Fatty acid derivatives for treating non-alcoholic steatohepatitis

Abstract

To provide compounds for use in therapeutic and / or prophylactic treatment of non-alcoholic steatohepatitis (NASH) and / or alcoholic steatohepatitis (ASH).SOLUTION: The compound for use according to the invention, is an unsaturated fatty acid with an oxygen incorporated in the β-position, and further comprising an α-substituent. More particularly, the invention provides a compound for use in treatment of NASH and / or ASH, and a method of use thereof, where the compound is of Formula (II), and this compound may be administered alone or in combination with an additional active agent. In the Formula, R1 is selected from C10-C22 alkenyls having 3 to 6 double bonds, R2 and R3 are the same or different, and are a hydrogen atom, a hydroxy group, an alkyl group, a halogen atom, an alkoxy group, an acyloxy group etc., and X is a carboxylic acid or a derivative thereof.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims the benefit of priority of Norwegian Patent Application No. 20171944, filed on Dec. 6, 2017; Norwegian Patent Application No. 20171945, filed on Dec. 6, 2017; and U.S. Provisional Patent Application No. 62 / 734,013, filed on Oct. 9, 2018. All of the foregoing applications are hereby incorporated by reference in their entirety. ; and U.S. Provisional Patent Application No. 62 / 734,013, filed on Oct. 9, 2018. All of the foregoing applications are hereby incorporated by reference in their entirety. This disclosure relates to methods of treating non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis (ASH), and other liver disorders characterized by fibrosis and / or liver inflammation in subjects in need thereof. Further, this disclosure relates to compounds and compositions containing compounds for use in the treatment of non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis (ASH), and other liver disorders characterized by fibrosis and / or liver inflammation in subjects in need thereof. The compounds for use according to the invention are unsaturated fatty acids having oxygen incorporated at the β-position and further comprising an α-substituent. This disclosure relates to methods of treating non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis (ASH), and other liver disorders characterized by fibrosis and / or liver inflammation in subjects in need thereof. Further, this disclosure relates to compounds and compositions containing compounds for use in the treatment of non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis (ASH), and other liver disorders characterized by fibrosis and / or liver inflammation in subjects in need thereof. The compounds for use according to the invention are unsaturated fatty acids having oxygen incorporated at the β-position and further comprising an α-substituent.

[0002]

[0003]

Background Art

[0003] Long-chain omega-3 fatty acids, such as (5Z,8Z,11Z,14Z,17Z)-icos-5,8,11,14,17-pentaenoic acid (EPA) and (4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (DHA), have a wide range of biological effects and affect plasma lipid levels, cardiovascular function and immune function, insulin action, neurodevelopment, and visual function. High-dose EPA / DHA is currently used for severe hyper- ,8,11,14,17-pentaenoic acid (EPA) and (4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (DHA), have a wide range of biological effects and affect plasma lipid levels, cardiovascular function and immune function, insulin action, neurodevelopment, and visual function. High-dose EPA / DHA is currently used for severe hyper- ,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (DHA), have a wide range of biological effects and affect plasma lipid levels, cardiovascular function and immune function, insulin action, neurodevelopment, and visual function. High-dose EPA / DHA is currently used for severe hyper- ,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (DHA), have a wide range of biological effects and affect plasma lipid levels, cardiovascular function and immune function, insulin action, neurodevelopment, and visual function. High-dose EPA / DHA is currently used for severe hyper- ,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid (DHA), have a wide range of biological effects and affect plasma lipid levels, cardiovascular function and immune function, insulin action, neurodevelopment, and visual function. High-dose EPA / DHA is currently used for severe hyper-​ It is prescribed for the treatment of hypertriglyceridemia (HTG). These effects are at least This is mediated in part by effects on fatty acid metabolism in the liver.

[0004] The use of EPA and DHA for the treatment of non-alcoholic steatohepatitis (NASH) The use of mega-3 compounds has been proposed in the prior art. For example, Mochida et al. WO2014 / 057522 discloses a method for treating or alleviating the symptoms of NASH. The present invention relates to a composition containing ethyl icosapentate for the purpose of preventing and treating skin irritation.

[0005] Dignity Science LTD (WO2014 / 118097) is a non-alcoholic 1 for treating fatty liver disorders such as non-alcoholic fatty liver disease (NAFLD) and NASH Modified omega-3 compounds such as 5-hydroxyeicosapentaenoic acid (15-OHEPA) Krisani Biosciences (WO2014 / 04 5293) also explores the use of modified omega-3 compounds to treat various diseases, including NASH. More recently, Pronova Biopharma AS (WO2 016173923A1) describes the use of sulfur-containing structurally modified fatty acids for the treatment of NASH. I suggested.

[0006] NAF encompasses a much broader range of liver diseases, including isolated hepatic steatosis (histologic >5% hepatocytes). Although LD encompasses non-alcoholic fatty liver disease (NAFLD) and non-alcoholic fatty liver disease (NAFLD), Nonalcoholic steatohepatitis (NASH) is often used synonymously. Hepatic steatosis is a condition characterized by an inflammatory response and When not accompanied by cellular damage, it is most likely a relatively benign disorder. Subgroups of LD patients have hepatocyte injury and inflammation in addition to hepatic steatosis, which is a condition known as non-alcoholic steatohepatitis (NASH). NASH is histologically almost indistinguishable from alcoholic steatohepatitis (ASH). Simple steatosis seen in NAFLD is not correlated with an increased short-term morbidity or mortality, but NASH dramatically increases the risk of cirrhosis, liver failure and hepatocellular carcinoma (HCC). Cirrhosis due to NASH is becoming an increasingly frequent reason for liver transplantation. Morbidity and mortality due to liver causes are greatly increased in patients with NASH, and they are even more strongly correlated with morbidity and mortality due to cardiovascular disease.

[0007] Uniform criteria for the diagnosis and staging of NASH are still under discussion (see later section for details). The key histological elements of NASH are steatosis, ballooning of hepatocytes and lobular inflammation, and fibrosis is not part of the histological definition of NASH. However, the degree (stage) of fibrosis in liver biopsy is useful for predicting prognosis, while the degree of inflammation and necrosis (grade) in liver biopsy is not useful.

[0008] Regarding various histological elements, when treatment is established at an early stage of the disease, treatment with omega-3 fatty acids has been shown to effectively reduce hepatic steatosis in patients with NAFLD (Scorletti E, et al., Effects of purifi ed eicosapentaenoic and docosahexanoic a cids in non-alcoholic fatty liver diseas e: Results from the *WELCOME study, Hepat ology.2014 Oct;60(4):1211 - 2nd, this will probably delay the progression to a more severe stage of the disease. However, it is doubtful whether omega - 3 fatty acids are potent enough to treat and / or reverse NASH in which significant histological / inflammatory changes have occurred (Sanyal AJ, et al; EPE - A Stud y Group, Gastroenterology.2014 Aug;147(2) :377 - 84.e1). The modest efficacy of omega - 3 fatty acids in the treatment of NASH may be secondary to their mild effects on other pathways underlying the etiology of NASH. Studies in both human and animal models of NASH have convincingly shown that, in contrast to simple steatosis, there are multiple factors involved in the development of steatohepatitis and fibrosis. These factors include insulin resistance, oxidative stress, inflammation, gut - derived endotoxins as well as excessive hepatic cholesterol and bile acids. All of these factors have been shown to play important contributing roles in genetically susceptible individuals and, therefore, drugs targeting these pathways are under development for the treatment of NASH.

[0009] Similar to NASH, alcoholic liver disease (ALD) can be grouped into histological stages representing the transition from fatty liver or simple steatosis to alcoholic hepatitis (i.e., ASH) and ultimately to chronic hepatitis with liver fibrosis or cirrhosis. Thus, although the origins of ASH and NASH may be different, the responses to each type of chronic injury

[0010] ​​​​​​​​​​​​Liver reactions involve macrophage activation and cytokine production and the resulting activation of stellate cells, i.e., proliferative myofibroblasts, and share many similarities, such as the inflammatory and fibrogenic cascades they participate in. (See, for example, Friedman, SL; Alcoholism: Clinical and Experimental Research. 1999 May; 23(5): 904-910.) One of the challenges in developing drugs targeting NASH and ASH is the lack of preclinical models that individually represent these diseases in humans. Rodent models that can better represent the metabolic abnormalities typically associated with NASH, such as dyslipidemia and insulin resistance, are characterized by very mild liver inflammation and fibrosis. At the opposite extreme are chemically induced fibrosis models, such as carbon tetrachloride (CCl4) or thioacetamide-induced fibrosis, which cause more severe fibrosis but cannot bridge the gap to humans in terms of both metabolic factors such as insulin resistance and obesity. Therefore, multiple preclinical models that address both ends of the disease spectrum (metabolic disruption - inflammatory response - fibrosis) are necessary for the identification of potential NASH and ASH drugs. Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models m: Clinical and Experimental Research.19 99 May;23(5):904-910 are referred to.)

[0011] In the development of drugs targeting NASH and ASH, the problem is that there are no preclinical models that individually represent these diseases in humans. Rodent models that can better represent the metabolic abnormalities typically associated with NASH, such as dyslipidemia and insulin resistance, are characterized by very mild liver inflammation and fibrosis. At the opposite extreme are chemically induced fibrosis models, such as carbon tetrachloride (CCl4) or thioacetamide-induced fibrosis, which cause more severe fibrosis but cannot bridge the gap to humans in terms of both metabolic factors such as insulin resistance and obesity. Therefore, multiple preclinical models that address both ends of the disease spectrum (metabolic disruption - inflammatory response - fibrosis) are necessary for the identification of potential NASH and ASH drugs. Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models 、carbon tetrachloride (CCl4) or thioacetamide-induced fibrosis, which cause more severe fibrosis but cannot bridge the gap to humans in terms of both metabolic factors such as insulin resistance and obesity. Therefore, multiple preclinical models that address both ends of the disease spectrum (metabolic disruption - inflammatory response - fibrosis) are necessary for the identification of potential NASH and ASH drugs. Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models fibrosis) are necessary for the identification of potential NASH and ASH drugs. fibrosis) are necessary for the identification of potential NASH and ASH drugs.

[0012] Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models Another important factor to consider when comparing rodent liver fibrosis and human liver fibrosis is the site and functional relevance of the induced fibrosis. For example, a more short-term severe fibrosis model may represent only large portal tracts with dense collagen rather than the parenchymal collagen deposition that is more highly functionally relevant and represents most of the liver collagen. Therefore, the biochemical (e.g., hydroxyproline) aspects of fibrosis in rodent models and used in combination with both histological (e.g., Sirius Red morphometry) evaluations is optimal with respect to the amount, location, and functional relevance of hepatic collagen deposits.

[0013] Liver fibrosis can progress to cirrhosis, and thus a significant increase in morbidity and mortality associated therewith, making it very important to identify drugs that target the fibrotic elements of NASH and ASH. This also represents a major hard endpoint in clinical studies of chronic liver disease. For example, emerging data suggest that fibrosis, rather than NASH itself, is the most important histological predictor of both liver-related and non-liver-related death. In addition, cirrhosis is a potent cofactor for primary liver cancer. Therefore, new drugs under development for the treatment of NASH and ASH must be potent enough to prevent the development of established fibrosis and / or reverse its progression.

[0014] WO2016173923 A1 of Pronova Biopharma Norge AS discloses that sulfur-containing structured modified fatty acids such as 2-ethyl-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5, 8,11,14,17-pentaenylthio)butanoic acid (Compound N) may be useful in the treatment of NASH. This was based on the finding that Compound N was superior to rosiglitazone, a PPAR-gamma agonist, in the prevention of diet-induced liver fibrosis. Compound N was also shown to prevent the influx of inflammatory cells into the liver. In APOE 3Leiden.CETP mice, fibrosis * The effectiveness of reducing the symptoms (measured by the hydroxyproline / proline ratio) of compound N has been shown. Importantly, APOE * 3 Leiden.CETP mice only exhibit a mild liver fibrosis reaction (a 20 - 30% increase in the extracellular matrix), as measured by a biochemical assay evaluating hydroxyproline (HYP) content. Furthermore in a specific model described in WO2016173923A1 (the content of which is incorporated herein by reference), the amount of liver fibrosis measured by Sirius Red (SR) morphometry is much less than that seen in five previous studies using similar experimental settings. In those studies, fibrosis measured by SR morphometry was approximately 4 - 5% after 20 weeks and 7 - 8% after 25 - 30 weeks, compared to 1.5% in the study shown in WO20161 73923A1. Regarding the measurement of fibrosis, SR morphometry is more sensitive than biochemical analysis (HYP), which may underestimate the collagen that is mainly parenchymal and has a higher functional relevance to the portal vein collagen that is less functionally relevant but quantitatively dominant.

[0015] Therefore, based on the need for powerful drugs to treat NASH and ASH, such as for prophylactically treating or reversing liver fibrosis, several new studies were conducted to identify compounds that can be used in the treatment of these aspects of liver disease and to confirm the effects based on new evaluations and better models for higher levels of fibrosis.

[0016] For prophylactically treating or reversing liver fibrosis, such as for treating NASH and ASH Based on the need for powerful drugs to do so, identify compounds that can be used in the treatment of these aspects of liver disease and confirm the effects based on new evaluations and better models, several new studies have been conducted.

SUMMARY OF THE INVENTION

[0017] The present disclosure relates to a method for treating non-alcoholic steatohepatitis and / or alcoholic steatohepatitis in a subject in need thereof, the method comprising administering a pharmaceutically effective amount of a compound of formula (II):

[0018]

Chemical Formula

[0019] An equivalent aspect of the present disclosure is for use in the therapeutic and / or prophylactic treatment of non-alcoholic steatohepatitis and / or alcoholic steato hepatitis, a compound of formula (II)

[0020] [Chemical formula] (wherein R1 is selected from C 10 -C 22 alkenyl having 3 to 6 double bonds , R2 and R3 are the same or different and are selected from the group of substituents consisting of a hydrogen atom, a hydroxy group, an alkyl group, a halo gen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an a ryl group, an alkylthio group, an alkoxycarbonyl group, a carboxy group, an alkylsulfi nyl group, an alkylsulfonyl group, an amino group, and an alkylamino group, where R2 and R3 can be linked to form a cycloalkane such as cyclopropane, cyclobutane, cyclopentane or cyclohexane, X is a carboxylic acid or a derivative thereof, and the derivative is a carboxylate such as a carboxylic acid ester; a glyceride; an anhydride; a carboxamide; a phospholipid; or a hydroxym ethyl; or a prodrug thereof) or a pharmaceutically acceptable salt, solvate, or solvate of such a salt of the compound . The present disclosure also provides a compound for use as a monotherapy or one or more ​​It is provided that the compound may be administered in combination with an additional active agent of the type.

[0021] In at least one embodiment, R2 and R3 are the same or different and are hydrogen atoms. The substituents may be selected from the group consisting of alkyl, alkoxy, and alkenyl groups. or R2 and R3 are cyclopropane, cyclobutane, cyclopentane or cycloalkyl; can be linked to form cycloalkanes such as cyclohexane, X represents a carboxylic acid or a derivative thereof, and the derivative may be a carboxylic acid ester, a glycerin, or the like. lipids or phospholipids, or a pharmaceutically acceptable salt, solvate, or solvate of such a salt thereof.

[0022] In particular, the present disclosure relates to a method for treating nonalcoholic steatohepatitis and / or a method for treating alcoholic steatohepatitis, comprising administering a pharmaceutically effective amount of a compound of formula (I):

[0023] [ka] wherein R2, R3 and X are defined as in formula II. In particular, R2 and R3 are hydrogen atoms or straight-chain, branched-chain and / or cyclic C1-C6 alkyl groups. independently selected from the alkyl group, X is a carboxylic acid or a derivative thereof, and the derivative is a carboxylic acid ester, a glycerin lipids or phospholipids) or a pharmaceutically acceptable salt, solvate, or solvate of such a salt and administering to a subject.

[0024] The present disclosure also provides a compound of formula (I) for use in the treatment of non-alcoholic steatohepatitis. :

[0025] [Chemical formula] (wherein, R2, R3 and X are defined in the same manner as in Formula II. In particular, R2 and R3 are independently selected from a hydrogen atom or a linear, branched and / or cyclic C1-C6 alkyl group, X is a carboxylic acid or a derivative thereof, and the derivative is a carboxylic acid ester, glyceride or phospholipid) of the compound or a pharmaceutically acceptable salt, solvate, or solvate of such a salt thereof is provided.

[0026] The present disclosure also relates to a method for treating non-alcoholic steatohepatitis and / or alcoholic steatohepatitis in a subject in need thereof, the method comprising administering to the subject a pharmaceutically effective amount of 2-(((5Z,8 Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1- yl)oxy)butanoic acid (Compound A):

[0027] [Chemical formula] or a pharmaceutically acceptable salt or ester thereof to the subject. is also provided.

[0028] The present disclosure also provides 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5 ,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (Compound A) or a pharmaceutically acceptable salt or ester thereof for use in the treatment of non-alcoholic steatohepatitis and / or alcoholic steatohepatitis. is also provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0029]

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Mode for Carrying Out the Invention

[0030] It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents. It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents. It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents. It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents. It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents. It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents. It should be noted that the embodiments and features described in the context of one aspect of the present disclosure apply to other aspects of the present invention. In particular, embodiments applicable to a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure also apply to aspects directed to a compound or a composition comprising the compound for use in the treatment of all non-alcoholic steatohepatitis or alcoholic steatohepatitis according to the present disclosure. In some embodiments, the compound or the composition comprising the compound is administered in combination with one or more additional active agents.

[0031] Certain aspects of the present disclosure will be described in more detail below. As used in this application, the terms and definitions set forth herein are intended to represent their meaning within the present disclosure .

[0032] The singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0033] The terms “about” and “approximately” mean substantially the same as the stated number or value. When used herein, the terms “about” and “approximately” are generally understood to encompass ±5% of the specified quantity, frequency, or value.

[0034] The terms “treat,” “treating,” and “treatment” include any therapeutic or prophylactic application that may confer a benefit to a human or non-human mammalian animal. Both human treatment and veterinary treatment are within the scope of the present disclosure. Treatment can be responsive to an existing condition or prophylactic, i.e., preventive .

[0035] As used herein, the terms “administer,” “administration,” and “administering” refer to (1) providing, giving, dosing, and / or treating a compound or composition according to the present disclosure by or under the direction of a physician or one of their agents, and (2) introducing, ingesting, or consuming a compound or composition according to the present disclosure by a human patient or the patient themselves or a non-human mammalian animal.

[0036] "Preventing and / or treating" and "therapeutic and / or prophylactic treatment" can be used synonymously. Typically, a compound of formula (I) or formula (I I) is used for the treatment of NASH or ASH, i.e., for therapeutic treatment. However, sometimes, for example, when a patient has one or more risk factors associated with NASH or ASH, it is also foreseeable that a compound of formula (I) or formula (II) will be used for the prophylactic treatment of NASH or ASH.

[0037] The terms "administered in combination" and "co - administration" or "coadministration" are used synonymously and refer to the administration of (a) a compound of formula (I) or (II), or a pharmaceutically acceptable salt, solvate thereof, or a solvate of such a salt; and (b) in conjunction, at least one additional active agent. For example, co - administration can be simultaneous administration, sequential administration, repeated administration, intermittent administration , continuous administration or combinations thereof. The method of administration may differ for the compound and the additional agent(s) (one or more), and co - administration can include any method of administration such as oral, subcutaneous, sublingual, transmucosal , parenteral, intravenous, intra - arterial, intraperitoneal, buccal, sublingual, topical, vaginal, rectal, ocular, otic, nasal, inhalational and transdermal, or combinations thereof. Examples of parenteral administration include intravenous (IV) administration, intra - arterial administration, intramuscular administration, subcutaneous administration, intraosseous administration, intrathecal administration or combinations thereof, but are not limited thereto. The compound of formula (I) or (II) and the additional active agent can be administered independently, for example, orally or parenterally. It can be administered. In one embodiment, the compound of formula (I) or (II) is administered orally, and the additional active agent is administered parenterally. Parenteral administration can be performed by injection or infusion. In some embodiments, the methods and / or uses of the present disclosure are directed to the therapeutic and / or prophylactic treatment of NASH or ASH using at least two different active agents, a compound of formula (I) or (II), and an additional active agent, respectively. At least two active agents can be considered as a "combination product" where the agents are, for example, packaged separately and both agents are required to achieve the optimal desired effect.

[0038] The term "pharmaceutically effective amount" means an amount sufficient to achieve the desired pharmacological and / or therapeutic effect, i.e., the amount of the disclosed compound effective for its intended purpose. The needs of individual subjects / patients can vary, but the determination of the optimal range of the effective amount of the disclosed compounds is within the skill of those in the art. In general, a dosing regimen for treating a disease and / or condition with the compounds disclosed in the present invention can be determined according to various factors such as the type, age, weight,

[0039] sex, diet, etc. of the subject / patient and / or the medical condition. The term "pharmaceutical composition" means a compound according to the present disclosure in any

[0040] form suitable for medical use. The compounds of formula (I) and (II) can exist in various stereoisomers, including enantiomers, diastereomers, or mixtures thereof. It will be understood that the present invention encompasses all​ Therefore, compounds of formulas (I) and (II) that exist as diastereomers, racemates and / or enantiomers are within the scope of the present disclosure.

[0041] In one embodiment, the compound for use according to the invention is of formula (II)

[0042] [Chemical formula] (wherein R1 is selected from C 10 -C 22 alkenyl having 3 to 6 double bonds , R2 and R3 are the same or different and are selected from the group of substituents consisting of a hydrogen atom, a hydroxy group, an alkyl group, a halo gen atom, an alkoxy group, an acyloxy group, an acyl group, an alkenyl group, an alkynyl group, an a ryl group, an alkylthio group, an alkoxycarbonyl group, a carboxy group, an alkylsulfi nyl group, an alkylsulfonyl group, an amino group and an alkylamino group, where R2 and R3 may be linked to form a cycloalkane such as cyclopropane, cyclobutane, cyclopentane or cyclohexane and X represents a carboxylic acid or a derivative thereof, and the derivative is a carboxylate such as a carboxylic acid ester; a glyceride; an anhydride; a carboxamide; a phospholipid; or a hydroxymeth yl; or a prodrug thereof) or a pharmaceutically acceptable salt, solvate, or solvate of such a salt thereof

[0043] In one embodiment, R1 is C having 3 to 6 double bonds such as 5 or 6 double bonds 18 -C 22It is alkenyl, preferably with one double bond at the omega-3 position of C 18 -C 22 alkenyl.

[0044] R2 and R3 are more preferably independently selected from a hydrogen atom or a linear, branched and / or cyclic C1-C6 alkyl group. In one embodiment, at least one of R2 and R 3 is a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, and an isopropyl group, a butyl group or a pentyl group.

[0045] X preferably represents a carboxylic acid or a carboxylic acid ester; or a pharmaceutically acceptable salt, solvate, or solvate of such a salt thereof.

[0046] The compound of formula (II) for use can be administered as a monotherapy or in combination with one or more additional active agents.

[0047] In particular, the present disclosure provides a method for treating non-alcoholic steatohepatitis or alcoholic steatohepatitis in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a compound of formula (I):

[0048] [Chemical formula] (wherein R2, R3 and X are defined as in formula (II)) including administering the compound to the subject.

[0049] Preferably, for the compound of formula (I), R2 and R3 are independently selected from a hydrogen atom or a linear, branched and / or cyclic C1-C6 alkyl group, X is a carboxylic acid or a carboxylic acid ester; or a pharmaceutically acceptable salt thereof , a solvate, or a solvate of such a salt.

[0050] The compounds of formula (I) may be used as monotherapy or in combination with one or more additional active agents. They can be administered in combination.

[0051] In some embodiments, the present disclosure provides a method for treating nonalcoholic steatohepatitis or alcoholic steatohepatitis. 2. A compound of formula (I):

[0052] [ka] (wherein R2, R3 and X are defined as in formula (II)). The present invention provides a compound of the formula:

[0053] Preferably, for the compounds of formula (I), R2 and R3 are hydrogen atoms or straight-chain, branched independently selected from branched and / or cyclic C1-C6 alkyl groups; X is a carboxylic acid or carboxylic acid ester; or a pharmaceutically acceptable salt thereof , a solvate, or a solvate of such a salt.

[0054] When R2 and R3 are different, the compounds of formula (I) and formula (II) exist in stereoisomeric forms. The present invention relates to all optical isomers of the compounds of formula (I) and formula (II). It will be understood that this encompasses both hydroxybenzoates and hydroxybenzoates, as well as mixtures thereof.

[0055] In at least one embodiment, R2 and R3 are selected from the group consisting of a hydrogen atom, a methyl group, an ethyl group, and the like. independently selected from the group consisting of an n-propyl group, an isopropyl group, a butyl group, and a pentyl group; will be done.

[0056] In at least one embodiment, R2 and R3 are independently selected from the group consisting of a hydrogen atom, a methyl group, and an ethyl group.

[0057] In at least one embodiment, one of R2 and R3 is a hydrogen atom, and the other of R2 and R3 is selected from C1-C3 alkyl groups. In one embodiment one of R2 and R3 is a hydrogen atom, and the other of R2 and R3 is selected from the group consisting of a methyl group and an ethyl group, and most preferably, one of R2 and R 3 is a hydrogen atom, and the other is an ethyl group.

[0058] For both the compounds of Formula I and Formula II, R2 and R3 are, in some embodiments independently C1-C6 alkyl groups. In some embodiments, both R2 and R 3 are C1-C3 alkyl groups. In some embodiments, R2 and R3 are the same or different and each is independently selected from a methyl group, an ethyl group, an n-propyl group or an isopropyl group. In some embodiments, R2 and R3 are the same and are selected from a pair of methyl groups, a pair of ethyl groups, a pair of n-propyl groups or a pair of i sopropyl groups. In at least one preferred embodiment, R2 and R3 are ethyl groups. In some embodiments, one of R2 and R3 is a me thyl group and the other is an ethyl group. In some embodiments, one of R2 and R 3 is an ethyl group and the other is an n-propyl group.

[0059] In at least one embodiment, the compound is an enantiomer (R or S), a diastereo They exist as various stereoisomers, such as stereomers or mixtures thereof. At least In at least one embodiment, the compound exists as a racemate.

