WO2024169938A1

WO2024169938A1 - Compositions and methods for treating liver diseases

Info

Publication number: WO2024169938A1
Application number: PCT/CN2024/076992
Authority: WO
Inventors: Liping Liu; Ru BAI
Original assignee: Shenzhen Hightide Biopharmaceutical Ltd
Current assignee: Shenzhen Hightide Biopharmaceutical Ltd
Priority date: 2023-02-16
Filing date: 2024-02-08
Publication date: 2024-08-22
Anticipated expiration: 2025-08-16
Also published as: CN120659618A; TW202438509A

Abstract

The invention provides compositions and methods for treating, reducing or preventing liver diseases, in particular alcoholic liver disease (e.g., alcoholic hepatitis, alcoholic liver failure) and drug-induced liver disease, or a related disease and disorder.

Description

COMPOSITIONS AND METHODS FOR TREATING LIVER DISEASES

Priority Claims and Related Patent Applications

This application claims the benefit of priority to U.S. Provisional Application Serial No. 63/446,087, filed February 16, 2023, and Chinese Patent Application No. 2024101689759 filed on February 6, 2024, all of which are incorporated herein by reference in their entireties.

Technical Field of the Invention

The invention generally relates to pharmaceuticals and therapeutic uses thereof. More particularly, the invention provides compositions and methods for treating liver diseases, especially for treating alcoholic liver disease (e.g., alcoholic hepatitis, alcoholic liver failure) and drug-induced liver disease, and related diseases and disorders.

Background of the Invention

Liver diseases, or hepatic diseases, refer to any of a number diseases of the liver. Some liver diseases can be long-lasting, or chronic, while other liver diseases can come on rapidly, or acute, and require urgent care. Liver diseases generally are categorized into those that are caused by viral infections, such as hepatitis A, hepatitis B and hepatitis C; those that are caused by chemicals, drugs, poisons, or overconsumption of alcohol; and those that are caused by genetics or inheritance, such as hemochromatosis and Wilson disease. Many of these diseases can lead to hepatitis, cirrhosis, cancer and/or liver failure. ( "Liver Diseases" MedlinePlus. https: //medlineplus. gov/liverdiseases. html; Williams 2006 "Global challenges in liver disease" Hepatology 522; Blachier, et al. 2013 "The burden of liver disease in Europe: A review of available epidemiological data" J Hepatology 58: 593-608. )

Drug-induced liver injury or disease is common as nearly all classes of medications can cause liver injury or disease. Drug-induced hepatotoxicity is the most frequent cause of acute liver failure in the US and can be categorized as acute or chronic, and either as hepatitis, cholestatic, or a mixed pattern of injury. (David, et al. 2011 "Drug-induced Liver Injury" US Gastroenterol Hepatol Rev. 6: 73-80; Ostapowicz, et al. 2002 "Results of a prospective study of acute liver failure at 17 tertiary care centers in the United States" Ann Intern Med. 137: 947-54. )

Alcoholic liver disease (ALD) , a.k.a. alcohol-related liver disease, is a common disease due to heavy use of alcohol. ALD includes three general stages of alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. Heavy alcohol drinkers can progress through these 3 types over time. Overconsumption of alcohol causes development of large fatty globules (macro-vesicular steatosis) throughout the liver. Alcoholic fatty liver is the build-up of fat inside the liver cells, which leads to an enlarged liver. Alcoholic hepatitis refers to inflammation of the hepatocytes. Death of liver cells over time can lead to progression of fibrosis and cirrhosis due to permanent scarring and destruction of normal liver tissue. Cirrhosis is a serious late-stage liver disease marked by inflammation, fibrosis and damaged membranes preventing detoxification of chemicals in the body, ending in scarring and necrosis. (O'Shea, et al. 2010 "Alcoholic liver disease: AASLD Practice Guidelines" Hepatology 51 (1) : 307-28; Basra 2011. "Definition, epidemiology and magnitude of alcoholic hepatitis" World Journal of Hepatology 3 (5) : 108-113; Seitz, et al. 2018 "Alcoholic liver disease" Nat. Rev. Disease Primers 4, 16. )

ALD can progress from alcoholic fatty liver (AFL) to alcoholic steatohepatitis (ASH) , which is characterized by hepatic inflammation. Chronic ASH can eventually lead to fibrosis and cirrhosis and in some cases hepatocellular cancer (HCC) . In addition, severe ASH (with or without cirrhosis) can lead to alcoholic hepatitis, which is an acute clinical presentation of ALD that is associated with liver failure and high mortality (Seitz, et al. 2018 "Alcoholic liver disease" Nat. Rev. Disease Primers 4, 16. )

