Pharmaceutical hydroxynidone formulations for diseases

Abstract

The present invention relates to a method of treating a subject suffering from liver fibrosis and / or liver cirrhosis. The present disclosure provides a pharmaceutical composition comprising hydroxynidone or a salt thereof that may be used to treat a subject suffering from liver fibrosis and / or liver cirrhosis. Liver hardness measurements and / or Ishak scores may be used to identify subjects for treatment of liver fibrosis and / or liver cirrhosis.

Description

Background Art

[0001] Liver fibrosis is a life-threatening disease with significant morbidity and mortality. Currently, there are no FDA-approved treatments for liver fibrosis in the United States, and better treatments are needed in many areas of the world. Summary of the invention

[0002] In one aspect, the present disclosure provides a method of treating a subject having liver fibrosis, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

[0003] In another aspect, the present disclosure provides a method of treating a subject having liver cirrhosis, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

[0004] In another aspect, the present disclosure provides a method of treating a subject having advanced hepatitis B virus infection, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

[0005] In another aspect, the present disclosure provides a method of treating a subject having NASH fibrosis, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

[0006] In another aspect, the disclosure provides a method of treating a subject, the method comprising identifying a subject having a liver stiffness measurement of at least 4 kilopascals (kPa) and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

[0007] In another aspect, the present disclosure provides a method of treating a subject, the method comprising identifying a subject having an Ishak value of about 1 to 6; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

[0008] In another aspect, the disclosure provides a method of treating a subject, the method comprising identifying a subject that is substantially free of hepatitis B virus protein; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

[0009] In another aspect, the present disclosure provides a method of treating a subject, the method comprising identifying a subject, wherein the subject tests positive for hepatitis viral DNA after receiving one or more courses of antiviral therapy; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

[0010] In another aspect, the present disclosure provides a pharmaceutical composition comprising about 20% to about 90% by weight of hydroxynidone or a salt thereof and one or more pharmaceutically acceptable excipients. In some cases, the weight percentage of hydroxynidone or a salt thereof is about 25% to about 35%. In some cases, the one or more pharmaceutically acceptable excipients are selected from lactose, sucrose, magnesium stearate, glucose, vegetable cellulose, calcium carbonate, zinc stearate, calcium stearate, stearic acid, palmitic acid, myristic acid, glyceryl dibehenate and talc.

[0011] Other aspects and advantages of the present disclosure will become clear to those skilled in the art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be appreciated, the present disclosure is capable of other and different embodiments, and its several details are capable of modification in various obvious aspects, all without departing from the present disclosure. Therefore, the drawings and description are to be regarded as illustrative rather than restrictive in nature. Incorporated by Reference

[0012] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent that publications and patents or patent applications incorporated by reference conflict with the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such conflicting material. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] The novel features of the present invention are particularly set forth in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by referring to the following detailed description, which sets forth illustrative embodiments using the principles of the present invention and the accompanying drawings (also referred to herein as "figures"), and in which:

[0014] Figure 1 The Ishak score analysis for each drug group is shown.

[0015] Figure 2 Analysis showing changes in Ishak scores within groups.

[0016] Figure 3 Analysis of patients with significant fibrosis is shown.

[0017] Figure 4 Combined analysis of the 270 mg and 360 mg groups is shown.

[0018] Figure 5 Displays the combinations of individual drug groups.

[0019] Figure 6Shown is the change in Ishak score after 52 weeks in patients with baseline HBeAg(+).

[0020] Figure 7 Shown is the change in Ishak score after 52 weeks in patients with baseline HBeAG(-).

[0021] Figure 8 Shows changes in liver stiffness measurement (LSM).

[0022] Fig. 9 The combinations of drug groups used for LSM analysis are shown.

[0023] Fig.10 Shown is an analysis of HBV DNA(+) patients after 52 weeks of entecavir therapy.

[0024] Fig.11 The effects of hydroxynidone on the NASH model induced in mice are shown.

[0025] Fig.12 H&E staining is shown for the effect of hydroxynidone on the induced mouse NASH model. DETAILED DESCRIPTION

[0026] Although various embodiments of the present invention have been shown and described herein, it is apparent to those skilled in the art that these embodiments are provided only as examples. Without departing from the present invention, those skilled in the art may expect many variations, changes and replacements. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed.

[0027] Whenever the term "at least", "greater than", or "greater than or equal to" precedes the first value in a series of two or more values, the term "at least", "greater than", or "greater than or equal to" applies to each value in the series. For example, greater than or equal to 1, 2, or 3 is equivalent to greater than or equal to 1, greater than or equal to 2, or greater than or equal to 3.

[0028] Whenever the term "not more than," "at most," "less than," or "less than or equal to" precedes the first value in a series of two or more values, the term "not more than," "at most," "less than," or "less than or equal to" applies to every value in the series. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1. Overview

[0029] Liver cirrhosis or hepatic cirrhosis is a disease in which the liver cannot function properly due to long-term damage. During the process of liver tissue repair, scar tissue formation called fibrosis is formed. The presence of significant fibrosis from chronic liver damage and subsequent repair over time may lead to sclerosis. Liver fibrosis is usually the first stage of liver scarring. Various factors may cause liver fibrosis of the liver, for example, any process that may cause liver homeostasis to be impaired (for example, inflammation, toxic damage, changes in liver blood flow and liver infection (virus, bacteria, fungi and parasites) etc.).

[0030] The antifibrotic effects of hydroxynidone were observed in the in vivo models of liver fibrosis of different etiologies described herein. The antifibrotic effects of hydroxynidone were evaluated in several animal models of liver fibrosis, such as human serum albumin (HSA)-induced rat liver fibrosis, dimethylnitrosamine (DMN)-induced rat liver fibrosis, and carbon tetrachloride (CCl4)-induced rat liver fibrosis. 4 )-induced liver fibrosis in mice. In these animal models, hydroxynidone exhibited potent antifibrotic effects, for example, significantly reducing the level of fibrosis and improving biochemical and pathological indicators of liver fibrosis, such as hydroxyproline content and the levels of liver enzymes such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST). 4 In induced liver fibrosis in mice, the effect of hydroxynidone is most obvious at a dose of 3-10 mg / kg / day, but some effects are also shown at a dose of 1 mg / kg / day. In some embodiments, this dose range corresponds to a human equivalent dose (HED) range of 15-50 mg. In some embodiments, these doses are 7-24 times lower than the human maximum daily dose of 360 mg (120 mg three times a day [TID]) administered to subjects with liver fibrosis due to chronic hepatitis B infection. In rats, according to the model, the anti-fibrotic effect of hydroxynidone is shown in a dose range of 10-250 mg / kg / day, which is comparable or more effective than silymarin extract (a known hepatoprotective drug) used as a positive control. These rat doses have HEDs in the range of 100-2,400 mg, which were all tested as safe in Phase I clinical trials of hydroxynidone, and in an ongoing Phase II clinical study of the efficacy and safety of hydroxynidone in China, the 100 mg dose is approximately one-third lower than the maximum daily dose of 360 mg (administered as 120 mg TID) administered to subjects with liver fibrosis due to chronic hepatitis B. Treatment

