Drugs for treating fatty liver disease

Abstract

To provide a therapeutic agent for fatty liver disease.SOLUTION: The therapeutic agent for fatty liver disease comprises a compound represented by the formula (1) in the figure or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: Figure 1A

Description

[Technical Field]

[0001] The present invention relates to a therapeutic agent for fatty liver disease. [Background technology]

[0002] Nonalcoholic fatty liver disease (NAFLD), a type of fatty liver disease, is a liver disease caused by the accumulation of excess lipids in the liver due to metabolic syndrome, such as obesity, diabetes, dyslipidemia, and hypertension. In certain populations, NAFLD progresses to nonalcoholic steatohepatitis (NASH), which causes liver fibrosis due to inflammation and hepatocyte death. This then leads to cirrhosis and may ultimately lead to liver cancer or cardiovascular disease (Non-Patent Document 1).

[0003] A definitive diagnosis of NASH is made by pathological examination. The activity of NASH is classified based on the total score of fatty degeneration, inflammatory cell infiltration, and ballooning, which is called the NAS (NAFLD activity score). The stage of the disease is also classified based on the degree of liver fibrosis: specifically, stage 1 is pericentral fibrosis, stage 2 is further periportal fibrosis, stage 3 is bridging fibrosis, and stage 4 is cirrhosis.

[0004] According to current FDA guidance (Non-Patent Document 2), the endpoint of phase 2 or later clinical trials must demonstrate a histological improvement effect, i.e., either an improvement in NAS or an improvement in liver fibrosis.

[0005] Thiazolidinedione drugs, which are PPARγ agonists, and vitamin E are recommended as treatments for NASH. However, because these drugs have an indirect effect associated with diabetes treatment, their effectiveness in preventing the development of liver cirrhosis and liver cancer has not been proven, and there are concerns about side effects from long-term administration. Therefore, no established treatments for NAFLD or NASH have been established. Therefore, various drugs, such as FXR agonists (e.g., obeticholic acid, tropifexor), SCD1 inhibitors (aramchol), ASK1 inhibitors (selonsertib), and PPARα / γ agonists (elafibranor), are currently under development, and new therapeutic agents are needed. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Hepatology. 2018 Jan;67(1):328-357. [Non-patent document 2] Noncirrhotic Nonalcoholic Steatohepatitis With Liver Fibrosis: Developing Drugs for Treatment Guidance for Industry Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a therapeutic agent for fatty liver disease. [Means for solving the problem]

[0008] As a result of extensive research, the present inventors have found that compounds having a specific structure can treat fatty liver disease, and have thus completed the present invention.

[0009] The present invention includes the following embodiments. [1] The following formula (1): [ka] A therapeutic agent for fatty liver disease, comprising a compound represented by the formula: [2] The therapeutic agent according to [1], wherein the pharmaceutically acceptable salt is a hydrochloride salt. [3] The therapeutic agent according to [1] or [2], wherein the fatty liver disease is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). [4] The therapeutic agent according to [3], wherein the fatty liver disease is NAFLD. [5] The therapeutic agent described in [4], wherein the NAFLD is accompanied by liver fibrosis. [6] The therapeutic agent described in [4], wherein the NAFLD is accompanied by liver inflammation. [7] The therapeutic agent according to [3], wherein the fatty liver disease is NASH. [8] The therapeutic agent described in [7], wherein the NASH is accompanied by liver fibrosis. [9] The therapeutic agent described in [7], wherein the NASH is accompanied by liver inflammation.

[0010] The above embodiment can also be expressed as follows. [A] A method for treating fatty liver disease, comprising administering to a patient in need of treatment a therapeutically effective amount of a compound represented by formula (1) as defined in [1] above or a pharmaceutically acceptable salt thereof. [B] A compound represented by formula (1) as defined in [1] above or a pharmaceutically acceptable salt thereof for use in the treatment of fatty liver disease. [C] Use of a compound represented by formula (1) as defined in [1] above or a pharmaceutically acceptable salt thereof for use in the treatment of fatty liver disease. [D] Use of a compound represented by formula (1) defined in [1] above or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating fatty liver disease. [Effects of the Invention]

[0011] According to the present invention, a therapeutic agent for fatty liver disease can be provided. [Brief explanation of the drawings]

[0012] [Figure 1A] FIG. 1A shows plasma ALT at 12 weeks after administration of the administered substances. [Figure 1B] FIG. 1B shows plasma ALT at 18 weeks after administration of the administered substances. [Figure 1C] FIG. 1C shows plasma AST at 18 weeks after administration of the administered substances. [Figure 2] FIG. 2 shows plasma Tcho at 18 weeks after administration of the administered substances. [Figure 3] FIG. 3 shows the TG in the liver 18 weeks after administration of the administered substances. [Figure 4] FIG. 4 shows the Sirius Red positive area rate in the liver 18 weeks after administration of the administered substances. [Figure 5] FIG. 5 shows the number of CD68-positive hCLS determined by immunostaining in the liver 18 weeks after administration of the administered substance. DETAILED DESCRIPTION OF THE INVENTION

[0013] Hereinafter, embodiments of the present invention will be described in detail, but the present invention is not limited to these and various modifications are possible without departing from the gist of the present invention.