ALD is a major cause of liver disease in the world. The prognosis of ALD depends on the liver histology and cofactors such as concomitant chronic viral hepatitis. ALD tends to be diagnosed at later stages than other liver diseases. Approximately 25%to a third of chronic heavy drinkers develop alcoholic hepatitis. Among patients with alcoholic hepatitis, progression to liver cirrhosis occurs at 10-20%per year, and approximately 70%will eventually develop cirrhosis. Despite cessation of alcohol use, only 10%will have normalization of histology and serum liver enzyme levels. In most cases, cirrhosis will eventually lead to liver failure. In the US, ALD mortality rate is 7.3 per 100,000. (Shah, et al. 2019 "Alcohol-Related Liver Disease Is Rarely Detected at Early Stages Compared with Liver Diseases of Other Etiologies Worldwide" Clin Gastroenterol Hepatol 17: 2320; Barrio, et al. 2004 "Liver Disease in Heavy Drinkers with and Without Alcohol Withdrawal Syndrome" Alcoholism: Clinical&Experimental Research 28 (1) : 131-136; Menon, et al. 2001 "Pathogenesis, diagnosis, and treatment of alcoholic liver disease" Mayo Clin. Proc. 76 (10) : 1021-9; Dunn, et al. 2005 "MELD accurately predicts mortality in patients with alcoholic hepatitis" Hepatology 41 (2) : 353-8; Signal, et al. 2018 "ACG Clinical Guideline: Alcoholic Liver Disease" Am. J. Gastroenterol. 113 (2) : 175-194; Peery, et al. 2022 "Burden and Cost of Gastrointestinal, Liver, and Pancreatic Diseases in the United States: Update 2021" Gastroenterol. 162, 2: 621-644. )

Currently, pharmacological treatment options for ALD in general and for alcoholic hepatitis in particular are very limited. Corticosteroids, despite conflicting study results in its efficacy and risk of side effects for long-term use, remain the current standard of care for patients with severe alcoholic hepatitis in the US and the EU. (Saberi, et al. 2016 "Current Management of Alcoholic Hepatitis and Future Therapies" J Clin Transl Hepatol. 4 (2) : 113-122. ) Other drugs besides corticosteroids (e.g., magnesium isoglycyrrhizinate injection, metadoxine) are approved in China although with inadequate efficacy. Challenges posted by high mortality rate, heterogeneity, pathophysiology, along with difficulties in recruiting and retaining patients have all contributed to the current lack of safe and effective treatment options for ALD in general and alcoholic hepatitis in particular.

There is clear unmet medical need for novel pharmacological treatment for ALD and alcoholic hepatitis.

Summary of the Invention

The invention relates to the herein disclosed analogue of the active center of Reg3α(e.g., Ac-IGLHDPSHGTLPAGS) for use in effectively treating or reducing ALD associated disease or conditions (e.g., alcoholic hepatitis and acute liver failure) .

In one aspect, the invention generally relates to a method for treating, reducing or preventing a liver disease or a related disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound that is an analogue of the active center of Reg3α.

In another aspect, the invention generally relates to a method for treating, reducing or preventing a drug-induced liver disease or a related disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound that is an analogue of the active center of Reg3α.

In yet another aspect, the invention generally relates to a method for treating, reducing or preventing alcoholic liver disease, or a related disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound that is an analogue of the active center of Reg3α.

In another aspect, the invention generally relates to a pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient, carrier, or diluent, suitable for treating alcoholic liver disease, or a related disease or condition.

In yet another aspect, the invention generally relates to a unit dosage form comprising the pharmaceutical composition.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating a liver disease, or a related disease or condition.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic liver disease, or a related disease or condition.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic fatty liver, or a related disease or condition.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic steatohepatitis, or a related disease or condition.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic hepatitis.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating moderate alcoholic hepatitis.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating severe alcoholic hepatitis.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic liver cirrhosis.

In yet another aspect, the invention generally relates to use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating acute liver failure associated with alcoholic hepatitis.

In yet another aspect, the invention generally relates to a method for reducing the severity of acute liver disease, comprising administering to a subject in need thereof a pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.

In yet another aspect, the invention generally relates to a method for reducing the mortality rate of alcoholic hepatitis, comprising administering to a subject in need thereof a pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.

