, dents, and double caps. Check the material balance; it should be between 98.00% and 100.00%.

[0066] 6) Inner packaging: Bottle and label, 63 capsules / bottle. Check material balance, which should be between 98.00% and 100.00%.

[0067] 7) Outsourcing.

[0068] Example of effect 1. Experimental Design 1.1. Overall Experimental Design This trial is a randomized, double-blind, placebo-controlled, multicenter, dose-exploratory phase II clinical study with baseline treatment. In addition to entecavir capsule antiviral therapy, this clinical trial established three different dose groups and a placebo control group, with a planned sample size of 60 patients in each group, for a total of 240 patients. The three dose groups were 180 mg, 270 mg, and 360 mg. The allocation ratio of each experimental group to the control group was 1:1:1:1, and cases were randomly assigned.

[0069] 1.2. Subject Selection Selection criteria: (1) Age 18-65 years old, gender not limited (2) History of chronic hepatitis B, HBsAg positive for ≥6 months (3) ALT (alanine aminotransferase) < 8 times ULN (upper limit of standard) (4) Significant liver fibrosis confirmed by liver biopsy (Ishak score ≥3, excluding decompensated cirrhosis) (5) HBeAg positive patients, HBV DNA > 2.0 × 10 4 IU / mL (10 5 (copy / mL); HBeAg-negative patients with HBV DNA > 2.0 × 10⁻⁶ 3 IU / mL (10 4 (copy / mL) (6) Have not received interferon and / or nucleoside analog antiviral therapy within 3 months prior to enrollment. (7) No anti-inflammatory or liver-protective drugs were taken within one month prior to selection. (8) Have not taken any medications with anti-fibrotic effects within the three months prior to selection. (9) Has the ability to understand and sign informed consent forms before conducting the research. Exclusion criteria: (1) Anyone who does not meet any of the inclusion criteria (2) Those who have had massive upper gastrointestinal bleeding within 3 months prior to enrollment (3) TBiL (total bilirubin) > 3 times ULN, or 3ULN < ALT < 8ULN and TBiL > 2 times ULN. (4) Although there are no signs of liver cancer, AFP (alpha-fetoprotein) >100 μg / L (5) PLT (platelet count) ≤ 60 × 10 9 / L (6) PTA (prothrombin activity) < 50% (7) Ultrasound showed a significant space-occupying lesion in the liver, suggesting a tumor. (8) The portal vein width is ≥1.2cm according to B-ultrasound examination. (9) Those with a Body Mass Index (BMI) > 30 (10) Patients with decompensated liver cirrhosis and liver tumors (11) Patients with alcoholic, drug-induced, hereditary, immune-related, other viral and non-viral chronic hepatitis (12) Patients with serious cardiovascular, pulmonary, renal, endocrine, nervous and hematological diseases, as well as mental patients. (13) Active peptic ulcer (14) Pregnant and lactating women (15) Individuals who have participated in other drug trials in the past 3 months (16) Patients deemed unsuitable for participation in this study by other researchers Exit criteria: (1) Subjects voluntarily withdraw or withdraw informed consent. (2) Loss to follow-up of subjects (3) The researchers decided to terminate the treatment. (4) The subject is pregnant or decides to become pregnant. (5) Any of the following applies to the subject: ①ALT>8ULN; ②ALT > 5ULN for 2 weeks; ③ALT > 3ULN and (TBiL > 3ULN or INR (International Normalized Ratio) > 1.5ULN); ④ ALT > 3 ULN accompanied by progressively worsening fatigue, nausea, vomiting, right upper quadrant pain or tenderness, fever, rash, and / or eosinophilia (> 5%). (6) The subject developed decompensated cirrhosis. (7) The subject developed liver cancer. (8) Patients who are allergic to this drug, experience serious adverse events, develop complications, or have special physiological changes are not suitable to continue the trial. Exclusion criteria: (1) Mistakenly included (2) Those who have never taken medication (3) Those with no testing records (4) The efficacy and safety cannot be evaluated due to the use of a prohibited drug. (5) Although the experiment was completed, the dosage was not within 80%-120% of the recommended dosage. (6) Discontinuation of investigational drug for a cumulative period of 4 weeks or more (7) Cases of unblinding or emergency unblinding 1.3. Test Drugs Both hydroxynidone capsules and placebo capsules were developed and supplied by Shanghai Ruixing Gene Technology Co., Ltd., and they are completely identical in appearance. Hydroxynidone capsule specification: 30mg / capsule. Investigational drug batch numbers: 20150201, 20170101.

[0070] Storage of the investigational drug: Store at room temperature. The shelf life of the drug is 36 months.

[0071] All clinical trial drugs were prepared by Beijing Contini Pharmaceutical Co., Ltd. in a GMP-compliant workshop and tested to meet the quality standards approved by the National Medical Products Administration.

[0072] Entecavir capsules (0.5mg / capsule, 7 capsules / blister / box, shelf life 24 months, batch numbers: 1411102, 1506204, 1606201, 1606202, 1602205, 1612205, 1703202, 3031802, 2031824), a basic antiviral treatment drug, were developed and manufactured by Fujian Guang Sheng Tang Pharmaceutical Co., Ltd., and provided by Shanghai Ruixing Gene Technology Co., Ltd.

[0073] Storage of entecavir capsules: Protect from light, seal tightly, and store in a dry place below 25°C.