[0060] When the compound according to formula (I) is a salt of a counterion having at least one stereocenter or an ester of an alcohol having at least one stereocenter, the compound may have multiple stereocenters. In those situations, the compounds of the present disclosure may exist as diastereomers . Thus, in at least one embodiment, the compounds of the present disclosure exist as at least one diastereomer.

[0061] In at least one embodiment, the compound of the present disclosure is 2-(((5Z,8Z,11 Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy )butanoic acid (Compound A):

[0062]

Chemical formula

[0063] In at least one embodiment, the compound of the present disclosure has the formula:

[0064]

Chemical formula

[0065] In some embodiments, 2-(((5Z,8Z,11Z,14Z,17Z)-i (5Z,8Z,11Z,14Z,17Z)-Icosapentaen-1-yl)oxy)butanoic acid (Compound A ) is administered as a monotherapy. In some embodiments, 2-(((5Z,8Z ,11Z,14Z,17Z)-icosapentaen-1-yl )oxy)butanoic acid (Compound A) is administered in combination with one or more additional active agents .

[0066] As described above, multiple independent and interdependent metabolic, inflammatory, and ultimately fibrotic factors converge in the development of human NASH. The success of any treatment will likely need to address multiple aspects of NASH, preferably through upstream metabolic / inflammatory targets . However, since fibrosis development is associated with clinical outcomes, an ideal NASH therapy should target inflammatory factors and, more ideally, reduce or prophylactically treat fibrosis development and reverse existing fibrosis. Examples include the surprising and unexpectedly potent anti-inflammatory and anti-fibrotic effects of Compound A and any oxygen-containing structurally modified fatty acids . These findings demonstrated in multiple preclinical NASH models support the use of the oxygen-containing compounds of the present disclosure in the therapeutic and prophylactic treatment of NASH and ASH in human subjects . For example, as measured by a biochemical assay evaluating hydroxyproline content in the development of fibrosis (2 - 400% increase in ECM compared to a 20 - 30% increase in ECM in the diet-induced fibrosis mouse model (more moderate APOE 3Leiden.CETP mouse model) such as CDAA (defined choline-deficient l-amino acid), the development of fibrosis is reduced . . . .

[0067] . . * . . as measured by a biochemical assay evaluating hydroxyproline content in the development of fibrosis (2 - 400% increase in ECM compared to a 20 - 30% increase in ECM in the diet-induced fibrosis mouse model (more moderate APOE​​​​​​​​​​ Compound A has a significantly high activity of treating or prophylactically treating and improving liver fibrosis, which was surprising to find.

[0068] New findings in the CDAA-induced NASH model further show that compound A also reduces liver fibrosis measured histologically using Sirius Red morphometry (SR morphometry). The SR morphometry quantifies perisinusoidal collagen deposition, which is more functionally relevant, compared to the biochemical assessment of hydroxyproline (HYP) content that measures collagen in large blood vessels.

[0069] Considering the central importance of fibrosis in NASH-related morbidity and mortality, the results in the CDAA-induced NASH model support the idea that oxygen-containing structural modified fatty acids such as compound A are effective in treating NASH-related complications. This finding is also supported by changes in fibrosis-related liver gene expression (Col1a1) and inflammatory response (hepatic TNF-a gene expression). The significant decrease in perisinusoidal collagen with higher functional relevance measured

[0070] by SR morphometry also supports the testing and use of compound A in human NASH. Despite the decisive importance of fibrosis in clinical outcomes, regulatory approval of new effective drugs for the treatment of NASH is related to the recovery of NASH without worsening of fibrosis. Improvement of NAFLD score elements by new compounds is also important. Therefore, in addition to the desired improvement of fibrosis, steatosis,

[0071] New findings in the STAM mouse model, another NASH model, were that in addition to the improvement of steatosis and inflammation, Compound A improved ballooning of hepatocytes. Ballooning of hepatocytes is usually defined as cell expansion 1.5 - 2 times the normal hepatocyte diameter, accompanied by cytoplasmic dilution, correlates with fibrosis, and has been shown to be associated with liver injury.

[0072] (Cell expansion, defined as hepatocyte hypertrophy) was also prevented by Compound A in APOE * 3L.CETP double transgenic mice. The definition of hypertrophy relative to ballooning is related to histological differences specific to rodent hepatocytes compared to human hepatocytes.

[0073] These new findings related to hepatocyte ballooning / hypertrophy, in combination with the described anti-inflammatory effects, improve all NAS score elements and thus highlight the potential usefulness of Compound A in contributing to positive results in clinical development and subsequent regulatory approval.

[0074] As shown above and in the examples, the effectiveness of Compound A in modulating both inflammatory and fibrotic elements in multiple NASH rodent models of varying severity was surprisingly found.

[0075] Since the interplay of adaptive and innate immune cell recruitment, activation, differentiation, and proliferation in NASH is complex, it is speculated that simultaneous measurement of fibrosis is required to interpret the functional significance of such readouts due to the dependence on any one inflammatory parameter. Therefore, the anti-fibrotic effect of Compound A in the CDAA model may enhance the clinically relevant anti-inflammatory effect observed with oxygen-substituted structured lipids. ​

[0076] Regulatory approval of new and effective drugs for the treatment of NASH is dependent on the recovery of NA SH without worsening of fibrosis. Therefore, the improvement of ballooning / swelling of hepatocytes observed in the treatment with Compound A in two separate NASH rodent models is also an important and novel finding (ballooning / swelling not measured in the CDAA mouse model).

[0077] In combination with the described anti-inflammatory effects, these novel findings related to ballooning / swelling of hepatocytes improve all NAS score elements and thus highlight the potential usefulness of Compound A in contributing to positive results in clinical development and subsequent regulatory approval.

[0078] Importantly, the ability of compounds such as Compound A to improve all NAS score elements as well as fibrosis related to both the portal tract and the parenchyma in multiple preclinical NASH models with different disease etiologies strongly supports its testing in human NASH subjects.

[0079] In addition, since a significant proportion of NASH patients also have type 2 diabetes, it is important to investigate the late effects of treatment with agents such as Compound A on fatty liver, inflammation and fibrosis in the diet-induced model of NASH as well as its effect on blood glucose control. The anti-inflammatory, anti-fibrotic and steatosis-reducing effects of Compound A described above were further demonstrated in the diet-induced NASH model (ob / ob AMLN high-fat-fed mice). Unlike thiazolidinediones (e.g., pioglitazone) which have an adverse effect on body weight, Compound A has a positive effect on blood glucose control without affecting body weight in this model .[[]END]] ​​​possessed. Further, Compound A reduced the plasma levels of alanine aminotransferase (ALT) and aspartate transaminase (AST). This indicates a decrease in liver cell damage and / or injury.

[0080] Based on these findings, the compounds of formula (II) or preferably the compounds of formula (I) can be administered to treat and / or ameliorate non-alcoholic steatohepatitis (NASH), or other liver disorders characterized by fibrosis, inflammation and / or ballooning of hepatocytes. In some embodiments, the treatment of NASH can be prophylactic. Further, the compounds can be administered to treat at least one disease, condition or risk factor associated with NASH. In some embodiments, the treatment of at least one disease, condition or risk factor associated with NASH can be prophylactic.

[0081] Given the similarities in the inflammation-promoting and fibrosis-promoting mechanisms between NASH and alcoholic steatohepatitis (ASH), the anti-inflammatory and anti-fibrotic effects of the disclosed compounds herein in NASH models and in vitro experiments are relevant to the treatment and / or amelioration of ASH, particularly the prevention of progression and the induction of regression of advanced ASH and associated fibrosis. For example, the inhibitory effect of Compound A in vitro on the proliferation of isolated LX-2 (human stellate) cells was independent of paracrine signaling from parenchymal and / or Kupffer cells. This suggests that the anti-fibrotic effect of Compound A could be achieved regardless of whether the upstream stimuli were derived from NASH-related or ASH-related liver insults. ​​​​​​​​​​​​​​

[0082] Thus, the compound of formula (II) or preferably the compound of formula (I) can be administered to treat and / or reverse ASH. In some embodiments , the treatment of ASH can be prophylactic. Furthermore, the compound can be administered to treat at least one disease, condition or risk factor associated with ASH. In some embodiments , the treatment of at least one disease, condition or risk factor associated with ASH can be prophylactic.

[0083] Thus, the present disclosure encompasses methods for reducing the onset of liver fibrosis or prophylactically treating and reducing existing liver fibrosis. By this method, fibrosis is treated by any of a reduction in the area of fibrosis, a decrease in the amount of fibrosis or a reduction in the severity of fibrosis. In at least one embodiment , the method achieves a decrease in the surface area percentage of liver fibrosis, e.g., a significant decrease. In at least one embodiment, the method achieves a reduction in the composite NAS score, e.g., a significant reduction in the NAS score. Furthermore, in addition to improving the fibrotic state, the method includes a decrease in liver inflammation such as lobular inflammation; a decrease in ballooning hepatocyte enlargement; and a decrease in steatohepatitis.

[0084] In some embodiments, the compounds of the present disclosure reduce the liver fibrosis area determined by Sirius Red morphometry by 20%, 25%, 30%, 35%, 40%, 45%, 50% , 55%, 60%, 65% or 70%. In some embodiments, the compounds of the present disclosure reduce the liver fibrosis area by 20 - 30%, 30 - 40%, 10 - 40%, 40 - 50% , 40 - 60%, 50 - 60%, 50 - 70% or 60 - 70%. In some embodiments In embodiments, the compounds of the present disclosure reduce the liver fibrosis amount determined by liver hydroxyproline content by 20%, 25%, 30%, 35% or 40%. In some embodiments the compounds of the present disclosure reduce the liver fibrosis amount by 20-30%, 20-25%, 24-30% or 30-40%, 30-35% or 35-40%. In some embodiments the compounds of the present disclosure reduce the liver collagen content by 20%, 25%, 30%, 35% or 40%. In some embodiments, the compounds of the present disclosure reduce the liver collagen content by 20-30%, 20-25%, 25-30%, 30-40%, 30-35% or 3 5-40%. In some embodiments, the compounds of the present disclosure reduce the liver α-SMA content area by 3% compared to the pre-treatment level.

[0085] In some embodiments, the compounds of the present disclosure reduce steatosis by 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or 90%. In some embodiments, the compounds of the present disclosure reduce steatosis by 40-50%, 50-60%, 60-70%, 50-70%, 70-80%, 60-80%, 70-90% or 80- 90%. In some embodiments, the compounds of the present disclosure reduce the total liver lipid content by 2 0%, 25%, 30%, 35%, 40%, 45% or 50%. In some embodiments the compounds of the present disclosure reduce the total liver lipid content by 20-30%, 30-40% or 40-50%.

[0086] In some embodiments, the compounds of the present disclosure reduce ballooning of hepatocytes by 30%, 35 Reduce by %, 40%, 45% or 50%. In some embodiments, the chemical compound of the present disclosure reduces ballooning hepatocyte by 30 - 40%, 30 - 35%, 35 - 40%, 40 - 50 %, 40 - 45% or 45 - 50%. In some embodiments, the chemical compound of the present disclosure reduces hepatocyte hypertrophy by 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80%. In some embodiments, the chemical compound of the present disclosure reduces hepatocyte hypertrophy by 40 - 50%, 50 - 60%, 60 - 70% or 70 - 80%. In some embodiments, the chemical compound of the present

[0087] disclosure reduces the NAS score by 30%, 35%, 40 %, 45%, 50%, 55%, 60%, 65% or 70%. In some embodiments, the chemical compound of the present disclosure reduces the NAS score by 30 - 40%, 40 - 50%, 30 - 5 0%, 50 - 60%, 60 - 70% or 50 - 70%. In some embodiments, the chemical compound of the present disclosure reduces liver inflammation determined by galectin (Gal - 3) level by 20%, 30% or 40%. In some embodiments, The compound reduces plasma aspartate transaminase (AST) levels by 10%, 15%, or 20%. In some embodiments, the compounds of the present disclosure reduce AST levels by 10 - 15%, 10 - 20%, or 15 - 20%. In some embodiments, the compounds of the present disclosure and / or treatment with the compounds of the present disclosure, e.g., compound A, improve the NAS score from a liver biopsy, MRI Live rMultiScan - evaluated PDFF and cT1, one or more liver function tests, HOMA - IR, and / or one or more biomarkers of inflammation and fibrosis (including hsCRP, Pro - C3, ELF panel, and other suitable biomarkers) compared to a control.

[0088] In some embodiments, the compounds of the present disclosure and / or treatment with the compounds of the present disclosure, e.g., compound A, lead to an improvement, e.g., a decrease in ballooning (e.g., score = 0), e.g., an improvement without worsening of fibrosis with a lobular inflammation score of 0 or 1 compared to a control. In some embodiments, the compounds of the present disclosure and / or treatment with the compounds of the present disclosure, e.g., compound A, lead to a change, e.g., an improvement from baseline in the NAS score from a liver biopsy, in the individual histological scores of steatosis, ballooning, inflammation, and fibrosis, in liver enzymes, in imaging parameters, and / or in biomarkers (including hsCRP, Pro - C3, ELF panel, cytokines, and other suitable biomarkers) compared to a control.

[0089]

[0090] ​​​​​​​​​​​​​​​In some embodiments, the safety and tolerability of the compounds of the present disclosure, such as Compound A, in patients can be evaluated by monitoring adverse events, monitored clinical laboratory values (hematology, biochemistry, and urinalysis), vital signs (blood pressure, pulse rate, and body temperature), hsCRP, and / or resting 12-lead ECG in patients compared to controls. The pharmacokinetics of Compound A in patients can be investigated, for example, by comparing the mean trough plasma concentration of Compound A at steady state measured at regular intervals. For example, the mean trough plasma concentration can be measured at 8-week intervals to determine the pharmacokinetics of Compound A. Patients who meet one or more of the following criteria may show an improved response to treatment with Compound A compared to patients who do not meet the same one or more criteria: histological diagnosis of NASH, fibrosis score of 1 or more and 3 or less (F1 with an upper limit of 30%), PDFF>10% by MRI, compensated liver disease according to the following hematological and biochemical criteria for inclusion in the protocol: ALT<5×ULN, AST>30, hemoglobin>11 g / dL (for women) and >12 g / dL (for men), white blood cells (WBC)>2.5 K / μL, neutrophil count>1.5 K / μL, platelets>100 K / μL, total bilirubin<35 μmol / L (however, in the case of unconjugated bilirubin within the setting of Gilbert's syndrome, patients with bilirubin>35 μmol / L can be included), albumin>36 g / L, international normalized ratio (INR)<1.4, serum creatinine<1.3 mg / dL (for men) or<1.1 (for women) or estimated glomerular filtration rate ≧60 mL / min / 1.73 m2, chronic liver disease (e.g.,

[0091] For example, autoimmune diseases, primary biliary cholangitis, HBV, HCV, Wilson's disease, alpha-1-albuminuria, No other causes of leukemia (antitrypsin deficiency, hemochromatosis) and / or If applicable, stable type 2 diabetes (HgbA1c<9.5% and fasting glucose <10mcg) mol / L, no change in medication in the last 6 months, and / or no change in medication in the last 3 months (defined as no new symptoms related to decompensated diabetes).

[0092] Conversely, patients who meet one or more of the following criteria are considered to be at higher risk for developing the same one or more criteria: may not respond to treatment with Compound A compared with patients lacking history of excessive alcohol consumption, unstable metabolic state (e.g., in the past 3 months prior to screening) weight gain or loss of more than 5 kg in the past six months, and diabetes with poor glycemic control (Hgb A1c > 9.5%) or introduction of antidiabetic or antiobesity medication / malabsorption or (defined as limited bariatric (weight loss) surgery), gastrointestinal malabsorption bariatric surgery within the last 5 years History of surgery or corticosteroids, high-dose estrogen, or methotrexate in the last 6 months known to cause fatty liver, including serotonin, tetracycline, or amiodarone history of drug use, HB antigen > 0, HCV PCR > 0 (patients with a history of HCV infection were (can be included if HCV PCR is negative for more than 1 year) or HIV infection , insulin-treated type 1 or type 2 diabetes, diabetic ketoacidosis, pulmonary embolism Abdominal triglycerides >300 mg / dL, impaired hemostasis or current treatment with anticoagulants, Well-controlled hypertension, including history or current arrhythmia and / or myocardial infarction Cardiovascular disease history, excluding those in the patient, and any clinically significant ECG abnormalities, and or / and the intake of anti-diabetic drugs known to have activity against NASH, such as pi oglitazone, and GLP-1 receptor agonists.

[0093] The compounds of formula (I) and formula (II) can be prepared, for example, as described in PCT application WO2009 / 06120 8, WO2010 / 128401, WO2011 / 089529, WO2016 / 156 912, and according to the following examples. In addition, compound A can be prepared, for example, as described in PCT WO2010 / 128401 and WO2014 / 132135 and according to Example 2 below.

[0094] The examples described below are illustrative, and those skilled in the art will understand how to apply these general methods to reach other compounds within the scope of formula (I) and formula (II). The compounds of the present disclosure can be in the form of pharmaceutically acceptable salts or esters. For example, the compounds of formula (I) and formula (II) can be in the form of esters such as phospholipids, glycerides or C1-C6-alkyl esters. In at least one embodiment, the ester is selected from glycerides or C1-C6-alkyl esters. In at least one embodiment, the ester is selected from triglycerides, 1,2-diglycerides, 1,3-di glycerides, 1-monoglycerides, 2-monoglycerides, methyl esters, ethyl esters propyl esters, isopropyl esters, n-butyl esters and tert-but yl esters. In at least one embodiment, the compound of formula (I) is , methyl ester, ethyl ester, isopropyl ester, n-butyl ester or exist as tert-butyl ester, for example, as methyl ester or ethyl ester. Typically, the ester represented by formula (I) (e.g., ethyl ester) is hydrolyzed in the digestive tract.

[0095] Suitable salts for the present disclosure include NH 4+ ; Li + , Na + , K + , Mg 2+ or Ca 2+ any metal ion; protonated primary amines such as tert-butylammonium, (3S,5S,7S)-adamantan- 1-ammonium, 1,3-dihydroxy-2-(hydroxymethyl)propane-2-ammonium, protonated aminopyridine (e.g., pyridin-2-ammonium), etc.; protonated secondary amines such as diethylammonium, 2,3,4,5,6-pentahydroxy -N-methylhexane-1-ammonium, N-ethylnaphthalene-1-ammonium, etc.; protonated tertiary amines such as 4-methylmorpholin-4-ium; protonated quaternary amines such as 2-hydroxy-N,N,N-trimethylethane-1-aminium and protonated guanidines such as amino((4-amino-4-carboxybutyl)amino)methanim nium or protonated heterocyclic salts such as 1H-imidazol-3-ium, but are not limited thereto. Additional examples of suitable salts include salts of diprotonated diamines such as ethane -1,2-diammonium or piperazine-1,4-diium. Other salts according to the present disclosure include protonated chitosan:

[0096]

Chemical formula

[0097] In at least one embodiment, the salt is selected from sodium salts, calcium salts, and choline salts. In one embodiment, the salt is a sodium salt or a calcium salt. There is.

[0098] The present disclosure is a method of treating NASH or ASH in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a compound of formula (I) or formula (II). is provided. The subject can be a human or a non-human mammal. The compounds disclosed in the present invention can be administered as a medicine, such as a medicine in a pharmaceutical composition. In one aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), such as compound A, for use in the treatment of non-alcoholic steatohepatitis. a compound of formula (II). The compositions disclosed in the present invention comprise at least one compound of the disclosure and may also comprise at least one inactive pharmaceutical ingredient, i.e., an excipient. The inactive ingredients are such that the formulation active ingredients can be solubilized, suspended, thickened, diluted, emulsified, stabilized, preserved, protected, colored, flavored, and / or shaped into an applicable and effective formulation so that they can be safe, convenient, and / or otherwise acceptable for use. Examples of excipients include, but are not limited to, solvents, carriers, diluents, binders, fillers, sweeteners, fragrances, pH adjusters, viscosity adjusters, antioxidants, bulking agents, water retention agents, disintegrants, dissolution retardants, absorption promoters, wetting agents, absorbents, lubricants, colorants, dispersants, and preservatives. The excipient is one ​​It may have a role or function exceeding that, or may be classified into more than one group; the classification is for illustrative purposes only and not for limitation. In some embodiments it is for illustrative purposes and not for limitation. In some embodiments , for example, at least one excipient may be selected from corn starch, lactose, glucose, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, citric acid, tartaric acid, water, ethanol, glycerol, sorbitol, polyethylene glycol, propylene glycol , cetylstearyl alcohol, carboxymethylcellulose and fatty substances such as hard fat or a suitable mixture thereof. In some embodiments the compositions disclosed in the present invention are of formula (II) such as one of formula (I) at least one compound and at least one pharmaceutically acceptable antioxidant, for example alpha-tocopherol, beta-tocopherol, gamma-tocopherol and delta tocopherol or a mixture thereof, 2-tert-butyl -4-hydroxyanisole and 3-tert-butyl-4-hydroxyanisole such as BHA or a mixture thereof and BHT (3,5-di-tert-butyl-4 -hydroxytoluene) or a mixture thereof.

[0099] The compositions disclosed in the present invention can be formulated into oral dosage forms, for example, tablets or soft or hard gelatin capsules. The dosage form can be of any shape suitable for oral administration, such as spherical, oval, elliptical, cubic, regular and / or irregular shapes. Compounds according to the present disclosure can be formulated using conventional formulation techniques known in the art . It can be in the form of gelatin capsules or tablets. It can be.

[0100] A suitable daily dosage of the disclosed compounds, such as the compound of formula (I) or the compound of formula (II), can range from about 5 mg to about 2 g, such as from about 5 mg to about 4 g. For example, in some embodiments, the daily dose is from about 10 mg to about 1.5 g, from about 50 mg to about 1 g, from about 1 00 mg to about 1 g, from about 150 mg to about 900 mg, from about 50 mg to about 800 mg, from about 100 mg to about 800 mg, from about 100 mg to about 600 mg, from about 150 to about 550 m g or from about 200 to about 500 mg. In at least one embodiment, the daily dose ranges from about 200 mg to about 600 mg. In at least one embodiment, the daily dose is about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 m g, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 55 0 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg or about 900 mg. The (one or more) compounds can be administered, for example, once, twice, or three times a day.

[0101] In at least one embodiment, the compound of formula (I) is administered in an amount ranging from about 200 mg per dose to about 800 mg. In at least one embodiment, the compound of formula (I) is administered once a day. In at least one embodiment, the compound of formula (I) is administered once a day at a dose of 750 mg. In some embodiments, the compound of formula (I) is administered once a day at a dose of 600 mg. In some embodiments, In some embodiments, the compound of formula (I) is administered once daily at a dose of 500 mg. In some embodiments, the compound of formula (I) is administered once daily at a dose of 300 mg. In some embodiments, the compound of formula (I) is administered once daily at a dose of 250 mg. Preferably, the compound of formula (I) is administered once daily at a dose of 300 mg or 600 mg. In at least one embodiment, the compound of formula (II) is administered in an amount in the range of about 200 mg to about 800 mg per dose. In at least one embodiment, the compound of formula (II) is administered once daily. In at least one embodiment, the compound of formula (II) is administered once daily at a dose of 750 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 600 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 500 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 300 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 250 mg. Preferably, the compound of formula (II) is administered once daily at a dose of 300 mg or 600 mg.

[0102] In at least one embodiment, the compound of formula (II) is administered in an amount in the range of about 200 mg to about 800 mg per dose. In at least one embodiment, the compound of formula (III) is administered once daily. In at least one embodiment, the compound of formula (III) is administered once daily at a dose of 750 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 600 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 500 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 300 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 250 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 300 mg. In some embodiments, the compound of formula (II) is administered once daily at a dose of 250 mg. Preferably, the compound of formula (II) is administered once daily at a dose of 300 mg or 600 mg.

[0103] According to the present disclosure, the compound of formula (I) or (II) can be administered as a monotherapy or in combination with one or more additional active agents. With respect to co-administration, the additional active agent is preferably a therapeutically active agent such as a drug, preferably for NASH. In some embodiments, the additional active agent is preferably a therapeutically active agent such as a drug, preferably for NASH. Preferably, the additional active agent is a therapeutically active agent such as a drug, preferably for NASH. or one or more of the factors involved in the onset and / or exacerbation of ASH, such as having a therapeutic effect on NASH or ASH. Preferably, the combination product, i.e., the use of a compound of formula (I) or (II) in combination with one or more additional active agents, has a synergistic effect on the prevention and / or treatment of NASH or ASH. In one embodiment, the one or more additional therapeutic agents are allosteric acetyl-CoA carboxylase (ACC) inhibitors, angiotensin II receptor antagonists, angiotensin-converting enzyme (ACE) inhibitors, apoptosis signal regulatory kinase-1 (ASK1) inhibitors, caspase inhibitors, cathepsin B inhibitors, CCR 2 chemokine antagonists, CCR5 chemokine antagonists, chloride channel stimulators, cholesterol solubilizers, diacylglycerol O-acyltransferase 1 (DGAT1) inhibitors, dipeptidyl peptidase IV (DPP IV) inhibitors, fibroblast growth factor (FGF)-21 agonists, farnesoid X receptor (FXR) agonists, anti-CD3 mAb, galectin-3 inhibitors, glucagon-like peptide 1 (GLP1) agonists, glutathione precursors, hepatitis C virus NS3 protease inhibitors, HMG CoA reductase inhibitors, 11β-hydroxysteroid dehydrogenase (11β -HSDI) inhibitors, heat shock protein (Hsp) 47 inhibitors, IL-Iβ antagonists, IL-6 antagonists, IL-10 agonists, IL-17 antagonists, ileal sodium bile acid cotransporter inhibitors, leptin analogs, 5-lipoxygenase inhibitors LPL gene stimulators, lysyl oxidase homolog 2 (LOXL2) inhibitors, lysophospho lipids, Fatty acid 1 (LPA1) receptor antagonist, omega-3 fatty acid, PDE3 inhibitor , PDE4 inhibitor, phospholipase C (PLC) inhibitor, PPARa agonist, PPA Ry agonist, PPAR5 agonist, recombinant human pentraxin-2 protein (P RF-1), Rho-associated protein kinase 2 (ROCK2) inhibitor, semicarbazide-sensitive amine oxidase (SSAO) inhibitor, sodium glucose transporter-2 (SGL T2) inhibitor, stearoyl-CoA desaturase-1 inhibitor, thyroid hormone receptor beta agonist, tumor necrosis factor alpha (TNFα) ligand inhibitor, transglutaminase inhibitor agent, transglutaminase inhibitor precursor and small interfering RNA (saRNA) selected independently from the group is selected.