In yet another aspect, the invention generally relates to a method for reducing the mortality rate of acute liver failure (e.g., associated with alcoholic liver disease or alcohol consumption) , comprising administering to a subject in need thereof a pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.

Brief Description of the Drawings

FIG. 1 shows exemplary experimental timeline.

FIG. 2 shows exemplary data on survival rates.

FIG. 3 shows exemplary experimental timeline.

FIG. 4 shows exemplary data on survival rates.

FIG. 5 shows exemplary data on survival rate.

Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

As used in the present disclosure, the following words and phrases are generally intended to have the meanings as set forth below unless expressly indicated otherwise or the context in which they are used indicates otherwise.

In this specification and the appended claims, the singular forms "a, " "an, " and "the" include plural reference, unless the context clearly dictates otherwise.

The term “and/or” is used in this disclosure to mean either “and” or “or” unless the context clearly dictates otherwise.

As used herein, “at least” a specific value is understood to be that value and all values greater than that value.

Applicant’s disclosure is described herein in preferred embodiments with reference to the Figures, in which like numbers represent the same or similar elements. Reference throughout this specification to “one embodiment, ” “an embodiment, ” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases “in one embodiment, ” “in an embodiment, ” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

The term “comprising” , when used to define compositions and methods, is intended to mean that the compositions and methods include the recited elements, but do not exclude other elements. The term “consisting essentially of” , when used to define compositions and methods, shall mean that the compositions and methods include the recited elements and exclude other elements of any essential significance to the compositions and methods. For example, “consisting essentially of” refers to administration of the pharmacologically active agents expressly recited and excludes pharmacologically active agents not expressly recited. The term consisting essentially of does not exclude pharmacologically inactive or inert agents, e.g., pharmaceutically acceptable excipients, carriers or diluents. The term “consisting of” , when used to define compositions and methods, shall mean excluding trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

Throughout the description, where compositions and kits are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions and kits of the present invention that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present invention that consist essentially of, or consist of, the recited processing steps.

In the application, where an element or component is said to be included in and/or selected from a list of recited elements or components, it should be understood that the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,1%, 0.5%, 0.1%, 0.05%, or 0.01%of the stated value. Unless otherwise clear from context, all numerical values provided herein can be modified by the term about.

At various places in the present specification, variables or parameters are disclosed in groups or in ranges. It is specifically intended that the description include each and every individual subcombination of the members of such groups and ranges. For example, a range of 1 to 16 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

As used herein, “pharmaceutical composition” refers to the combination of a therapeutically active agent with one or more pharmaceutically acceptable excipients, carriers, or diluents, making the composition especially suitable for diagnostic or therapeutic use in vivo or ex vivo.

As used herein, the term “pharmaceutically acceptable excipient, carrier, or diluent” refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate, magnesium stearate, and polyethylene oxide-polypropylene oxide copolymer as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

As used herein, the term “subject” refers to any animal (e.g., a mammal) , including, but not limited to humans, non-human primates, rodents, and the like, which is to be the recipient of a particular treatment. Typically, the terms “subject” and “patient” are used interchangeably herein in reference to a human subject. A “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult) ) and/or a non-human animal, e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys) , cattle, pigs, horses, sheep, goats, rodents, cats, and/or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal.

As used herein, “administering” means oral administration, administration as a pulmonary, suppository, intramuscular administration, intrathecal administration, intranasal administration or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or) . Parenteral administration includes, e.g., intramuscular and subcutaneous. Other modes of delivery include, but are not limited to, the use of liposomal formulations, etc. By “co-administer” it is meant that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies (e.g., therapeutic agent, chemotherapeutic, or treatment for a neurodegenerative disease) . The compound of formula (I) can be administered alone or can be co-administered to the patient. Co-administration is meant to include simultaneous or sequential administration of the compound individually or in combination (more than one compound or agent) . Thus, the preparations can also be combined, when desired, with other active substances (e.g., to reduce metabolic degradation) .

The terms “disease, ” “disorder, ” and “condition” are used interchangeably herein.