[0074] Route of administration and dosage settings: Based on the results of animal pharmacodynamic experiments and Phase I clinical trials, the dosage of the test drug in each group of this experiment was determined as follows: 180mg group: Hydroxynidone capsules, 60mg orally three times a day, for 52 consecutive weeks; 270mg group: Hydroxynidone capsules, 90mg orally three times a day, for 52 consecutive weeks; 360mg group: Hydroxynidone capsules, 120mg orally three times a day, for 52 consecutive weeks; Placebo group (control group): Placebo capsules, 4 capsules each time, three times a day, orally, for 52 consecutive weeks; Basic treatment: Entecavir capsules, 0.5 mg once daily, orally, for 52 weeks; Both the placebo group and each dose group of hydroxyketone received basic entecavir treatment concurrently.

[0075] 1.4. Test Procedure 1.4.1. Screening Period / Baseline Period ● Demography: including date of birth, age, sex, ethnicity, height, weight, and occupation.

[0076] ● Present medical history, past medical history (cardiovascular, respiratory, digestive, genitourinary, nervous, mental systems, etc.), medication history, and habits such as smoking and drinking.

[0077] ● Clinical symptom record.

[0078] ● Sign the informed consent form.

[0079] ● Assign filter numbers.

[0080] ● Physical examination.

[0081] ● Routine laboratory tests: complete blood count, urinalysis, urine pregnancy test (for women of childbearing age), blood glucose, etc.

[0082] ● Serological tests: alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), total protein (TP), albumin (A), globulin (G), total bilirubin (TBil), direct bilirubin (DBil), blood urea nitrogen (BUN), creatinine (Cr), triglycerides (TG), cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), prothrombin time (PT), prothrombin activity (PTA), alpha-fetoprotein (AFP).

[0083] ● Transient elastography of the liver (LSM) (kPa).

[0084] ● Virological tests: HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, HBV DNA, anti-HCV.

[0085] ● Electrocardiogram: 12-lead electrocardiogram.

[0086] ● Ultrasound examination: to examine the liver, gallbladder, spleen, portal vein width and / or blood flow velocity, etc.

[0087] ● Percutaneous liver biopsy (results within 3 months prior to screening can be used as baseline data).

[0088] ● Fill in the CRF.

[0089] 1.4.2. Treatment Period 1.4.2.1. At week 0 ● Assign drug numbers.

[0090] ● Distribute experimental drugs.

[0091] ● Record any accompanying medications.

[0092] ● Record adverse events.

[0093] ● Fill in the CRF.

[0094] 1.4.2.2. 1 week (±1 day) ● Record the administration of the experimental drug.

[0095] ● Record any accompanying medications.

[0096] ● Record adverse events.

[0097] ● Fill in the CRF.

[0098] 1.4.2.3. At 4 weeks (±5 days) ● Recover and distribute experimental drugs.

[0099] ● Record the administration of the experimental drug.

[0100] ● Record any accompanying medications.

[0101] ● Physical examination.

[0102] ● Record clinical symptoms.

[0103] ● Routine laboratory tests: complete blood count, urinalysis, and urine pregnancy test (for women of childbearing age).

[0104] ● Serological tests: ALT, AST, GGT, ALP, TP, A, G, TBil, DBil, BUN, Cr, TG, TC, LDL-C, HDL-C, PT, PTA, AFP.

[0105] ● Virological test: HBV DNA.

[0106] ● Record adverse events.

[0107] ● Fill in the CRF.

[0108] 1.4.2.4. At 8 weeks (±5 days) ● Recover and distribute experimental drugs.

[0109] ● Record the administration of the experimental drug.

[0110] ● Record any accompanying medications.

[0111] ● Physical examination.

[0112] ● Record clinical symptoms.

[0113] ● Serological tests: ALT, AST, GGT, ALP, TP, A, G, TBil, DBil, TG, TC, LDL-C, HDL-C.

[0114] ● Virological test: HBV DNA.

[0115] ● Record adverse events.

[0116] ● Fill in the CRF.

[0117] 1.4.2.5. At 12 weeks (±5 days) ● Recover and distribute experimental drugs.

[0118] ● Record the administration of the experimental drug.

[0119] ● Record any accompanying medications.

[0120] ● Physical examination.

[0121] ● Record clinical symptoms.

[0122] ● Routine laboratory tests: complete blood count, urinalysis, urine pregnancy test (for women of childbearing age), blood glucose, etc.

[0123] ● Serological tests: ALT, AST, GGT, ALP, TP, A, G, TBil, DBil, BUN, Cr, TG, TC, LDL-C, HDL-C, PT, PTA, AFP.

[0124] ● Virological test: HBV DNA.

[0125] ● Electrocardiogram (ECG).

[0126] ● Record adverse events.

[0127] ● Fill in the CRF.

[0128] 1.4.2.6. At 16 weeks (±5 days) ● Recover and distribute experimental drugs.

[0129] ● Record the administration of the experimental drug.

[0130] ● Record any accompanying medications.

[0131] ● Physical examination.

[0132] ● Record clinical symptoms.

[0133] ● Serological test: ALT.

[0134] ● Record adverse events.

[0135] ● Fill in the CRF.

[0136] 1.4.2.7. At 20 weeks (±5 days) ● Recover and distribute experimental drugs.

[0137] ● Record the administration of the experimental drug.

[0138] ● Record any accompanying medications.

[0139] ● Physical examination.

[0140] ● Record clinical symptoms.

[0141] ● Serological test: ALT.

[0142] ● Record adverse events.

[0143] ● Fill in the CRF.

[0144] 1.4.2.8. At 24 weeks (±5 days) ● Recover and distribute experimental drugs.

[0145] ● Record the administration of the experimental drug.

[0146] ● Record any accompanying medications.

[0147] ● Physical examination.

[0148] ● Record clinical symptoms.

[0149] ● Routine laboratory tests: complete blood count, urinalysis, urine pregnancy test (for women of childbearing age), blood glucose, etc.