[0014] One embodiment of the present invention is a compound represented by the following formula (1): [ka] or a pharmaceutically acceptable salt thereof.

[0015] The compound represented by formula (1) or a pharmaceutically acceptable salt thereof can be easily synthesized by a person skilled in the art, for example, by referring to the synthesis methods described in WO 2015 / 137407 and WO 2015 / 137408.

[0016] The pharmaceutically acceptable salt of the compound represented by formula (1) is not particularly limited as long as it is usable as a pharmaceutical, and examples thereof include inorganic acid salts such as hydrochloride, sulfate, nitrate, hydrobromide, and phosphate, and organic acid salts such as fumarate, maleate, malate, tartrate, citrate, succinate, methanesulfonate, p-toluenesulfonate, lactate, acetate, and palmitate.

[0017] The compound represented by formula (1) or a pharmaceutically acceptable salt thereof may form a solvate such as a hydrate. In this specification, solvates are included in the compound represented by formula (1) or a pharmaceutically acceptable salt thereof.

[0018] The therapeutic agent of this embodiment may contain only the compound represented by formula (1) or a pharmaceutically acceptable salt thereof, or may further contain other ingredients. The other ingredients vary depending on the dosage form, etc., but may include, for example, excipients, binders, lubricants, disintegrants, surfactants, suspending agents, emulsifiers, preservatives, colorants, flavoring agents, sweeteners, flavoring agents, stabilizers, and thickeners.

[0019] The therapeutic agent of this embodiment can be administered orally or parenterally. Examples of dosage forms for oral administration include tablets, pills, granules, powders, capsules, syrups, emulsions, and suspensions. Examples of dosage forms for parenteral administration include injections, infusions, drip infusions, eye drops, and suppositories.

[0020] The therapeutic agent of this embodiment can treat fatty liver disease. In this specification, "treatment" includes preventing the onset of fatty liver disease, suppressing the progression of fatty liver disease, alleviating the symptoms of fatty liver disease, curing fatty liver disease, etc.

[0021] Specific examples of fatty liver disease include NAFLD, NASH, metabolically associated fatty liver disease (MAFLD), and fatty liver. Although not particularly limited, the therapeutic agent of the present embodiment can be suitably used to treat NAFLD and / or NASH.

[0022] The therapeutic agent of this embodiment, when treating fatty liver disease, can improve one or more symptoms selected from the group consisting of liver damage (e.g., increased plasma ALT and / or AST), dyslipidemia (e.g., increased plasma total cholesterol), hepatic steatosis (e.g., increased triglycerides in the liver), hepatic fibrosis (e.g., increased Sirius Red-positive area rate), and hepatic inflammation (e.g., increased hCLS count). [Example]

[0023] The present invention will be described in more detail below using examples, but the technical scope of the present invention is not limited to these examples.

[0024] In this example, the effects of test drugs were evaluated using a high-fat diet / high-fructose-induced NASH model (Fast Food Diet model: FFD model). For details of the FFD model, see BMC Gastroenterol. 2020; 20: 210. and Am J Physiol Gastrointest Liver Physiol. 2013 Oct 1; 305(7): G483-95.

[0025] <Administered substance> 0.5% (w / v) methylcellulose solution (hereinafter referred to as "0.5% MC") Test drug The test drug was prepared by adding the hydrochloride salt of the compound represented by the above formula (1) (hereinafter referred to as "Compound 1") to 0.5% MC and crushing it by ultrasonication.

[0026] <Experimental animals> Six-week-old C57BL / 6J Jms Slc male mice were fed a standard diet (D09100304, Research Diet) or a pathological FFD diet (D09100310N, Research Diet) for 20 weeks. Afterward, mice were divided into groups based on plasma alanine aminotransferase (ALT) levels, body weight, and fasting blood glucose levels, as shown in Table 1.

[0027] <Administration method> According to the dosage and administration method shown in Table 1, the substance was orally administered to each group for 18 weeks by force. [Table 1]

[0028] <Evaluation method> ALT was measured by tail vein blood sampling at week 12 of treatment. Plasma aspartate aminotransferase (AST), plasma total cholesterol (Tcho), and liver triglyceride (TG) were measured at necropsy at week 18 of treatment. In addition, Sirius Red-positive area rate and hepatic crown-like structure (hCLS) count were measured by liver pathological analysis.

[0029] <Statistical analysis> Statistical analysis was performed using GraphPad Prism 6 (GraphPad Software, Inc.). A t-test or Mann-Whitney's test was performed for the normal group and vehicle group. A Dunnett's multiple test or Dunn's multiple test was performed for the vehicle group and compound 1 group. An appropriate test method was selected depending on the distribution of each parameter. The significance level for all tests was set at p = 0.05.