[0031] The present disclosure provides a method for treating a subject with liver fibrosis. The method may include administering hydroxynidone to a subject in need thereof. The method may include administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

[0032] The present disclosure also provides a method of treating a subject with cirrhosis. The method may include administering hydroxynidone to a subject in need thereof. The method may include administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

[0033] The present disclosure also provides a method for treating a subject with NASH fibrosis. The method may include administering hydroxynidone or a salt thereof to a subject in need thereof. The method may include administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

[0034] The present disclosure also provides a method of treating a subject having advanced hepatitis B virus infection. The method may include administering a pharmaceutical composition comprising hydroxynidone or a salt to the subject.

[0035] The present disclosure also provides a method for treating a subject, the method comprising identifying a subject with a liver stiffness measurement value and administering a pharmaceutical composition comprising hydroxynidone or a salt thereof to the subject. The pharmaceutical composition comprising hydroxynidone or a salt thereof may be as described elsewhere herein. In some cases, the liver stiffness measurement value may be obtained by transient elastography. In some cases, the liver stiffness measurement value may be obtained by FibroTouch and / or FibroScan. In some cases, the liver stiffness measurement value may be calibrated against a reference standard. The reference standard may be, for example, a liver biopsy. The liver stiffness measurement value may be about 4 kilopascals (kPa) to 13kPa. The liver stiffness measurement value may be about 4kPa to 8kPa. The liver stiffness measurement value may be about 8kPa to 13kPa. The liver stiffness measurement value may be about 8kPa to 75kPa. The liver stiffness measurement value may be at most about 13kPa. The liver stiffness measurement value may be at most about 75kPa.

[0036] The present disclosure also provides a method for treating a subject, the method comprising identifying a subject having an Ishak value and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof. The pharmaceutical composition comprising hydroxynidone or a salt thereof may be as described elsewhere herein. The Ishak value may be about 1 to 6. The Ishak value may be 1 to 3. The Ishak value may be 3 to 6.

[0037] In some embodiments, an Ishak score of 0 may indicate no fibrosis. In some embodiments, an Ishak score of 1 may indicate fibrous expansion of some portal areas, with the presence or absence of short fiber septa. In some embodiments, an Ishak score of 2 may indicate fibrous expansion of most portal areas, with the presence or absence of short fiber septa. In some embodiments, an Ishak score of 3 may indicate fibrous expansion of most portal areas, with occasional portal-portal bridging. In some embodiments, an Ishak score of 4 may indicate fibrous expansion of portal areas, with the presence of significant bridging (e.g., portal-portal bridging and portal-central bridging). In some embodiments, an Ishak score of 5 may indicate significant bridging (e.g., portal-portal bridging and / or portal-central bridging), with occasional nodules (e.g., incomplete sclerosis). In some embodiments, an Ishak score of 6 may indicate possible or definite sclerosis.

[0038] In some embodiments, the Ishak value can be converted to another scoring system. The scoring system can be, for example, METAVIR score, Knodell score, etc.

[0039] In some embodiments, the method for treating a subject may include identifying a subject with a METAVIR score and administering a pharmaceutical composition comprising hydroxynidone or its salt to the subject. In some cases, a METAVIR score of F0 may indicate no fibrosis. In some cases, a METAVIR score of F1 may indicate portal fibrosis without the presence of a diaphragm. In some cases, a METAVIR score of F2 may indicate portal fibrosis with a rare diaphragm. In some cases, a METAVIR score of F3 may indicate a lot of diaphragms without sclerosis. In some cases, a METAVIR score of F4 may indicate sclerosis. In some cases, a METAVIR score of F0 may indicate F4, F1 to F4, F2 to F4, F3 to F4, F1 to F3, F1 to F2, or F2 to F3.

[0040] The present disclosure also provides a method for treating a subject, the method comprising identifying a subject that is substantially free of hepatitis B virus protein and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

[0041] The present disclosure also provides a method for treating a subject, the method comprising identifying a subject, wherein the subject tests positive for hepatitis virus DNA after receiving one or more courses of antiviral therapy; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof. In some cases, the hepatitis virus DNA is selected from hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, and hepatitis E virus. In some cases, the hepatitis virus DNA is hepatitis B. Pharmaceutical composition

[0042] The present disclosure provides a pharmaceutical composition. The pharmaceutical composition may include a compound shown in formula (I) The compound of formula (I) may be referred to as hydroxynidone and / or N-(4-hydroxyphenyl)-5-methyl-2-pyridone. The hydroxynidone or pharmaceutical composition described herein may be used for the treatment of liver fibrosis associated with chronic liver disease. The hydroxynidone or pharmaceutical composition described herein may be used for the treatment of liver cirrhosis associated with chronic liver disease.

[0043] The pharmaceutical composition may include hydroxynidone. The pharmaceutical composition may include a certain weight percentage of hydroxynidone or a salt thereof. The weight percentage of hydroxynidone or a salt thereof in the pharmaceutical composition may be about 10% to 90%, 10% to 50%, 10% to 40%, 10% to 30%, 20% to 90%, 20% to 50%, 20% to 40%, 20% to 30%, 30% to 90%, 30% to 50% or 30% to 40%. The weight percentage of hydroxynidone in the pharmaceutical composition may be at least about 20%, 25%, 30%, 35% or 40%. The weight percentage of hydroxynidone in the pharmaceutical composition may be up to about 40%, 35%, 30%, 25% or 20%. The weight percentage of hydroxynidone in the pharmaceutical composition may be about 30%.