[0150] ● Serological tests: ALT, AST, GGT, ALP, TP, A, G, TBil, DBil, BUN, Cr, TG, TC, LDL-C, HDL-C, PT, PTA, AFP.

[0151] ● Transient elastography of the liver (LSM) (kPa).

[0152] ● Virological test: HBV DNA.

[0153] ● Electrocardiogram (ECG).

[0154] ● Ultrasound examination: to examine the liver, gallbladder, spleen, portal vein width and / or blood flow velocity, etc.

[0155] ● Record adverse events.

[0156] ● Fill in the CRF.

[0157] 1.4.2.9. 28 weeks (±5 days) ● Recover and distribute experimental drugs.

[0158] ● Record the administration of the experimental drug.

[0159] ● Record any accompanying medications.

[0160] ● Physical examination.

[0161] ● Record clinical symptoms.

[0162] ● Record adverse events.

[0163] ● Fill in the CRF.

[0164] 1.4.2.10. At 32 weeks (±5 days) ● Recover and distribute experimental drugs.

[0165] ● Record the administration of the experimental drug.

[0166] ● Record any accompanying medications.

[0167] ● Physical examination.

[0168] ● Record clinical symptoms.

[0169] ● Record adverse events.

[0170] ● Fill in the CRF.

[0171] 1.4.2.11.36 weeks (±5 days) ● Recover and distribute experimental drugs.

[0172] ● Record the administration of the experimental drug.

[0173] ● Record any accompanying medications.

[0174] ● Physical examination.

[0175] ● Record clinical symptoms.

[0176] ● Routine laboratory tests: complete blood count, urinalysis, urine pregnancy test (for women of childbearing age), blood glucose, etc.

[0177] ● Serological tests: ALT, AST, GGT, ALP, TP, A, G, TBil, DBil, BUN, Cr, TG, TC, LDL-C, HDL-C, PT, PTA, AFP.

[0178] ● Virological test: HBV DNA.

[0179] ● Electrocardiogram (ECG).

[0180] ● Record adverse events.

[0181] ● Fill in the CRF.

[0182] 1.4.2.12. At 40 weeks (±5 days) ● Recover and distribute experimental drugs.

[0183] ● Record the administration of the experimental drug.

[0184] ● Record any accompanying medications.

[0185] ● Physical examination.

[0186] ● Record clinical symptoms.

[0187] ● Record adverse events.

[0188] ● Fill in the CRF.

[0189] 1.4.2.13. At 44 weeks (±5 days) ● Recover and distribute experimental drugs.

[0190] ● Record the administration of the experimental drug.

[0191] ● Record any accompanying medications.

[0192] ● Physical examination.

[0193] ● Record clinical symptoms.

[0194] ● Record adverse events.

[0195] ● Fill in the CRF.

[0196] 1.4.2.14. At 48 weeks (±5 days) ● Recover and distribute experimental drugs.

[0197] ● Record the administration of the experimental drug.

[0198] ● Record any accompanying medications.

[0199] ● Physical examination.

[0200] ● Record clinical symptoms.

[0201] ● Record adverse events.

[0202] ● Fill in the CRF.

[0203] 1.4.2.15. At 52 weeks (±5 days) ● Recover the experimental drug.

[0204] ● Record the administration of the experimental drug.

[0205] ● Record any accompanying medications.

[0206] ● Physical examination.

[0207] ● Record clinical symptoms.

[0208] ● Routine laboratory tests: complete blood count, urinalysis, urine pregnancy test (for women of childbearing age), blood glucose, etc.

[0209] ● Serological tests: ALT, AST, GGT, ALP, TP, A, G, TBil, DBil, BUN, Cr, TG, TC, LDL-C, HDL-C, PT, PTA, AFP.

[0210] ● Transient elastography of the liver (LSM) (kPa).

[0211] ● Virological tests: HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, HBV DNA.

[0212] ● Electrocardiogram (ECG).

[0213] ● Ultrasound examination: to examine the liver, gallbladder, spleen, portal vein width and / or blood flow velocity, etc.

[0214] ● Percutaneous liver biopsy.

[0215] ● Record adverse events.

[0216] Fill in the CRF.

[0217] 1.4.3. Additional Follow-up 1.4.3.1. Additional Follow-up Conditions Additional follow-up should be performed if any of the following three conditions occur. ● If a subject stops taking the investigational drug for any reason, but is still able to be followed up on schedule.

[0218] ● Case dropout: Subjects stop taking the medication and follow up.

[0219] ● Follow-up should be increased after adverse events occur in the subjects.

[0220] 1.4.3.2. Additional Follow-up Content ● Record the reasons and timing for any additional follow-up visits.

[0221] ● Record the administration of the experimental drug.

[0222] ● Record any accompanying medications.

[0223] ● Record clinical symptoms as needed, and conduct physical examinations, routine laboratory and biochemical tests, virological tests, electrocardiograms, ultrasounds, and other auxiliary examinations.

[0224] ● Record adverse events.

[0225] ● Fill in the CRF.

[0226] 1.5. Observation Indicators The main observation indicator is the proportion of patients whose Ishak score for liver fibrosis decreased by more than 1 point after treatment compared to before treatment.

[0227] Secondary observation indicators: (1) The seroconversion rate of HBV DNA after treatment (HBV DNA < 20 copies / mL) and the magnitude of the decrease.

[0228] (2) The percentage decrease in transient elastography LSM (kPa) value after treatment compared to before treatment.

[0229] (3) The proportion of liver tissue inflammation grade decreased by grade I or higher after treatment and fibrosis did not worsen.