[0030] <Plasma ALT and AST> Figure 1A shows the plasma ALT at the 12th week of administration, Figure 1B shows the plasma ALT at the 18th week of administration, and Figure 1C shows the plasma AST at the 18th week of administration. The Vehicle group showed a statistically significant increase in plasma ALT and AST compared to the Normal group. Also, the Compound 1 (60) group showed a statistically significant inhibitory effect on the increase in plasma ALT and AST compared to the Vehicle group. From these results, it was confirmed that Compound 1 improves liver damage.

[0031] <Plasma Tcho> Figure 2 shows the plasma Tcho value at the 18th week of administration. The Vehicle group showed a statistically significant increase in plasma Tcho compared to the Normal group. Also, the Compound 1 (60) group showed a statistically significant inhibitory effect on the increase in plasma Tcho compared to the Vehicle group. From these results, it was confirmed that Compound 1 improves lipid metabolism.

[0032] <Liver TG> Liver samples were homogenized with phosphate buffer, methanol and chloroform were added and extracted overnight, and then chloroform and distilled water were further added. The mixture was centrifuged to separate the chloroform layer, and the solvent was dried. The residue was dissolved in isopropanol / TritonX-100, used as a measurement sample, and measured using Triglyceride E-Test Wako (FUJIFILM Wako Pure Chemical Corporation).

[0033] Figure 3 shows the liver TG value at the 18th week of administration. The Vehicle group showed a statistically significant increase in liver TG compared to the Normal group. Also, the Compound 1 (30) group and the Compound 1 (60) group showed a statistically significant inhibitory effect on the increase in liver TG compared to the Vehicle group. From these results, it was confirmed that Compound 1 has an inhibitory effect on liver steatosis.

[0034] <Sirius Red positive area ratio> The excised liver was fixed with 10% neutral buffered formalin solution, paraffin-embedded according to a conventional method, and Sirius Red-stained specimens were prepared. Using the Sirius Red-stained specimens, the positive area ratio was calculated. The average positive area ratio of the Normal group was used as the background.

[0035] Figure 4 shows the Sirius Red positive area ratio of the liver at 18 weeks after administration. The Vehicle group showed a statistically significant increase in the Sirius Red positive area ratio of the liver compared to the Normal group. Also, the Compound 1 (60) group showed a statistically significant inhibitory effect on the increase in the Sirius Red positive area ratio of the liver compared to the Vehicle group. From these results, it was confirmed that Compound 1 has an anti-fibrotic effect on the liver.

[0036] <Number of hCLS> CD68 immunostained specimens (rabbit anti-CD68 polyclonal antibody [ab125212], abcam) were prepared and the number of hCLS was counted. Figure 5 shows the number of CD68 positive hCLS by liver immunostaining at 18 weeks after administration. The Vehicle group showed a statistically significant increase in the number of hCLS in the liver compared to the Normal group. Also, the Compound 1 (10) group and the Compound 1 (60) group showed a statistically significant inhibitory effect on the increase in the number of hCLS in the liver compared to the Vehicle group. From these results, it was confirmed that Compound 1 has an anti-inflammatory effect on the liver.

[0037] <Pathological analysis> The findings of the pathological tissues of the liver are shown in Table 2. Also in the pathological analysis, it was confirmed that Compound 1 has an improving effect on liver steatosis and fibrosis.

Table 2

[0038] <Comprehensive evaluation> Compound 1 suppressed the increase in plasma ALT / AST / Tcho and liver TG, thereby significantly improving indicators of fatty liver disease (liver enzymes and lipid metabolism). Furthermore, Compound 1 significantly suppressed the Sirius Red-positive area rate and the number of hCLS, improving indicators of liver fibrosis and inflammation. Histopathological findings also showed results supporting these improving effects. These results confirmed the efficacy of Compound 1 in the NASH model.

Claims

1. The following formula (1): 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein the therapeutic agent ameliorates one or more symptoms selected from the group consisting of liver damage, dyslipidemia, hepatic steatosis, hepatic fibrosis, and hepatic inflammation.

2. the symptom is liver damage, The liver damage is evaluated by measuring an increase in plasma ALT and / or AST. The therapeutic agent according to claim 1.

3. the symptom is dyslipidemia or hepatic steatosis, The lipid metabolism disorder or hepatic steatosis is indexed by an increase in plasma total cholesterol or an increase in hepatic triglycerides. The therapeutic agent according to claim 1.

4. the symptom is liver fibrosis, The liver fibrosis is evaluated by an increase in the Sirius Red positive area ratio. The therapeutic agent according to claim 1.

5. the symptom is liver inflammation, The liver inflammation is indexed by an increase in the number of hCLS. The therapeutic agent according to claim 1.

6. The therapeutic agent according to any one of claims 1 to 5, wherein the pharmaceutically acceptable salt is a hydrochloride salt.

7. The following formula (1): 【Chemistry 2】 A therapeutic agent for fatty liver, comprising a compound represented by the formula:

8. The therapeutic agent of claim 7, wherein the pharmaceutically acceptable salt is a hydrochloride salt.

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