As used herein, the term “treating” , “reducing” , or “preventing” a disease or disorder refers to ameliorating such a condition before or after it has occurred. As compared with an equivalent untreated control, such reduction or degree of prevention is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100%as measured by any standard technique. The terms “treat, ” “treating” and “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition ( “therapeutic treatment” ) , and also contemplates an action that occurs before a subject begins to suffer from the specified disease, disorder or condition ( “prophylactic treatment” ) . In one embodiment, the compounds provided herein are contemplated to be used in methods of therapeutic treatment wherein the action occurs while a subject is suffering from the specified disease, disorder or condition and results in a reduction in the severity of the disease, disorder or condition, or retardation or slowing of the progression of the disease, disorder or condition. In an alternate embodiment, the compounds provided herein are contemplated to be used in methods of prophylactic treatment wherein the action occurs before a subject begins to suffer from the specified disease, disorder or condition and results in preventing a disease, disorder or condition, or one or more symptoms associated with the disease, disorder or condition, or preventing the recurrence of the disease, disorder or condition.

As used herein, the term “effective amount” of an active agent refers to an amount sufficient to elicit the desired biological response. As will be appreciated by those of ordinary skill in this art, the effective amount of a compound of the invention may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the patient.

Compounds of the present invention are, subsequent to their preparation, preferably isolated and purified to obtain a composition containing an amount by weight equal to or greater than 95% ( “substantially pure” ) , which is then used or formulated as described herein. In certain embodiments, the compounds of the present invention are more than 99%pure.

Detailed Description of the Invention

The invention is based in part on the surprising discovery that the herein disclosed analogue of the active center of Reg3α (e.g., Ac-IGLHDPSHGTLPAGS) can be effectively used to reduce or treat ALD associated disease or conditions, in particular for alcoholic hepatitis and acute liver failure.

In certain embodiments, the amino acid sequence of the active center of Reg3αis H-IGLHDPSHGTLPNGS-OH.

In certain embodiments, the compound is Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.

In certain embodiments, the subject is suffering from a liver disease, or a related disease or condition caused or induced by at least a chemical.

In certain embodiments, the subject is suffering from a chronic liver disease, or a related disease or condition.

In certain embodiments, the subject is suffering from an acute liver disease, or a related disease or condition.

In certain embodiments, the subject is suffering from steatohepatitis, or a related disease or condition.

In certain embodiments, the subject is suffering from liver cirrhosis, or a related disease or condition.

In certain embodiments, the subject is suffering from acute liver injury.

In certain embodiments, the subject is suffering from acute liver failure.

In certain embodiments, the chemical is an industrial chemical or a drug.

In certain embodiments, the industrial chemical is selected from alcohol, CCl₄, CHCl₃, nitrite, pesticides, poisons, and food additives.

In certain embodiments, the subject is suffering from alcoholic liver disease.

In certain embodiments, the subject is suffering from alcoholic hepatitis.

In certain embodiments, the subject is suffering from a drug-induced chronic liver disease, or a related disease or condition.

In certain embodiments, the subject is suffering from a drug-induced acute liver disease, or a related disease or condition.

In certain embodiments, the subject is suffering from drug-induced fatty liver, or a related disease or condition.

In certain embodiments, the subject is suffering from drug-induced steatohepatitis, or a related disease or condition.

In certain embodiments, the subject is suffering from drug-induced moderate steatohepatitis.

In certain embodiments, the subject is suffering from drug-induced severe steatohepatitis.

In certain embodiments, the subject is suffering from liver cirrhosis.

In certain embodiments, the subject is suffering from acute liver injury associated with a drug.

In certain embodiments, the subject is suffering from acute liver failure associated with a drug.

In certain embodiments, the subject survives drug-induced acute liver failure after treatment.

Drugs that may cause a liver disease (e.g., liver injury, liver failure, hepatitis) include, but is not limited to, antibiotics, anti-cancer drugs, analgesics, antipyretics, antituberculotic, anti-epileptic drugs, anti-depressant drugs, lipid-lowering drugs, traditional Chinese medicine or dietary supplements.

In certain embodiments, the subject is suffering from acetaminophen (N-acetyl-p-aminophenol, APAP or) induced liver injury.

In certain embodiments, the subject is suffering from alcoholic fatty liver, or a related disease or condition.

In certain embodiments, the subject is suffering from alcoholic steatohepatitis, or a related disease or condition.

In certain embodiments, the subject is suffering from severe alcoholic steatohepatitis.

In certain embodiments, the subject is suffering from alcoholic hepatitis.

In certain embodiments, the subject is suffering from moderate alcoholic hepatitis.

In certain embodiments, the subject is suffering from severe alcoholic hepatitis.

In certain embodiments, the subject survives acute liver failure after treatment.

In certain embodiments, the subject is suffering from moderate alcoholic steatohepatitis.

In certain embodiments, the subject is suffering from alcoholic liver cirrhosis.

In certain embodiments, the subject is suffering from acute liver failure associated with alcoholic hepatitis.