[0230] (4) Improvement of ALT levels in liver function.

[0231] Safety indicators: vital signs, laboratory indicators, adverse events / adverse reactions, etc.

[0232] 1.6. Statistical Protocol and Sample Size Determination ● Full Analysis Set (FAS) Based on the intention-to-treat principle, all randomized cases that had used the study drug at least once and had post-treatment evaluation data constituted the FAS (Follow-up Observational Study) in this study. Missing data in the efficacy-related portion of the FAS were carried forward using the Last Observation Carry Forward (LOCF) method. The FAS represents the primary population for efficacy evaluation.

[0233] ● Compliant with Protocol Set (PPS) The PPS population will be finalized during blinded data review and will be considered a secondary population for efficacy evaluation, based on at least the following criteria: 1) Meets the inclusion criteria specified in the test protocol.

[0234] 2) Complete all planned visits.

[0235] 3) No drugs or treatments that could affect the evaluation of efficacy were used during the trial.

[0236] 4) Good compliance (dosage rate between 80% and 120%).

[0237] ● Safety Set (SS) dataset for safety evaluation All cases that were randomly assigned, used the study drug at least once, and had post-treatment safety evaluation data constituted the safety population for this study.

[0238] ● Handling missing data and outliers For missing, unused, incorrect, or unreasonable data, please first verify the original data table with your center and supplement or correct it. Data that cannot be supplemented or corrected, and for which the main efficacy indicators are completely missing, incorrect, or contradictory, will not be included in the statistical analysis.

[0239] For missing data in the primary efficacy endpoints, the Last Observation Carryover (LOCF) method was used to fill in the missing data. Secondary efficacy endpoints and safety endpoints were not filled in.

[0240] ● General principles of statistical analysis All statistical analyses were performed using SAS 9.4 or later, a professional statistical analysis software.

[0241] For the primary efficacy endpoint, the superiority test was used to compare the experimental group with the placebo control group; a p-value ≤ 0.05 was considered a valid measure of superiority. All other tests were two-tailed. P A difference ≤0.05 will be considered statistically significant.

[0242] Quantitative indicators are described using mean, standard deviation, median, minimum, and maximum values; categorical indicators are described using frequency and percentage.

[0243] For quantitative indicators, the Wilcoxon signed-rank test was used to compare differences within each group with baseline values ​​during the screening period, and the Kruskal-Wallis H test was used to compare differences between groups before and after treatment. Changes in key efficacy indicators relative to baseline were compared between groups using an analysis of covariance model, with baseline as a covariate and the effects of centering and grouping considered.

[0244] For count data, CMH- The groups were compared using either the test or Fisher's exact probability method.

[0245] ● Shedding analysis Summarize the number of enrolled and completed cases at each center, calculate the dropout / rejection rate and reasons for dropout at each center, and compile a list of dropout cases.

[0246] The comparison of the total shedding rate and the shedding rate due to adverse events in each group will be conducted using CMH- Test or Fisher's exact probability method.

[0247] ● Demographic data and baseline characteristics analysis Statistical descriptions of patients' demographic characteristics (age, height, vital signs, etc.) were performed, and analysis of variance or Kruskal-Wallis H test was used. Tests or Fisher's exact probability method can be used to compare demographic data with other baseline indicators such as vital signs, medical history, and basic treatment to measure the balance between groups.

[0248] ● Compliance Analysis Adherence was categorized into good and poor for each group, and the adherence rates of each group were compared and analyzed. Good adherence was defined as 80% ≤ actual dosage / prescribed dosage ≤ 120%.

[0249] ● Validity Analysis The primary time point for efficacy evaluation is the end of 12 months (for PPS) or the time of discontinuation (for FAS). Efficacy at other follow-up time points will also be described.

[0250] For the primary efficacy endpoint, the superiority test was used to compare the experimental group with the placebo control group. A p-value ≤ 0.05 was considered to indicate superiority.

[0251] Changes in efficacy endpoints relative to baseline were assessed. For quantitative endpoints, changes before and after treatment within each group were evaluated using the signed-rank test. Between-group comparisons were performed using analysis of covariance, with baseline as a covariate, considering the center effect, and calculating the least squares mean (LSMEAN) and its 95% CI. For categorical endpoints, the CMH-χ² test was used, considering the center effect, and calculating the 95% CI of the difference between the two rates. Logistic regression was used for evaluation when necessary.

[0252] ● Therapeutic effect analysis ● Key therapeutic indicators The proportion of patients whose Ishak scores for liver fibrosis decreased by 1 point or more after treatment compared with those before treatment was assessed. The CMH-χ2 test, which takes into account the center effect, was used to compare the differences between groups. Confounding factors that were difficult to control or were not controlled before treatment, such as factors that were unbalanced between groups before treatment and might affect the evaluation of efficacy, were used as covariates. Logistic regression model was used to evaluate the differences in efficacy between groups, taking into account the center effect.

[0253] ● Secondary efficacy indicators 1. The HBV DNA seroconversion rate after treatment was compared between groups using the CMH-χ2 test or Fisher's exact test.

[0254] 2. The Kruskal-Wallis H test was used to compare the decrease in liver tissue elasticity between groups before and after treatment.

[0255] 3. The Kruskal-Wallis H test was used to compare the changes in liver tissue inflammation grade scores before and after treatment between groups; the CMH-χ2 test or Fisher's exact test was used to compare the proportion of patients whose liver tissue inflammation grade scores decreased by one grade or more after treatment.

[0256] 4. The CMH-χ2 test or Fisher's exact test was used to compare the rates of normalization of liver function indicators between groups after treatment. The analysis of variance or Kruskal-Wallis H test was used to compare the measured values ​​of each indicator between groups.