[0104] In particular, in some embodiments, the one or more additional active agents are glucagon like peptide 1 (GLP-1) agonist; dipeptidyl peptidase inhibitor (DPP- 4 antagonist) and omega-3 (n-3) fatty acids selected from the group.

[0105] GLP-1 receptor agonist or glucagon-like peptide-1 receptor agonist, also known as incretin mimetics, is an agonist of the GLP-1 receptor. This class of drugs is typically used for the treatment of type 2 diabetes. A non-limiting list of examples of GLP-agonists includes exenatide, liraglutide, lixisenatide, albiglutide , dulaglutide, taspoglutide and semaglutide.

[0106] In some embodiments, the additional active agent is a dipeptidyl peptidase inhibitor (D PP-4 antagonist). DPP-4 antagonists are a class of oral hypoglycemic agents that block DPP-4 (DPP- IV). They can be used to treat type 2 diabetes mellitus. Glucagon raises blood glucose levels, and DPP-4 inhibitors lower glucagon and blood glucose levels. The mechanism of action of DPP-4 inhibitors is to increase incretin levels ( GLP-1 and GIP), which inhibits glucagon release, and thus increases insulin secretion, decreases gastric emptying, and lowers blood glucose levels. A non-limiting list of dipeptidyl peptidase inhibitors includes sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, dutogliptin. In some embodiments, the additional agent is an omega-3 fatty acid. When the additional active agent is an omega-3 fatty acid, the omega-3 fatty acid is typically a long-chain polyunsaturated omega-3 fatty acid (LC n-3 PUFA). Preferably, this includes at least one of (all-Z omega-3 )-5,8,11,14,17-eicosapentaenoic acid (EPA) and (all-Z omega- 3)-4,7,10,13,16,19-docosahexaenoic acid (DHA) or derivatives thereof. N-3 PUFAs such as EPA and DHA can be in different forms, free fatty acid form; esterified forms such as C1-C4 alkyl esters, preferably ethyl esters; phospholipids; mono / di / triglycerides; and salts thereof, provided by at least one of them. The omega-3 fatty acid is provided in a composition such as an orally administered composition. A non-limiting list of dipeptidyl peptidase inhibitors includes sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, dutogliptin. A non-limiting list of dipeptidyl peptidase inhibitors includes sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, dutogliptin. A non-limiting list of dipeptidyl peptidase inhibitors includes sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, dutogliptin. A non-limiting list of dipeptidyl peptidase inhibitors includes sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, dutogliptin.

[0107] In some embodiments, the additional agent is an omega-3 fatty acid. When the additional active agent is an omega-3 fatty acid, the omega-3 fatty acid is typically a long-chain polyunsaturated omega-3 fatty acid (LC n-3 PUFA). When the additional active agent is an omega-3 fatty acid, the omega-3 fatty acid is typically a long-chain polyunsaturated omega-3 fatty acid (LC n-3 PUFA). -3 fatty acid (LC n-3 PUFA). Preferably, this is (all-Z omega-3 )-5,8,11,14,17-eicosapentaenoic acid (EPA) and (all-Z omega- 3)-4,7,10,13,16,19-docosahexaenoic acid (DHA) or at least one of their derivatives. N-3 PUFAs such as EPA and DHA can be in different forms, free fatty acid form; esterified forms such as C1-C4 alkyl esters, preferably ethyl esters; phospholipids; mono / di / triglycerides; and salts thereof, provided by at least one of them. N-3 PUFAs such as EPA and DHA can be in different forms, free fatty acid form; esterified forms such as C1-C4 alkyl esters, preferably ethyl esters; phospholipids; mono / di / triglycerides; and salts thereof, provided by at least one of them. Preferably, it is an ethyl ester; phospholipids; mono / di / triglycerides; and salts thereof, provided by at least one of them. The omega-3 fatty acid is provided in a composition such as an orally administered composition. The omega-3 fatty acid is provided in a composition such as an orally administered composition. can be provided in the form of. Such a composition may contain at least 50%, 60%, 70% or 80% etc., at least 40% of active omega-3 fatty acids. In some embodiments the additional active agent is a composition that contains at least one of EPA and DHA, preferably is in the ethyl ester form and contains at least 70% concentration.

[0108] In some embodiments, the one or more additional active agents are acetylsalicy lic acid, alipogene tiparvovec, alamchol, atorvastatin, BI 1467 335, BLX-1002, BMS-986036, BMS-986020, cenicriviroc , cobiprostone, colesevelam, emricasan, enalapril, foramulab ( foramulab), GFT-505, GR-MD-02, GS-0976, GS-9 674, hydrochlorothiazide, icosapent ethyl ester (eicosapentaenoic acid eth yl ester, EPA ethyl ester), IMM-124E, IVA337, K-877, KD- 025, linagliptin, liraglutide, mercaptamine, MGL-3196, ND-L 02-s0201, obeticholic acid, olesoxime, peg-ilotekin, pioglitazone , PRM-151, PX-102, remogliflozin etabonate, seronesertib , simtuzumab, SHP-626, solithromycin, tipepidast, TRX-318, ursodeoxycholic acid and are independently selected from the group of VBY-376. The first element of the combination product , i.e., the compound of formula (I) or (II), can be administered or formulated by any of the methods described above. The second element of the combination product, the additional active agent, is of the type of the drug and the second element of the combination product, the additional active agent, is of the type of the drug It can be formulated to be suitable and depends on several factors including the method of drug administration. Additional The dosage of the active agent depends on the type of agent selected and should follow the amount approved for the particular agent. As described in the examples, the use of a CDAA (defined choline-deficient l-amino acid) diet provides a model of both liver fibrosis and inflammation. As supported by the examples, the combination of a GLP-1 agonist and a compound of formula (I) or (II) such as compound A has excellent effects for NASH-related fibrosis and inflammation compared to treatment with GLP-1 alone. Since liver fibrosis is mostly a response to the inflammatory response, this also shows the superiority of the oxygen-containing structurally modified fatty acid (compound A) in combination with a GLP-1 agonist over GLP-1 alone, as demonstrated by the shown changes in inflammation-related liver gene expression (TNF-α). Overall, the data suggest that a combination of a GLP-1 agonist (or alternatively a DPP-4 antagonist) and the compounds of the present disclosure provides a synergistic effect for the treatment of both liver inflammation and fibrosis.

[0109] Although fatty liver may be a benign condition in itself, it can make the liver sensitive to "second attacks" from other factors that induce an inflammatory response. Thus, weight loss of steatosis is desirable with respect to both its contribution as a potential "priming" factor for other or continuous liver insults and the recovery of NASH. The novel and significant improvement in liver lipids achieved by the combination of high-dose omega-3 ethyl ester and low-dose oxygen-containing structurally modified fatty acid (compound A) strongly suggests that a synergistic effect is obtained, especially since neither compound affects ApoE significantly. of either compound *In 3L-CETP transgenic mice, the significant reduction in hepatic triglyceride (TG) was not achieved alone. This is because

[0110] Remarkably, compound A significantly upregulates or downregulates more than 1094 gene expression probes relative to less than 10 full-length omega-3 ethyl esters in the livers of APOE * 3.CETP transgenic mice (data not shown). Without being limited by theory, these pleiotropic effects on the hepatic transcriptome may suggest that the observed synergistic effect is secondary to the targeting of various pathways rather than a cumulative dose effect. The significant effect on hepatic cholesterol content may also have a positive effect on the inflammatory response associated with the pathogenesis of NASH and ASH. Overall, the data in APOE 3.CETP mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. Without being limited by theory, these pleiotropic effects on the hepatic transcriptome may suggest that the observed synergistic effect is secondary to the targeting of various pathways rather than a cumulative dose effect. The significant effect on hepatic cholesterol content may also have a positive effect on the inflammatory response associated with the pathogenesis of NASH and ASH. Overall, the data in APOE 3.CETP mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. Without being limited by theory, these pleiotropic effects on the hepatic transcriptome may suggest that the observed synergistic effect is secondary to the targeting of various pathways rather than a cumulative dose effect. The significant effect on hepatic cholesterol content may also have a positive effect on the inflammatory response associated with the pathogenesis of NASH and ASH. Overall, the data in APOE 3.CETP mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. Overall, the data in APOE * 3.CETP mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. Overall, the data in APOE 3.CETP mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. Overall, the data in APOE

[0111] The compound of formula (II) or preferably the compound of formula (I) can be administered as a monotherapy or in combination with at least one additional active agent for treating and / or ameliorating non-alcoholic steatohepatitis (NASH) or alcoholic steatohepatitis (ASH). In some embodiments, the at least one additional active agent is a GLP -1 agonist. Additionally, the data in APOE 3.CETP transgenic mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. -1 agonist. Additionally, the data in APOE * 3.CETP transgenic mice support the combination of omega-3 PUFA formulations such as EPA ethyl ester with oxygen-containing structurally modified fatty acids of formula (I) or (II) as a potent therapy for reducing fatty liver and thus for the treatment of NASH and / or ASH. Based on the findings herein, the compound of formula (II) or preferably the compound of formula (I) is used for treating and / or preventing fatty liver (triglyceride and / or cholesterol content) and can be administered in combination with at least one additional active agent to treat and / or ameliorate it. In some embodiments, the at least one additional active agent is an omega-3 fatty acid such as omega-3 PUFA ethyl ester.

[0112] The above examples highlight the potential of oxygen-containing structure-modified fatty acids in combination with several other active agents for the treatment of NASH and / or ASH. These combinations can not only improve the results related to efficacy against monotherapy, but also improve safety. As an example of the latter, compound A has been shown to significantly decrease CETP expression in APOE 3.CETP mice and improve the lipid profile in both rodents and humans. This may be advantageous compared to a combination with obeticholic acid (FXR agonist), which increases CETP expression and worsens the lipid profile in both APOE 3.CETP mice and humans. * 3.CETP mice and improve the lipid profile in both rodents and humans. * 3.CETP mice and humans.

[0113] The inventors have found that compounds of formula (I) such as 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8, 11,14,17-pentaen-1-yl)oxy)butanoic acid have significantly good pharmaceutical activities. Surprisingly, the compounds of formula (I) disclosed in the present invention exhibit improved biological activities for treating NASH-related liver fibrosis and inflammation compared to glucagon-like peptide 1 receptor (GLP-1R) agonists. When administered in combination with additional active agents, the compounds of formula (I) may perhaps exhibit a synergistic effect.

Examples

[0114] The present disclosure is further illustrated by the following non-limiting examples, in which, where appropriate, standard techniques known to those skilled in the art and techniques similar to those described in these examples may be used. It is understood that those skilled in the art will envision additional embodiments consistent with the present disclosure described in this specification. It is understood.

[0115] Unless otherwise stated, the reactions were carried out at room temperature, typically in the range between 18 and 25 °C under anhydrous conditions using HPLC grade solvents. Evaporation was carried out by rotary evaporation in vacuo Column chromatography was carried out by the flash method using silica gel 40 - 63 μm (Merck), or by Armen Spotfla sh using prepacked silica gel columns "MiniVarioFlash" ", "SuperVarioFlash", "SuperVarioPrep" or "EasyVarioPrep" (Merck). Nuclear magnetic resonance (NMR) shift values were recorded on a Bruker Avance DPX 200 or 300 instrument using the peak multiplicities described below: s, singlet; d, doublet; dd, doublet of doublets; t, triplet; q, quartet; p, pentet; m, multiplet ; br, broad. Mass spectra were recorded using an LC / MS spectrometer. Separation was carried out by gradient elution on an Eclipse XDB-C 18 2.1×150 mm column using an Agilent 1100 series module. As the eluent, 0.01% T ; br, broad. Mass spectra were recorded using an LC / MS spectrometer. Separation was carried out by gradient elution on an Eclipse XDB-C 18 2.1×150 mm column using an Agilent 1100 series module. As the eluent, 0.01% T ; br, broad. Mass spectra were recorded using an LC / MS spectrometer. Separation was carried out by gradient elution on an Eclipse XDB-C 5 - 95% acetonitrile in a buffer containing trifluoroacetic acid or 0.005% sodium formate A gradient of toluene was used. Mass spectra were recorded using a G1956A mass spectrometer (electrospray, 3000 V) by switching between positive and negative ionization modes. The reported yields are illustrative and do not necessarily represent the maximum achievable yields.

[0116] Preparation of Compounds

Example

[0117] Preparation of tert-butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8, 11,14,17-pentaen-1-yloxy)butanoate

[0118]

Chemical Structure

Example

[0119] 2 - ((5Z,8Z,11Z,14Z,17Z)-Icosa - 5,8,11,14,17 - pentaenyloxy)butanoic acid (Compound A) Preparation

[0120]

Chemical formula

Example

[0121] Calcium 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11 ,14,17-pentaen-1-yl)oxy)butanoate Preparation

[0122]

Chemical formula

Example

[0123] Sodium 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11 ,14,17-pentaen-1-yl)oxy)butanoate Preparation

[0124]

Chem.

Example

[0125] ​ 2-Hydroxy-N,N,N-trimethylethan-1-aminium 2-(((5Z,8 Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaene-1- Preparation of yl)oxy)butanoate

[0126] [ka] A scintillation flask containing approximately 2.5 mL of MTBE and 7.5 mL of n-heptane was used. Choline hydroxide in water (327.7 μL) was pipetted into the nitrogen chamber. Within, 2-(((5Z,8Z,11Z,14Z,17Z)-equalizer-5,8,11,1 4,17-pentaen-1-yl)oxy)butanoic acid (500 mg, 95.8%) Approximately 1.0 mL of water was slowly added to the vial under stirring in a nitrogen chamber. The vial was then sealed. The reaction mixture was stirred for about 30 minutes. The resulting 2- (((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pe Choline salt of (butanoic acid) benzoyl-1-en-1-yloxy is a hard gel-like material, which is The wet material on the filter was washed three times with 1 mL of MTBE. The washed material appeared as a hard gel-like solid. [Example]

[0127] (4S,5R)-3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-I Cosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5 -phenyloxazolidin-2-one and (4S,5R)-3-((R)-2-((5 (5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaenyl Preparation of ((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5-phenyloxazolidin-2-one

[0128] [Chemical formula] DMAP (1.10 g, 8.90 mmol) and DCC (1.90 g, 9.30 mmol) were added to a mixture of 2-((5Z,8 Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy )butanoic acid (3.20 g, 8.50 mmol) in dry dichloromethane (100 mL) maintained at 0 °C under nitrogen. The resulting mixture was stirred at 0 °C for 20 minutes. (4S,5R)-4-Methyl-5-phenyloxazolidin-2- one (1.50 g, 8.50 mmol) was added and the resulting turbid mixture was stirred at ambient temperature for 5 days . The mixture was filtered and concentrated under reduced pressure to give a crude product containing the desired product as a mixture of two di astereomers. The residue was purified by flash chromatography on silica gel using 15% ethyl acetate in heptane as the eluent . The two diastereomers were separated and the appropriate fractions were concentrated. (4S,5R)-3-((S)-2 -((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5-phenyloxazolidin-2-one eluted first and was obtained as an oil, 1.1 g (40% yield). (4S,5R)-3- ((R)-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,1 4,17-pentaenyloxy)butanoyl)-4-methyl-5-phenyloxazolidin e-2-one eluted later and was obtained as an oil, 1.1 g (40% yield). 0.95 g (34% yield) of thion-2-one was obtained as an oil.

[0129] (4S,5R)-3-((S)-2-((5Z,8Z,11Z,14Z,17Z)-I Cosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5 -phenyloxazolidin-2-one (E1): 1 H-NMR (300 MHz, CDCl3): δ 0.90 (d, 3H), 1.00 (t, 3H), 1.07 (t, 3H), 1.45-1.57 (m, 2H), 1.62-1.76 (m, 3H), 1.85-1.9 5 (m, 1H), 2.05-2.15 (m, 4H), 2.87 (m, 8H), 3.39 (m, 1H), 3.57 (m, 1H), 4.85-4.9 2 (m, 2H), 5.30-5.45 (m, 10H), 5.75 (d, 1H), 7.32 (m, 2H), 7.43 (m, 3H).

[0130] (4S,5R)-3-((R)-2-((5Z,8Z,11Z,14Z,17Z)-I Cosa-5,8,11,14,17-pentaenyloxy)butanoyl)-4-methyl-5 -phenyloxazolidin-2-one (E2): 1 H-NMR (300 MHz, CDCl3): δ 0.98 (d, 3H), 0.99 (t, 3H), 1.08 (t, 3H), 1.40-1.52 (m, 2H), 1.55-1.75 (m, 3H), 1.80-1.9 0 (m, 1H), 2.05-2.15 (m, 4H), 2.84 (m, 8H), 3.39 (m, 1H), 3.56 (m, 1H), 4.79 (five double line, 1H), 4.97 (dd, 1H), 5.30-5.45 (m, 10H), 5.71 (d, 1H), 7.33 (m, 2H), 7.43 ( m, 3H).

Example

[0131] (S)-2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,1 Preparation of 4,17-pentaenyloxy)butanoic acid

[0132]

Chem.

Example

[0133] (R)-2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,1 4,17-pentaenyloxy)butanoic acid preparation

[0134]

Chemical formula

Example

[0135] Ethyl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,1 4,17-pentaen-1-yl)oxy)butanoate Preparation

[0136]

Chemical formula

Example

[0137] Preparation of isopropyl 2 - (((5Z,8Z,11Z,14Z,17Z)-icosa - 5,8,1 1,14,17 - pentaen - 1 - yl)oxy)butanoate

[0138]

Chemical formula

Example

[0139] Methyl 2 - ((((5Z,8Z,11Z,14Z,17Z)-icosa - 5,8,11,1 4,17 - pentaen - 1 - yl)oxy)butanoate Preparation

[0140]

Chemical formula

Example

[0141] Butyl 2 - (((5Z,8Z,11Z,14Z,17Z)-icosa - 5,8,11,1 4,17 - pentaen - 1 - yl)oxy)butanoate Preparation

[0142]

Chemical formula

Example

[0143] 2,3-Dihydroxypropyl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate Preparation Step a) Preparation of (2,2-dimethyl-1,3-dioxolan-4-yl)methyl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate (2,2-Dimethyl-1,3-dioxolan-4-yl)methyl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate (5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-penta en-1-yl)oxy)butanoate Preparation

[0144]

Chemical Structure

[0145] Step b) Preparation of 2,3 - Dihydroxypropyl 2 - ((((5Z,8Z,11Z,14Z, 17Z) - Icosa - 5,8,11,14,17 - pentaen - 1 - yl)oxy)butanoate

[0146]

Chemical Structure

Example

[0147] 1,3-Dihydroxypropan-2-yl 2-(((5Z,8Z,11Z,14Z,1 7Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate Preparation Step a) Preparation of oxiran-2-ylmethyl 2-(((5Z,8Z,11Z,14Z,1 7Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoate

[0148]

Chemical formula

[0149] Step b) Preparation of 2 - ((2 - (((5Z,8Z,11Z,14Z,17Z)-icosa - 5 ,8,11,14,17 - pentaen - 1 - yl)oxy)butanoyl)oxy)prop ane - 1,3 - diyl bis(2,2,2 - trifluoroacetate)

[0150] Trifluoroacetic anhydride (TFAA) (0.55 mL, 3. 96 mmol) in dry DCM (3 mL) was added portionwise to a pre - cooled solution of oxiran - 2 - yl methyl 2 - (((5Z,8Z,11Z,14Z,17Z)-icosa - 5,8,11, 14,17 - pentaen - 1 - yl)oxy)butanoate (286 mg, 0.66 mmol ) in dry DCM (3 mL) at - 20 °C under N2 atmosphere. The cooling bath was removed and the mixture was stirred at ambient temperature for 19 hours and then the reaction mixture was ​​, passed through a pad of silica (6.5 g) while eluting with toluene (150 mL). Concentrated in vacuo to give 357 mg (84% yield) of the title compound as an oil. 1 H NMR (400 MHz, CDCl3) δ 0.95 (2xt, 6H), 1.38 - 1.45 (m, 2H), 1.57 - 1.63 (m, 2H), 1.66 - 1.78 ( m, 2H), 2.09 - 2.02 (m, 4H), 2.78 - 2.84 (m, 8H), 3.27 - 3.33 (m, 1H), 3.51 - 3.56 (m, 1 H), 3.77 (dd, 1H), 4.30 - 4.53 (m, 2H), 4.60 - 4.69 (m, 2H), 5.17 - 5.43 (m, 10H), 5.4 3 - 5.55 (m, 1H). MS (electrospray): 661.1 [M+Na] + .

[0151] Step c) Preparation of 1,3 - dihydroxypropan - 2 - yl 2 - (((5Z,8Z,11Z ,14Z,17Z) - icosa - 5,8,11,14,17 - pentaen - 1 - yl)oxy )butanoate

[0152]

Chemical Structure

Example

[0153] 3 - Hydroxypropane - 1,2 - diylbis(2 - (((5Z,8Z,11Z,14 Z,17Z) - icosa - 5,8,11,14,17 - pentaen - 1 - yl)oxy)but anoate) Preparation Step a) Preparation of tert - butyl((2,2 - dimethyl - 1,3 - dioxolan - 4 - yl methyl)methoxy)dimethylsilane

[0154]

Chem.

[0155] Step b) Preparation of 3-((tert-butyldimethylsilyl)oxy)propane-1,2-diol

[0156]

Chemical formula

[0157] Step c) Preparation of 3-((tert-butyldimethylsilyl)oxy)propane-1,2- diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11 ,14,17-pentaen-1-yl)oxy)butanoate)

[0158]

Chemical Structure

[0159] Step d) Preparation of 3-Hydroxypropane-1,2-diylbis(2-(((5Z,8Z, 11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl )oxy)butanoate)

[0160] [ka] 3-((tert-butyldimethylsilyl)oxy)propane in dioxane (100 mL) Propane-1,2-diylbis(2-(((5Z,8Z,11Z,14Z,17Z)-isopropyl 5,8,11,14,17-pentaen-1-yl)oxy)butanoate) (2. To a solution of 26 g (2.46 mmol) of HCl (37% (w / w, 2 mL) was added aq. The mixture was stirred at ambient temperature under nitrogen for 3 hours and then concentrated in vacuo. Flash chromatography on silica gel using 5% ethyl acetate as the eluent The residue was purified by HPLC. The appropriate fractions were concentrated to give 0.83 g (42% yield) of the title compound. The mixture was obtained as an oil. 1 H NMR (300 MHz, CDCl3) δ 0.96-1.03 (m, 12H), 1.40-1.53 (m, 4H), 1.58-1.68 (m, 4H), 1.70-1.85 (m, 4H), 1.87-2.01 (m, 1H), 2.05-2.15 (m, 8H), 2.75-2.95 (m, 16H), 3.28-3.41 (m, 2H), 3.56-3.65 (m, 2H), 3.73-3.85 (m, 4H) ), 4.24-4.37 (m, 1H), 4.42-4.53 (m, 1H), 5.14-5.23 (m, 1H), 5.26-5.51 (m, 20H). MS (electrospray): 827.5 [M+Na] + . [Example]

[0161] 2-Hydroxypropane-1,3-diylbis(2-(((5Z,8Z,11Z,14 ((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)but anoate) Preparation Step a) 2-Oxopropane-1,3-diylbis(2-(((5Z,8Z,11 Z,14Z,17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)o xy)butanoate)

[0162]

Chemical Structure

[0163] Step b) 2-Hydroxypropane-1,3-diylbis(2-(((5Z,8Z, 11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl )oxy)butanoate)

[0164]

Chemical formula

Example

[0165] Preparation of propane - 1,2,3 - triyltris(2 - (((5Z,8Z,11Z,14Z,1 7Z) - icosa - 5,8,11,14,17 - pentaen - 1 - yloxy)butanoate)

[0166]

Chemical formula

Example

[0167] tert-Butyl 2-((2-((5Z,8Z,11Z,14Z,17Z)-icosa- 5,8,11,14,17-pentaen-1-yloxy)butanoyl)oxy)benzoate Preparation

[0168]

Chemical formula

Example

[0169] 2 - ((2 - ((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,1 4,17 - pentaen - 1 - yloxy)butanoyl)oxy)benzoic acid Preparation

[0170]

Chem.