[0044] The pharmaceutical composition may comprise hydroxynidone and one or more excipients. The one or more excipients may include, for example, lactose, sucrose, glucose, vegetable cellulose, calcium carbonate, magnesium stearate, zinc stearate, calcium stearate, stearic acid, palmitic acid, myristic acid, glyceryl dibehenate and talc, etc. The one or more excipients may include magnesium stearate and lactose.

[0045] The one or more excipients can account for a certain weight percentage of the pharmaceutical composition. The one or more excipients can be, for example, lactose. The weight percentage of lactose in the pharmaceutical composition can be about 10% to 90%, 50% to 90%, 60% to 90%, 50% to 80%, 50% to 80%, 60% to 80%, 65% to 75%, 67% to 72% or 68% to 71%. The weight percentage of lactose in the pharmaceutical composition can be at least about 50%, 60%, 65%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 75% or 80%. The weight percentage of lactose in the pharmaceutical composition can be up to about 80%, 75%, 73%, 72%, 71%, 69%, 68%, 67%, 65%, 60% or 50%. The weight percentage of lactose in the pharmaceutical formulation can be about 70%.

[0046] The one or more excipients can be, for example, magnesium stearate. The weight percentage of magnesium stearate in the pharmaceutical composition can be about 0.01% to 2%, 0.1% to 1.5%, 0.2% to 1.0%, 0.25% to 0.5%, 0.15% to 0.30%, 0.2% to 0.3% or 0.21% to 0.29%. The weight percentage of magnesium stearate in the pharmaceutical composition can be at least about 0.01%, 0.05%, 0.1%, 0.15%, 0.20%, 0.25%, 0.30%, 0.35% or 0.40%. The weight percentage of one or more lubricants in the pharmaceutical composition can be up to about 0.40%, 0.35%, 0.30%, 0.25%, 0.20%, 0.15%, 0.10%, 0.05% or 0.01%.

[0047] The pharmaceutical composition may include hydroxynidone, lactose and magnesium stearate. The weight percentages of hydroxynidone, lactose and magnesium may be about 30%, 69.8% and 0.2%, respectively. The weight percentages of hydroxynidone, lactose and magnesium may be about 30%, 69.7% and 0.3%, respectively.

[0048] In certain embodiments, provided herein are compositions comprising a therapeutically effective amount of any compound or salt of hydroxynidone (also referred to herein as a "pharmaceutical agent").

[0049] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers (including excipients and adjuvants) that aid in processing the pharmaceutical agent into a pharmaceutically useful preparation. Appropriate formulations depend on the selected route of administration. Overviews of pharmaceutical compositions are found, for example, in Remington: The Science and Practice of Pharmacy, 19th Edition (Easton, Pa., Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition (Lippincott Williams & Wilkins, 1999).

[0050] The compositions and methods of the present disclosure can be used to treat individuals in need. In certain embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal such as a human, the composition or the pharmaceutical agent is preferably administered as a pharmaceutical composition comprising, for example, a pharmaceutical agent and a pharmaceutically acceptable carrier or excipient. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions, such as water or physiologically buffered saline; or other solvents or vehicles, such as glycols, glycerol, oils (such as olive oil) or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive administration routes (e.g., routes that avoid transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipient can be selected, for example, to achieve delayed release of the agent or selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in the form of dosage units such as tablets, capsules, granules, lyophiles for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system (eg, a skin patch).The composition may also be present in a solution suitable for topical administration, such as eye drops.

[0051] Pharmaceutically acceptable excipients may contain physiologically acceptable agents that act, for example, on stable compounds such as pharmaceutical agents, increase the solubility of the compounds, or increase the absorption of the compounds. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; or other stabilizers or excipients. The selection of pharmaceutically acceptable excipients including physiologically acceptable agents depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix into which, for example, the compound of the present invention may be incorporated. Liposomes (e.g., including phospholipids or other lipids) are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0052] The pharmaceutical composition (formulation) can be administered to a subject by any of a variety of routes of administration, including, for example, orally, for example, as a drench in an aqueous or non-aqueous solution or suspension, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, and pastes for application to the tongue.

[0053] The pharmaceutical composition can be a sterile aqueous or non-aqueous solution, suspension or emulsion (e.g., microemulsion). The excipients described herein are examples and are by no means limiting. An effective amount or therapeutically effective amount refers to an amount of one or more pharmaceutical agents administered to a subject as a single dose or as part of a series of doses that is effective to produce the desired therapeutic effect.

[0054] Typically, the subject can be monitored for therapeutic effectiveness using assays and methods suitable for the condition being treated, which assays will be familiar to those skilled in the art and described herein. The pharmacokinetics of a drug agent or one or more metabolites thereof administered to a subject can be monitored by determining the level of the drug agent or metabolite in a biological fluid from the subject (e.g., in blood, a blood fraction (e.g., serum) and / or in urine) and / or in other biological samples or biological tissues. The level of a drug agent or metabolite during the course of treatment can be measured using any method practiced in the art and described herein for detecting an agent.

[0055] The dosage of the pharmaceutical agents described herein for treating a disease or disorder may depend on the condition of the subject, i.e., the stage of the disease, the severity of the symptoms caused by the disease, the overall health status, and the age, sex and weight, as well as other factors obvious to the medical technician. The pharmaceutical composition may be administered in a manner suitable for the disease to be treated as determined by the medical technician. In addition to the factors related to the use of the pharmaceutical agent for treating a disease or disorder described herein and above, the appropriate duration and frequency of administering the pharmaceutical agent may also be determined or adjusted according to factors such as the patient's condition, the type and severity of the patient's disease, the specific form of the active ingredient, and the method of administration. The optimal dose of the agent may generally be determined using experimental models and / or clinical trials. The optimal dose may depend on the subject's body mass, body weight, or blood volume. It is generally preferred to use the minimum dose sufficient to provide effective treatment. The design and execution of preclinical and clinical studies for pharmaceutical agents described herein (including when administered for preventive benefit) are within the skill of the technicians in the relevant field. When administering two or more pharmaceutical agents to treat a disease or disorder, the optimal dose of each pharmaceutical agent may be different, such as less than the dose when any one agent is administered alone as a single agent therapy. In some specific embodiments, two kinds of drug agents of combination can act synergistically or additively, and any medicament can be used with the amount less than the situation of being used alone.The amount of the drug agent that can be used every day can be for example between about 0.01mg / kg and 100mg / kg body weight, for example, between about 0.1 to 1mg / kg, between about 1 to 10mg / kg, between about 10-50mg / kg, between about 50-100mg / kg.In other embodiments, the amount of the drug agent that can be used every day is between about 0.01mg / kg and 1000mg / kg body weight, between about 100-500mg / kg or between about 500-1000mg / kg.The optimum dose every day or each course of treatment can be different for the disease to be treated or obstacle, and can also change according to route of administration and treatment regimen.