[0257] ● Security Analysis Safety indicators include adverse events / adverse reactions, laboratory indicators, physical examinations, etc.

[0258] 1. Using CMH- χ Compare the incidence of adverse events / adverse reactions in each group using the 2-test or Fisher's exact probability method, and describe in a table the frequency and severity of all adverse events in this trial, their relationship with the drug, treatment measures, and outcomes. 2. List and describe any normal / abnormal changes in physical examination results before and after the experiment.

[0259] 3. List and describe the normal / abnormal changes and abnormal exacerbations of laboratory indicators before and after the experiment, and evaluate their relationship with the investigational drug. Statistical descriptions of measurement indicators at each measurement time point were provided. Within-group comparisons were performed using the Wilcoxon signed-rank test, and between-group comparisons were performed using the Kruskal-Wallis H test.

[0260] 2. Research Results: A total of 168 subjects were randomly enrolled in this trial, with 21 dropouts / cancellations (21 / 168, 12.50%). The full analysis set (FAS) included 167 subjects (43 in the placebo control group, 42 in the 180mg hydroxynidone capsule group, 41 in the 270mg hydroxynidone capsule group, and 41 in the 360mg hydroxynidone capsule group), the protocol compliance set (PPS) included 147 subjects (42 in the placebo control group, 36 in the 180mg hydroxynidone capsule group, 35 in the 270mg hydroxynidone capsule group, and 34 in the 360mg hydroxynidone capsule group), and the safety set (SS) included 168 subjects (43 in the placebo control group, 42 in the 180mg hydroxynidone capsule group, 42 in the 270mg hydroxynidone capsule group, and 41 in the 360mg hydroxynidone capsule group).

[0261] This study involved 8 trial centers and 168 participants, including 121 males and 47 females. At baseline, the mean age of the participants was 39.55 ± 10.67 years. In the FAS (Frequency Assay), there were no statistically significant differences in demographic data and general characteristics among the groups, except for systolic blood pressure. P >0.05), indicating good balance between groups.

[0262] Main therapeutic effects: The primary efficacy endpoint of this study was the proportion of subjects whose Ishak score for liver fibrosis decreased by 1 point or more after treatment (52 weeks of treatment) compared to baseline.

[0263] 2.1 Analysis of the temporal changes and sensitivity analysis of Ishak scores for liver fibrosis As mentioned above, the pathological evaluation of liver biopsy centers in this trial revealed that although some subjects met the inclusion criteria of the trial, their liver biopsy results showed an Ishak score ≥3, as determined independently by the research unit at the time of enrollment, their pathological evaluation at the liver biopsy center did not meet the Ishak score ≥3. This included subjects with Ishak scores ≥2 and Ishak scores ≥1.

[0264] The inventors evaluated all enrolled subjects, including those with Ishak scores ≥1, ≥2, and ≥3. The primary efficacy endpoint was the proportion of subjects whose liver fibrosis Ishak scores decreased by ≥1 point after treatment (52 weeks of treatment) compared to baseline. The results of FAS and PPS analyses are shown in Table 1.

[0265] The FAS analysis results in Table 1 showed that there was no statistically significant difference in the baseline Ishak scores for liver fibrosis among the four groups. P=0.8610). After 52 weeks of treatment, a certain proportion of subjects in each group showed a decrease of ≥1 point in their Ishak score for liver fibrosis compared to baseline. The changes in Ishak score for liver fibrosis before and after treatment were statistically significant in the hydroxynidone group. P <0.05), there was no statistically significant difference in Ishak scores for liver fibrosis before and after treatment in the placebo control group. P =0.3205), see Figure 1 The proportions of subjects whose Ishak scores for liver fibrosis decreased by ≥1 point from baseline in each group were as follows: placebo control group: 11 cases (25.58%), 180mg hydroxynidone capsule group: 17 cases (40.48%), 270mg hydroxynidone capsule group: 23 cases (56.10%), and 360mg hydroxynidone capsule group: 18 cases (43.90%). The differences between the four groups were statistically significant. P =0.0245). The difference in the proportion of subjects with a ≥1-point decrease in Ishak score for liver fibrosis between the placebo group and the 270mg hydroxynidone group, and its 95% CI, was -30.52% (-48.12%, -9.50%), indicating that 270mg / day hydroxynidone capsules significantly reversed the degree of liver fibrosis compared to placebo, and the reversal effect in the 270mg hydroxynidone capsule group was better than that in the 180mg and 360mg hydroxynidone groups. (See...) Figure 3 .

[0266] The PPS analysis results in Table 1 showed that there was no statistically significant difference in the baseline Ishak scores for liver fibrosis among the four groups. P =0.7361). At 52 weeks of treatment, a certain proportion of subjects in each group experienced a decrease of ≥1 point in their Ishak score for liver fibrosis compared to baseline. The differences in Ishak scores for liver fibrosis before and after treatment were statistically significant in the hydroxynidone group. P <0.05), there was no statistically significant change in Ishak score for liver fibrosis before and after treatment in the placebo control group. P =0.3205), see Figure 2 The proportions of subjects whose Ishak scores for liver fibrosis decreased by ≥1 point from baseline in each group were as follows: placebo control group: 11 cases (26.19%), 180mg hydroxynidone capsule group: 17 cases (47.22%), 270mg hydroxynidone capsule group: 23 cases (65.71%), and 360mg hydroxynidone capsule group: 18 cases (52.94%). The differences between the four groups were statistically significant. P=0.0058). Among them, the differences in the proportion of subjects with a decrease of ≥1 point in Ishak score for liver fibrosis between the placebo control group and the 270mg hydroxynidone capsule group and the 360mg hydroxynidone capsule group, and their 95% CIs were -39.52% (-56.83%, -17.26%), -26.75% (-45.78%, -4.75%), respectively. This indicates that the 270mg / day hydroxynidone capsule and 360mg / day hydroxynidone capsules significantly reversed the degree of liver fibrosis compared to placebo, with the 270mg hydroxynidone capsule group showing the best reversal effect. (See...) Figure 4 .