Example

[0171] Preparation of 2-((tert-butoxycarbonyl)amino)ethyl 2-((5Z,8Z,11Z ,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy )butanoate

[0172] ​ [Chemical formula] O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyl Uronium hexafluorophosphate (HATU) (800 mg, 2.1 mmol) and TEA (0.56 mL, 4 mmol) were added to a solution of 2-((5Z,8 Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1- yloxy)butanoic acid (748 mg, 2 mmol) in DCM (10 mL), and the reaction mixture was stirred for 20 minutes. A solution of tert-butyl N-(2-hydroxyethyl)carbamate (340 mg, 2.1 mmol) in DCM (1 mL) was added, and the resulting mixture was stirred overnight at room temperature. Et2O (100 mL) was added, and the mixture was washed with water, 1M HCl(aq), saturated NaHCO3(aq) and brine, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash chromatography using a gradient of 10 - 20% EtOAc in heptane as the eluent. The appropriate fractions were concentrated to give 780 mg (76% yield) of the title compound. H NMR (300 MHz, CDCl3): δ 0.98 (t, 6H), 1.40 - 1.55 (m, 2H), 1.45 (s, 9H), 1.60 - 1.85 (m, 4H), 2.05 - 2.20 (m, 4H), 2.75 - 2.90 (m, 8H), 3.30 - 3.45 (m, 3H), 3.55 - 3.65 (m, 1H), 3.75 - 3.80 (m, 1H), 4.20 - 4.25 (m, 2H), 4.76 (br 1 s, 1H), 5.30 - 5.45 (m, 10H). MS (ESI): 540 [M+Na] . . . + . [Example]

[0173] 2-(2-acetoxybenzamido)ethyl 2-((5Z,8Z,11Z,14Z,1 7Z)-Icosa-5,8,11,14,17-pentaen-1-yloxy)butanoate Preparation of

[0174] [ka] Acetyl chloride (1 mL) was dissolved in 2-(tert-butoxy)-2-methylpropional in MeOH (5 mL) at 0 °C. (Dicarbonyl)amino)ethyl 2-((5Z,8Z,11Z,14Z,17Z)-icosapentaenoic acid -5,8,11,14,17-pentaen-1-yloxy)butanoate (300mg The mixture was added to a solution of 1,000 mg of HCl (0.58 mmol), and the reaction mixture was stirred at room temperature for 1 hour. Concentrated in 2-aminoethyl 2-((5Z,8Z,11Z,14Z,17Z)-isothiazolinone -5,8,11,14,17-pentaen-1-yloxy) butanoate HCl salt ( TEA (109 μL, 0.78 mmol) was added to DC Add to a solution of acetylsalicylic acid (137 mg, 0.76 mmol) in M (10 mL) The mixture was cooled to 0° C. Ethyl chloroformate (75 μL, 0.78 mmol) was added dropwise. The reaction mixture was stirred for 2 h. The solution was then diluted with DCM (10 mL) and TEVA 2-aminoethyl 2-((5Z,8Z,11Z,14Z,17Z) Z)-Icosa-5,8,11,14,17-pentaen-1-yloxy)butanoate The resulting mixture was stirred at room temperature for 3.5 hours, and then , water was added. The two resulting phases were separated, and the aqueous phase was extracted twice with DCM. The combined organic phase was washed with 1 M HCl(aq), saturated NaHCO3(aq), and water, dried (Na2S O4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography using 20% EtOAc in heptane as the eluent. The appropriate fractions were concentrated to give 90 mg (23% yield) of the title compound. 1 H NMR (300 MHz, CDCl3): δ 0.95 - 1 .05 (m, 6H), 1.35 - 1.45 (m, 2H), 1.55 - 1.70 (m, 2H), 1.70 - 1.85 (m, 2H), 2.05 - 2.15 (m, 4H), 2.35 (s, 3H), 2.80 - 2.90 (m, 8H), 3.30 - 3.40 (m, 1H), 3.55 - 3.65 (m, 1H), 3.70 - 3.85 (m, 3H), 4.33 (t, 2H), 5.30 - 5.45 (m, 10H), 6.61 (br s, 1H), 7.10 - 7.15 (m, 1H), 7.25 - 7.35 (m, 1H), 7.45 - 7.50 (m, 1H), 7.70 - 7.75 (m, 1H). MS (ESI); 602 [M + Na] + .

Example

[0175] 2-(2-Hydroxybenzamide)ethyl 2-((5Z,8Z,11Z,14Z,1 7Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanoate Preparation

[0176]

Chemical formula

Example

[0177] N-Benzyl-2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8, 11,14,17-pentaen-1-yl)oxy)butanamide Preparation

[0178]

Chem.

Example

[0179] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 7-pentaen-1-yl)oxy)-N,N-dimethylbutanamide Preparation

[0180]

Chemical

Example

[0181] N-Ethyl-2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,1 1,14,17-pentaen-1-yl)oxy)butanamide Preparation

[0182]

Chem.

Example

[0183] tert-Butyl (2-(2-(((5Z,8Z,11Z,14Z,17Z)-icosa -5,8,11,14,17-pentaen-1-yl)oxy)butanamide)ethyl) carbamate preparation

[0184]

Chemical formula

Example

[0185] N-(2-Aminoethyl)-2-(((5Z,8Z,11Z,14Z,17Z)-icosa Preparation of tert-butyl (2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanamide)

[0186] [Chemical formula] To a solution of tert-butyl (2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanamide)ethyl)carbamate (774 mg, 1.5 mmol) in DCM (8 mL) at ambient temperature under a N2 atmosphere was added TFA (2 mL), and the reaction mixture was stirred at room temperature for 30 minutes and then concentrated in vacuo. Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen -1-yl)oxy)butanamide)ethyl)carbamate (774 mg, 1.5 mmol) l) was added TFA (2 mL), and the reaction mixture was stirred at room temperature for 30 minutes and then concentrated in vacuo. Diethyl ether (50 mL) and NaOH (aq, 1 M, 50 mL) were added to the residue, and the mixture was stirred vigorously for 30 minutes. The phases were separated, the organic phase was washed with brine, dried (Na2SO4), filtered, and concentrated in vacuo to give 560 mg (90%) of the title compound. The phases were separated, the organic phase was washed with brine, dried (Na2SO4), filtered, and concentrated in vacuo to give 560 mg (90%) of the title compound. Obtained. 1 1H NMR (300 MHz, CDCl3): δ 0.85 - 1.05 (2xt, 6H), 1.35 - 1.50 (m, 2H), 1.5 5 - 1.85 (m, 4H), 2.05 - 2.20 (m, 4H), 2.75 - 2.95 (m, 8H), 3.10 - (m, 1H), 3.30 - 3.80 (m, 6H), 4.05 - 4.20 (br m, 1H), 4.25 - 4.75 (br s, 2H), 5.30 - 5.5 0 (m, 10H). MS (electrospray); 417 [M+H] + , 439 [M+Na] + [Example]

[0187] N-(2-Hydroxyethyl)-2-(((5Z,8Z,11Z,14Z,17Z)- Preparation of ((5Z,8Z,11Z,14Z,17Z)-icos-5,8,11,14,17-pentaen-1-yl)oxy)butanamide Preparation

[0188] [Chemical formula] To a solution of butanoic acid (4.5 g, 12 mmol) in DCM (100 mL) at room temperature under N2 atmosphere ,17Z)-icos-5,8,11,14,17-pentaen-1-yl)oxy)but anoic acid, oxalyl chloride (8.4 mL, 100 mmol ) was added, followed by 2 drops of DMF. The mixture was stirred at room temperature for 30 minutes and concentrated in vacuo . The residue was dissolved in DCM (100 mL) under N2 atmosphere. Triethylamine (3. 34 mL, 24 mmol), followed by ethanolamine (1.08 mL, 18 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. Water (300 mL) was added and the mixture was extracted twice with dichloro methane. The combined organic phases were dried (Na2SO4), filtered, and concentrated in vacuo . A polarity-increasing mixture of heptane and ethyl acetate (50:50 → 20:80) was used as the eluent to purify the residue by flash chromatography on silica gel . The appropriate fractions were concentrated to give 4.6 g (92% yield) of the title compound. 1 1H NMR ( 300 MHz, CDCl3): δ 0.90 - 1.05 (m, 6H), 1.40 - 1.55 (m, 2H), 1.60 - 1.90 (m, 4H) , 2.05 - 2.20 (m, 4H), 2.75 - 2.90 (m, 8H), 3.05 (br s, 1H), 3.40 - 3.55 (m, 4H) , 3.70 - 3.80 (m, 3H), 5.30 - 5.45 (m, 10H), 7.04 (br s, 1H). MS (electrospray) 440 [M+Na] + [Example]

[0189] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 Preparation of 7-pentaen-1-yl)oxy)-N-isopropylbutanamide

[0190] [ka] 2-(((5Z,8Z,11Z,14Z, 17Z)-Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butane acid (300 mg, 0.801 mmol) and o-benzotriazol-1-yl-N, N,N',N'-Tetramethyluronium tetrafluoroborate (TBTU) (284 A solution of 123 μL of triethylamine (123 μL, 0.884 mmol) was added to the solution of 123 μL of triethylamine (123 μL, 0.884 mmol). After stirring at room temperature for 15 minutes under a N2 atmosphere, isopropylamine (76 μL, 0.884 mmol) was added and the reaction mixture was stirred at room temperature overnight. 5 mL) was added and the mixture was extracted with tert-butyl methyl ether (2 x 50 mL). The organic phases were combined, dried (Na2SO4), filtered and evaporated in vacuo. The chromatogram was analyzed using increasingly polar mixtures of ethyl acetate and ethyl acetate (95:5 → 50:50) as eluents. The residue was purified by flash chromatography on silica gel. Concentration gave 0.207 g (59% yield) of the title compound as an oil.1 1H NMR (400 MHz z, CDCl3) δ 0.83 - 0.93 (m, 3H), 0.95 (t, J = 7.5, 3H), 1.14 (t, J = 6.1, 6H), 1 .39 - 1.47 (m, 2H), 1.56 - 1.73 (m, 3H), 1.73 - 1.83 (m, 1H), 2.02 - 2.11 (m, 4 H), 2.80 - 2.84 (m, 8H), 3.43 (t, J = 6.4, 2H), 3.60 (dd, J = 6.6, 4.4, 1H), 4.0 2 - 4.11 (m, 1H), 5.66 - 5.01 (m, 10H), 6.33 (d, J = 5.9, 1H).

Example

[0191] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 7-pentaen-1-yl)oxy)-N-methylbutanamide preparation

[0192]

Chem.

Example

[0193] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 7-pentaen-1-yl)oxy)-1-(piperidin-1-yl)butan-1-one Preparation

[0194]

Chem.

Example

[0195] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 7-pentaen-1-yl)oxy)butanamide Preparation

[0196]

Chem.

Example

[0197] N-(tert-Butyl)-2-(((5Z,8Z,11Z,14Z,17Z)-icosa- 5,8,11,14,17-pentaen-1-yl)oxy)butanamide Preparation

[0198]

Chemical formula

Example

[0199] (R)-2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11, 14,17-pentaen-1-yl)oxy)-N-((R)-1-phenylethyl)but anamide Preparation

[0200]

Chemical formula

Example

[0201] (2S)-Ethyl 2-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5 ,8,11,14,17-pentaen-1-yloxy)butanamido)-4-methylpe ntanoate Preparation

[0202]

Chem.

Example

[0203] (2S)-2-(2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8, 11,14,17-pentaen-1-yloxy)butanamide)-4-methylpentane acid preparation

[0204]

Chemical formula

Example

[0205] tert-Butyl 2-(((2S)-2-(2-((5Z,8Z,11Z,14Z,1 7Z)-Icosa-5,8,11,14,17-pentaen-1-yloxy)butanam do)-4-methylpentanoyl)oxy)benzoate Preparation

[0206]

Chemical

Example

[0207] 2 - (((2S) - 2 - (2 - ((5Z,8Z,11Z,14Z,17Z) - icosa - 5,8,11,14,17 - pentaen - 1 - yloxy)butanamide) - 4 - methyl pentanoyl)oxy)benzoic acid Preparation

[0208]

Chem.

Example

[0209] ​tert-Butyl (2-(2-((5Z,8Z,11Z,14Z,17Z)-icosa- 5,8,11,14,17-pentaen-1-yloxy)butanamido)ethyl)carb Preparation of

[0210]

Chemical formula

Example

[0211] 2-Hydroxy-N-(2-(2-((5Z,8Z,11Z,14Z,17Z)-icosa -5,8,11,14,17-pentaen-1-yloxy)butanamido)ethyl) Preparation of benzamide

[0212]

Chem.

Example

[0213] 2-(2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14 ,17-pentaen-1-yloxy)butanamido)ethyl 2-acetoxybenzoate Preparation

[0214]

Chem.

[0215] 2-(2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14 ,17-pentaen-1-yloxy)butanamide)ethyl 2-hydroxybenzoate Preparation of

[0216] [Chemical formula] Ammonia (aq, 28%, 3 mL) was added dropwise to a solution of ethyl 2-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,1 1,14,17-pentaen-1-yloxy)butanamide) 2-acetoxybenzoate (390 mg, 0.67 mmol) in 2-propanol (27 mL) and water (9 mL), and the mixture was stirred for 10 minutes . Water was added, and the resulting mixture was extracted twice with Et2O. The combined organic phases were washed with brine, dried (Na2SO4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography using a gradient of 0 - 20% EtO Ac in heptane as the eluent . The appropriate fractions were concentrated to give 63 mg (18% yield) of the title compound. 1 H NMR (30 0 MHz, CDCl3): δ 0.91 (t, 3H), 0.97 (t, 3H), 1.35 - 1.45 (m, 2H), 1.55 - 1.90 (m, 4 H), 2.00 - 2.15 (m, 4H), 2.80 - 2.90 (m, 8H), 3.44 (t, 2H), 3.65 - 3.80 (m, 3H), 4.40 - 4.50 (m, 2H), 5.30 - 5.45 (m, 10H), 6.85 - 6.95 (m, 2H), 6.95 - 7.05 (m, 1H), 7.40 - 7.5 0 (m, 1H), 7.80 - 7.85 (m, 1H), 10.63 (s, 1H). MS (ESI): 560 [M+Na]​+ .

Example

[0217] Methyl 2-hydroxy-5-(2-((5Z,8Z,11Z,14Z,17Z)-icosa -5,8,11,14,17-pentaen-1-yloxy)butanamide)benzo -ate Preparation

[0218]

Chem.

Example

[0219] 2-Hydroxy-5-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5 ,8,11,14,17-Pentaen-1-yloxy)butanamide)benzoic acid Preparation

[0220] [Chemical formula] 2M NaOH (aq, 6 mL) was added to a solution of methyl 2-hydroxy- 5-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14, 17-pentaen-1-yloxy)butanamide)benzoate (320 mg, 0.6 1 mmol), and the reaction mixture was heated at 50 °C overnight. The mixture was cooled to ambient temperature and acidified to pH ~2 with 5M HCl (aq). The resulting mixture was extracted with EtOAc , the organic phase was dried (Na2SO4), filtered, and concentrated in vacuo. Flash chromatography using a gradient of 1 ~2% MeOH in EtOAc as the eluent was used to purify the residue. The appropriate fractions were concentrated to give 85 mg (27% yield) of the title compound . . 1 H NMR (300 MHz, CDCl3): δ 0.90 - 1.05 (t, 3H), 1.20 - 1.30 (m, 1H), 1.45 - 1.60 (m , 2H), 1.65 - 1.75 (m, 2H), 1.80 - 1.95 (m, 2H), 2.05 - 2.20 (m, 4H), 2.80 - 2.90 (m, 8H ), 3.50 - 3.65 (m, 2H), 3.85 - 3.95 (m, 1H), 5.30 - 5.45 (m, 10H), 6.90 - 7.00 (m, 1H), 7.58 (br s, 1H), 8.11 (br s, 1H), 8.40 (s, 1H). MS (ESI): 508 [M-H] - . [Example]

[0221] (2S)-Ethyl 2-(2-((5Z,8Z,11Z,14Z,17Z)-icosa-5 ,8,11,14,17-pentaen-1-yloxy)butanamido)-4-methylpe ntanoate Preparation

[0222] [Chemical Structure] DCC (1.13 g, 5.5 mmol) and HOBt (0.74 g, 5.5 mmol ) were added successively to a solution of 2- ((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-penta en-1-yloxy)butanoic acid (1.87 g, 5.0 mmol) in THF (20 mL), and the mixture was stirred for 10 minutes. L-Leucine ethyl ester hydrochloride (0.89 g, 4.6 mmo l) was added, and the resulting mixture was stirred for 2 hours. EtOAc (100 mL) was added, and the mixture was washed with water, 1 M HCl(aq), saturated NaHCO3(aq) and brine. The organic phase was dried (Na2SO4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography using a gradient of 10 - 15% Et OAc in heptane as the eluent to give 1.84 g (79% yield) of the title compound. The appropriate fractions were concentrated to afford 1.84 g (79% yield) of the title compound. 1 1H NMR (300 MHz, CDCl3): δ 0.85 - 1.05 (m, 12H), 1.25 - 1.35 (m, 3H), 1.40 - 1.85 (m, 9H), 2.05 - 2.20 (m, 4H), 2.80 - 2.90 (m, 8H), 3.45 - 3.75 (m, 3H), 4.15 - 4.25 (m, 2H), 4.55 -4.75 (m, 1H), 5.30 - 5.45 (m, 10H), 6.80 - 6.95 (m, 1H). MS (ESI): 538 [M+Na] + .

Example

[0223] (2S)-2-(2-((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8, 11,14,17-pentaen-1-yloxy)butanamido)-4-methylpentane Preparation of acid

[0224]

Chem.

Example

[0225] Methyl 2-hydroxy-5-((2S)-2-(2-((5Z,8Z,11Z,14 ,17Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butan amide)-4-methylpentanamide)benzoate Preparation

[0226]

Chem.

Example

[0227] 2-Hydroxy-5-((2S)-2-(2-((5Z,8Z,11Z,14Z,17 Z)-Icosa-5,8,11,14,17-pentaen-1-yloxy)butanamide ))-4-methylpentanamide)benzoic acid preparation

[0228]

Chemical

Example

[0229] Methyl 2-hydroxy-5-(((2-(2-((5Z,8Z,11Z,14Z,17 Z)-icos-5,8,11,14,17-pentaen-1-yloxy)butanamide )ethoxy)carbonyl)amino)benzoate Preparation

[0230]

Chem.

Example

[0231] 2-Hydroxy-5-(((2-(2-((5Z,8Z,11Z,14Z,17Z)- Icosa-5,8,11,14,17-pentaen-1-yloxy)butanamide)eth oxy)carbonyl)amino)benzoic acid Preparation

[0232]

Chemical formula

Example

[0233] 2 - ((5Z,8Z,11Z,14Z,17Z) - Icosa - 5,8,11,14,17 - pentaen - 1 - yloxy) - N - (2 - isocyanatoethyl)butanamide Preparation

[0234]

Chem.

Example

[0235] Methyl 2-hydroxy-5-(3-(2-(2-((5Z,8Z,11Z,14Z,1 7Z)-icosa-5,8,11,14,17-pentaen-1-yloxy)butanamido)ethyl)ureido)benzoate Preparation

[0236]

Chemical formula

Example

[0237] 2 - Hydroxy - 5 - (3 - (2 - (2 - ((5Z,8Z,11Z,14Z,17Z) - Icosa - 5,8,11,14,17 - pentaen - 1 - yloxy)butanamido)ethyl)ureido)benzoic acid Preparation 1M LiOH(aq, 2.9 mL) was added dropwise to a solution of methyl 2 - hydroxy

[0238]

Chemical formula

Example

[0239] tert-Butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8, 11,14,17-pentaenyloxy)propanoate Preparation

[0240]

Chemical

Example

[0241] 2 - ((5Z,8Z,11Z,14Z,17Z)-Icosa - 5,8,11,14,17 - pentaenyloxy)propanoic acid preparation

[0242]

Chemical formula

Example

[0243] tert-Butyl 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8, 11,14,17-pentaenyloxy)-2-methylpropanoate Preparation

[0244]

Chemical

Example

[0245] Preparation of 2 - ((5Z,8Z,11Z,14Z,17Z) - icosa - 5,8,11,14,17 - pentaenyloxy) - 2 - methylpropanoic acid

[0246]

Chemical formula

Example

[0247] tert-Butyl 2-ethyl-2-((5Z,8Z,11Z,14Z,17Z)-icosa- 5,8,11,14,17-pentaen-1-yloxy)butanoate Preparation

[0248]

Chemical formula

Example

[0249] 2 - Ethyl - 2 - ((5Z,8Z,11Z,14Z,17Z) - icosa - 5,8,11 ,14,17 - pentaen - 1 - yloxy)butanoic acid preparation

[0250]

Chemical formula

Example

[0251] tert-Butyl 2-(((3Z,6Z,9Z,12Z)-pentadeca-3,6,9, 12-tetraen-1-yl)oxy)butanoate Preparation Step a) (8Z,11Z,14Z,17Z)-5,6-Dihydroxyicosa-8, 11,14,17-tetraenoic acid

[0252]

Chemical formula

[0253] Step b) (3Z,6Z,9Z,12Z)-Pentadeca-3,6,9,12-tetra en-1-ol

[0254] [Chemical formula] Sodium periodate (9.34 g, 43.7 mmol) was added to a solution of (8Z,11Z,14Z,17Z)-5,6-dihydroxyicosa- 8,11,14,17-tetraenoic acid (9.8 g, 29.1 mmol) in THF:water (150 mL) at 0 °C under nitrogen, and the mixture was stirred for 1 hour. The reaction mixture was extracted with EtOAc (300 mL). The organic layer was separated, washed with dilute brine (200 mL), dried (Na2SO4), filtered, and concentrated in vacuo. The residue was dissolved in MeOH (100 ml) and cooled to 0 °C. NaBH4 (3.31 g, 8 7 mmol) was added portionwise carefully, and the mixture was stirred at 0 °C for 30 minutes. Water (300 m L) was added carefully, and the mixture was extracted with EtOAc (2200 mL). The organic phases were combined, washed with water (200 mL), dried (Na2SO4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using 20% ethyl acetate in heptane as the eluent. The appropriate fractions were concentrated to give 4.06 g (63 %) of the title compound as an oil. The organic phases were combined, washed with water (200 mL), dried (Na2SO4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel using 20% ethyl acetate in heptane as the eluent. The appropriate fractions were concentrated to give 4.06 g (63 %) of the title compound as an oil. %, yield) of the title compound as an oil. %) of the title compound as an oil. 1H NMR (400 MHz, CDCl3) δ 0.91 (t, 3H), 1.97 - 2.05 (m, 2H), 2.27 - 2.32 (m, 2H) , 2.73 - 2.81 (m, 6H), 3.57 - 3.62 (m, 2H), 5.17 - 5.41 (m, 7H), 5.42 - 5.56 (m, 1H).

[0255] Step c) tert-Butyl 2-(((3Z,6Z,9Z,12Z)-pentadeca- 3,6,9,12-tetraen-1-yl)oxy)butanoate

[0256]

Chemical formula

Example

[0257] 2-(((3Z,6Z,9Z,12Z)-Pentadeca-3,6,9,12-tetraene -1-yl)oxy)butanoic acid preparation

[0258]

Chemical formula

Example

[0259] 2-(((5Z,8Z,11Z,14Z,17Z)-Nonadeca-5,8,11,14, Preparation of (17 - pentaen - 1 - yloxy) butanoic acid

[0260] [Chemical formula] Step 1: A solution of deca - 2,5,8 - trien - 1 - ol (0.94 g, 6.43 mmol) in THF (100 ml) cooled to 0 °C under nitrogen was added to methanesulfonyl chloride (72 0 μL, 9.24 mmol), followed by dropwise addition of TEA (1.97 mL, 14.13 mmol ) over 15 minutes. The mixture was stirred at 0 °C for 2 hours. Then, the mixture was poured into water: ice 1:1 (200 grams) and extracted with diethyl ether (2 × 100). The combined organic layers were washed with sat.(aq.) NaHCO3 (200 mL), water (200 mL) and brine (200 mL), dried (Na2SO4), filtered, and evaporated in vacuo to give the crude product, which was further purified by dry flash over silica gel. Fractions 1 - 3 were collected using heptane , fractions 4 - 6 were collected using EtOAc:heptane 2.5:100, fractions 7 - 10 were collected using EtOAc:heptane 5:100, and fraction 11 was collected using EtOAc . The appropriate fractions were pooled and concentrated to give 1.07 g (4.77 mmo l, 74.1% yield) of nona - 2,5,7 - trien - 1 - yl methanesulfonate .

[0261] Step 2: A solution of propargyl alcohol (0.35 mL, 5.99 m mol) was added portionwise over 20 minutes to a surrounding solution of ethylmagnesium bromide (4.00 mL, 1 2.00 mmol) diluted with dry THF (25 mL). After refluxing the reaction mixture for 90 minutes, 0 °C​​ It was cooled to 30 minutes. A catalytic amount of bromo(dimethyl sulfide)copper(I) (330 mg , 1.605 mmol) was added and the cooling bath was removed. After stirring for 15 minutes, it was heated to dissolve nona-2,5,7-triyn-1-ylmethanesulfonate (1.05 g, 4.68 mmol) dissolved in THF (5 ml) over 5 minutes dropwise. The reaction mixture was heated to reflux and stirred overnight while refluxing. The reaction was cooled to ambient temperature and stopped by carefully adding 50 mL of sat.aq.N H4Cl. The resulting mixture was transferred to a separatory funnel containing diethyl ether (100 mL). The organic phase was separated and the aqueous phase was extracted twice with diethyl ether (2×100 mL). The combined organic phases were washed with brine (50 mL) and dried (Na2SO4). The residue was concentrated in vacuo and then purified by flash chromatography on silica gel. The appropriate fractions were pooled and concentrated to give 0.49 g (2.6 5 mmol, 56.6% yield) of dodeca-2,5,8,10-tetrayn-1-ol .

[0262] Step 3: To a solution of dodeca-2,5,8,10-tetrayn-1-ol (465 mg, 2,52 mmol) in THF (30 ml) cooled to 0 °C under nitrogen, methanesulfonyl chloride (0.256 ml, 3.28 mmol) was added, followed by dropwise addition of TEA (0.704 ml, 5. 05 mmol) over 15 minutes. The mixture was stirred at 0 °C for 1 hour. Then the mixture was poured into water:ice 1:1 (100 grams) and extracted with diethyl ether (2×100 mL) . The combined organic layers were washed with sat.(aq.)NaHCO3 (200 mL), water (2 ​​​​washed with water (100 mL) and brine (200 mL), dried (Na2SO4), filtered, and evaporated in vacuo to give 622 mg of the crude product, which was further purified by dry flash chromatography. Fractions 1-3 were eluted with heptane, fractions 4-6 were eluted with EtOAc:heptane 2.5:100, fractions 7-10 were eluted with EtOAc:heptane 5:100, and fraction 11 was eluted with EtOAc. Appropriate fractions were pooled and concentrated to give 466 mg (1.78 mmol, 70.4% yield) of trideca-2,5,8,11-tetrayl methanesulfonate. Step 4: To a solution / suspension of cesium carbonate (553 mg, 1.696 mmol), sodium iodide (280 mg, 1.866 mmol), and copper(I) iodide (323 mg, 1.696 mmol) in dry DMF (15 mL) was added ethyl hex-5-ynoate (262 mg, 1.866 mmol) in dry DMF (5 mL). The mixture was stirred at room temperature under a nitrogen atmosphere for 40 minutes. Trideca-2,5,8,11-tetrayl methanesulfonate (445 mg, 1.696 mmol) in dry DMF (5 mL) was added and the mixture was stirred at room temperature overnight. Saturated NH4Cl (200 mL) was added and the mixture was extracted with EtOAc:heptane (1:1, 3 x 150 mL). The combined organic phases were washed with brine (200 mL), dried (MgSO4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel starting with a 2% EtOAc in heptane solution and using a gradient of increasing polarity of heptane and ethyl acetate as the eluent. Appropriate

[0263] fractions were pooled and concentrated. ​​​​​​​​​​​​​​The sad fractions were pooled and concentrated to give 51 mg (0.166 mmol, 9.8% yield) of ethyl nonadeca-5,8,11,14,17-pentynoate.