[0056] Pharmaceutical compositions comprising pharmaceutical agents can be formulated in a manner suitable for the delivery method using techniques routinely practiced in the art. The compositions can be in the form of solids (e.g., tablets, capsules), semisolids (e.g., gels), liquids, or gases (e.g., aerosols).

[0057] Pharmaceutically acceptable excipients are well known in the pharmaceutical field and are described, for example, in Rowe et al., Handbook of Pharmaceutical Excipients: A Comprehensive Guide to Uses, Properties, and Safety, 5th edition, 2006; and Remington: The Science and Practice of Pharmacy (Gennaro, 21st edition Mack Pub. Co., Easton, PA (2005)). Exemplary pharmaceutically acceptable excipients include sterile saline and phosphate buffered saline at physiological pH. Preservatives, stabilizers, dyes, buffers, etc. can be provided in the pharmaceutical composition. In addition, antioxidants and suspending agents can also be used. Generally, the type of excipient is selected based on the mode of administration and the chemical composition of one or more active ingredients. Alternatively, the compositions described herein can be formulated as lyophilized products. The compositions described herein can be lyophilized or otherwise formulated as lyophilized products using one or more appropriate excipient solutions for dissolving and / or diluting one or more pharmaceutical agents of the composition when administered. In other embodiments, the pharmaceutical agent may be encapsulated within a liposome using techniques known and practiced in the art. In certain specific embodiments, the pharmaceutical agent is not formulated within a liposome for application to a stent for treating an artery that is highly occluded but not completely occluded. The pharmaceutical composition may be formulated for any suitable mode of administration described herein and in the art.

[0058] Pharmaceutical compositions (e.g., for oral administration) can be in liquid form. Liquid pharmaceutical compositions can include, for example, one or more of the following: sterile diluents, such as water, saline solutions (preferably physiological saline), Ringer's solution, isotonic sodium chloride, fixed oils that can be used as solvents or suspension media, polyethylene glycol, glycerol, propylene glycol or other solvents; antibacterial agents; antioxidants; chelating agents; buffers and agents for adjusting tonicity, such as sodium chloride or dextrose. Parenteral compositions can be packaged in ampoules, disposable syringes or multi-dose vials made of glass or plastic. It is preferred to use physiological saline, and the injectable pharmaceutical composition is preferably sterile. In another embodiment, for the treatment of ophthalmic conditions or diseases, liquid pharmaceutical compositions can be applied to the eye in the form of eye drops. Liquid pharmaceutical compositions can be delivered orally.

[0059] For oral formulations, at least one pharmaceutical agent as described herein can be used alone or in combination with suitable additives for preparing tablets, powders, granules or capsules, and if desired, in combination with diluents, buffers, wetting agents, preservatives, coloring agents and flavoring agents. Pharmaceutical agents can be formulated with buffers to provide protection for the compound at the low pH of the gastric environment, and / or formulated with enteric coatings. The pharmaceutical agents included in the pharmaceutical composition can be formulated with flavoring agents for oral delivery, for example, in liquid, solid or semisolid formulations, and / or formulated with enteric coatings.

[0060] Pharmaceutical compositions comprising any of the pharmaceutical agents described herein can be formulated for sustained release or slow release, also referred to as delayed release or controlled release. Such compositions can be prepared using well-known techniques, and are administered, for example, by oral, rectal, intradermal or subcutaneous implantation or by implantation at a desired target site. Sustained release formulations can contain compounds dispersed in a carrier matrix and / or contained in a reservoir surrounded by a rate-controlling membrane. The excipients used in such formulations are biocompatible and can also be biodegradable; preferably, the formulation provides a relatively constant level of active ingredient release. The amount of the pharmaceutical agent contained in the sustained release formulation depends on the implantation site, the rate of release and the expected duration, and the nature of the condition, disease or disorder to be treated or prevented.

[0061] Polymer formulations can also be used to provide controlled release or sustained release. Bioadhesive polymers described in the art can be used. For example, sustained release gels and compounds can be incorporated into polymeric matrices, such as hydrophobic polymer matrices. Examples of polymeric matrices include microparticles. The microparticles can be microspheres, and the core can be a material different from the polymeric shell. Alternatively, the polymer can be cast into a sheet or film, a powder or gel such as a hydrogel produced by grinding or other standard techniques. The polymer can also be in the form of a coating or a part of a bandage, stent, catheter, artificial blood vessel or other device to facilitate the delivery of a pharmaceutical agent. The matrix can be formed by solvent evaporation, spray drying, solvent extraction and other methods known to those skilled in the art.

[0062] Kits are provided having unit doses (generally oral or injectable doses) of one or more of the agents described herein. Such kits may include a container containing the unit dose, an informative package insert describing the use of the drug in the treatment of the disease and the attendant benefits, and optionally an apparatus or device for delivering the composition. Example

[0063] Example 1: In vitro human metabolism of hydroxynidone - Phase 1

[0064] The in vitro phase I metabolism of hydroxynidone was evaluated by incubating human liver microsomes with hydroxynidone for 60 minutes. The sensitivity of the assay was demonstrated by incubating human liver microsomes with dextromethorphan (positive control) for 20 minutes, resulting in the formation of the metabolites dextrorphan and 3-methoxymorphinan. After 60 minutes of incubation of hydroxynidone in the human liver microsome reaction system, no metabolite formation was observed, indicating that CYP450 does not mediate the metabolic reaction of hydroxynidone to form M2 metabolites.

[0065] Example 2: In vitro human metabolism of hydroxynidone - Phase 2

[0066] The generation of M4 metabolite was detected after 30 minutes of incubation of hydroxynidone in the reaction system of human liver microsomes, indicating that the metabolic reaction of hydroxynidone to M4 is most likely mediated by uridine 5'-diphospho-glucuronosyltransferase (UDP-glucuronosyltransferase, UGT).