[0267] Table 1. Historical changes in Ishak scores for liver fibrosis

[0268]

[0269]

[0270] - Between-group comparisons take into account the center effect. CMH - test; - The confidence interval for the rate difference was calculated using the Newcombe-Wilson method; - The LOCF method was used to fill in the missing Ishak Central Laboratory imaging scores in the full analysis set (FAS) at week 52 of treatment. - Subjects with missing baseline Ishak central laboratory MRI scores were treated as invalid; - "Baseline - 52 weeks of treatment" refers to the change in Ishak score obtained by subtracting the Ishak score after 52 weeks of treatment from the baseline Ishak score.

[0271] Table 2 Sensitivity analysis of the chronological changes in Ishak scores for liver fibrosis after 52 weeks of treatment.

[0272] - Ishak score progression: Ishak score of liver fibrosis increased by ≥1 point from baseline after 52 weeks of treatment; Ishak score stability: Ishak score of liver fibrosis remained unchanged from baseline after 52 weeks of treatment; Ishak score reversal: Ishak score of liver fibrosis decreased by ≥1 point from baseline after 52 weeks of treatment. - The LOCF method was used to fill in the missing Ishak Central Laboratory imaging scores in the full analysis set (FAS) at week 52 of treatment. - Subjects with missing baseline Ishak central laboratory imaging scores were treated as invalid, meaning their Ishak scores were considered stable and there was no change in Ishak scores for liver fibrosis after 52 weeks of treatment compared to baseline.

[0273] - [1] Indicates intergroup differences in the distribution of subjects with stable Ishak scores; [2] This indicates intergroup differences in the distribution of subjects progressing on the Ishak score.

[0274] Table 2 shows the statistical results of Ishak score progression of liver fibrosis after 52 weeks of treatment compared to baseline (Ishak score increase ≥1). Figure 3 , Figure 4 The results showed that more subjects in the placebo group had worsened Ishak scores for liver fibrosis than in the other three groups, while there was no statistically significant difference in FAS scores among the groups. P =0.4929), the proportion of subjects whose Ishak scores for liver fibrosis worsened from baseline in each group was as follows: placebo group 9 (20.93%), 180mg hydroxynidone capsule group 7 (16.67%), 270mg hydroxynidone capsule group 4 (9.76%), 360mg hydroxynidone capsule group 5 (12.20%). The PPS results were similar to FAS, with no statistically significant difference between groups (P=0.6545), the proportion of subjects whose Ishak scores for liver fibrosis progressed from baseline in each group was as follows: placebo control group 9 (21.43%), 180mg hydroxynidone capsule group 7 (19.44%), 270mg hydroxynidone capsule group 4 (11.43%), 360mg hydroxynidone capsule group 5 (14.71%).

[0275] Table 2 shows the statistical results of no change in Ishak score for liver fibrosis compared to baseline after 52 weeks of treatment. Figure 1 , Figure 2 The results showed that the placebo control group had a higher number of subjects with stable Ishak scores for liver fibrosis than the other three groups, while there was no statistically significant difference in FAS scores among the groups. P =0.3634), the proportions of subjects whose Ishak scores for liver fibrosis remained unchanged from baseline in each group were as follows: placebo control group: 23 cases (53.49%), 18 cases (42.86%) in the 180mg hydroxynidone capsule group, 14 cases (34.15%) in the 270mg hydroxynidone capsule group, and 18 cases (43.90%) in the 360mg hydroxynidone capsule group. There was no statistically significant difference among the PPS groups. P = 0.0511), the proportions of subjects whose Ishak scores for liver fibrosis did not change from baseline in each group were as follows: placebo control group 22 (52.38%), 180mg hydroxynidone capsule group 12 (33.33%), 270mg hydroxynidone capsule group 8 (22.86%), 360mg hydroxynidone capsule group 11 (32.35%).

[0276] In conclusion, after 52 weeks of treatment, the hydroxynidone group showed a greater ability to reverse liver fibrosis compared to the placebo group, with the 270 mg / day hydroxynidone capsules showing the best effect in reversing liver fibrosis.

[0277] 2.2 Statistical analysis results of patients with Ishak score >= 2 The inventors also evaluated the efficacy of hydroxynidone in liver fibrosis patients with a baseline Ishak score ≥2. Data are shown in Table 3.

[0278] Table 3. Results Analysis of *Baseline ISHAK score >= 2, B / C / D combination and compared with group A, ISHAK score decrease >= 1*

[0279] Note: "Screening period - 15-year follow-up" refers to the change in Ishak score between the screening period and the 15-year follow-up period.

[0280] 2.3 Evaluation of primary treatment efficacy in patients with significant fibrosis Statistical analysis results of patients with Ishak scores ≥3 The pathological evaluation of liver biopsies in this trial revealed that some subjects, although their liver biopsy results met the Ishak score ≥3 at enrollment (as independently determined by the research unit, meeting the trial's inclusion criteria), did not meet the Ishak score ≥3 in the central liver biopsy evaluation. This included subjects with Ishak scores ≥2 and ≥1. After review and confirmation in a blinded meeting, the central pathological score was only used for primary efficacy assessment. Therefore, the results of this trial will be more consistent with real-world conditions, allowing for observation of the therapeutic effects of the investigational drug on liver fibrosis at various stages.