[0264] Step 5: To a solution of ethyl nonadeca-5,8,11 ,14,17-pentynoate (60 mg, 0.196 mmol) in heptane (10 mL) and toluene (15 mL) was bubbled with N2 gas, and then quinoline (10 μL, 0.084 mmol) and Lindlar catalyst (50 m g, 0.023 mmol) were added under an N2 atmosphere. The suspension was stirred for 15 h under an H2 atmosphere (1 atm). The reaction mixture was filtered through Celite and evaporated in vacuo. The crude material was purified by flash chromatography on silica gel using a mixture of 2% EtOAc in heptane as the eluent. The appropriate fractions were pooled and concentrated to give (5Z,8Z,11Z,14Z,17Z)-ethyl nonadeca-5,8,11,14,17- pentaenoate (24.6 mg, 0.078 mmol, 39.7% yield) as an oil. 1H NMR (400 MHz, chloroform-d) δ 1.27 (t, 2H), 1.59 - 1.70 (m, 2H), 1

[0265] Step 6: To a solution of (5Z,8Z,11Z,14Z, 17Z)-ethyl nonadeca-5,8,11,14,17-pentaenoate (22 mg, 0.070 mmol) in tetrahydrofuran (5 mL) cooled to 0 °C was added LAH (8 mg, 。The reaction mixture was stirred at 0 °C for 30 minutes and then at ambient temperature for 90 minutes. The reaction mixture was poured into saturated NH4Cl(aq.) (10 mL) and ice (10 g), and the mixture was acidified to pH ~1-2 using 10% HCl(aq. ). The mixture was extracted twice with Et2O (25 ml), and the combined organic layers were washed with water (25 mL) and brine (25 mL), then dried (M gSO4), filtered, and concentrated in vacuo to give (5Z,8Z,11Z,14Z,17Z)- nonadeca-5,8,11,14,17-pentaen-1-ol (15.5 mg, 0.0 56 mmol, 81% yield) as an oil.

[0266] Step 7: 2-Bromobutyric acid (100 mg, 0.599 mmol) was added to a solution of (5Z,8Z,11Z,14Z,17Z)-nonadeca-5,8,11,14,1 7-pentaen-1-ol (8 mg, 0.029 mmol) in tetrahydrofuran (10 m l). The mixture was cooled to 5 °C, and then a 17% (w / w) solution of sodium te rt-butoxide (500 μl, 0.678 mmol) in tert-butyl methyl ether (30 ml) was added dropwise over 5 minutes. After 3 hours, additional sodium tert-butoxide (100 μL 0 ,136 mmol) was added and the reaction mixture was stirred for an additional 3 hours. The reaction was quenched by adding formic acid (0.100 ml , 2.61 mmol) and water (10 ml), and the resulting mixture was extracted with tert-butyl methyl ether (30 ml). The organic phase was washed 4 times with water (4 × 1 0 ml), dried (MgSO4), filtered, and evaporated in vacuo. The concentrate was dissolved in Me-THF (50 ml) and washed 4 times with saturated aqueous NH4Cl (20 ml). The organic phase was dried (MgSO4), filtered, and evaporated in vacuo. The concentrate was dissolved in Me-THF (50 ml) and washed 4 times with saturated aqueous NH4Cl (20 ml). The organic phase was dried (MgSO4), filtered, and evaporated in vacuo. The concentrate was dissolved in Me-THF (50 ml) and washed 4 times with saturated aqueous NH4Cl (20 ml). The The machine phase was dried (MgSO4), filtered, and evaporated in vacuo to give 2-(((5Z,8Z,1 1Z,14Z,17Z)-nonadeca-5,8,11,14,17-pentaen-1-yl )oxy)butanoic acid (6 mg, 0.017 mmol, 57.1%) as an oil. HRMS (electrospray), [M-H] - : Calculated: 359.2586, Found: 359.2578.

Example

[0267] 2-(((6Z,9Z,12Z,15Z)-octadeca-6,9,12,15-tetra en-1-yl)oxy)butanoic acid Preparation

[0268]

Chemical formula

Example

[0269] 2 - (((4Z,7Z,10Z,13Z,16Z)-Nonadeca-4,7,10,13, 16 - pentaen-1-yl)oxy)butanoic acid preparation

[0270]

Chemical formula

[0271] Step 2: A mixture of tert-butyl 2-((4Z,7Z,10Z,13Z, 16Z)-nonadeca-4,7,10,13,16-pentaenyloxy)butanoate ( 373 mg, 0.895 mmol) in HCOOH (10 mL) was vigorously stirred for 2.5 hours. The mixture was concentrated and, after a total reaction time of 3 hours, diluted with ethyl acetate (50 mL) and washed with water (3×50 mL) to neutralize the pH of the aqueous phase. The organic phase was dried (MgSO4), filtered, and concentrated in vacuo . Purification by dry flash using silica gel (8 g), eluting with heptane:EtoAc 90:10 (3×30 m L), followed by 80:20 (7×50 mL). Fractions 4 - 6 were collected and concentrated in vacuo to give 2-((4Z,7Z,10Z,13Z,16Z)- nonadeca-4,7,10,13,16-pentaenyloxy)butanoic acid (122 mg, 0 .338 mmol, yield 37.8%). 1H NMR (300 MHz, chloroform-d) δ 1. 00 (td, 2.6 Hz, 6H), 1.62 - 1.79 (m, 2H), 1.79 - 1.92 (m, 2H), 1.99 - 2.13 (m, 2 H), 2.19 (dd, 2H), 2.85 (dtd, 8H), 3.47 - 3.67 (m, 2H), 3.88 (dd, 1H), 5.31 - 5. 47 (m, 10H). HRMS (Electrospray), [M-H] - : Calculated: 359.2586, Found: 359.25 90.

Example

[0272] 2-(((8Z,11Z,14Z)-Octadeca-8,11,14,17-tetraene -1-yl)oxy)butanoic acid Preparation

[0273]

Chemical formula

[0274] Step 2: A suspension of Lindlar catalyst (1.063 g, 0.499 mmol) in heptane (15 ml) under N2 atmosphere was added to a solution of methyl octadeca-17-ene-8,11,14-triynoate (1 .42 g, 4.99 mmol) in quinoline (0.177 ml, 1.498 mmol) and heptane (5 ml). The mixture was stirred overnight under H2 atmosphere (1 atm). The mixture was filtered and concentrated in vacuo. By flash chromatography (heptane / EtOAc 98.5 / 1.5), (8Z,11Z,14Z)-methyl octadeca-8,11,14,17-tetraenoate (0.830 g, 2.86 mmol, yield 57.2%) was obtained as an oil. 1H NMR (400 MHz, CDCl3) δ 5.84 - 5.65 (m, 1H) ​​, 5.43 - 5.19 (m, 6H), 5.05 - 4.85 (m, 2H), 3.60 (s, 3H), 2.81 - 2.68 (m, 6H), 2 .23 (t, 2H), 2.02 - 1.92 (m, 2H), 1.59 - 1.52 (m, 2H), 1.34 - 1.18 (m, 6H). MS ( Electrospray): 313.2 [M+Na]+.

[0275] Step 3: (8Z,11Z,14Z)-Methyloctadeca-8,11,14 ,17-tetraenoate (830 mg, 2.86 mmol) in THF (2 ml) was added dropwise to a cooled (0 °C) suspension of LAH (114 mg, 3.00 mmol) in THF (10 ml) under a N2 atmosphere . The mixture was stirred at 0 °C for 40 minutes and carefully poured into cold saturated NH4Cl (20 mL). The aqueous layer was acidified to pH ~2 with HCl (2M). The phases were separated and the aqueous phase was extracted with heptane (3 × 20 ml). The combined organic phases were washed with brine (20 mL), dried (Na2SO4), filtered, and concentrated in vacuo to give (8Z,11Z,14Z)-octadeca-8 ,11,14,17-tetraene-1-ol (730 mg, 2.78 mmol, 97% yield) as an oil. 1H NMR (400 MHz, CDCl3) δ 5.84 - 5.66 (m, 1H), 5.43 - 5.20 (m, 6H), 5.03 - 4.82 (m, 2H), 3.57 (t, 2H), 2.84 - 2.62 (m, 6H), 2.06 - 1. 90 (m, 4H), 1.58 - 1.42 (m, 3H), 1.33 - 1.20 (m, 6H). MS (Electrospray): 2 85.1 [M+Na]+.

[0276] ​​​Step 4: A solution of NaOH (2 ml, 37.9 mmol) was slowly added to a vigorously stirred solution of (8Z,11Z,14Z)-octadeca-8,11,14,17-tetraen-1-ol (720 mg, 2.74 mmol), tert-butyl 2-bromobutanoate ( (8Z,11Z,14Z)-octadeca-8,11,14,17-tetraen-1-ol (918 mg, 4.12 mmol) and tetrabutylammonium hydrogen sulfate (279 mg , 0.823 mmol) in toluene (8 ml) under a N2 atmosphere. The mixture was heated at 40 °C overnight. Additional tert-butyl 2-bromobutanoate (306 mg, 1.372 mmol) was added twice, after 5 h and 24 h. After 48 h, the mixture was cooled to 5 - 10 °C and saturated NH4Cl (20 ml) was added. The phases were separated and the aqueous phase was extracted with EtOAc ( 50 ml). The combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo. Flash chromatography (heptane / EtOAc 99 / 1) gave tert-butyl 2-(((8Z,11Z,14Z)-octadeca-8,11,14,17 -tetraen-1-yl)oxy)butanoate (675 mg, 1.668 mmol, yield 60.8%) as an oil. 1H NMR (400 MHz, CDCl3) δ 5.86 - 5.65 (m, 1H) , 5.41 - 5.21 (m, 6H), 5.04 - 4.83 (m, 2H), 3.57 - 3.46 (m, 2H), 3.30 - 3.19 (m, 1H), 2.81 - 2.68 (m, 6H), 2.02 - 1.89 (m, 4H), 1.71 - 1.60 (m, 2H), 1.54 - 1.50 (m, 2H), 1.41 (s, 12H), 1.32 - 1.20 (m, 8H), 0.89 (t, 3H). tert-butyl 2-(((8Z,11Z,14Z)-octadeca-8,11,14,17 -tetraen-1-yl)oxy)butanoate (675 mg, 1.668 mmol, yield 60.8%) as an oil. 1H NMR (400 MHz, CDCl3) δ 5.86 - 5.65 (m, 1H) , 5.41 - 5.21 (m, 6H), 5.04 - 4.83 (m, 2H), 3.57 - 3.46 (m, 2H), 3.30 - 3.19 (m, 1H), 2.81 - 2.68 (m, 6H), 2.02 - 1.89 (m, 4H), 1.71 - 1.60 (m, 2H), 1.54 - 1.50 (m, 2H), 1.41 (s, 12H), 1.32 - 1.20 (m, 8H), 0.89 (t, 3H).

[0277] Step 5: A solution of tert-butyl 2-(((8Z, 11Z,14Z)-octadeca-8,11,14,17-tetraen-1-yl)oxy )butanoate (675 mg, 1.668 mmol) in HCOOH (6.75 ml, 179 mmol) was stirred at 40 °C for 21 hours under a N2 atmosphere. The reaction mixture was evaporated under reduced pressure and purified by preparative HPLC eluting with CH3C N:H20 - 90:10 containing 0.02% HCOOH. The preparative fractions containing the pure product were evaporated under reduced pressure to remove CH3CN. To the residue was added tert-butylme thyl ether and brine, and the phases were separated. The organic phase was dried (Na2SO4), filtered and evaporated under reduced pressure to give 2-((8Z,11Z,14Z)-octadeca-8,11,1 4,17-tetraen-1-yloxy)butanoic acid (163 mg, 0.464 mmol, 27.8% yield) as an oil. 1H NMR (400 MHz, CDCl3) δ 5.89 - 5.70 (m, 1 H), 5.51 - 5.22 (m, 6H), 5.00 (dd, 2H), 3.84 (t, 1H), 3.62 - 3.38 (m, 2H), 2.79 (m, 6H), 2.12 - 1.95 (m, 2H), 1.88 - 1.75 (m, 2H), 1.64 - 1.58 (m, 2H), 1.31 (s, 8H), 0.96 (t, 3H). HRMS (electrospray) [M-H] : Calculated: 347.2586, Found: - 347.2589. 347.2589.

Example

[0278] 2-(((5Z,8Z,11Z,14Z)-nonadeca-5,8,11,14-tetraene Preparation of (5Z,8Z,11Z,14Z)-nonadeca-5,8,11,14-tetraen-1-yl 2-bromobutanoate

[0279]

Chem.

[0280] Step 2: To a solution of (5Z,8Z,11Z,14Z)-nonadeca-5,8,11 ,14-tetraen-1-ol (17.9 g, 64.7 mmol) in THF (350 ml) was added 2-bromo butanoic acid (10.35 ml, 97 mmol). The reaction mixture was cooled to 8 - 10 °C​ Cooled. While maintaining the temperature between 8 and 10 °C, sodium tert-butoxide (17.4 2 g, 181 mmol) (17% in MTBE (120 mL)) was added over 25 minutes . The reaction mixture was stirred at 8 - 10 °C for 30 minutes. Additional sodium tert-butoxide ( 2.489 g, 25.9 mmol) (17% in MTBE (25 mL)) was added over 5 minutes . The mixture was stirred at 8 - 10 °C for 30 minutes, and then 2-bromobutyric acid (6.90 mL , 64.7 mmol) was added over 5 minutes. Then the mixture was stirred at 8 - 10 °C for 30 minutes . Additional sodium tert-butoxide (4.98 g, 51.8 mmol) (17% in MT BE (40 mL)) was added over 5 minutes. The mixture was stirred at 8 - 10 °C for 30 minutes . Additional sodium tert-butoxide (2.489 g, 25.9 mmol) (17% in MT BE (25 mL)) was added over 5 minutes. The mixture was stirred at 8 - 10 °C for 30 minutes . HCOOH (3 mL) and then 2 M HCl(aq) were added to adjust the pH to ~2 . The phases were separated and the aqueous phase was extracted with MTBE (2 × 300 mL). The combined organic phases were washed with water (3 00 mL), saturated NaHCO3 (4 × 200 mL) and brine (300 mL), dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash chromatography (heptane / EtOAc / HCOOH 90 / 10 / 0.5), and the appropriate fractions were pooled and concentrated in vacuo. Then, 88% MeCN (containing 0.02% HCOOH) in water (containing 10% MeCN and 0.02% HCOOH) was used as the eluent to further purify the residue by preparative HPLC. The solvent was removed to give 10.1 g 6g of 2-(((5Z,8Z,11Z,14Z)-nonadeca-5,8,11,14-tetra en-1-yl)oxy)butanoic acid (26.8 mmol, yield 41.5%) was generated as an oil and 1H NMR (300 MHz, CDCl3) δ 9.93 (s, 1H), 5.51 - 5.21 (m, 8H), 3.84 - 3.80 (m, 1H), 3.62 - 3.54 (m, 1H), 3.50 - 3.39 (m, 1H), 2.96 - 2.61 (m, 6H), 2.12 - 2.01 (m, 4H), 1.90 - 1.71 (m, 2H), 1.69 - 1.57 (m, 2H), 1.49 - 1.37 (m, 2 H), 1.36 - 1.23 (m, 4H), 0.97 (t, 3H), 0.91 - 0.84 (m, 3H). MS (electrospray ionization): 361.2 [M-H]-.

Example

[0281] 2-(((8Z,11Z,14Z)-octadeca-8,11,14-trien-1-yl oxy)butanoic acid preparation

[0282]

Chemical formula

[0283] Step 2: To a stirred solution of nickel(II) acetate tetrahydrate (7.67 mL, 55. 4 mmol) in anhydrous EtOH (150 mL) at room temperature, solid NaBH4 (2.097 g , 55.4 mmol) was added under an H2 atmosphere (1 atm). The resulting suspension was stirred for 30 minutes, then ethylenediamine (12.38 mL, 185 mmol) was added. After complete addition, the suspension was stirred for an additional 15 minutes and then ethyl octadeca - 8,11,14 - triynoate (11.1 g, 36.9 mmol) in anhydrous EtOH (50 mL) was added. The reaction mixture was stirred at room temperature under H2 (1 atm) for three nights. Diethyl ether (500 mL) was added to dilute the reaction mixture, and then the resulting mixture was passed through a short silica gel column. The filtrate was concentrated in vacuo. Flash column chromatography (heptane / EtOAc 98 / 2) was used to purify the residue ​​ Ethyl (8Z,11Z,14Z)-octadeca-8,11,14-trienoate The compound (10 g, 32.6 mmol, 88% yield) was obtained as an oil. 1H NMR (300 MHz, CDC l3) δ 5.46 - 5.23 (m, 6H), 4.10 (q, 2H), 2.87 - 2.67 (m, 3H), 2.26 (t, 2H), 2.0 7 - 1.97 (m, 4H), 1.66 - 1.55 (m, 2H), 1.41 - 1.18 (m, 11H), 0.91 - 0.84 (m, 3H) .

[0284] Step 3: Ethyl (8Z,11Z,14Z)-octadeca-8,11,1 in THF (50 ml) A solution of 4-trienoate (24.6 g, 80 mmol) was dissolved in dry THF (30 A cooled (0°C) suspension of LAH (3.05 g, 80 mmol) in 0 mL of LAH was added over 15 min. The mixture was stirred at 0°C for 1 hour. The mixture was then added dropwise to cold saturated NH4Cl (300 mL The aqueous layer was acidified with HCl (2M) to pH 2. The phases were separated and the aqueous The organic phase was extracted with heptane (2 x 250 mL). The combined organic phase was washed with brine (300 mL). Wash, dry (Na2SO4), filter, and concentrate in vacuo to give (8Z, 11Z, 14Z). -Octadeca-8,11,14-trien-1-ol (20.7 g, 78 mmol, yield The product was obtained as an oil in 98% yield. 1H NMR (300 MHz, CDCl3) δ 5.49 - 5.23 (m, 6H), 3. 61 (t, 2H), 2.86 - 2.67 (m, 4H), 2.11 - 1.92 (m, 4H), 1.59 - 1.49 (m, 2H), 1.43 - 1.21 (m, 10H), 0.94 - 0.83 (m, 3H).

[0285] Step 4: To a solution of (8Z,11Z,14Z)-octadeca-8,11,14-trien-1-ol (20.7 g, 78 mmol) in THF (350 mL) was added 2-bromobutanoic acid (1 2.51 mL, 117 mmol). The reaction mixture was cooled to 8 - 10 °C. While maintaining the temperature between 8 - 10 °C, a solution of sodium tert-butoxide (21.06 g, 219 mmol) (17% in MTBE (130 mL)) was added over 15 minutes. The reaction mixture was stirred at 8 - 10 °C for 30 minutes. Additional sodium tert-butoxide (3.01 g, 31.3 mmol) (17% in MTBE (25 mL)) was added over 5 minutes. The mixture was stirred at 8 - 10 °C for 30 minutes, followed by the addition of 2-bromobutyric acid (8.34 mL, 78 mmol ) over 5 minutes. Then the mixture was stirred at 8 - 10 °C for 30 minutes. Additional sodium tert-butoxide (6.02 g, 62.6 mmol) (17% in MTBE (40 mL)) was added over 5 minutes. The mixture was stirred at 8 - 10 °C for 30 minutes. Additional sodium tert-butoxide (3.01 g, 31.3 mmol) (17% in MTBE (25 mL)) was added over 5 minutes. The mixture was stirred at 8 - 10 °C for 30 minutes.

[0286] HCOOH (3 mL) and then 2 M HCl(aq) were added to adjust the pH to ~2. The phases were separated and the aqueous phase was extracted with MTBE (2×300 mL). The combined organic phases were washed with water (300 m L), saturated NaHCO3 (4×200 ml) and brine (300 ml), dried (Na2SO4), filtered, and concentrated in vacuo. The residue was purified by flash chromatography( ​​​​Purified by heptane / EtOAc / HCOOH 90 / 10 / 0.5), and appropriate fractions were pooled and concentrated under vacuum. Then, 88% MeCN (containing 0.02% HCOOH) in water (containing 10% MeCN and 0.02% HCO OH) was used as the eluent, and the residue was further purified by preparative HPLC. The solvent was removed to give 10.55 g of 2-(((8Z,11Z,14Z)-octadeca-8,11,14-trien-1-yl )oxy)butanoic acid (28.8 mmol, 36.8% yield) as an oil. 1H NMR (300 MHz, CDCl3) δ 9.70 (s, 1H), 5.47 - 5.25 (m, 6H), 3.85 - 3.81 (m, 1H), 3.60 - 3.52 (m, 1H), 3.49 - 3.42 (m, 1H), 2.87 - 2.68 (m, 4H), 2.10 - 1.94 (m, 4H), 1.89 - 1.69 (m, 2H), 1.66 - 1.54 (m, 2H), 1.43 - 1.24 (m, 10H), 0.96 (t, 3H ), 0.89 (t, 3H). MS (electrospray): 349.2 [M-H]-.

Example

[0287] Preparation of 2-(((9Z,12Z,15Z)-octadeca-9,12,15-trien-1-yl )oxy)butanoic acid

[0288]

Chemical formula

[0289] HCOOH (3 mL) was added followed by 2 M HCl (aq) to bring the pH to 2. The aqueous phase was separated and extracted with MTBE (2 x 300 mL). The combined organic phase was diluted with water (300 mL). L), washed with saturated NaHCO3 (4 x 200 ml) and brine (300 ml), and dried (N The residue was purified by flash chromatography on silica gel. Purification by chromatography (heptane / EtOAc / HCOOH 90 / 10 / 0.5) The appropriate fractions were pooled and concentrated in vacuo to give 2-(((9Z,12Z,15Z)- Octadeca-9,12,15-trien-1-yl)oxy)butanoic acid (28g, 78m mol, yield 69.9%) was produced as an oil. 1H NMR (300 MHz, CDCl3) δ 10.07 (s, 1H), 5.48 - 5.16 (m, 6H), 3.84 - 3.80 (m, 1H), 3.60 - 3.53 (m, 1H), 3.48 - 3.40 (m, 1H), 2.87 - 2.70 (m, 4H), 2.12 - 1.97 (m, 4H), 1.91 - 1.70 (m, 2H), 1.6 6 - 1.53 (m, 2H), 1.38 - 1.27 (m, 10H), 0.99 - 0.93 (m, 6H). MS (electrospray ionization): 373.3 [M+Na]+.

Example

[0290] 2-(((6Z,9Z,12Z)-Octadeca-6,9,12-trien-1-yl) oxy)butanoic acid preparation

[0291]

Chemical formula

[0292] 1H), 5.46 - 5.22 (m, 6H), 3.84 - 3.80 (m, 1H), 3.61 - 3.54 (m, 1H), 3.47 - 3.40 (m, 1H), 2.86 - 2.71 (m, 4H), 2.10 - 1.99 (m, 4H), 1.91 - 1.70 (m, 2H), 1.65 - 1 L), saturated NaHCO3 (4 × 200 mL) and brine (300 mL), dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (heptane / EtOAc / HCOOH 90 / 10 / 0.5). Appropriate fractions were pooled and concentrated in vacuo to yield 2-((6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy)butanoic acid (30.2 g, 83 mmol, 74.1%) as an oil. 1H NMR (300 MHz, CDCl3) δ 9.56 (s, a2SO4) and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (heptane / EtOAc / HCOOH 90 / 10 / 0.5). Appropriate fractions were pooled and concentrated in vacuo to yield 2-((6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy)butanoic acid (30.2 g, 83 mmol, 74.1%) as an oil. 1H NMR (300 MHz, CDCl3) δ 9.56 (s, purified by flash chromatography on silica gel (heptane / EtOAc / HCOOH 90 / 10 / 0.5). Appropriate fractions were pooled and concentrated in vacuo to yield 2-((6Z,9Z,12Z)-octadeca-6,9,12-trien-1-yloxy)butanoic acid (30.2 g, 83 mmol, 74.1%) as an oil. 1H NMR (300 MHz, CDCl3) δ 9.56 (s, 1H), 5.46 - 5.22 (m, 6H), 3.84 - 3.80 (m, 1H), 3.61 - 3.54 (m, 1H), 3.47 - 3.40 l, yield 74.1%) as an oil. 1H NMR (300 MHz, CDCl3) δ 9.56 (s, 1H), 5.46 - 5.22 (m, 6H), 3.84 - 3.80 (m, 1H), 3.61 - 3.54 (m, 1H), 3.47 - 3.40 (m, 1H), 2.86 - 2.71 (m, 4H), 2.10 - 1.99 (m, 4H), 1.91 - 1.70 (m, 2H), 1.65 - 1 .56 (m, 2H), 1.45 - 1.18 (m, 10H), 0.97 (t, 3H), 0.87 (t, 3H). MS (Electrospray): 373.2 [M+Na]+. (Electrospray): 373.2 [M+Na]+.