[0067] Example 3: In vitro human metabolism of hydroxynidone - interaction with CYP450

[0068] In vitro studies investigated whether hydroxynidone is an inhibitor of human cytochrome P450 (CYP450). The effect of different hydroxynidone concentrations was determined by the fluorescence intensity of the metabolic substrates of CYP1A2, 2C9, 2C19, 2D6 and 3A4 produced by the action of the relevant enzymes. If inhibition of hydroxynidone was observed, the corresponding IC 50 Values. Known CYP450 inhibitors were used as positive controls to show the sensitivity of the assay. The results showed that hydroxynidone had no significant in vitro inhibitory activity against human CYP 1A2 (IC50 value > 100 μM, while the positive control furaphylline had an IC50 value of 2.59 μM), 2C9 (IC50 value > 100 μM, while the positive control sulfaphenazole had an IC50 of 0.49 μM), 2C19 (IC50 value > 100 μM, while the positive control tranylcypromine had an IC50 of 4.9 μM), 2D6 (IC50 value > 100 μM, while the positive control quinidine had an IC50 of 0.01 μM), and 3A4 enzymes (IC50 value > 100 μM, while the positive control ketoconazole had an IC50 value of 0.027 μM). It can be concluded that hydroxynidone lacks significant interactions with human CYP450.

[0069] Example 4: In vitro human metabolism-pharmacokinetics of hydroxynidone Drug-drug interactions

[0070] The potential for drug interaction of hydroxynidone with two antiviral drugs, entecavir and lamivudine, relevant to the indication of chronic hepatitis B infection in China, was evaluated in vitro using human liver microsomes. In an ongoing Phase II trial of efficacy and safety conducted in China, entecavir was used concomitantly with hydroxynidone in subjects with liver fibrosis due to chronic hepatitis B infection. In the in vitro drug interaction experiments, entecavir and lamivudine were used concomitantly with hydroxynidone at their respective concentrations of 1.5 mM, or alone. The results obtained in the drug interaction studies indicated that: a) hepatic metabolism is not the major metabolic pathway of hydroxynidone, and b) entecavir and lamivudine did not exhibit drug interactions with hydroxynidone, as there were no significant differences in residual drug concentrations between the monotherapy group and the combination therapy group (P>0.05).

[0071] Example 5: Summary of Phase I Clinical Pharmacology Study of Hydroxynidone

[0072] Example 6: Single and Multiple Dose Double-Blind, Placebo-Controlled, Inpatient Study of Hydroxynidone

[0073] The primary objectives of this study were to evaluate the tolerability of single and multiple ascending doses of hydroxynidone in healthy subjects to support further Phase 1 studies and to provide supportive information for dosing regimens in Phase 2 clinical trials.

[0074] This was a randomized, double-blind, placebo-controlled, inpatient study of single and multiple ascending doses of hydroxynidone in healthy Chinese male and female subjects aged 18-45 years who met all inclusion and exclusion criteria. Hydroxynidone 100 mg and 200 mg capsule dose strengths were used in the study. In each cohort, subjects were randomized to hydroxynidone or placebo in a 3:1 ratio.

[0075] The single-dose component of the trial included 30 subjects who received 100 mg, 200 mg, 400 mg, 600 mg and 800 mg of hydroxynidone (6 subjects / cohort), respectively. Post-dose observations were performed 24 h after administration. All subjects completed the trial. Hydroxynidone was safe and well tolerated at all doses tested. No deaths or serious adverse events (SAEs) were reported during the trial. One subject in each of the 800 mg hydroxynidone and placebo cohorts reported mild transient dizziness after administration of the test product. The dizziness resolved on its own with no sequelae. In both the hydroxynidone and placebo groups, the post-dose values ​​of some laboratory parameters were abnormal. However, these changes were considered clinically insignificant.

[0076] The multiple-dose component of the trial included 20 subjects who were assigned to receive either 600 mg hydroxynidone (8 subjects) (total daily dose of 1800 mg) three times daily (TID) or 800 mg hydroxynidone (12 subjects) (total daily dose of 2400 mg) TID for ten (10) consecutive days. Due to several AEs of hypertriglyceridemia observed in the hydroxynidone 600 mg TID dose group (4 of 8 subjects), subjects in the hydroxynidone TID 800 mg dose group (n=8) were divided into two subgroups, each consisting of 4 subjects. A staggered enrollment approach was applied: if no SAE was observed in the first subgroup, enrollment of subjects in the second subgroup could proceed at the same dose of hydroxynidone 800 mg TID. If an SAE was observed in the first subgroup, further analysis was to be performed by the principal investigator to determine the dose level for the second subgroup. Due to an inadvertent oversight of blood glucose testing in the first subgroup (all four subjects) of the hydroxynidone 800 mg TID dose group at the final study visit, enrollment was extended to include four (4) new subjects at the discretion of the principal investigator, increasing the total number of subjects in this dose group to 12. Safety evaluations were performed prior to dosing, on day 5 of dosing, and 24 h after the last dose of the 10-day dosing period.

[0077] No deaths or SAEs were reported in this part of the trial. All subjects in the 600 mg TID cohort completed the study. In the 800 mg TID cohort, all subjects completed the study with the exception of one subject who discontinued the study prematurely on Day 6 based on investigator perceived risk due to abnormal liver function parameters. This subject was assigned to hydroxynidone treatment. In the 600 mg TID cohort, seven (7) AEs were reported: 3 cases of hypertriglyceridemia, 1 case each of chest tightness, abdominal distension, rash, and oral ulcers; in the placebo group, one case of rash and one case of hyperlipidemia were reported. Increased lipid parameters (triglyceridemia) were judged to be clinically significant in three subjects in this group (subject numbers 1, 2, and 6); the increased triglyceride levels returned to their normal values ​​after one week without any treatment. In the 800 mg TID cohort and placebo group, 14 and 5 AEs were reported, respectively. In the 800 mg hydroxynidone group, adverse events included liver dysfunction and hyperlipidemia in three cases each, and headache, dizziness, constipation, flatulence, nausea, heartburn, rash, and eosinophilia in one case each. In the placebo group, adverse events included hyperlipidemia in two cases, diarrhea in two cases, and abnormal liver function in one case.