[0281] In this study, Ishak pathological scores of liver tissue showed that the 270mg hydroxynidone group had a significant reversal effect on liver fibrosis. Simultaneously, transient elastography of the liver also reflected that the 270mg hydroxynidone group showed the best trend in reversing liver stiffness, consistent with the pathological scores. None of the hydroxynidone groups had any effect on reducing viral titers with entecavir or on ALT normalization.

[0282] The inventors further analyzed the primary efficacy evaluation of patients with a liver biopsy center pathological Ishak score ≥3 (i.e., significant fibrosis), as shown in Table 4. The results were consistent with the analysis above, with the hydroxynidone 270mg group showing the most significant reversal effect on liver fibrosis.

[0283] Table 4. Analysis of outcomes where subjects with a baseline Ishak score ≥3 experienced a decrease in Ishak score ≥1 after 52 weeks of treatment.

[0284]

[0285] Note: "Screening period - 15-year follow-up" refers to the change in Ishak score between the screening period and the 15-year follow-up period.

[0286] The inventors compared the effects of a combination of medium- and high-dose oxynisone (270 mg + 360 mg) and placebo in patients with liver fibrosis and a baseline Ishak score ≥3 (i.e., significant fibrosis). The results showed that in patients with liver fibrosis and an Ishak score ≥3, 4, or =6, medium- and high-dose oxynisone significantly reversed fibrosis. P <0.05). This suggests that hydroxynidone is effective in treating significant fibrosis, advanced fibrosis, and cirrhosis. Efficacy data are shown in Table 5.

[0287] Table 5. Analysis of outcomes in subjects with a baseline Ishak score ≥3 who experienced a ≥1 decrease in Ishak score after 52 weeks of treatment with the combination of medium- and high-dose therapy.

[0288] Note: "Baseline - 52 weeks after treatment" refers to the change in Ishak score obtained by subtracting the Ishak score after 52 weeks of treatment from the baseline Ishak score.

[0289] Secondary therapeutic effects: Liver transient elastography LSM (kPa) decrease rate There was no statistically significant difference in baseline transient liver elastography (LSM) (kPa) values ​​among the four groups. P =0.6906). After 52 weeks of treatment, the transient liver elastography (LSM) (kPa) values ​​decreased in all groups, and the intra-group changes in LSM (kPa) values ​​before and after treatment were statistically significant in each group. P <0.01). After 52 weeks of treatment, the mean decrease rate of transient liver elastography (LSM) (kPa) in the hydroxynidone group was higher than that in the control group, with the 270 mg hydroxynidone capsule group showing the largest mean decrease rate, consistent with the pathological score results. However, there was no statistically significant difference in FAS between the groups. P =0.7311). The mean (standard deviation) of the decrease rate of liver transient elastography LSM (kPa) in each group were as follows: placebo control group 10.16% (63.19), 180 mg hydroxynidone capsule group 18.30% (29.65), 270 mg hydroxynidone capsule group 24.62% (31.23), and 360 mg hydroxynidone capsule group 18.98% (33.96). The PPS analysis results were similar to those of FAS.

[0290] Improvement of liver tissue inflammation There was no statistically significant difference in baseline liver tissue inflammation grading scores among the four groups (FAS: P =0.9123, PPS: P =0.7984). After 52 weeks of treatment, liver tissue inflammation decreased in all groups, and the changes in liver tissue inflammation scores before and after treatment were statistically significant within each group. P <0.001). The proportions of subjects with improved liver tissue inflammation in each group were as follows: placebo control group: 22 cases (52.38%), 180mg hydroxynidone capsule group: 21 cases (58.33%), 270mg hydroxynidone capsule group: 20 cases (57.14%), 360mg hydroxynidone capsule group: 19 cases (55.88%). Improvement was observed in the hydroxynidone group, but the difference among the four groups was not statistically significant. P =0.9570). PPS and FAS treatment had the same results after 52 weeks.

[0291] HBV DNA seroconversion rate There was no statistically significant difference in the baseline HBV DNA positivity rate among the four groups. P =0.1474). After 52 weeks of treatment, HBV DNA levels in all groups were essentially negative. Specifically, the negative conversion rate was 95.24% in the placebo control group (n=40), 94.44% in the 180mg hydroxynidone capsule group (n=34), 97.14% in the 270mg hydroxynidone capsule group (n=34), and 94.12% in the 360mg hydroxynidone capsule group (n=32). There was no statistically significant difference in the negative conversion rate among the four groups. P =0.9569). The PPS analysis results are similar to those of FAS.

[0292] ALT normalization rate FAS analysis showed no statistically significant differences in baseline ALT values ​​among the four groups. P =0.3058). After 52 weeks of treatment, ALT levels decreased in all groups, and the intra-group changes in ALT before and after treatment were statistically significant in each group. P <0.001). Mean (standard deviation) of ALT levels from baseline after 52 weeks of treatment: 64.15 (73.63) in the placebo control group, 44.71 (53.50) in the 180 mg hydroxynidone capsule group, 66.47 (78.13) in the 270 mg hydroxynidone capsule group, and 43.58 (59.87) in the 360 ​​mg hydroxynidone capsule group; however, there was no statistically significant difference in changes between the groups. P =0.5132). The PPS analysis results are similar to those of FAS.