Example

[0293] 2-(((6Z,9Z,12Z,15Z,18Z)-Heneicosa-6,9,12,15 ,18-pentaen-1-yl)oxy)butanoic acid Preparation

[0294]

Chemical formula

Example

[0295] 2-(((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10, 13,16,19-hexaen-1-yl)oxy)butanoic acid preparation

[0296] [Chemical] (4Z,7Z,10Z,13Z ,16Z,19Z)-docosa-4,7,10,13,16,19-hexaen-1-ol (30 g, 95 mmol) in toluene (11 ml) and THF (400 ml) was added 2-bromobutyric acid (17.28 ml, 162 mmol ). Subsequently, a solution of sodium -t-butoxide (25.7 g, 267 mmol) in THF (160 ml) was added over 35 minutes while maintaining the temperature between 8 - 10 °C. The reaction mixture was stirred at 8 - 10 °C for 40 minutes. Sodium -t-butoxide (4.17 g, 43.4 mmol) in THF (60 ml) was added over 10 minutes. The resulting mixture was stirred at 8 - 10 °C for 40 minutes, then 2-bromobutyric acid (10.17 ml, 95 mmol) was added one more time. The mixture was stirred at 8 - 10 °C for 1 hour, then while keeping the temperature below 12 °C, sodium -t-butoxide (7.33 g, 76 mmol) in THF (100 ml) was added over 10 minutes. The reaction mixture was stirred at 10 °C for 60 minutes. Sodium -t-butoxide (4.17 g, 43.4 mmol) was added in one portion, then stirred for an additional 1 hour. 2 -bromobutyric acid (8.5 ml, 50 mmol) and sodium -t-butoxide (4.8 g, 50 mmol) were further added and the mixture was stirred at 8 - 10 °C for 1 hour. 2-bromobutyric acid (8.5 ml, 50 mmol) and sodium -t-butoxide (4.8 g, 50 m mol) were further added and the mixture was stirred at 8 - 10 °C for 1 hour. THF (200 ml) was further added and the reaction mixture was stirred at room temperature overnight. 2-bromobutyric acid (17 ml, 100 mm ol) ol) and sodium - t - butoxide (9.6 g, 100 mmol) were further added, The reaction mixture was stirred at 8 - 10 °C for 1 hour. Then, formic acid (1 ml) was added to the reaction, and the resulting mixture was stirred at 8 - 10 °C for 10 minutes, and then 6M HCl was added to adjust the pH to ~2. Water (100 mL) was added, and the aqueous and organic phases were separated. The aqueous phase was extracted twice with diethyl ether (2 ×500 mL). The combined organic phases were washed with water (3×1000 ml), sat. NaHCO3 (3×500 ml) and brine (1×100 ml), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (EtOAc / heptane / 5% HCOOH) (90 / 10) to give 2 - ( ((4Z,7Z,10Z,13Z,16Z,19Z)-docosa - 4,7,10,13,1 6,19 - hexaen - 1 - yl)oxy)butanoic acid (10.3 g, 25.07 mmol ((4Z,7Z,10Z,13Z,16Z,19Z)-docosa - 4,7,10,13,1 6,19 - hexaen - 1 - yl)oxy)butanoic acid (10.3 g, 25.07 mmol, yield 26.3%) as an oil. 1H NMR (300 MHz, CDCl3) δ 5.49 - 5.20 (m, 12H), 3.90 - 3.75 (m, 1H), 3.65 - 3.52 (m, 1H), 3.51 - 3.35 (m, 1H), 2.91 - 2.6 9 (m, 10H), 2.21 - 1.98 (m, 4H), 1.91 - 1.59 (m, 4H), 1.05 - 0.88 (m, 6H). MS ( electrospray): 423.3 [M + Na]+.

Example

[0297] 2 - ((((8Z,11Z,14Z,17Z)-icosa - 8,11,14,17 - tetra ene - 1 - yl)oxy)butanoic acid Preparation

[0298] [Chemical] Step 1: A heated suspension of magnesium (0.217 g, 8.93 mmol) in THF (3 ml) was added with 1,2-dibromoethane (2 drops), followed by a solution of 2-((5-bromopentyl)oxy)tetrahydro-2H-pyran (2.07 g, 8 .24 mmol) in THF (15 ml) under a nitrogen atmosphere dropwise. The mixture was refluxed for 1 hour and cooled to ambient temperature. A solution of (3Z,6Z,9Z,12Z)-pentadeca-3,6,9,12 -tetraenal (1.5 g, 6.87 mmol) in THF (10 ml) was added dropwise, and the mixture was stirred at ambient temperature for 1 hour. 1 M HCl was added, and the mixture was extracted with EtOAc (2 × 50 mL). The combined organic phases were washed with brine (50 mL), dried (Na2SO4), filtered, and concentrated in vacuo. Flash chromatography on silica gel (heptane / EtOAc 80 / 20 - 75 / 25) gave (8Z,11Z,14Z,17Z)-1-((tetra hydro-2H-pyran-2-yl)oxy)icosa-8,11,14,17-tetraene -6-ol (0.74 g, 1.895 mmol, 27.6% yield) as an oil. 1H NMR (400 MHz, CDCl3) δ 5.59 - 5.49 (m, 1H), 5.49 - 5.25 (m, 7H), 4.56 - 4. 53 (m, 1H), 3.88 - 3.82 (m, 1H), 3.77 - 3.67 (m, 1H), 3.65 - 3.59 (m, 1H), 3.52 - 3.43 (m, 1H), 3.40 - 3.34 (m, 1H), 2.85 - 2.77 (m, 6H), 2.23 (t, 2H), 2.13 - 1 59 (m, 1H), 1.95 - 1.83 (m, 2H), 1.77 - 1.63 (m, 2H), 1.60 - 1.48 (m, 2H), 1.45 - 1.30 (m, 2H), 1.30 - 1.15 (m, 2H), 1.15 - 0.95 (m, 3H), 0.95 - 0.85 (m, 3H), 0.85 - 0.75 (m, 3H), 0.75 - 0.65 (m, 3H), 0.65 - 0.55 (m, 3H), 0.55 - 0.45 (m, 3H), 0.45 - 0.35 (m, 3H), 0.35 - 0.25 (m, 3H), 0.25 - 0.15 (m, 3H), 0.15 - 0.05 (m, 3H). .97 (m, 2H), 1.84 - 1.77 (m, 1H), 1.73 - 1.67 (m, 1H), 1.63 - 1.34 (m, 12H), 0.9 6 (t, 3H). MS (Electrospray): 413.3 [M+Na]+.

[0299] Step 2: (8Z,11Z,14Z,17Z)-1-((Tetrahydro- 2H-pyran-2-yl)oxy)icosa-8,11,14,17-tetraen-6-ol (0.74 g, 1.895 mmol) in THF (20 ml), TEA (0.528 ml, 3.79 mmol) was added, followed by methanesulfonyl chloride (0.192 ml, 2.463 mmol) . The mixture was stirred at ambient temperature for 3 h under a nitrogen atmosphere. Citric acid (5%, aq, 5 0 mL) was added and the mixture was extracted with EtOAc (2 × 50 mL). The combined organic phases were washed with saturated NaHCO3 (50 mL) and brine (50 mL), dried (Na2SO4), filtered, and concentrated in vacuo to give 0.82 g of the intermediate mesylate. The intermediate mesylate was dissolved in Et2O (10 ml) and added dropwise to a stirred suspension of LAH (0.288 g, 7. 58 mmol) in Et2O (25 ml). The mixture was stirred at ambient temperature for 1 h. 1 M H Cl (50 mL) was carefully added. The mixture was extracted with MTBE (2 × 50 mL). The combined organic phases were washed with saturated NaHCO3 (50 mL) and brine (50 mL), dried (N a2SO4), filtered, and concentrated in vacuo. Flash chromatography on silica gel (heptane / EtOAc 97 / 3) gave 2-(((8Z,11Z,14Z, 17Z)-icosa-8,11,14,17-tetraen-1-yl)oxy)tetrahydro RO-2H-pyran (0.46 g, 1.228 mmol, yield 64.8%) was obtained as an oil. 1H NMR (400 MHz, CDCl3) δ 5.46 - 5.20 (m, 8H), 4.57 - 4.54 (m, 1H), 3.88 - 3.82 (m, 1H), 3.74 - 3.68 (m, 1H), 3.51 - 3.44 (m, 1H), 3.39 - 3.33 (m, 1H), 2 .84 - 2.77 (m, 6H), 2.14 - 1.96 (m, 4H), 1.85 - 1.77 (m, 1H), 1.73 - 1.65 (m, 1H ), 1.62 - 1.46 (m, 6H), 1.37 - 1.24 (m, 8H), 0.96 (t, 3H). MS (electrospray -): 397.3 [M+Na]+.

[0300] Step 3: To a solution of 2-(((8Z,11Z,14Z,17Z)-icosa-8,1 1,14,17-tetraen-1-yl)oxy)tetrahydro-2H-pyran (0.4 6 g, 1.228 mmol) in EtOH (5 ml) was added pyridinium-p-toluenesulfonate (PP TS, 0.309 g, 1.228 mmol), and the mixture was heated at 50 °C for 2 h. The mixture was cooled to ambient temperature. Saturated NaHCO3 (50 mL) was added and the mixture was extracted with EtO Ac (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried (N a2SO4), filtered, and concentrated in vacuo. Flash chromatography on silica gel (heptane / EtOAc 80 / 20) gave (8Z,11Z,14Z,17Z )-icosa-8,11,14,17-tetraen-1-ol (0.26 g, 0.895 mmol, yield 72.9%) was obtained as an oil. 1H NMR (400 MHz, CDCl3) δ 5.41 - 5.17 (m, 8H), 3.57 (t, 2H), 2.84 - 2.68 (m, 6H), 2.06 - 1.94 (m, 4H), 1.54 - 1.4 6 (m, 2H), 1.34 - 1.21 (m, 8H), 1.20 - 1.13 (m, 1H), 0.91 (t, 3H).

[0301] Step 4: To a solution of (8Z,11Z,14Z,17Z)-icosa-8,11,14 ,17-tetraen-1-ol (93 mg, 0.32 mmol) in toluene (3 mL) was added tert- butyl 2-bromobutanoate (143 mg, 0.64 mmol), tetrabutylammo nium hydroxide (TBAOH) (5 mg, 0.02 mmol) and NaOH (50 w / w%, 1 mL), and the mixture was heated to 45 °C. The mixture was stirred at 45 °C for 1.5 h After that, 71.4 mg (0.32 mmol) of tert-butyl 2-bromobutanoate was added to the reaction mixture. After 3 h, 71.4 mg (0. 32 mmol) of tert-butyl 2-bromobutanoate was added to the reaction mixture. After 8 h, 143 mg (0.64 mmol) of tert-butyl 2-bromobutano ate was added to the reaction mixture. After 24 h, the mixture was cooled to room temperature and water was added (40 mL). The resulting mixture was extracted twice with Et2O (25 + 40 mL) , and the combined organic extracts were washed with water (4 × 25 mL) and brine (25 mL), dried (MgSO4), filtered, and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (heptane → heptane:acetone - 5:1). Appropriate Pooled and concentrated the fractions to obtain 15 mg of tert-butyl 2-(((8Z,11Z,14 Z,17Z)-icosa-8,11,14,17-tetraen-1-yl)oxy)butano ate. This intermediate was dissolved in formic acid (5 mL), and the resulting mixture was stirred at room temperature for 1 hour and 45 minutes. The solvent was removed under vacuum, and EtOAc (30 mL) was added to the residue. The solution was washed with water (4 × 25 mL) and brine (25 mL), dried (MgSO4), filtered, and concentrated in vacuo under air. The residue was purified by preparative HPLC using 85% MeCN in water as the eluent. Appropriate fractions were pooled and concentrated to obtain 15 mg (0.04 mmol, yield 12%) of 2-(((8Z,11Z,14Z,17Z)-icosa-8,11,14,1 7-tetraen-1-yl)oxy)butanoic acid. 1H NMR (400 MHz, CDCl3) δ 1.0 0 (td, 6H), 1.49 - 1.20 (m, 8H), 1.64 (t, 2H), 1.96 - 1.73 (m, 2H), 2.08 (dt, 4H ), 2.84 (q, 6H), 3.55 - 3.43 (m, 1H), 3.65 - 3.54 (m, 1H), 3.87 (dd, 1H), 5.49 - 5.24 (m, 8H). HRMS (electrospray), [M-H] : calculated: 375.2899, found: 375. - 2892. 289.

Example

[0302] Preparation of 2-(((5Z,8Z,11Z,14Z)-icosa-5,8,11,14-tetraene -1-yl)oxy)butanoic acid

[0303]

Chemical formula

[0304] Step 2: To a solution of (5Z,8Z,11Z,14Z)-icos-5,8,11,1 4-tetraen-1-ol (349 mg, 1.20 mmol) in toluene (5 mL) were added tert-butyl 2-bromobutanoate (535 mg, 2.40 mmol), tetrabutylammonium hydroxide (TBAOH) (156 mg, 0.24 mmol, 40% w / w) and ​​NaOH (50 w / w%, 2 mL) was added, and the mixture was heated to 45 °C. After 12 hours , the mixture was cooled to room temperature, and ice water (25 mL) and Et2O (20 mL) were added. The phases were separated, and the aqueous phase was extracted with Et2O (20 mL). The combined organic extracts were washed with water (25 mL L) and brine (25 mL), dried (MgSO4), filtered, and evaporated in vacuo . The residue was dissolved in EtOH (20 mL) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (heptane:acetone - 1:20 → heptane:acetone - 1:10). Appropriate fractions were pooled and concentrated to give 360 mg (0. 83 mmol, 69% yield) of the intermediate tert-butyl 2-(((5Z,8Z,11Z, 14Z)-icosa-5,8,11,14-tetraen-1-yl)oxy)butanoate . 342 mg (0.79 mmol) of this intermediate was dissolved in formic acid (7 mL), and the resulting mixture was stirred at room temperature for 3 hours. The solvent was removed under vacuum, and the residue was taken up in EtOAc (75 mL ). The solution was washed with water (3 × 50 mL), dried (MgSO4), filtered, and concentrated in vacuo . The residue was purified by flash chromatography on silica gel (acetone:heptane :HOAc - 10:10:1 → acetone:heptane:HOAc - 30:70:1) . Appropriate fractions were pooled and concentrated to give 181 mg (0.48 mmol, 58% yield ) of 2-(((5Z,8Z,11Z,14Z)-icosa-5,8,11,14- tetraen-1-yl)oxy)butanoic acid .

Example

[0305] 2-(((7Z,10Z,13Z,16Z,19Z)-docosa-7,10,13,16 Preparation of (7Z,10Z,13Z,16Z,19Z)-docosa-7,10,13,16,19-pentaen-1-yl)oxy)butanoic acid

[0306]

Chem.

[0307] Step 2: To a solution of (7Z,10Z,13Z,16Z,19Z)-docosa-7,1 0,13,16,19-pentaen-1-ol (1.16 g, 3.66 mmol) in THF (50 ml) was added 2-bromobutanoic acid (0.918 g, 5.50 mmol). The reaction mixture was ​It was cooled to 8 - 10 °C. A solution of sodium tert - butoxide ( 0.986 g, 10.26 mmol) in MTBE (15 mL) was added over 5 minutes. The reaction mixture was stirred at 8 - 10 °C for 30 minutes. Additional sodium tert - butoxide (0.141 g, 1.466 mmol) in MTBE (4 mL) was added. The mixture was stirred at 8 - 10 °C for 30 minutes , and then 2 - bromobutanoic acid (0.612 g, 3.66 mmol) was added over 5 minutes . The mixture was then stirred at 8 - 10 °C for 30 minutes. Additional sodium tert - butoxide (0.282 g, 2.93 mmol) in MTBE (8 mL) was added. The mixture was stirred at 8 - 10 °C for 30 minutes. Additional sodium tert - butoxide (0.141 g, 1.466 mmol) in MTBE (4 mL) was added. The mixture was stirred at 8 - 10 °C for 30 minutes. HCOOH (0.5 mL) and then 1 M HCl(aq) were added to adjust the pH to ~ 2. The phases were separated and the aqueous phase was extracted with MTBE (2×50 mL). The combined organic phases were washed with water (50 mL), saturated NaHCO3 (4×30 ml) and brine (30 ml), dried (Na2SO4), filtered and concentrated in vacuo. 2 - (((7Z,10Z,13Z,16Z, 19Z) - docosa - 7,10,13,16,19 - pentaen - 1 - yl)oxy)butanoic acid (1 g, 2.439 mmol, yield 66.6%) was obtained as an oil by preparative HPLC using 90% MeCN (containing 0.02% HCOOH) in water (containing 10% MeCN and 0. 02% HCOOH) as the eluent. 1H NMR (400 MHz, CDCl3) δ 5.44 - 5.24 (m, 10H), 3.84 - 3.81 (m, 1H), 3.59 - 3.64 (m, 1H), 3 ​​.47 - 3.41 (m, 1H), 2.90 - 2.73 (m, 8H), 2.11 - 1.98 (m, 4H), 1.90 - 1.71 (m, 2H ), 1.64 - 1.57 (m, 2H), 1.41 - 1.26 (m, 6H), 0.98 - 0.93 (m, 6H). HRMS (electro spray), [M] + : Calculated: 402.3134, Found: 402.3141.

Example

[0308] (5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-penta en-1-yl 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8, 11,14,17-pentaen-1-yl)oxy)butanoate Preparation

[0309]

Chemical formula

Example

[0310] (2R)-2-((2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5, 8,11,14,17-pentaen-1-yl)oxy)butanoyl)oxy)butanoic acid Preparation

[0311]

Chemical formula

[0312] Step 2: (R)-1-(tert-Butoxy)-1-oxobutan-2-yl 2-(((5Z, 8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1 -yl)oxy)butanoate (161 mg, 0.312 mmol) was dissolved in formic acid (4 mL), and the reaction mixture was stirred at room temperature for 17 hours. EtOAc (10 mL) was added to the mixture, and the resulting mixture was washed with water (10 mL) and brine (10 mL). The organic phase was dried (Na 2SO4), filtered, and concentrated in vacuo. 60 mg (0.130 mmol, 41. 8% yield) of (2R)-2-((2-(((5Z,8Z,11Z,14Z,17Z)-icosa -5,8,11,14,17-pentaen-1-yl)oxy)butanoyl)oxy)butanoic acid was obtained. anoic acid was obtained. Tannic acid was isolated as an oil. MS (electrospray): 459.3 [M-H]-.

Example

[0313] (2S,3S,4S,5R,6S)-3,4,5-Trihydroxy-6-((2-(( (5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,17-penta en-1-yl)oxy)butanoyl)oxy)tetrahydro-2H-pyran-2-car Preparation of acid

[0314]

Chem.

[0315] Step 2: (4-Methoxycyclohexyl)methyl (2S,3S,4S,5R,6S)-3,4, 5-trihydroxy-6-((2-(((5Z,8Z,11Z,14Z,17Z)-icosa -5,8,11,14,17-pentaen-1-yl)oxy)butanoyl)oxy) tetrahydro-2H-pyran-2-carboxylate (1.20 g, 1.79 mmol) was treated with a solution of 10% TFA in DCM (8 mL) at 0 °C under a nitrogen atmosphere. After stirring at ambient temperature for 3 h, the solvent was removed in vacuo. Flash chromatography (DCM / MeOH / HCOOH - 95 / 5 / 0.2) gave 500 mg of an impure product. The product was further purified by preparative HPLC using water (containing 10% MeCN and 0.02% HCOOH) in 85% MeCN (0 .02% HCOOH) as the eluent. The appropriate fractions were pooled and concentrated to give (2S,3S,4S,5R,6S)- 3,4,5-trihydroxy-6-((2-(((5Z,8Z,11Z,14Z,17Z )-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoyl) )oxy)tetrahydro-2H-pyran-2-carboxylic acid (80 mg, 0.139 mmol , yield 7.8%) as a solid. 1H NMR (400 MHz, MeOD) δ 5.60 - 5.56 (m, 1H), 5.51 - 5.27 (m, 10H), 3.99 - 3.88 (m, 2H), 3.71 - 3.63 (m, 1H), 3.62 - 3.54 (m, 1H), 3.53 - 3.38 (m, 3H), 2.93 - 2.83 (m, 8H), 2.19 - 2.07 (m, 4H), 1.93 - 1.83 (m, 4H), 1.66 - 1.56 (m, 4H), 1.49 - 1.37 (m, 4H), 1.33 - 1.21 (m, 4H), 1.19 - 1.07 (m, 4H), 0.91 - 0.80 (m, 3H), 0.80 - 0.68 (m, 3H), 0.68 - 0.56 (m, 3H), 0.56 - 0.44 (m, 3H), 0.44 - 0.32 (m, 3H), 0.32 - 0.20 (m, 3H), 0.20 - 0.08 (m, 3H), 0.08 - 0.00 (m, 3H), 0.00 - 0.08 (m, 3H), 0.08 - 0.20 (m, 3H), 0.20 - 0.32 (m, 3H), 0.32 - 0.44 (m, 3H), 0.44 - 0.56 (m, 3H), 0.56 - 0.68 (m, 3H), 0.68 - 0.80 (m, 3H), 0.80 - 0.91 (m, 3H), 1.07 - 1.19 (m, 4H), 1.21 - 1.33 (m, 4H), 1.37 - 1.49 (m, 4H), 1.56 - 1.66 (m, 4H), 1.83 - 1.93 (m, 4H), 2.07 - 2.19 (m, 4H), 2.83 - 2.93 (m, 8H), 3.38 - 3.53 (m, 3H), 3.54 - 3.62 (m, 1H), 3.63 - 3.71 (m, 1H), 3.88 - 3.99 (m, 2H), 5.27 - 5.51 (m, 10H), 5.56 - 5.60 (m, 1H). (m, 1H), 1.81 - 1.72 (m, 1H), 1.69 - 1.60 (m, 2H), 1.53 - 1.45 (m, 2H), 1.04 - 0.98 (m, 6H). MS (Electrospray): 549.3 [M-H]-.

Example

[0316] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 7-pentaen-1-yl)oxy)butan-1-ol Preparation

[0317]

Chemical

[0318] 2-(((5Z,8Z,11Z,14Z,17Z)-Icosa-5,8,11,14,1 Preparation of 7-pentaen-1-yl)oxy)butyl acetate

[0319] [ka] 2-(((5Z,8Z,11Z,14Z,17Z)- in dry CH2Cl2 (5 ml) Icosa-5,8,11,14,17-pentaen-1-yl)oxy)butan-1-ol (300.9 mg, 0.834 mmol), triethylamine (140 ml, 1.00 A mixture of acetic anhydride (9 mmol) and DMAP (4.7 mg, 0.038 mmol) was The reaction mixture was stirred at room temperature under a N2 atmosphere for 80 minutes. Water (10 ml) was added and the mixture was extracted with heptane (2 x 50 ml) and brine (20 ml). The mixture was washed with water (Na2SO4), dried (Na2SO4), filtered and concentrated under reduced pressure. Rush chromatography eluted with heptane-EtOAc (100:1) to obtain 0.3 Obtained 19 g (95%) of the title compound as an oil. 1H NMR (400 MHz, CDCl3) δ 5.59 - 5.14 (m, 10H), 4.10 (dd, 1H), 4.02 (dd, 1H), 3.54 (dt, 1H), 3.43 (dt, 1H), 3.3 8 - 3.29 (m, 1H), 2.92 - 2.65 (m, 8H), 2.14 - 1.95 (m, 4H), 2.05 (s, 3H), 1.63 - 1.47 (m, 4H), 1.45 - 1.38 (m, 2H), 0.96 (t, 3H), 0.92 (t, 3H). MS (ESI, cationic) 425 [M+Na]+.

[0320] Biological Examples Evaluation of Compound A in a Diet-Induced NASH Mouse Model (CDAA / High-Fat Diet) In preclinical drug development, methionine and choline-deficient mouse models (MCD and CDAA, respectively) for studying the onset and treatment of NASH have been well established. As a precursor for phosphatidylcholine synthesis, insufficient intake of methionine / choline from diet results in the inability to synthesize hepatic lipoproteins for triglyceride excretion, leading to severe fatty liver, inflammation, and fibrosis. The widely used methionine-choline-deficient (MCD) diet consistently reproduces severe NASH-like liver inflammation and fibrosis in mice but is also associated with severe weight loss (decrease in both skeletal muscle and body fat mass).

[0321] This is associated with an increased risk of death and is a major problem in long-term fibrosis experiments. By adding suboptimal doses of methionine (0.17%), the CDAA diet model overcomes these problems and successively induces steatohepatitis, liver fibrosis, and liver cancer in both mice and rats without severe weight loss, mimicking human NASH. ​​​

[0322] In Biological Examples 1 to 7, investigations were conducted in male C56BL / 6J mice (9 weeks old). They were given either a choline-supplemented (CS) diet or a choline-deficient high-fat diet (31% total calories; "CDAA / high-fat"). Rather than a preventive approach, the investigation was designed as a treatment / recovery setting, and a NASH-inducing diet was started 6 weeks before the start of treatment.

[0323] In Biological Examples 1 to 5, mice were divided into 6 experimental groups (n = 9 per group) and given either the CS diet or the CDAA / high-fat diet. Six weeks after either the CS diet or the CDAA diet, compound A or comparative compound N was orally administered to the mice at two doses, 0.15 mmol / kg bw / day (low dose, LD ) and 0.3 mmol / kg bw / day (high dose, HD). The test substances were orally administered as mixtures in the high-fat diet. Both hydroxyproline (HYP) content and Sirius Red (SR) morphometry were measured to evaluate liver fibrosis. Transcript levels related to fibrosis, inflammation, and metabolism were measured by quantitative real-time polymerase chain reaction (qPCR).

[0324] In Biological Examples 6 and 7, the mice were given either the CS diet or the CDAA / high-fat diet as described above. In Example 6, 6 weeks after either CS or the CDAA / high-fat diet, compound A at 56 mg / kg bw / day delivered orally was administered to the mice, or the long-acting GLP-1R agonist Bydureon (trademark ) (exenatide), which is approved for the treatment of type 2 diabetes, was subcutaneously injected into the mice at 0.4 mg / kg / week for 6 weeks. The vehicle control ​Isolation from the group given saline (n = 8), compound A (n = 9) and GLP-1R (n = 8) In liver samples, the effects on liver collagen fiber number, thickness and length were determined by a non-linear multi-photon optical imaging system.

[0325] In biological example 7, after 6 weeks on either CS or CDAA / high-fat diet, 112 mg / kg bw / day of compound A, or an equimolar dose of eicosapentaenoic acid (EPA) delivered orally was administered to mice. Liver lipidomic analysis was performed on isolated liver samples from the CS (n = 9) group, compound A (n = 9) group and E PA (n = 8) group.