[0078] In the 800mg TID hydroxynidone dose group and the placebo group, two (2) subjects each had low-density lipoprotein cholesterol (LDL-CHOL) values ​​slightly above the upper limit of normal (ULN). These increases were considered clinically insignificant and returned to normal values ​​after stopping treatment with the study drug. The AEs of abnormal liver function in the subjects of the 800mg TID hydroxynidone dose group included two moderate but clinically significantly increased liver parameters. In one subject in the group, on the fifth day of administration, the ALT and AST values ​​were 79U / L and 47U / L, respectively. When the test was repeated on the 6th day of the study, its ALT and AST values ​​were 101U / L and 61U / L, respectively, with heartburn, rash, headache and other symptoms. Due to the progressive increase in liver enzymes, the investigator terminated the subject due to the perceived risk to the subject's health. The subject was monitored in the following week. Transaminases, without treatment, recovered their normal pre-dose values ​​within one week after the study drug was discontinued. Although this AE met the criteria for an SAE, the investigator classified it as an important adverse event rather than an SAE because no specific treatment was assigned to this subject.

[0079] In summary, the most common AEs associated with 10 days of dosing with 600 mg TID (total daily dose of 1800 mg) or 800 mg TID (total daily dose of 2400 mg) hydroxynidone were hyperlipidemia (triglyceridemia) and increased liver transaminases. These AEs were transient and resolved without treatment. Because these AEs were observed in both the hydroxynidone and placebo groups, it is unclear whether these AEs are specifically related to hydroxynidone treatment.

[0080] Example 7: PK study of single ascending oral doses

[0081] This was a randomized, open-label, 4-period, crossover PK study of single ascending oral doses of hydroxynidone in healthy volunteers.

[0082] The single oral dose of 200mg, 400mg, 600mg and 800mg hydroxynidone was given to 12 qualified healthy male Chinese subjects aged 26-38 years old who met all the inclusion and exclusion criteria, and they were randomly divided into four dose groups, 3 subjects in each group, under fasting conditions. According to Williams design, the subjects in each group were treated with hydroxynidone in four stages with four different dosages. PK blood samples were collected at the following time: 0 (before administration), 10min, 20min, 30min, 40min, 1h, 1.5h, 2h, 3h, 4h, 5h, 6h, 8h, 10h, 12h, 16h, and 24h and 48h after administration. The subjects also provided PK urine samples at 0 (before administration), 5h, 12h and 24h after administration. After the PK sample collection, there was another 24h washout period, which amounted to 48h between the first dose level application and the next dose level application. Blood and urine samples were analyzed to determine plasma and urine concentrations of hydroxynidone and its metabolite (M3).

[0083] Table 1: Single oral dose PK parameters of hydroxynidone in healthy Chinese subjects (Example 7) [Mean (SD)] in Study SG-L-200801 *For Tmax, median value (Min-Max)

[0084] The amount of intact drug excreted via urine during 48 h was 82.25 ± 40.95 mg, 183.75 ± 142.14 mg, 218.93 ± 182.73 mg, and 234.72 ± 210.50 mg. The urine percentage of hydroxynidone was 0.634 ± 0.57%, 0.479 ± 0.22%, 0.476 ± 0.2%, 0.403 ± 0.25%, which shows that most of the orally administered hydroxynidone is not completely eliminated via urine. The percentage of M3 excreted via urine was 29.47%, 32.94%, 26.14%, and 21.04%, showing that hydroxynidone is converted and then excreted as a metabolite via urine.

[0085] Administration of single oral doses of 200 mg, 400 mg, 600 mg, and 800 mg of hydroxynidone was safe and well tolerated in healthy Chinese subjects, with no adverse events or SAEs reported.

[0086] Example 8: Crossover PK Study of the Effect of Food on Safety, Tolerability and PK of a Single Oral Dose

[0087] This is a randomized, open-label, 2-stage, crossover PK study of the effect of food on the safety, tolerability, and PK of a single oral dose of hydroxynidone in healthy Chinese subjects. The PK parameters of the M3 and M4 metabolites of hydroxynidone in the presence or absence of food were also determined.

[0088] In this inpatient study, twelve healthy Chinese subjects were enrolled, dosed and. They were randomly assigned to two groups / stages to receive a single oral 600mg (3x 200mg) dose of hydroxynidone under fasting or feeding (high fat) conditions. In each treatment stage, PK blood samples were collected at the following times: 0 (before administration), 10min, 20min, 30min, 40min, 1h, 1.5h, 2h, 3h, 4h, 5h, 6h, 8h, 10h, 12h, 16h, and 24h and 48h after administration. The washout period between the two stages in this test was 48 hours.

[0089] PK results showed that after oral administration of a single 600 mg dose of hydroxynidone, the presence of food (high-fat meal) delayed the absorption rate of hydroxynidone and its major metabolites M3 and M4, thereby reducing their Cmax values, however, did not change their exposure, as shown by the fact that their AUC values ​​were not affected. Therefore, and based on the AUC results, it was shown that eating had no effect on the exposure levels of hydroxynidone and its major metabolites M3 and M4. Hydroxynidone was safe and well tolerated, with no SAEs or AEs reported.

[0090] Example 9: Open-label, parallel-group PK study of multiple ascending oral doses

[0091] This was a randomized, open-label, parallel-group PK study of multiple ascending oral doses of hydroxynidone in healthy Chinese subjects.

[0092] In this study, increasing oral doses of 200 mg, 400 mg and 600 mg TID hydroxynidone were administered to 27 healthy subjects (15 males, 12 females) divided into 3 groups (9 subjects / group) for four (4) consecutive days. This was an inpatient study. Hydroxynidone was administered under fasting conditions. Blood and urine samples were collected for PK evaluation. PK evaluations were performed after single (after the first dose) and multiple doses. Standard safety evaluations were also performed, including physical examination, ECG, hematology, serum biochemistry and urinalysis.

[0093] PK results show that hydroxynidone is rapidly absorbed, with a Tmax of approximately 1 hour. After a single dose, both the parent drug and the major metabolite were undetectable 36 h after administration. The exposure levels of hydroxynidone and metabolites in human plasma were in the following order from high to low: hydroxynidone-M4>hydroxynidone-M3>hydroxynidone-M2; after a single hydroxynidone administration, the total recovery of the complete drug and M2, M3 and M4 metabolites in urine was 82.7%, 85.5% and 84.0%, respectively. The PK results obtained from single and multiple dosing regimens show that hydroxynidone has no obvious accumulation. In the dose levels administered, male subjects were exposed lower than female subjects. Exposure increases with dose, but due to high variability, there is no strict dose proportionality.