Any suitable route of administration may be utilized in methods of the invention. In certain embodiments, the administration is subcutaneous, intravenous, intramuscular, or inhaled administration.

In certain embodiments, the administration is subcutaneous injection.

In certain embodiments, the compound is administered at a daily dosage in the range of about 10 mg to about 2,000 mg.

In certain embodiments, the compound is administered at a daily dosage in the range of about 10 mg to about 1,000 mg.

In certain embodiments, the compound is administered for a time period of about 1 day to about 6 months.

In certain embodiments, the compound is administered for a time period of about 1 day to about 30 days.

In certain embodiments, the compound is administered at a daily dosage in the range of about 10 mg to about 2,000 mg for a time period of about 1 to about 30 days.

In certain embodiments, the compound is administered at a daily dosage in the range of about 10 mg to about 1,000 mg for a time period of about 1 to about 30 days.

In certain embodiments, the first dose of the compound is administered within 48 hours from the onset or diagnose of alcoholic hepatitis.

In certain embodiments, the first dose of the compound is administered within 24 hours from the onset or diagnose of alcoholic hepatitis.

In certain embodiments, the first dose of the compound is administered within 12, 6, 4, 3, 2 or 1 hour (s) from the onset or diagnose of the alcoholic hepatitis.

In certain embodiments, the first dose of the compound is administered within 24 hours from the onset or diagnose of acute liver failure.

In certain embodiments, the first dose of the compound is administered within 12, 6, 2 or 1 hours from the onset or diagnose of acute liver failure associated with alcoholic liver disease or alcohol consumption.

In certain embodiments, the method of the invention further comprises administering to the subject a second therapeutic agent.

In certain embodiments, the second therapeutic agent is a corticosteroid.

In certain embodiments, the subject had been previously treated with corticosteroids.

In certain embodiments, the pharmaceutical composition is an aqueous formulation suitable for subcutaneous injection.

In certain embodiments, the aqueous formulation is stable at a temperature between about 2 ℃ to about 8 ℃ for at least 48 months.

In certain embodiments, the subject suffers from alcoholic hepatitis or acute liver injury associated with alcoholic liver disease, alcoholic consumption, or an industrial chemical or a drug.

Examples

The following examples are meant to be illustrative of the practice of the invention and not limiting in any way.

Materials, Instruments and Methods

Carbon tetrachloride (CCl₄) (Sigma)

Test compound: Ac-IGLHDPSHGTLPAGS (Shenzhen Hightide Biopharmaceutical Ltd. )

DUR-928: (Shenzhen Hightide Biopharmaceutical Ltd. )

Vehicle control: 0.9%NaCl injection (Guizhou Kelun Pharmaceutical Co., Ltd. )

DUR-928 (Larsucosterol) is an endogenous sulfated oxysterol and an epigenetic regulator. It primarily targets DNA Methyltransferases (DNMTs) and epigenetically modulates the expression of multiple clusters of master genes that are involved in many important cell signaling pathways, through which it stabilizes mitochondria, reduces lipotoxicity, regulates inflammatory or stress responses, and promotes cell survival. DUR-928 is currently under investigation by DURECT for the treatment of alcoholic hepatitis and non-alcoholic steatohepatitis. (https: //classic. clinicaltrials. gov/ct2/show/NCT04563026; Hassanein, et al. 2024 Am J Gastroenterol. 119 (1) : 107-115. )

Example 1. Efficacy Study in CCl4 Induced Rats Acute Liver Injury Model.

Animal Model

Sprague-Dawley rats (6～10 weeks old with body weight about 220-350g, SPF grade) . The SPF grade animal room was maintained at a target temperature of 20℃ to 26℃ and a relative humidity of 40%to 70%throughout the study. The actual room temperature was 20.1℃to 26.8℃; the actual relative humidity was 36%to 78.55%. Fluorescent lighting provided illumination 12 hours per day. Certified rodent diet and filtered water were provided ad libitum during the quarantine and study periods except there are any requirement for the study operation. After acclimated for 7 days, acute liver failure was induced by i.p. injection of 50% (v/v) of CCl4 in Olive oil with dosing volume of 4mL/kg.

Group Design and Treatment

The model rats were randomly divided into 2 groups: the vehicle control group (G1, n=22) and the test article treatment group (G2, n=21) . One hour after model induction, the animals from G1 and G2 received one dose of Normal Saline (4 mL/kg) as vehicle control or Test compound (1.25 mg/kg) through S.C. injection, respectively. All animals were observed continuously during the study period for signs of, but not be limited to, mortality, morbidity, mental status, behavior, respiration, secretion, feces, and for the availability of food and water intake. The detailed study design is summarized in Table 1 below.