[0326] In biological examples 21 - 24, the effects of compound A alone and compound A in combination with exenatide, a GLP-1 agonist, were investigated. Male C56BL / 6J mice (9 weeks old ) were investigated. As described above, mice were divided into 6 experimental groups (n = 9 per group) containing either the CS diet or the CDAA / high-fat diet. To evaluate treatment / amelioration rather than prevention, the NASH-inducing CDAA / high-fat diet was started 6 weeks before the start of treatment. Thus, after 6 weeks on either CS or CDAA / high-fat diet, (A) oral compound A at only 0.3 mmol / kg bw / day, (B) GLP-1 agonist alone delivered i.p. at 0.4 mg / kg once a week, or (C) compound A at 0.3 mmol / kg bw / day in combination with a GLP-1 agonist delivered i.p. at 0.4 mg / kg once a week (reduced to 0.1 mg / kg for the last 3 weeks due to excessive weight loss) were used to treat the mice. Compound A was administered orally as a mixture in the high-fat diet, while the GLP-1 a g onist was delivered i.p. For the last 3 weeks, the dose was reduced to 0.1 mg / kg to avoid excessive weight loss). Compound A was administered orally as a mixture in the high-fat diet, while the GLP-1 a Gonist was delivered by intraperitoneal injection (i.p.). Transcript levels related to fibrosis, inflammation and metabolism were measured by quantitative real-time polymerase chain reaction (qPCR) in isolated liver tissues. In Biological Example 8, pathogen-free pregnant day 15 C57BL / 6 mice were obtained from Charles River Laboratories Inc. (Kanagawa, Japan). NASH was established in male mice by a single subcutaneous injection of streptozotocin (STZ) (Sigma, USA) after birth. After 4 weeks of age (day 28 ± 2), a high-fat diet (HFD; CLEA, Japan) was given ad libitum.

[0327] Evaluation of Compound A in a Streptozotocin-Injected / High-Fat Diet-Induced NASH Model (STAM Model) Evaluation of the Effect of Compound A in a Diet-Induced NASH Mouse Model (APOE Treatment: On the day before starting the vehicle (saline), compound A and telmisartan (positive control), 6-week-old mice were randomly divided into 3 groups of 8 mice. After treatment with compound A at a dose of 37 mg / kg bw / day for 6 weeks by oral gavage in combination with a high-fat diet, the mice were sacrificed. The NAFLD activity score (NAS) was calculated according to standardized criteria, and the fibrosis area was evaluated by Sirius Red staining.

[0328] 3Leiden.CETP Double Trans * genic Mice) Evaluation of the Effect of Compound A in an Obesity Diet-Induced NASH Mouse Model (ob / ob AMLN Model) APOE * APOE3Leiden.CETP double transgenic mice express APOE3Leiden, a mutant of human apolipoprotein E3 (APOE3), in addition to human apolipoprotein C1 (APOC1) and CETP. * APOE * 3Le ​​​​​​​​​iden. CETP double transgenic mice show elevated plasma cholesterol and triglyceride levels mainly limited to the VLDL / LDL size lipoprotein fraction. By increasing the cholesterol content of the diet in this model, all the features of human NASH appear. In Biological Example 9, APOE3 Leiden.CETP mice were investigated with high-fat diets (24% fat w / w) with various cholesterol contents (0.25 - 1% cholesterol w / w). In one study (using 1% cholesterol w / w), after a 3-week lead-in period, low-responding mice (20% of the total) were excluded from the study, and the remaining mice were subdivided into 4 groups of 12 mice each ( + 5 mice in the control group) with matched plasma cholesterol, triglycerides, blood glucose, body weight, and age (t = 0), and treatment was initiated.

[0329] Mice were gavaged daily between 07:00 and 10:00 with either 0.3 mmol / kg bw / day of Compound A, the comparative compound Compound N (0.3 mmol / kg bw / day), rosiglitazone (13 mg / kg bw / day), or control (corn oil). After 20 weeks of treatment, the mice were sacrificed by CO2 asphyxiation, the livers were harvested, and the level of hepatocyte hypertrophy was graded. * 3Leiden.CET In Biological Example 25, the effects of Compound A alone and Compound A in combination with omega-3 fatty acids were investigated. APOE3 Leiden.CETP mice were given a semi-synthetic high-fat diet (24% fat w / w) with 0.25 cholesterol w / w. After a 4-week lead-in period, low-responding mice (20% of the total) were excluded from the study, and the remaining mice were subdivided into 4 groups of 12 mice each ( + 5 mice in the control group) with matched plasma cholesterol, triglycerides, blood glucose, body weight, and age (t = 0), and treatment was initiated. Mice were gavaged daily between 07:00 and 10:00 with either 0.3 mmol / kg bw / day of Compound A, the comparative compound Compound N (0.3 mmol / kg bw / day), rosiglitazone (13 mg / kg bw / day), or control (corn oil). .3 mmol / kg bw / day of Compound A, the comparative compound Compound N (0.3 mmol / kg bw / day), rosiglitazone (13 mg / kg bw / day), or control (corn oil). After 20 weeks of treatment, the mice were sacrificed by CO2 asphyxiation, the livers were harvested, and the level of hepatocyte hypertrophy was graded. Mice were gavaged daily between 07:00 and 10:00 with either 0.3 mmol / kg bw / day of Compound A, the comparative compound Compound N (0.3 mmol / kg bw / day), rosiglitazone (13 mg / kg bw / day), or control (corn oil). After 20 weeks of treatment, the mice were sacrificed by CO2 asphyxiation, the livers were harvested, and the level of hepatocyte hypertrophy was graded. After 20 weeks of treatment, the mice were sacrificed by CO2 asphyxiation, the livers were harvested, and the level of hepatocyte hypertrophy was graded.

[0330] In Biological Example 25, the effects of Compound A alone and Compound A in combination with omega-3 fatty acids were investigated. APOE3 Leiden.CETP mice were given a semi-synthetic high-fat diet (24% fat w / w) with 0.25 cholesterol w / w. * 3Leiden.CETP mice were given a semi-synthetic high-fat diet (24% fat w / w) with 0.25 cholesterol w / w. After a 4-week lead-in period, low-responding mice (20% of the total) were excluded from the study, and the remaining mice were subdivided into 4 groups of 12 mice each ( + 5 mice in the control group) with matched plasma cholesterol, triglycerides, blood glucose, body weight, and age (t = 0), and treatment was initiated. Excluding the responder mice from the study, plasma cholesterol, triglyceride, blood glucose, body weight and age (t = 0) were matched, and the remaining mice were subdivided into groups of 8 mice each and treatment was initiated. Sunflower oil was added to facilitate compound mixing, resulting in a diet with a total oil content of 10 mL / kg. Compound A was administered at 0.3 mmol / kg bw / day, while omega-3 fatty acid ethyl ester (85% w / w EPA / DHA ethyl ester) was administered at 3.0 mmol / kg bw / day. Both Compound A and omega-3 ethyl ester were administered orally as a mixture in a high-fat diet. After 4 weeks of treatment, the mice were sacrificed by CO2 asphyxiation, the livers were harvested, and after extraction and separation by high-performance thin-layer chromatography, the hepatic contents of free cholesterol, cholesterol ester and triglyceride were

[0331] In Vitro Evaluation of the Effect of Compound A on Human Hepatic Stellate Cells Biological Example 1. Effect of Compound A and Compound N on Hepatic Fibrosis Measured by Total Hydroxyproline (HYP) Content per Liver Cholesterol (2%), 40% fat (including 18% trans fatty acids) and 20% fructose are added to the high-calorie diet (i.e., AMLN high-fat diet), and obese diet-induced B 6.V-Lepob / Jrj mice (ob / ob) are consistently prone to fibrosis (liver). Ob / ob mice fed the AMLN diet (referred to as "ob / ob AMLN mice") develop steatohepatitis and fibrosis within a shorter time frame (within 12 weeks) compared to wild-type C57BL / 6 mice fed the same diet (AMLN mice). Ob / ob AMLN mice provide an obese diet-induced model of NASH, which also allows investigation of the effects on blood glucose control. The development of NASH in the ob / ob AMLN model is confirmed by biopsy. ​

[0332] In Biological Examples 10 to 12, 15, and 19, male ob / ob mice (5 weeks old) were fed an AMLN high-fat diet for 15 weeks, then randomly divided into 3 groups (n = 10 per group), and for an additional 4 weeks, received via diet either 11 2 mg / kg bw / day of Compound A, 30 mg / kg bw / day of pioglitazone, or no treatment (vehicle). An oral glucose tolerance test (OGTT) was performed at week 3. Liver fibrosis was evaluated by measuring hepatic fibrogenesis / fibrolysis gene expression by RNA sequencing over 4 weeks.

[0333] In Biological Examples 10, 13 to 14, and 16 to 18, male ob / ob mice (5 weeks old) were fed an AMLN high-fat diet for 18 weeks, then randomly divided into 6 groups (n = 10 per group), and for an additional 8 weeks, received via diet Compound A at 3 different daily doses (45 mg / kg, 90 mg / kg, or 135 mg / [[ID=2I]] kg), 30 mg / kg bw / day of obeticholic acid (OCA), or no treatment (vehicle). At the end of the study, NASH-related parameters and fibrosis were evaluated by quantitative immunohistochemistry (IHC) measurements.

[0334] For clinical bridging, the doses of Compound A used here (45 mg / kg, 90 mg / kg, and 135 mg / kg in the 8-week study and 112 mg / kg in the 4-week study), when corrected for interspecies differences, are approximately 250 mg / day, 500 mg / day, and 750 mg / day (and 600 mg / day in the 4-week study) for a 70-kg human. ​​​​​​​corresponds to the human dose (mg / day). For guidance on interspecies dose conversion, see Nair, A.B . and Jacob, S., J Basic Clin Pharm, 2016;7(2 ):27-31.

[0335] Biological Example 2. Effect of Compound A and Compound N on Hepatic Fibrosis Measured by Relative Hydroxyproline (HYP) Content (μg HYP per 100 mg of Liver) Stellate cells are resident hepatocytes and are the major cell type involved in the initiation and progression of liver fibrosis . In Biological Example 20, an in vitro investigation was performed on cultured human stellate cells (LX-2 cells) to evaluate the direct effect of Compound A on these cells. The cells were treated with Compound A (10 μM, 25 μM, 50 μM or 75 μM) or oleic acid (OA; 75 μM) for 24 hours, and bromodeoxyuridine (BrdU) incorporation was evaluated to determine the effect on proliferation . The MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3 -carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium ) assay was used to determine cell viability.

[0336] Biological Example 3. Effect of Compound A and Compound N on Hepatic Inflammatory Gene Expression (TNF-α mRNA Expression) Both low dose (LD) and high dose (HD) of Compound A administered to CDAA / high fat diet mice induced a significant reduction in liver fibrosis (HYP content per liver) compared to the control (p < 0.01 and 0.001 * respectively ** ). No significant effect was observed for the comparative Compound N . The effect of Compound A on liver HYP content is shown in Table 1.

[0337]

Table 1

[0338] Biological Example 4. Effect of Compound A and Compound N on Fibrosis-Related Gene Expression (Col1a1 mRNA Expression) Both low dose (LD) and high dose (HD) of compound A administered to CDAA / high-fat diet-fed mice reduced hepatic fibrosis (HYP content per liver), whereas no effect was observed for compound N. The effect of compound A on relative HYP content is shown in Table 2.

[0339] [Table 2]

[0340] Biological Example 5. Effect of Compound A on Hepatic Fibrosis Measured by Sirius Red Morphometry Both low dose (LD) and high dose (HD) of compound A administered to CDAA / high-fat diet-fed mice induced a significant decrease in hepatic inflammation (TNF-a mRNA) compared to the control ( both p<0.001 *** ). A significant decrease (p<0001) was also observed for compound N, but not for the high dose (HD). The effect of compound A on hepatic TNF-a is shown in Table 3.

[0341] [Table 3]

[0342] Biological Example 6. Effect of Compound A and Bydureon (Registered Trademark ) on Collagen Fibers Both low dose (LD) and high dose (HD) of compound A administered to CDAA / high-fat diet-fed mice induced a significant reduction in hepatic fibrosis-related gene expression (Col1a1 mRNA) compared to the control (both p<0.001 *** ). For compound N, a significant ​​​No effect was observed. The effect of compound A on hepatic Col1a1 is shown in Table 4.

[0343]

Table 4

[0344] Biological Example 7. Effect of Compound A and EPA on Hepatic Lipid Metabolism Biological Example 8. Effect of Compound A on Hepatocyte Ballooning, Lobular Inflammation, Steatosis, Composite NAS Score, and Fibrosis Area To more accurately quantify the amount, location, and functional relevance of hepatic fibrosis, Sirius Red (SR) morphometry was performed. For Examples 1 and 2 that used biochemical evaluation of hydroxypro line (HYP) content to measure collagen in large blood vessels, SR morphometry quantifies periportal collagen deposition with higher functional relevance. Using a magnification of 100x ten regions per slide were measured. Both low dose ( LD) and high dose (HD) of compound A administered to CDAA / high-fat diet mice induced a significant decrease in the surface area percentage of hepatic fibrosis compared to the control (p < 0.001 *** and 0.05 * ), respectively). The effect of compound A on hepatic fibrosis measured by SR morpho metry is shown in Table 5.

[0345]

Table 5

[0346] Biological Example 9. Effect of Compound A and Compound N on Hepatocyte Hypertrophy Biological Example 10. Effect of Compound A, Pioglitazone, and OCA When administered at a low dose (56 mg / kg) to CDAA / high-fat diet mice, compound A significantly decreased the number of collagen fibers by 51% (p = 0.01) (Figure 1A). This effect is the slight decrease in fiber thickness and length by compound A in Figure 1B and Figure 1C, respectively As reflected in the lack of increase, there was an almost complete disappearance of short (<8.5 μm) and thin (<3.5 μm) collagen fibers. Bydureon (trademark) (injected subcutaneously at 0.4 mg / kg weekly) had no significant effect.

[0347] on Liver Weight and Body Weight To further understand the effect of Compound A on hepatic lipid metabolism, lipidomic analysis of CS- and CDAA / high-fat diet-fed mice ("CD") was performed on isolated liver tissues. When administered at a high dose (112 mg / kg) to CDAA / high-fat diet-fed mice, Compound A significantly reduced the effects of choline deficiency as measured by hepatic triglyceride (TG) (Figure 2A), hepatic diglyceride (DG) (Figure 2B), hepatic free fatty acid (FFA )(Figure 2C) or cholesteryl ester (Figure 2D). In contrast, treatment with an equimolar amount of EPA did not reduce the increase in hepatic TG, DG, FFA or cholesteryl ester associated with choline-deficient diet. (Figure 2D). (Figure 2D).

[0348] Biological Example 11. Effect of Compound A and Piog litazone on Hepatic Fibrogenesis and Fibrolysis Gene Expression Compound A administered to STAM (model) mice reduced ballooning of hepatocytes in addition to steatosis and lobular inflammation. Both the composite NAS score (p<0.001 ) and the fibrosis area * (Sirius Red positive area) were reduced by Compound A. (Figure 2D).

[0349] [Table 6]

[0350] Biological Example 12. Effect of Compound A and Pioglitazone on Hepatic Extracellular Matrix Stability and Fibrolysis Gene Expression ApoE * ​Administered to 3L-CETP mice (0.25% cholesterol diet w / w) Compound A induced a significant decrease in hepatocyte hypertrophy (p<0.01 * ), a 83% decrease compared to the control (p<0.01). No effect was observed with Compound N.

[0351]

Table 7

[0352] Biological Example 13. Effect of Compound A and OCA on Hepatic Fibrosis Progression Biological Example 14. Effect of Compound A and OCA on Regression of Hepatic Fibrosis As shown in Fig. 3A, after 4 weeks, Compound A (112 mg / kg) had no effect on the body weight of ob / ob AMLN mice, while pioglitazone induced a significant 15 % increase in body weight (p<0.01). As shown in Fig. 3B, liver weight did not change in all groups at week 4. As shown in Fig. 3C, after 8 weeks, neither Compound A (45 mg / kg, 9 0 mg / kg and 135 mg / kg) nor OCA had a significant effect on body weight, and food intake was not affected in any of the groups. As shown in Fig. 3D, OCA significantly (p<0.05) decreased liver weight after 8 weeks, while Compound A had no significant effect at any dose compared to the vehicle.

[0353] Biological Example 15. Effect of Compound A and Pioglitazone on Inflammatory Gene Expression ​ The effects of 4-week treatment with either Compound A (112 mg / kg) or pioglitazone (30 mg / kg) on fibrosis and fibrolysis gene expression in ob / ob AMLN mouse isolated liver samples are shown in Figs. 4A - 4F. Compound A decreased platelet-derived growth ​​​​Platelet-derived growth factor-beta (PDGF-β) (Figure 4A), platelet-derived growth factor receptor-β (PDGF R-β) (Figure 4B), type I collagen α1 (col1A1) (Figure 4C), col3A1( Figure 4D), col4A1 (Figure 4E) and heat shock protein-47 (HSP47)( Figure 4F), significantly decreased the expression of classical genes that control liver fibrosis. Pioglitazone significantly decreased the expression of the collagen isoforms investigated (col1A1, col3A1, col4A 1), while HSP47 had no effect on the expression of PDGF-β or PDGFR-β .

[0354] ​ ​ As shown in Figures 5A to 5D, for genes that control extracellular matrix (ECM) stability or fibrinolysis, four-week treatment with compound A significantly decreased the hepatic expression of lysyl oxidase (LOX) (Figure 5A), LOX-like 2 (LOXL2) (Figure 5B) and L OXL1 (Figure 5C) as well as tissue inhibitor of metalloproteinase (TIMP1) (Figure 5D ). Pioglitazone also significantly reduced the transcript levels of these transcripts, but the effect was milder than that obtained with compound A.

[0355] ​ To evaluate the effect of 8-week administration of either compound A (45 mg / kg , 90 mg / kg or 135 mg / kg) or OCA (30 mg / kg) on the progression of liver fibrosis in ob / ob AMLN mice, quantitative immunohistochemistry (IHC) was performed to measure the col1A1 and α-smooth muscle actin (SM A) content. As shown in Figures 6A and 6B ​​As shown, at doses of 90 mg / kg and 135 mg / kg, compound A significantly reduced α-SMA, a marker of myofibroblasts (expressed as both total amount and percentage of fractional area content), while OCA showed no significant effect. Similarly, compound A administered at 90 and 135 mg / kg reduced the total col1A1 content (Figure 6C), while OCA had no significant effect. When expressed as a percentage of the total surface area (Figure 6D), a dose of 90 mg / kg induced a significant reduction in col1A1 levels, while the reductions at doses of 45 and 135 mg / kg were not statistically significant. To confirm the anti-fibrotic effect shown by quantitative IHC, as shown in Figures 6E and 6F, the hepatic concentration of hydroxyproline (HYP) was measured. Only compound A reduced the HYP content, expressed as either total content (p < 0.01 for both doses of 90 and 135 mg / kg) or relative content (p < 0.05 for both doses of 90 and 135 mg / kg). When performing baseline (pre-treatment) biopsies of ob / ob AMLN mice incorporating IHC measurements of both hepatic col1A1 and α-SMA, the final values of col1A1 and α-SMA were compared to the baseline (pre-treatment biopsy) values in the entire group to confirm whether the significant reduction relative to the vehicle observed by the inventors after treatment with compound A at the end of the study was associated with a reduction in fibrosis. As shown in Figure 7A, treatment with compound A at 135 mg / kg

[0356] significantly reduced α-SMA, a marker of myofibroblasts (expressed as both total amount and percentage of fractional area content), while OCA showed no significant effect. Similarly, compound A administered at 90 and 135 mg / kg reduced the total col1A1 content (Figure 6C), while OCA had no significant effect. When expressed as a percentage of the total surface area (Figure 6D), a dose of 90 mg / kg induced a significant reduction in col1A1 levels, while the reductions at doses of 45 and 135 mg / kg were not statistically significant. To confirm the anti-fibrotic effect shown by quantitative IHC, as shown in Figures 6E and 6F, the hepatic concentration of hydroxyproline (HYP) was measured. Only compound A reduced the HYP content, expressed as either total content (p < 0.01 for both doses of 90 and 135 mg / kg) or relative content (p < 0.05 for both doses of 90 and 135 mg / kg). When performing baseline (pre-treatment) biopsies of ob / ob AMLN mice incorporating IHC measurements of both hepatic col1A1 and α-SMA, the final values of col1A1 and α-SMA were compared to the baseline (pre-treatment biopsy) values in the entire group to confirm whether the significant reduction relative to the vehicle observed by the inventors after treatment with compound A at the end of the study was associated with a reduction in fibrosis. As shown in Figure 7A, treatment with compound A at 135 mg / kg significantly reduced α-SMA, a marker of myofibroblasts (expressed as both total amount and percentage of fractional area content), while OCA showed no significant effect. Similarly, compound A administered at 90 and 135 mg / kg reduced the total col1A1 content (Figure 6C), while OCA had no significant effect. When expressed as a percentage of the total surface area (Figure 6D), a dose of 90 mg / kg induced a significant reduction in col1A1 levels, while the reductions at doses of 45 and 135 mg / kg were not statistically significant. To confirm the anti-fibrotic effect shown by quantitative IHC, as shown in Figures 6E and 6F, the hepatic concentration of hydroxyproline (HYP) was measured. Only compound A reduced the HYP content, expressed as either total content (p < 0.01 for both doses of 90 and 135 mg / kg) or relative content (p < 0.05 for both doses of 90 and 135 mg / kg). When performing baseline (pre-treatment) biopsies of ob / ob AMLN mice incorporating IHC measurements of both hepatic col1A1 and α-SMA, the final values of col1A1 and α-SMA were compared to the baseline (pre-treatment biopsy) values in the entire group to confirm whether the significant reduction relative to the vehicle observed by the inventors after treatment with compound A at the end of the study was associated with a reduction in fibrosis. As shown in Figure 7A, treatment with compound A at 135 mg / kg significantly reduced α-SMA, a marker of myofibroblasts (expressed as both total amount and percentage of fractional area content), while OCA showed no significant effect.

[0357] ​ significantly reduced α-SMA, a marker of myofibroblasts (expressed as both total amount and percentage of fractional area content), while OCA showed no significant effect. Similarly, compound A administered at 90 and 135 mg / kg reduced the total col1A1 content (Figure 6C), while OCA had no significant effect. When expressed as a percentage of the total surface area (Figure 6D), a dose of 90 mg / kg induced a significant reduction in col1A1 levels, while the reductions at doses of 45 and 135 mg / kg were not statistically significant. To confirm the anti-fibrotic effect shown by quantitative IHC, as shown in Figures 6E and 6F, the hepatic concentration of hydroxyproline (HYP) was measured. Only compound A reduced the HYP content, expressed as either total content (p < 0.01 for both doses of 90 and 135 mg / kg) or relative content (p < 0.05 for both doses of 90 and 135 mg / kg). When performing baseline (pre-treatment) biopsies of ob / ob AMLN mice incorporating IHC measurements of both hepatic col1A1 and α-SMA, the final values of col1A1 and α-SMA were compared to the baseline (pre-treatment biopsy) values in the entire group to confirm whether the significant reduction relative to the vehicle observed by the inventors after treatment with compound A at the end of the study was associated with a reduction in fibrosis. As shown in Figure 7A, treatment with compound A at 135 mg / kg significantly reduced α-SMA, a marker of myofibroblasts (expressed as both total amount and percentage of fractional area content), while OCA showed no significant effect. The α-SMA content evaluated by immunohistochemical staining was decreased compared to the α-SMA content in the biopsy. The area ratio of was decreased. Compounds A at doses of 45 mg / kg and 90 mg / kg showed a tendency that was not significant. OCA had no significant effect. Similarly, all doses of compound A showed a tendency towards a decrease in col1A1 content compared to the pre-biopsy content, but only the dose of 9 0 mg / kg was statistically significant (p < 0.005) (Figure 7B). After 8 weeks of administration period, both vehicle treatment and OCA treatment showed a slight increase in hepatic col1A1 content although the increase was significant only for OCA (p < 0.05).

[0358] ​ Among the specific associations with the progression of fibrosis in ob / ob AMLN mice, 4-week treatment with compound A at 112 mg / kg significantly decreased the expression of inflammatory genes compared to vehicle-administered control mice. Specifically, compound A reduced the transcript levels of both transforming growth factor beta 1 (TGF-β1) (p < 0.05) and TGF-β1 receptor (TGFRβ) (p < 0.01) as shown in Figures 8A and 8B respectively . Pioglitazone decreased the expression of TGFRβ (p < 0.05) but did not decrease the expression of TGF- β1. As shown in Figures 8C, 8D and 8F, after compound A treatment , significant decreases (all p < 0.001) in CCR2, MAC-2 (data not shown), CD68 and CD14 were observed compared to the control . Compound A treatment also significantly reduced galectin-3 (Gal-3) levels compared to vehicle-administered control (Figure 8E). Pioglitazone decreased CCR2 (p < 0.01) (Figure 8C), CD68 (p < 0.01) (Figure 8D) and CD14 (p<0.001) (Fig. 8F). did not significantly reduce interleukin (IL)-1β or MCP-1 transcript levels. (data not shown).

[0359] Biological Example 16. Effects of Compound A and OCA on Liver Inflammation Measured by Liver Gal-3 Effect of A Compound A (45 mg / kg, 90 mg / kg) on hepatic inflammation in ob / ob AMLN mice mg / kg or 135 mg / kg) or OCA (30 mg / kg) To assess the effect of 8 weeks of treatment, IHC was performed to detect galectin-3 (Gal-3). As shown in Figures 9A and 9B, treatment with Compound A significantly increased the total or The level of Gal-3 expression measured either as a percentage of the area or as a function of the surface area was decreased in a dose-response manner. whereas OCA reduced Gal-3 expression levels when measured as total content. It was just reduced.

[0360] Biological Example 17. Effects of Compound A and OCA on Hepatocyte Injury Compound A and P1000 on hepatocellular injury or damage in ob / ob AMLN mice To assess the effect of oglitazone, plasma alanine aminotransferase was assessed after 8 weeks of treatment. ALT and aspartate transaminase (AST) were measured. As shown in Figure 10A, a marked and significant decrease in ALT was observed at doses of 90 mg / kg and 135 mg / kg. / kg Compound A (p<0.01 and p<0.001, respectively) At doses of 90 mg / kg and 135 mg / kg, the compound Substance A also significantly reduced AST (p<0.05 and p<0.01, respectively), but OC A showed no effect on AST levels (Fig. 10B).