[0094] All 27 subjects completed the study. In this study, hydroxynidone was safe and well tolerated within the single dose range of 200-600 mg TID. No deaths or SAEs were reported in this experiment. Two subjects from the 200 mg TID multiple dose group and five (5) subjects from the 400 mg TID multiple dose group reported seven (7) adverse events. AEs reported in this part of the study included one case each of dizziness, leukopenia and abdominal pain, and two cases each of abdominal distension and diarrhea. All AEs were considered possibly related to hydroxynidone treatment. The severity of the AEs was mild and all recovered spontaneously. All subjects with AEs completed the study.

[0095] Example 10: Open-label study of safety, tolerability and PK of single-dose, 7-day and 4-week multiple oral doses

[0096] This was a randomized, open-label study of the safety, tolerability, and PK of single, 7-day, and 4-week multiple oral doses of hydroxynidone in healthy Chinese subjects.

[0097] This study was conducted in three parts. In the first part, single increasing doses of hydroxynidone of 15 mg, 30 mg, 60 mg, 90 mg and 120 mg / day were administered to six (6), eight (8), eight (8), six (6) and six (6) healthy subjects, respectively, according to the Fibonacci increasing method (modified Fibonacci method). The age, height, weight, body mass index and gender composition ratio of the subjects in each dose group were comparable at baseline. Because no adverse experiences were reported at any dose level, the two highest single dose levels of 90 mg and 120 mg hydroxynidone were selected for the second part of the study and each was administered TID to 6 subjects for seven consecutive days. Therefore, the total daily dose of hydroxynidone in these two dose groups was 270 mg and 360 mg, respectively. All subjects completed the seven-day dosing regimen without early termination. No clinically significant abnormal changes were found in the vital signs, ECG and laboratory test results of each group of subjects. In the third part of the study, six (6) subjects were given oral hydroxynidone at a daily dose of 60 mg TID (total daily dose of 180 mg) for four (4) consecutive weeks. All subjects completed the seven-day dosing regimen without early discontinuation. No clinically significant abnormal changes were found in the vital signs, ECG and laboratory test results of each group of subjects.

[0098] PK assessments were performed in each part of the study. In a single part of the study, PK parameters also included food effect assessments.

[0099] In the single-dose portion of the study (Part I), the main pharmacokinetic parameters of hydroxynidone obtained from 36 healthy subjects orally administered single doses of 30, 60 and 120 mg hydroxynidone in the fasting state were as follows: Tmax was (0.60±0.49).

[0100] A single dose of hydroxynidone within the range of 15-120 mg was safe and well tolerated for all subjects in the study. All subjects completed all study procedures and observations. No death, SAE or AE occurred during the entire study, and there were no abnormal changes in the symptoms perceived by the subjects subjectively, and physical examinations were observed after administration. As compared with baseline and between different dose groups, there were no abnormalities in any vital signs at any time point after administration (p>0.05). Three subjects experienced an increase in their T-BIL and D-BIL post-administration values, two of whom were in the 30 mg dose group and one in the 120 mg dose group. When tested again, these values ​​were within their normal range. The researchers believe that none of these increases are clinically significant.

[0101] Hydroxynidone administration at 90 mg TID (270 mg / day) and 120 mg TID (360 mg / day) for 7 consecutive days was safe and well tolerated for all subjects in the study. There was no premature discontinuation and all subjects completed the study. No deaths, SAEs or AEs occurred during the entire study, and there were no abnormal changes in symptoms perceived subjectively by the subjects, and physical examinations were observed after administration. No clinically significant abnormal changes in vital signs, laboratory parameters, and ECG parameters were found in any dose group (p>0.05).

[0102] Hydroxynidone administration at 60 mg TID (180 mg / day) for 4 consecutive weeks was safe and well tolerated by all subjects in the study. There were no premature discontinuations and all subjects completed the study. No deaths or SAEs were reported.

[0103] Regarding AEs, no AEs were reported in the tolerability portion of the study; only two cases of mild transient dizziness occurred in two female subjects (out of 12 subjects) at 30 min and 1.0 h after dosing in the PK group of multiple doses of 60 mg TID hydroxynidone. These AEs resolved within 1-1.5 h without treatment.

[0104] No clinically significant abnormal changes in vital signs and ECG parameters were found in any dose group (p>0.05). The investigators considered the observed differences in laboratory parameters to be clinically insignificant. There was no abnormal prolongation of the QTc interval at any dose level.

[0105] In summary, within the single oral dose range of 15mg to 120mg, hydroxynidone is safe and well tolerated. The maximum tolerated single oral dose of hydroxynidone is 120mg. When 120mg TID is used for seven consecutive days, up to 360mg / day hydroxynidone is also safe and well tolerated. When 60mg TID is used to healthy subjects for four (4) consecutive weeks, at a dosage of 180mg / day, hydroxynidone is safe and well tolerated. There is no drug accumulation in the body, as indicated by the PK analysis of the main PK parameters at comparable hydroxynidone dosage levels according to single and multiple dosing regimens.

[0106] Example 11: Phase II Clinical Pharmacology Study of Hydroxynidone

[0107] This is a Phase II, randomized, double-blind, placebo-controlled, entecavir-based, multicenter, dose-ranging study of the efficacy and safety of hydroxynidone when administered to Chinese subjects with liver fibrosis due to chronic hepatitis B for 52 consecutive weeks.

[0108] The primary objective of this study is to explore the effective dose and assess the safety of hydroxynidone when co-administered with the antiviral drug entecavir to Chinese subjects with liver fibrosis due to chronic hepatitis B infection for 52 consecutive weeks. The secondary objective of this study is to evaluate the effect of hydroxynidone on the improvement of liver inflammation and liver function when co-administered with the antiviral drug entecavir to Chinese subjects with liver fibrosis due to chronic hepatitis B infection.