Table 1

FIGs. 1-2 show certain exemplary data regarding experimental timeline and survival rates.

Results are shown in FIG. 2. The animals began to die～3 h after ALF induction. It was evident from the results that the death was postponed in the Test compound treated group compared to the vehicle control. At the end of the study, the survival rate for the test article group was 38.1%whereas the survival rate for the model group was 27.3%.

Decrease in spontaneous motor activity, shortness of the breath, and torsion reaction caused by pain could be seen from cage-side observations in the animals after model induction. In addition to the increased survival rate in the test article treated group, the activity states of the animals also improved significantly. All those results demonstrated that the test article had beneficial effects on the liver injury.

Example 2. Efficacy Study in CCl4 Induced Rats Acute Liver Injury Model with Characteristics of Alcoholic Hepatitis.

Animal Model

Sprague-Dawley rats (6-10 weeks old with body weight about 220-350g, SPF grade) . The SPF grade animal room was maintained at a target temperature of 20℃ to 26℃ and a relative humidity of 40%to 70%throughout the study. The actual room temperature was 20.1℃to 26.8℃. The actual relative humidity was 36%to 78.55%. Fluorescent lighting provided illumination 12 hours per day. Certified rodent diet and filtered water were provided ad libitum during the quarantine and study periods except when required by the study. After acclimated for 7 days, acute liver failure was induced by i.p. injection of 50% (v/v) of CCl4 in olive oil with dosing volume of 4 mL/kg.

Grouping and treatment

The model rats were randomly divided into 3 groups: the vehicle control group (G1) and the test article treatment groups (G2 and G3) , with 18 rats in each group. One hour after model induction, animals from G1～G3 received one single dose of Normal Saline (4 mL/kg) as vehicle control or the test compound (1.25 mg/kg) through S.C. injection, respectively. All animals were observed continuously during the study period for signs of, but not be limited to, mortality, morbidity, mental status, behavior, respiration, secretion, feces, and for the availability of food and water intake. The detailed study design is summarized in Table 2 below.

Table 2

FIGs. 3-4 show certain exemplary data regarding experimental timeline and survival rates.

The results are shown in FIG. 4. The animals began to die～3.9 h after ALF induction. Death was postponed in the Test compound treated groups as compared to the vehicle control. Better efficacy was observed in the Test compound-1h group than the Test compound-2h group.

Example 3. Efficacy Study in APAP Induced C57 mice model with alcoholic hepatitis characterized by acute liver injury

C57 mice (12-week-old male, SPF grade) . The SPF grade animal room was maintained at a target temperature of 20℃ to 26℃ and a relative humidity of 40%to 70% throughout the study. The actual room temperature was 20.1℃ to 26.8℃. The actual relative humidity was 36%to 78.55%. Fluorescent lighting provided illumination 12 hours per day. Certified rodent diet and filtered water were provided ad libitum during the quarantine and study periods except when required by the study. After acclimated for 7 days, the mice were weight-pair assigned into several groups including the vehicle group, the control group and the test groups. Acute liver failure was induced by single i.p. injection of acetaminophen (APAP) in PBS solution for the control group and all test groups, while the vehicle group was designed to receive PBS in the same way. Two hours before, on, or after challenge with APAP or PBS solution, each group was assigned to receive the test article or the vehicle as designed. The frequency, interval, dose level, and the time period of administration for each group can be designed adaptively. All animals were observed continuously during the study period for signs of, but not be limited to, mortality, morbidity, mental status, behavior, respiration, secretion, feces, and for the availability of food and water intake. (General methods for establishing acute liver injury model and the associate effect study can also be referenced to Steinebrunner, et al. 2014 “Pharmacologic cholinesterase inhibition improves survival in acetaminophen-induced acute liver failure in the mouse. ” BMC Gastroenterol. 14: 148. )

Example 4. Protective effects of Ac-IGLHDPSHGTLPAGS (HTD4010) and DUR-928 in a lipopolysaccharides (LPS) -induced acute liver failure (ALF) mouse model.

Male C57BL/6 mice (8-9 weeks old, SPF grade) were acclimated for at least 1 week before randomization in to 4 groups, including model control group G0 and test groups of G1-G3, with 15 mice/group as shown in the table below. All mice were intravenously challenged with 40 mg/kg LPS to induce ALF model. Day 1 was defined as the day of LPS injection.