[0293] The FAS analysis showed that after 52 weeks of treatment, a certain number of subjects in each group had their clinically significant ALT levels change from abnormal (baseline) to normal. Specifically, this was observed in the placebo control group (n=21, 87.50%), the 180mg hydroxynidone capsule group (n=15, 78.95%), the 270mg hydroxynidone capsule group (n=18, 85.71%), and the 360mg hydroxynidone capsule group (n=16, 88.89%). The PPS analysis results were similar to those of the FAS analysis.

[0294] Security: Hydroxynesone has a good safety profile. During the trial, there were no statistically significant differences in the incidence of adverse events, adverse reactions, and SAEs among the four groups.

[0295] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. 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. Use of the compound of formula (I), its solvate, hydrate, prodrug, or pharmaceutically acceptable salt thereof in the preparation of medicaments for the treatment or prevention of chronic hepatitis B with liver fibrosis. （I） ， Subjects with chronic hepatitis B and liver fibrosis were excluded if their ALT (alanine aminotransferase) was < 8 times the ULN (upper limit of standard), while subjects with 3 ULN < ALT < 8 ULN and TBiL > 2 times the ULN were excluded.

2. The use according to claim 1, characterized in that, The chronic hepatitis B with liver fibrosis refers to significant liver fibrosis, progressive liver fibrosis, or cirrhosis. Preferably, the cirrhosis is compensated cirrhosis.

3. The use according to any one of claims 1-2, characterized in that, The significant liver fibrosis can be characterized by one, two, or more of the following: (1) The significant liver fibrosis described is characterized by the Ishak scoring system, which refers to liver fibrosis with an Ishak score ≥3 or ≥4; for example, an Ishak score of approximately 3-6. (2) The significant liver fibrosis described by the Scheuer scoring system refers to liver fibrosis with a Scheuer score of approximately ≥2 or ≥3; for example, a Scheuer score of approximately 2-4. (3) The significant liver fibrosis described by the METAVIR scoring system refers to liver fibrosis with a METAVIR score of approximately ≥2 or ≥3; for example, a METAVIR score of approximately 2-4. (4) The significant liver fibrosis described by the Knodell scoring system refers to liver fibrosis with a Knodell score of approximately ≥2 or ≥3; for example, a Knodell score of approximately 2-4; and / or (5) The significant liver fibrosis is characterized by liver stiffness measurement value, which refers to liver fibrosis with an LSM of approximately ≥5.8 kPa, for example, an LSM of approximately 5.8-12.4 kPa.

4. The use according to any one of claims 1-3, characterized in that, The advanced liver fibrosis can be characterized by one, two, or more of the following: (1) The advanced liver fibrosis is characterized by the Ishak scoring system, for example, liver fibrosis with an Ishak score ≥4 or ≥5; for example, an Ishak score of approximately 4-6; (2) The advanced liver fibrosis described is characterized by the Scheuer scoring system, which refers to liver fibrosis with a Scheuer score of approximately ≥3 or approximately 3-4. (3) The advanced liver fibrosis described is characterized by the METAVIR scoring system, which refers to liver fibrosis with a METAVIR score of approximately ≥3 or approximately 3-4; (4) The advanced liver fibrosis described is characterized by the Knodell scoring system, referring to liver fibrosis with a Knodell score of approximately ≥2 or ≥3, or approximately 2-4 or 2-3; and / or (5) The progressive liver fibrosis is characterized by liver stiffness measurement. When bilirubin is normal and ALT < 5 ULN, the LSM is approximately ≥ 9.0 kPa or ≥ 10.0 kPa of liver fibrosis. Or when bilirubin and ALT are normal, the LSM is approximately ≥ 6.0 kPa of liver fibrosis.

5. The use according to any one of claims 1-4, characterized in that, The cirrhosis can be characterized by one, two, or more of the following: (1) The cirrhosis described is characterized by the Ishak scoring system, which refers to cirrhosis with an Ishak score ≥5 or approximately 5-6; (2) The cirrhosis described is characterized by the Scheuer scoring system, which refers to cirrhosis with a Scheuer score of approximately ≥4; (3) The liver cirrhosis described by the METAVIR scoring system refers to liver cirrhosis with a METAVIR score of approximately ≥4; (4) The cirrhosis described is characterized by the Knodell scoring system, referring to cirrhosis with a Knodell score of approximately ≥4; and / or (5) The liver cirrhosis described is characterized by liver stiffness measurement values, which refers to liver cirrhosis with an LSM of approximately ≥12.0 kPa or approximately ≥13.0 kPa.

6. The use according to any one of claims 1-5, characterized in that, The patient had chronic hepatitis B with liver fibrosis that did not progress to liver cancer.

7. The use according to any one of claims 1-6, characterized in that, The dosage of the compound shown in formula (I) is 50-500 mg / day, for example 80-450 mg / day, 100-400 mg / day, 120-360 mg / day; 180 mg / day, 270 mg / day, 360 mg / day; Preferably, the drug is administered 1-5 times per day, for example, 3 times per day.

8. The use according to any one of claims 1-7, characterized in that, The pharmaceutically acceptable salts include those formed by the compound of formula (I) with an organic acid selected from propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, and citric acid, or an acidic amino acid selected from aspartic acid and glutamic acid, and then with an inorganic base, including sodium, potassium, calcium, aluminum, and ammonium salts; or salts formed with an organic base, including methylamine salts, ethylamine salts, and ethanolamine salts; or salts formed with a basic amino acid selected from lysine, arginine, and ornithine, and then with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; or salts formed with an organic acid selected from formic acid, acetic acid, picric acid, methanesulfonic acid, and ethanesulfonic acid.

9. The use according to any one of claims 1-8, characterized in that, The drug also includes one or more pharmaceutically acceptable carriers or excipients.

10. The use according to any one of claims 1-9, characterized in that, The dosage form of the drug is a capsule.

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