[0109] Approximately 240 eligible subjects (approximately 60 subjects per dose cohort) are planned to be enrolled in the study and treated with 30 mg, 60 mg or 120 mg TID oral doses of hydroxynidone (total daily doses of 180 mg, 270 mg and 360 mg, respectively) daily for 52 consecutive weeks, or with placebo, concomitantly administered with a baseline antiviral therapy of 0.5 mg daily dose of entecavir. In parallel, the PK of a group of 12 subjects will be evaluated.

[0110] Example 12: Ishak score analysis of drug groups (primary endpoint)

[0111] like Figure 1 As shown in , after 52 weeks of treatment, hydroxynidone was able to significantly reduce the Ishak score by ≥ 1. The 270 mg group appeared to be the best.

[0112] Example 13: Analysis of changes in Ishak scores within a group

[0113] like Figure 2 As shown in , after 52 weeks of treatment, all drug groups showed a statistically significant decrease in Ishak scores of ≥ 1. The placebo group did not show a statistically significant decrease in Ishak scores of ≥ 1.

[0114] Example 14: Analysis of patients with significant fibrosis

[0115] like Figure 3 As shown in the results of the primary endpoint analysis, the improvement in Ishak score was consistent with the results of the primary endpoint analysis. The 270 mg group appeared to be the best dose group.

[0116] Example 15: Combined analysis of 270 mg and 360 mg groups

[0117] like Figure 4 As shown in the literature, hydroxynidone can improve fibrosis in patients with significant fibrosis and cirrhosis.

[0118] Example 16: Combined analysis of all drug groups

[0119] like Figure 5 As shown in the literature, hydroxynidone can improve fibrosis in patients with significant fibrosis and cirrhosis.

[0120] Example 17: Changes in Ishak scores after 52 weeks in patients with baseline HBeAg(+)

[0121] like Figure 6 As shown in , hydroxynidone showed statistically significant changes when the dose groups were combined.

[0122] Example 18: Changes in Ishak scores after 52 weeks in patients with baseline HBeAg(-)

[0123] like Figure 7 As shown in , hydroxynidone showed statistically significant changes when the dose groups were combined. The improvement in HBeAg(-) patients appeared to be superior to that in HBeAg(+) patients.

[0124] Example 19: Changes in Liver Stiffness Measurement LSM

[0125] like Figure 8 As shown in Figure 3, after 52 weeks of treatment, all drug groups showed a decrease in LSM (kPa) (within-group P < 0.01).

[0126] Example 20: Combination of drug groups for LSM analysis

[0127] like Fig. 9 As shown in , in advanced fibrosis (Ishak ≥ 4), LSM score showed statistical significance.

[0128] Example 21: Analysis of HBV DNA+ patients after 52 weeks of entecavir therapy

[0129] like Fig.10 As shown in Figure 3, all patients (Ishak>3) responded to hydroxynidone therapy.

[0130] Example 22: Effect of Hydroxynidone on (CCl4+HFD)-induced NASH model in mice

[0131] like Fig.11 As shown in , various tests were performed to demonstrate the use of hydroxynidone at different doses. The total NASH score was lower at doses of 15 mg / kg (mpk) and 50 mpk.

[0132] Example 23: H&E staining with hydroxynidone

[0133] like Fig.12 As shown in Figure , the effect of different doses of hydroxynidone was shown by H&E staining. 4 Hydroxynidone at 15 mpk and 50 mpk significantly inhibited CCl4- and WE-induced fibrosis and cellular ballooning. Hydroxynidone was more effective than pirfenidone.

Claims

1. A method of treating a subject suffering from liver fibrosis, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

2. A method of treating a subject having liver cirrhosis, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

3. A method of treating a subject having advanced hepatitis B virus infection, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

4. A method of treating a subject having NASH fibrosis, the method comprising administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

5. The method according to any one of claims 1 to 4, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

6. The method according to any one of claims 1 to 5, further comprising identifying a subject having a liver stiffness value between at least two threshold values.

7. The method of any one of claims 1 to 5, further comprising identifying a subject whose Ishak score is between at least two thresholds.

8. A method of treating a subject, the method comprising identifying a subject having a liver stiffness measurement of at least 4 kilopascals (kPa) and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof.

9. The method of claim 8, wherein the liver stiffness measurement is about 4 kPA to 8 kPa.

10. The method of claim 8, wherein the liver stiffness measurement is above 8 kPa.

11. The method of claim 6 or 8, wherein the liver stiffness measurement is measured using a FibroTouch or FibroScan device.

12. A method of treating a subject, the method comprising identifying a subject having an Ishak value of about 1 to 6; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

13. The method of claim 12, wherein the Ishak value is about 1 to 3.

14. The method of claim 12, wherein the Ishak value is about 3 to 6.

15. A method of treating a subject, the method comprising identifying a subject that is substantially free of hepatitis B virus protein; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

16. A method of treating a subject, the method comprising identifying a subject, wherein the subject tests positive for hepatitis viral DNA after receiving one or more courses of antiviral therapy; and administering to the subject a pharmaceutical composition comprising hydroxynidone or a salt thereof and a pharmaceutically acceptable excipient.

17. The method of claim 16, wherein the hepatitis virus DNA is selected from the group consisting of hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus and hepatitis E virus.

18. The method of any one of claims 1 to 17, wherein the pharmaceutical composition comprises from about 20% to about 90% by weight of hydroxynidone or a salt thereof and one or more excipients.

19. The method of any one of claims 1 to 18, wherein the weight percentage of hydroxynidone or a salt thereof is about 30%.

20. The method according to any one of claims 1 to 19, wherein the pharmaceutical excipients comprise magnesium stearate and lactose.

21. A pharmaceutical composition comprising about 20% to about 90% by weight of hydroxynidone or a salt thereof and one or more pharmaceutically acceptable excipients.

22. The pharmaceutical composition of claim 21, wherein the weight percentage of hydroxynidone or a salt thereof is about 25% to about 35%.

23. The pharmaceutical composition of claim 22, wherein the weight percentage of hydroxynidone or a salt thereof is about 30%.

24. The pharmaceutical composition of claim 23, wherein the one or more pharmaceutically acceptable excipients are selected from lactose, sucrose, magnesium stearate, glucose, vegetable cellulose, calcium carbonate, zinc stearate, calcium stearate, stearic acid, palmitic acid, myristic acid, glyceryl dibehenate and talc.

25. The pharmaceutical composition of claim 24, wherein the one or more pharmaceutically acceptable excipients comprise magnesium stearate and lactose.

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