For group G0 on Day 1, the mice were subcutaneously injected with vehicle once at 1 h after LPS stimulation and once at 8 h after LPS injection, respectively; and from Day2 and thereafter, the mice were subcutaneously injected with vehicle twice a day.

For group G1 on Day 1, the mice were subcutaneously injected with HTD4010 (5mg/kg) once at 1 hour after LPS stimulation and once at 8 h after LPS injection, respectively; and from Day 2 and thereafter: the mice were subcutaneously injected with HTD4010 (5mg/kg) twice a day.

For group G2, the mice were pre-treated with DUR-928 (50 mg/kg) by intravenous injection 2 h before LPS injection on Day 1.

For group G3 1-hour after LPS injection, the mice were treated with DUR-928 (50mg/kg) by intravenous injection on Day 1.

The animals were closely monitored, including mental status, activity, excretion and mortality during the whole study period. The study was terminated on about Day 7 until no mortality is observed in 12 h for each group) .

Table 3. Grouping and Treatment

Note: 1. For G1-HTD4010 group, the dose levels of 10mg/kg/day and dosing method (sc, 1h after LPS injection, BID) used in the study were selected base on previous study results; 2. For G2-DUR-928 (pre) group, the dose level of 50mg/kg and dosing method (iv injection, two hours before LPS injection) were selected base on literature (Ning, et al. 2017 Metabolism 71: 83-93; Wang, et al. 2021 Cells 10 (11) : 3027) and clinical route of drug delivery (Am J Gastroenterol. 2023 May 8) ; 3. For G3-DUR-928 group, the dosing method of DUR-928 with 1 h after LPS injection was the same with HTD4010 also included.

Results

FIG. 5 shows exemplary data on survival rate of each group in LPS induced ALF mouse model (*P<0.05 as compared to G0 Model control with Log-rank (Mantel-Cox) test method. #P<0.05 as compared to G3 of DUR-928 group. )

DUR-928 when preventive treated (50 mg/kg, 2h before LPS injection) showed significant protective effects in this ALF mouse model (P<0.05) , which was consistent with reported data in the literature (Ning, et al. 2017 Metabolism 71: 83-93) and were stronger than given at 1 h after LPS injection (80%vs. 53%) .

HTD4010 at dose levels of 10 mg/kg/day showed significant protective effects when given after LPS injection (P<0.01) . The treatment effects of HTD4010, when given after LPS injection, were slightly stronger than the effects of DUR-928 when it was preventive treatment before LPS injection (87%vs. 80%) . The protective effects of HTD4010 groups were significantly stronger than DUR-928 when they were both treated after the LPS injection (87%vs. 53%) , with statistic difference achieved (P＜0.05) .

The described features, structures, or characteristics of Applicant’s disclosure may be combined in any suitable manner in one or more embodiments. In the description, herein, numerous specific details are recited to provide a thorough understanding of embodiments of the invention. One skilled in the relevant art will recognize, however, that Applicant’s composition and/or method may be practiced without one or more of the specific details, or with other methods, components, materials, and so forth. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the disclosure.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Methods recited herein may be carried out in any order that is logically possible, in addition to a particular order disclosed.

Incorporation by Reference

References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made in this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes. Any material, or portion thereof, that is said to be incorporated by reference herein, but which conflicts with existing definitions, statements, or other disclosure material explicitly set forth herein is only incorporated to the extent that no conflict arises between that incorporated material and the present disclosure material. In the event of a conflict, the conflict is to be resolved in favor of the present disclosure as the preferred disclosure.

Equivalents

The representative examples are intended to help illustrate the invention, and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including the examples and the references to the scientific and patent literature included herein. The examples contain important additional information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.