[0361] Biological Example 18. Effects of Compound A and OCA on Fatty Liver, Liver Lipid Levels, and Plasma Lipid Levels and OCA As shown in FIGS. 11A and 11B, both compound A and OCA significantly reduced adiposity (expressed as either percent area or total lipid content) in ob / ob AMLN mice after 8 weeks of administration and OCA showed efficacy equivalent to that of compound A at the highest dose (135 mg / kg) (both p<0.001). Similarly, as shown in FIGS. 11D and 11E, both OCA and compound A (at doses 90 mg / kg and 135 mg / kg) showed comparable reduction in plasma triglyceride levels and total cholesterol levels and OCA treatment, but not compound A treatment, showed a decrease in total hepatic cholesterol content (p<0.01) (FIG. 11C). After 3 weeks of administration of compound A (112 mg / kg), pioglitazone or vehicle control, an oral glucose tolerance test (OGTT) was performed in ob / ob AMLN mice As shown in FIG. 12A, both compound A and pioglitazone significantly reduced blood glucose fluctuations over the first 240 minutes after glucose challenge (AUC 0-240 minutes), but the effect of pioglitazone was more pronounced Similarly, a significant reduction in fasting plasma glucose levels was achieved by both compounds, but the effect of pioglitazone was more pronounced compared to compound A (FIG. 12B). Pioglitazone, but not compound A, decreased fasting insulin (p<0.001) (FIG. 12C).

[0362] Biological Example 19. Effects of Compound A and Pioglitazone on Blood Glucose Control

[0363] Biological Example 20. Effects of Compound A and Oleic Acid In Vitro on Stellate Cell Proliferation and Survival Rate and Oleic Acid ​​​​​​​​​​ LX-2 human hepatic stellate cells were treated with 10 - 75 μM of compound A or oleic acid (OA) (75 μM) for 24 hours and cell proliferation was evaluated by BrdU incorporation. Results are normalized mean ± S.E.M. of five independent experiments for eicosapentaenoic acid and two independent experiments for OA. As shown in Fig. 13A, LX-2 cells treated with 25 μM (p < 0.005) , 50 μM (p < 0.0001) and 75 μM (p < 0.005) of compound A were less proliferative compared to OA or vehicle. The decrease in proliferation by compound A was 25 - 34% at 24 hours, while OA had no effect. Comparisons were made by one-way ANOVA and Dunnett's correction for multiple comparisons. As shown in Fig. 13B, neither compound A nor OA had a significant effect on cell viability measured by MTS assay in two independent experiments

[0364] Biological Example 21. Effects of Compound A Alone and in Combination with GLP-1 Agonists (GLP-1a, Exenatide) on Liver Fibrosis-Related Gene Expression (Col1a1 mRNA Expression) Alone and in Combination of Compound A and GLP-1 Agonists (GLP-1a, Exenatide) Compound A, GLP-1a and compound A + GLP-1a combination administered to CDAA / high-fat diet mice induced a significant decrease in Col1a1 mRNA expression compared to CDAA-fed mice (all p < 0.001 )). The effects of compound A alone and compound A in combination with GLP-1a on hepatic Col1a1 mRNA are shown in Table 8 ***

[0365]

Table 8

[0366] Biological Example 22. Effects of Compound A Alone and in Combination with GLP-1 Agonists (GLP-1a, Exenatide) on Liver Fibrosis-Related Gene Expression (TIMP (Tissue Inhibitor of Matrix Metalloproteinase)-1a mRNA Expression) ​​​​​​​​​​Alone and in Combination of Compound A and GLP-1 Agonists (GLP-1a, Exenatide) Compound A, GLP-1a, and the compound A + GLP-1a combination administered to CDAA / high-fat diet-fed mice induced a significant decrease in hepatic TIMP-1a mRNA expression in CDAA diet-fed mice (all p < 0.001 ). The effects of compound A alone and compound A in combination with GLP-1a on hepatic TIMP-1a mRNA are shown in Table 9. *** )

[0367]

Table 9

[0368] Biological Example 23. Effects of Compound A Alone and in Combination with GLP-1 Agonists (GLP-1a, Exenatide) on Liver Fibrosis-Related Gene Expression (MMP (Matrix Metalloproteinase)-13 mRNA Expression) Alone and in Combination of Compound A and GLP-1 Agonists (GLP-1a, Exenatide) Compound A, GLP-1a, and the compound A + GLP-1a combination administered to CDAA / high-fat diet-fed mice induced a significant decrease in hepatic MMP-13 mRNA expression in CDAA diet-fed mice (all p < 0.001 ). The effects of compound A alone and compound A in combination with GLP-1a on hepatic MMP-13 mRNA are shown in Table 10. *** )

[0369]

Table 10

[0370] Biological Example 24. Effects of Compound A Alone and in Combination with GLP-1 Agonists (GLP-1a, Exenatide) on Liver Inflammation-Related Gene Expression (TNF-α mRNA Expression) Alone and in Combination of Compound A and GLP-1 Agonists (GLP-1a, Exenatide) Compound A, GLP-1a, and the compound A + GLP-1a combination administered to CDAA / high-fat diet-fed mice induced a significant ​​Induced reduction (for compound A alone or compound A combined with GLP-1a, p <0.001; for GLP-1a alone, p < 0.05). The effects of compound A alone and compound A combined with GLP-1a on hepatic TNF-α mRNA are shown in Table 11.

[0371]

Table 11

[0372] Biological Example 25. Effects of Compound A Alone and in Combination with High-Concentration Omega-3 Ethyl Ester on Liver Lipids (Cholesterol Ester-CE, Free Cholesterol-FC, Triglyceride-TG) Alone and in Combination of Compound A and High-Concentration Omega-3 Ethyl Ester A Western diet containing 0.25% cholesterol combined with high-dose (3 mmol / kg bw / day) 85% EPA / DHA ethyl ester omega -3 (HD) was administered to ApoE * 3L-CETP transgenic mice. Low-dose (0.3 mmol / kg bw / day) compound A (LD) induced significant reductions in FC (p < 0.01), CE (p < 0.001), and TG (p < 0.01). In this model, no compound significantly reduced hepatic TG as monotherapy.

[0373]

Table 12

[0374] Biological Example 26. Multicenter, Randomized, Double-Blind, Placebo-Controlled, Parallel-Group Trial of Compound A in Patients with Non-Alcoholic Steatohepatitis (NASH) Patients The goal of this clinical trial is to evaluate the efficacy of different doses of compound A in restoring NASH in patients without exacerbation of fibrosis, and to determine the safety and tolerability of compound A in patients with NASH.

[0375] ​​​​ Treatment Allocation Screen approximately 30 locations for eligibility in 600 patients with NASH and it is expected that approximately 264 will meet the selection criteria outlined below, and approximately 198 are expected to complete the full trial (giving 66 subjects per treatment arm assuming a 25% dropout of patients).

[0376] Investigation Design Randomly divide the patients determined to be eligible for the study into three parallel groups of 88 patients each. Administer placebo compound A to group 1, 300 mg of compound A to group 2, and 600 mg of compound A to group 3. Administer compound A or placebo once daily for 52 weeks (1 capsule (300 mg) or 2 capsules (600 mg)). F1 Stratification of F2 / F3.

[0377] Evaluate the efficacy of treatment with compound A by, for example, NAS score from liver biopsy, MRI LiverMultiScan evaluation of PDFF and cT1, liver function tests, HOMA-IR as well as biomarkers of inflammation and fibrosis (including hsCRP, Pro-C3, ELF panel and other suitable biomarkers). Evaluate safety and tolerability by monitoring adverse event reports, vital signs, physical examinations, hematology and biochemistry

[0378] (kidney, liver), urinalysis as well as resting 12-lead ECG monitoring. Monitor blood pressure by three seated blood pressure readings measured at each visit.

[0379]

[0380] Patient Inclusion Criteria: Surveys were conducted over 58 weeks and 10 clinical visits per patient, including screening, secondary This includes a double-blind treatment period and post-treatment follow-up. The procedures performed at each visit are as follows: These are explained below and summarized in Table 13. Visit 1 (1-42 days prior to the start of treatment administration): Follow the criteria outlined below for eligibility. Screen patients and obtain consent. This may be implemented. Visit 2: Patients are randomized to one of three treatment groups. 300 or 600 mg of Compound A or placebo will be administered daily for 52 weeks. Liver imaging (MRI-PDFF and cT1), vital signs, safety labs (baseline hematology , biochemistry and urinalysis), resting 12-lead ECG and biomarkers (HOMA-IR, A baseline assessment will be performed including hsCRP, Pro-C3 and ELF panel. Visit 3 (4 weeks post-treatment): Adverse events, vital signs, safety labs (hematology, biochemistry) Evaluate patients for clinical findings (e.g., urinalysis and urinalysis), resting 12-lead ECG, and trough PK samples. do. Visit 4 (10 weeks post-treatment): Adverse events, vital signs, safety labs (hematology, biopsy) Patients will be evaluated for chemistries and urinalysis, resting 12-lead ECGs, and trough PK samples. do. Visit 5 (after 16 weeks of treatment): Adverse events, liver imaging (MRI-PDFF and cT1) , vital signs, safety labs (hematology, biochemistry and urinalysis), resting 12-lead ECG, Biomarkers (HOMA-IR, hsCRP, Pro-C3 and ELF panel) and Patients will be evaluated for pharmacokinetic and trough PK samples. Visits 6, 7, and 8 (after 24, 32, and 40 weeks of treatment): Adverse events, vital signs In, safety laboratory (hematology, biochemistry, and urinalysis), 12-lead resting ECG, biomarkers - (hsCRP) and trough PK samples are used to evaluate the patients. · Visit 9 (after 52 weeks of treatment): liver biopsy, liver imaging (MRI-PDFF and cT1), vital signs, safety laboratory (hematology, biochemistry, and urinalysis), 12-lead resting ECG, biomarkers (HOMA-IR, hsCRP, Pro-C3, and ELF panel) and trough PK samples are used to evaluate the patients. · Visit 10 (14 - 21 days after discontinuation of investigational medication): The patients receive safety follow-up, adverse events, vital signs, safety laboratory (hematology, biochemistry, and urinalysis), 12-lead resting ECG, and trough PK samples are used to evaluate the patients.

[0381] Patients who drop out of the study before completion are evaluated for adverse events, vital signs, safety laboratory ( hematology, biochemistry, and urinalysis), 12-lead resting ECG, trough PK samples, and biomarkers (HOMA-IR, hsCRP, Pro-C3, and ELF panel). evaluate them.

[0382] Patient Exclusion Criteria Patients included in the study meet the following criteria. · Between 18 and 75 years old. · Women are not pregnant or use acceptable contraceptive methods. · If the female partner of male patients is at risk of pregnancy, male patients should voluntarily use contraception ( e.g., condoms). In addition, their female partners should use contraception (e.g., hormonal drugs, intrauterine devices, etc.). Double contraception should be used from the first dose of the compound drug to 90 days after the last dose of the investigational drug. · Written informed consent. · Optional fibroscan criteria before biopsy. · Histological diagnosis of NASH at screening or within 6 months of screening. · NAS score ≥ 4. · Fibrosis score 1 to 3 (F1 with a 30% upper limit). · Optional PDFF > 10% by MRI · Willingness to undergo liver biopsy after 52 weeks of treatment. · Compensated liver disease according to the following hematological and biochemical criteria for inclusion in the protocol: · ALT < 5 × ULN · AST > 30 · Hemoglobin > 11 g / dL (for women) and > 12 g / dL (for men) · White blood cells (WBC) > 2.5 K / μL · Neutrophil count > 1.5 K / μL · Platelets > 100 K / μL · Total bilirubin < 35 μmol / L. In the case of unconjugated bilirubin within the setting of Gilbert syndrome, patients with bilirubin > 35 μmol / L can be included. · Albumin > 36 g / L · International normalized ratio (INR) < 1.4 · Serum creatinine < 1.3 mg / dL (for men) or < 1.1 (for women) or estimated glomerular filtration rate ≥ 60 mL / min / 1.73 m2 · No other cause of chronic liver disease (autoimmune disease, primary biliary cholangitis, HBV, HCV, Wilson disease, α-1-antitrypsin deficiency, hemochromatosis, etc...). · If applicable, stable type 2 diabetes (defined as HgbA1c < 9.5% and fasting hyperglycemia < 10 mmol / L, no change in medication in the last 6 months, and no new symptoms related to decompensated diabetes in the last 3 months). · Have a stable weight after liver biopsy, where stable is defined as a decrease of 5% or less of the initial weight. Decrease.

[0383] Endpoint Criteria The patient does not meet any of the following exclusion criteria: · A history of persistent excessive alcohol intake as evaluated by an alcohol intake questionnaire. · An unstable metabolic state defined as follows: a weight gain or loss of more than 5 kg in the 3 months prior to screening, poorly controlled diabetes (HgbA1c > 9 .5%) in the past 6 months, or the introduction / absorption failure or restrictive obesity ( weight loss) surgery of an antidiabetic or anti-obesity drug. · A history of gastrointestinal malabsorption obesity surgery within less than 5 years or corticosteroids, high-dose estrogen, methotrexate, tetracycline, or amiodarone in the past 6 months. A history of taking drugs known to cause fatty liver. · In the opinion of the evaluator, congestive heart failure (AHA classes C and D) that would interfere with treatment with compound A, unstable coronary artery disease, cerebrovascular disease, lung disease, renal insufficiency, organ transplantation, severe systemic diseases or serious diseases other than liver diseases, including mental disorders or malignant tumors. · HB antigen > 0, HCV PCR > 0 (patients with a history of HCV infection can be included if HCV PCR has been negative for more than 3 years), or HIV infection. · Any other condition that, in the opinion of the evaluator, would interfere with competence or compliance, or would probably interfere with the completion of the investigation. · Body mass index (BMI) > 45 kg / m 2 2. · Type 1 or type 2 diabetes treated with insulin. · Diabetic ketoacidosis. · Fasting triglycerides > 300 mg / dL. · Current treatment with a hemostatic disorder or anticoagulant. · Contraindication to liver biopsy. · History of cardiovascular disease including arrhythmia history or current arrhythmia and / or myocardial infarction, excluding those in patients with well-controlled hypertension, and any clinically significant ECG abnormalities. · Participation in any other investigational drug study within the last 3 months or within 5 half-lives of the drug. · Known hypersensitivity to either the components or excipients of the IMP. · Intake of antidiabetic drugs known to have activity against NASH, such as pioglitazone , and GLP-1 receptor agonists.

[0384] Statistical Methods The primary endpoint of the efficacy of Compound A is the proportion of patients in whom NASH has recovered as defined by the disappearance of ballooning swelling (score = 0) without progression of fibrosis with a lobular inflammation score of 0 or 1.

[0385] The secondary endpoints of the efficacy of Compound A are changes from baseline in the NAS score, changes in the individual histological scores of steatosis, ballooning swelling, inflammation, and fibrosis, changes in liver enzymes, changes in imaging parameters, and changes in biomarkers (including hsCRP, Pro-C3, the ELF panel, and cytokines).

[0386] The secondary endpoints of safety / tolerability include reported adverse events, clinical laboratory values (hematology, biochemistry, and urinalysis), vital signs (blood pressure, pulse rate, and body temperature), and hsCRP.

[0387] Secondary endpoints of pharmacokinetics include comparison of mean trough plasma concentrations at steady state for three doses of Compound A measured at Visits 4, 5, 6, 7, and 8.

[0388] ​ A sample size of 88 patients per group provides 80% power to detect a 40% responder rate for the active agent versus placebo, assuming a 18% placebo response rate and a 25% dropout rate. There may be a re-estimation of the sample size after a 16-week interim analysis. An efficacy analysis is performed based on the modified intention to treat population, which consists of those with baseline and at least one post-baseline efficacy measurement. Comparisons are made between individual doses and placebo.

[0389]

Table 13

Claims

1. For use in the therapeutic treatment of non-alcoholic steatohepatitis in a subject having type 2 diabetes Formula (II) 【Chemical 1】 (wherein R １ is C having 3 to 6 double bonds １８ - C ２２ is selected from alkenyl, R ２ and R ３ are the same or different and are selected from the group consisting of a hydrogen atom and an alkyl group, and R ２ and R ３ can be linked to form a cycloalkane such as cyclopropane, cyclobutane, cyclopentane or cyclohexane. X is a carboxylic acid (C(O)OH), C １ -C ６ alkyl ester, or C(O)NH 2 , N-methylamide, N-dimethylamide, N-ethylamide, N-isopropylamide, N-(tert-butyl)amide, and N-(2-hydroxyethyl)-1-amide, or a pharmaceutically acceptable salt, solvate, or solvate of such a salt of a compound selected from the group consisting of carboxamides) and a pharmaceutical composition comprising the same.

2. The pharmaceutical composition according to claim 1, wherein the use alleviates or prophylactically treats the onset of liver fibrosis or reduces existing liver fibrosis.

3. Formula (II) for use in the therapeutic treatment of non-alcoholic steatohepatitis in a subject having liver fibrosis [Chemical Formula 2] (wherein, R １ is C having 3 to 6 double bonds １８ -C ２２ selected from alkenyl, R ２ and R ３ are the same or different and are selected from the group consisting of a hydrogen atom and an alkyl group, X is a carboxylic acid (C(O)OH), C １ -C ６ alkyl ester, or C(O)NH 2 , N-methylamide, N-dimethylamide, N-ethylamide, N-isopropylamide, N-(tert-butyl)amide, and N-(2-hydroxyethyl)-1-amide, and a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt, solvate, or solvate of such a salt thereof, wherein the liver fibrosis does not progress as compared to an untreated subject, said pharmaceutical composition.

4. The pharmaceutical composition according to claim 3, wherein the subject has type 2 diabetes.

5. The compound is of formula (I) [Chemical Formula 3] is a compound, R ２ , R ３ and X are as defined for formula (II), the pharmaceutical composition according to any one of claims 1 to 4.

6. R ２ and R ３ are each independently selected from a hydrogen atom or a linear and / or branched C １ -C ６ alkyl group wherein X is a carboxylic acid or a C １ -C ６ alkyl ester, and the pharmaceutical composition according to any one of claims 1 to 5.

7. R ２ and R ３ is independently selected from a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, and an isopropyl group, the pharmaceutical composition according to any one of claims 1 to 6.

8. R ２ and R ３ are each independently C １ -C ６ an alkyl group, the pharmaceutical composition according to any one of claims 1 to 6.

9. R ２ and R ３ The pharmaceutical composition according to any one of claims 1 to 6, wherein one of them is a hydrogen atom and the other is an ethyl group.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein X is carboxylic acid (C(O)OH).

11. X is C １ -C ６ The pharmaceutical composition according to any one of claims 1 to 9, which is an alkyl ester.

12. The pharmaceutical composition according to claim 11, wherein X is selected from methyl ester, ethyl ester, isopropyl ester, n-butyl ester and tert-butyl ester.

13. The pharmaceutical composition according to any one of claims 1 to 9, wherein X is selected from the group consisting of methyl ester and ethyl ester.

14. The pharmaceutical composition according to any one of claims 1 to 9, which exists in the form of an enantiomer, diastereomer or a mixture thereof.

15. The pharmaceutical composition according to claim 14, which exists as its R-form, its S-form or a racemate.

16. The compound is 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (Compound A) or a pharmaceutically acceptable salt or ester thereof, and the formula is [Chemical Formula 4] The pharmaceutical composition according to any one of claims 1 to 5.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein the use improves non-alcoholic steatohepatitis (NASH).

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein the use provides a reduction in liver inflammation; a reduction in ballooning hepatocyte swelling; and / or a reduction in steatohepatitis.

19. The pharmaceutical composition according to any one of claims 1 to 18, wherein the pharmaceutical composition is formulated for oral administration.

20. The pharmaceutical composition according to any one of claims 1 to 19, further comprising at least one binder, excipient, diluent or antioxidant or any combination thereof.

21. The pharmaceutical composition according to any one of claims 1 to 20, which is administered once a day.

22. The pharmaceutical composition according to any one of claims 1 to 21, which is administered at a dose between about 5 mg and about 4 g per dose.

23. The pharmaceutical composition according to any one of claims 1 to 21, wherein the pharmaceutical composition is administered daily at a dose of 600 mg.

24. The pharmaceutical composition according to any one of claims 1 to 21, wherein the pharmaceutical composition is administered daily at a dose of 300 mg.

25. The pharmaceutical composition according to any one of claims 1 to 24, which is administered as monotherapy.

26. The pharmaceutical composition according to any one of claims 1 to 24, which is co-administered with at least one additional active agent.

27. The one or more additional active agents are selected independently from an allosteric acetyl-CoA carboxylase (ACC) inhibitor, an angiotensin II receptor antagonist, an angiotensin converting enzyme (ACE) inhibitor, an apoptosis signal-regulating kinase-1 (ASK1) inhibitor, a caspase inhibitor, a cathepsin B inhibitor, a CCR2 chemokine antagonist, a CCR5 chemokine antagonist, a chloride channel stimulant, a cholesterol solubilizer, a diacylglycerol O-acyltransferase 1 (DGAT1) inhibitor, a dipeptidyl peptidase IV (DPP IV) inhibitor, a fibroblast growth factor (FGF)-21 agonist, a farnesoid X receptor (FXR) agonist, an anti-CD3 mAb, a galectin-3 inhibitor, a glucagon-like peptide 1 (GLP1) agonist, a glutathione precursor, a hepatitis C virus NS3 protease inhibitor, an HMG CoA reductase inhibitor, a 17β-hydroxysteroid dehydrogenase (17β-HSD) inhibitor, a heat shock protein (Hsp) 47 inhibitor, an IL-Iβ antagonist, an IL-6 antagonist, an IL-10 agonist, an IL-17 antagonist, an ileal sodium bile acid cotransporter inhibitor, a leptin analog, a 5-lipoxygenase inhibitor, an LPL gene stimulant, a lysyl oxidase homolog 2 (LOXL2) inhibitor, a lysophosphatidic acid 1 (LPA1) receptor antagonist, an omega-3 fatty acid, a PDE3 inhibitor, a PDE4 inhibitor, a phospholipase C (PLC) inhibitor, a PPARα agonist, a PPARγ agonist, a PPARβ / δ agonist, a recombinant human pentraxin-2 protein (PRF-1), a Rho-associated protein kinase 2 (ROCK2) inhibitor, a semicarbazide-sensitive amine oxidase (SSAO) inhibitor, a sodium glucose transporter-2 (SGLT2) inhibitor, a stearoyl-CoA desaturase-1 inhibitor, a thyroid hormone receptor β agonist, a tumor necrosis factor a (TNFα) ligand inhibitor, a transglutaminase inhibitor, a transglutaminase inhibitor precursor, and a small activating RNA (saRNA), the pharmaceutical composition according to claim 26. [

28. ] The pharmaceutical composition according to claim 26 or 27, wherein the one or more additional active agents are independently selected from glucagon-like peptide 1 (GLP-1) agonists, dipeptidyl peptidase (DPP IV) inhibitors, and omega-3 fatty acids (n-3 PUFAs).

29. The pharmaceutical composition according to claim 26, wherein the one or more additional active agents are independently selected from acetylsalicylic acid, aliposene tiparborbéc, aramchol, atorvastatin, BI 1467335, BLX-1002, BMS-986036, BMS-986020, cenicriviroc, cobiprostone, colesevelam, emricasan, enalapril, folarub, GFT-505, GR-MD-02, GS-0976, GS-9674, hydrochlorothiazide,icosapent ethyl ester (eicosapentaenoic acid ethyl ester), IMM-124E, IVA337, K-877, KD-025, linagliptin, liraglutide, mercaptamine, MGL-3196, ND-L02-s0201, obeticholic acid, olesoxime, peg-irodecakin, pioglitazone, PRM-151, PX-102, remogliflozin etabonate, serolsertib, simtuzumab, SHP-626, solithromycin, tipelcast, TRX-318, ursodeoxycholic acid and VBY-376.

30. The pharmaceutical composition according to any one of claims 26 to 29, wherein the co-administration is by simultaneous administration, sequential administration, repeated administration, intermittent administration, continuous administration or a combination thereof.

31. In a subject having type 2 diabetes, in the manufacture of a medicament for the therapeutic treatment of non-alcoholic steatohepatitis, formula (II)

32. 【Chemical Formula 5】 (wherein, R １ is C having 3 to 6 double bonds １８ -C ２２ selected from alkenyl, R ２ and R ３ are the same or different and are selected from the group consisting of a hydrogen atom and an alkyl group, X is a carboxylic acid (C(O)OH), C １ -C ６ alkyl ester, or C(O)NH 2 , N-methylamide, N-dimethylamide, N-ethylamide,, N-isopropylamide, N-(tert-butyl)amide, and N-(2-hydroxyethyl)-1-amide, or a pharmaceutically acceptable salt, solvate, or solvate of such a salt of a compound selected from the group consisting of carboxamides) for use in a pharmaceutical composition. The use according to claim 31, wherein the medicament reduces or prophylactically treats the onset of liver fibrosis or reduces existing liver fibrosis.

33. In the manufacture of a medicament for the therapeutic treatment of non-alcoholic steatohepatitis in a subject having liver fibrosis, formula (II)

34. 【Chemical Formula 6】 (wherein, R １ is C having 3 to 6 double bonds １８ -C ２２ selected from alkenyl, R ２ and R ３ are the same or different and are selected from the group consisting of a hydrogen atom and an alkyl group, X is a carboxylic acid (C(O)OH), C １ -C ６ alkyl ester, or C(O)NH 2 , N-methylamide, N-dimethylamide, N-ethylamide, N-isopropylamide, N-(tert-butyl)amide, and N-(2-hydroxyethyl)-1-amide, or a pharmaceutically acceptable salt, solvate, or solvate of such a salt of a compound selected from the group consisting of: the use of a pharmaceutical composition comprising: the progression of liver fibrosis does not progress compared to an untreated subject. The use according to claim 33, wherein the subject has type 2 diabetes.

35. The compound is 2-(((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaen-1-yl)oxy)butanoic acid (Compound A) or a pharmaceutically acceptable salt or ester thereof, and the formula is ​ [Chemical Formula 7] The use according to any one of claims 31 to 34, which is as claimed.

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