Claims

A method for treating, reducing or preventing a liver disease or a related disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound that is an analogue of the active center of Reg3α.
The method of claim 1, wherein the compound is Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.
The method of claim 1 or 2, wherein the subject is suffering from a liver disease, or a related disease or condition caused or induced by at least a chemical.
The method of any one of claims 1-3, wherein the subject is suffering from a chronic liver disease, or a related disease or condition.
The method of any one of claims 1-3, wherein the subject is suffering from an acute liver disease, or a related disease or condition.
The method of any one of claims 1-3, wherein the subject is suffering from steatohepatitis, or a related disease or condition.
The method of any one of claims 1-3, wherein the subject is suffering from liver cirrhosis, or a related disease or condition.
The method of any one of claims 1-3, wherein the subject is suffering from acute liver injury.
The method of any one of claims 1-3, wherein the subject is suffering from acute liver failure.
The method of any one claims 3-9, wherein the chemical is an industrial chemical or a drug.
The method of claim 1 or 2, wherein the subject is suffering from alcoholic liver disease.
The method of claim 1 or 2, wherein the subject is suffering from alcoholic hepatitis.
A method for treating, reducing or preventing a drug-induced liver disease, or a related disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound that is an analogue of the active center of Reg3α.
The method of claim 13, wherein the compound is Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.
The method of claim 13, wherein the subject is suffering from a drug-induced acute liver disease, or a related disease or condition.
The method of claim 13, wherein the subject is suffering from drug-induced fatty liver, or a related disease or condition.
The method of claim 13, wherein the subject is suffering from drug-induced steatohepatitis, or a related disease or condition.
The method of claim 13, wherein the subject is suffering from drug-induced moderate steatohepatitis.
The method of claim 13, wherein the subject is suffering from drug-induced severe steatohepatitis.
The method of claim 13, wherein the subject is suffering from liver cirrhosis.
The method of claim 13, wherein the subject is suffering from acute liver injury.
The method of claim 13, wherein the subject is suffering from acute liver failure.
The method of claim 22, wherein the subject survives drug-induced acute liver failure after treatment.
The method of any one of claims 13-23, wherein the drug is selected from antibiotics, anti-cancer drugs, analgesics, antipyretics, antituberculotic, anti-epileptic drugs, anti-depressant drugs, lipid-lowering drugs, traditional Chinese medicine, and dietary supplements.
A method for treating, reducing or preventing alcoholic liver disease, or a related disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound that is an analogue of the active center of Reg3α.
The method of claim 25, wherein the compound is Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.
The method of claim 25 or 26, wherein the subject is suffering from alcoholic fatty liver, or a related disease or condition.
The method of claim 25 or 26, wherein the subject is suffering from alcoholic steatohepatitis, or a related disease or condition.
The method of claim 25 or 26, wherein the subject is suffering from alcoholic hepatitis.
The method of claim 25 or 26, wherein the subject is suffering from moderate alcoholic hepatitis.
The method of claim 25 or 26, wherein the subject is suffering from severe alcoholic hepatitis.
The method of claim 25 or 26, wherein the subject is suffering from alcoholic liver cirrhosis.
The method of claim 25 or 26, wherein the related disease or condition is suffering from acute liver failure associated with alcoholic hepatitis.
The method of any one of claims 1-33, wherein the administration is subcutaneous, intravenous, intramuscular, or inhaled administration.
The method of claim 34, wherein the administration is subcutaneous injection.
The method of claim 34 or 35, wherein the compound is administered at a daily dosage in the range of about 10 mg to about 2,000 mg for a time period of about 1 day to about 6 months.
The method of claim 36, wherein the compound is administered at a daily dosage in the range of about 10 mg to about 1,000 mg for a time period of about 1 day to about 1 month.
The method of claim 34 or 35, wherein the compound is first administered within 12 hours from the onset of the alcoholic hepatitis or alcoholic liver failure.
The method of any one of claims 1-38, further comprising administering to the subject a second therapeutic agent.
The method of claim 39, wherein the second therapeutic agent is selected from corticosteroids.
The method of any one of claims 1-38, wherein the subject was previously treated with corticosteroids.
A method for reducing the mortality rate of alcoholic hepatitis, comprising administering to a subject in need thereof a pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.
A method for reducing the mortality rate of acute liver failure, comprising administering to a subject in need thereof a pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof.
A pharmaceutical composition comprising Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient, carrier, or diluent, suitable for treating alcoholic liver disease, or a related disease or condition.
A unit dosage form comprising a pharmaceutical composition according to claim 44.
The unit dosage form of claim 45, wherein the pharmaceutical composition is an aqueous formulation suitable for subcutaneous injection.
The unit dosage form of claim 46, wherein the aqueous formulation is stable at a temperature between about 2 ℃ to about 8 ℃ for at least 48 months.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating a liver disease, or a related disease or condition.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic liver disease, or a related disease or condition.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic fatty liver, or a related disease or condition.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic steatohepatitis, or a related disease or condition.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating alcoholic hepatitis.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating moderate alcoholic hepatitis.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating severe alcoholic hepatitis.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating liver cirrhosis.
Use of Ac-IGLHDPSHGTLPAGS, or a pharmaceutically acceptable form thereof, for treating acute liver failure.