Therapeutic agents for fatty liver disease

Abstract

An object of the present application is to provide a therapeutic drug for fatty liver disease. The above object can be achieved by a therapeutic drug for fatty liver disease, which comprises a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

Description

TECHNICAL FIELD

[0001] The present application relates to a therapeutic agent for fatty liver disease. BACKGROUND

[0002] Non-alcoholic fatty liver disease (NAFLD) is one of the fatty liver diseases caused by excessive accumulation of lipids in the liver due to metabolic syndrome such as obesity, diabetes, dyslipidemia, hypertension, etc. A certain population in NAFLD develops non-alcoholic steatohepatitis (NASH), and fibrosis of the liver is formed due to inflammation, hepatocyte death, etc. There are also cases where cirrhosis is caused thereafter, and eventually, liver cancer, cardiovascular disease (non-patent literature 1).

[0003] The diagnosis of NASH is made by pathological diagnosis. The activity is scored by fatty degeneration, inflammatory cell infiltration, and ballooning degeneration, respectively, and classified based on NAS (NAFLD activity score) which is the total score of these. In addition, the disease is classified by the degree of liver fibrosis, and specifically, stage 1 is fibrosis around the central vein, stage 2 is further fibrosis around the portal vein, stage 3 is bridging fibrosis, and stage 4 is cirrhosis.

[0004] According to the current FDA guidelines (non-patent literature 2), the endpoint after phase 2 of the clinical trial needs to show histological improvement effects, that is, either improvement of NAS or improvement of liver fibrosis.

[0005] As a therapeutic agent for NASH, a thiazolidine agent, vitamin E, which is a PPARγ agonist, is recommended, but this is an indirect effect accompanying diabetes treatment, and therefore, it has not been proven to have an effect of preventing the onset of cirrhosis and liver cancer, and rather, there are more concerns about side effects caused by long-term administration. Thus, the treatment method for NAFLD and NASH has not yet been established, and therefore, various agents such as FXR agonists (obeticholic acid, Tropifexor, etc.), SCD1 antagonists (Aramchol), ASK1 antagonists (Selonsertib), PPARα / γ agonists (Elafibranor) are still under development, and new therapeutic agents are expected.

[0006] PRIOR ART DOCUMENTS

[0007] NON-PATENT LITERATURE

[0008] Non-patent literature 1: Hepatology. 2018 Jan; 67(1): 328-357.

[0009] Noncirrhotic Nonalcoholic Steatohepatitis With Liver Fibrosis: Developing Drugs for Treatment Guidance for Industry SUMMARY

[0010] PROBLEMS TO BE SOLVED BY THE INVENTION

[0011] An object of the present application is to provide a therapeutic drug for fatty liver disease.

[0012] MEANS FOR SOLVING THE PROBLEMS

[0013] The present inventors and others have conducted intensive studies, and as a result, have found that a compound having a specific structure can treat fatty liver disease, thereby completing the present application.

[0014] The present application includes the following embodiments. [1]

[0016] A therapeutic drug for fatty liver disease, comprising a compound represented by the following formula (1), or a pharmaceutically acceptable salt thereof,

[0017] [Chemical Formula 1]

[0018] [2]

[0020] The therapeutic drug according to [1], wherein the pharmaceutically acceptable salt is a hydrochloride. [3]

[0022] The therapeutic drug according to [1] or [2], wherein the fatty liver disease is nonalcoholic fatty liver disease (NAFLD) or nonalcoholic steatohepatitis (NASH). [4]

[0024] The therapeutic drug according to [3], wherein the fatty liver disease is NAFLD. [5]

[0026] The therapeutic drug according to [4], wherein the NAFLD is accompanied by fibrosis of the liver. [6]

[0028] The therapeutic drug according to [4], wherein the NAFLD is accompanied by inflammation of the liver. [7]

[0030] The therapeutic drug according to [3], wherein the fatty liver disease is NASH. [8]

[0032] The therapeutic agent according to [7], wherein the NASH is accompanied by fibrosis of the liver. [9]

[0034] The therapeutic agent according to [7], wherein the NASH is accompanied by inflammation of the liver.

[0035] The above-described embodiments can also be expressed as follows.

[0036] [A]

[0037] 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.

[0038] [B]

[0039] 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.

[0040] [C]

[0041] Use of a compound represented by formula (1) as defined in [1] above or a pharmaceutically acceptable salt thereof for the treatment of fatty liver disease.

[0042] [D]

[0043] Use of a compound represented by formula (1) as defined in [1] above or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of fatty liver disease.

[0044] Effects of the Invention

[0045] According to the present application, it is possible to provide a therapeutic agent for fatty liver disease. BRIEF DESCRIPTION OF DRAWINGS

[0046] [ Figure 1A ] Figure 1A ALT in plasma at week 12 of administration with the administration substance.

[0047] [ Figure 1B ] Figure 1B ALT in plasma at week 18 of administration with the administration substance.

[0048] [ Figure 1C ] Figure 1C AST in plasma at week 18 of administration with the administration substance.

[0049] [ Figure 2 ] Figure 2 Tcho in plasma at week 18 of administration with the administration substance.

[0050] [ Figure 3 ] Figure 3 TG in the liver at week 18 of administration with the administration substance.

[0051] [ Figure 4 ] Figure 4 Sirius Red-positive area ratio of the liver at week 18 of administration with the administration substance.

[0052] [ Figure 5 ] Figure 5 CD68-positive hCLS number in the liver at week 18 of administration with the administration substance based on immunostaining. DETAILED DESCRIPTION

[0053] Hereinafter, an embodiment of the present application will be specifically described, but the present application is not limited thereto, and various modifications can be made within the scope of the gist of the present application.

[0054] One embodiment of the present application relates to a therapeutic drug for fatty liver disease, which comprises a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof,

[0055] [Chemical Formula 2]

[0056]

[0057] The compound represented by formula (1) or a pharmaceutically acceptable salt thereof can be synthesized easily by those skilled in the art, for example, with reference to the synthetic methods described in International Publication No. 2015 / 137407 and International Publication No. 2015 / 137408.

[0058] The pharmaceutically acceptable salt of the compound represented by formula (1) is not particularly limited as long as it is a salt that can be used as a pharmaceutical product, and examples 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.

[0059] The compound represented by formula (1) or a pharmaceutically acceptable salt thereof can also form a solvate such as a hydrate. In the present specification, the solvate is included in the compound represented by formula (1) or a pharmaceutically acceptable salt thereof.

[0060] The therapeutic agent of the present embodiment can contain only the compound represented by formula (1) or a pharmaceutically acceptable salt thereof, or can further contain other ingredients. As the other ingredients, it is possible to appropriately vary depending on the dosage form and the like, and examples include excipients, binders, lubricants, disintegrants, surfactants, suspending agents, emulsifying agents, preservatives, coloring agents, flavoring agents, sweetening agents, taste correctors, stabilizers, and thickening agents.

[0061] The therapeutic agent of the present embodiment can be administered orally or non-orally. As the dosage form for oral administration, examples include tablets, pills, granules, powders, capsules, syrups, emulsions, and suspensions. As the dosage form for non-oral administration, examples include injections, infusions, drops, eye drops, and suppositories.

[0062] The therapeutic agent of the present embodiment can treat fatty liver disease. In the present specification, "treatment" includes preventing the onset of fatty liver disease, inhibiting the progression of fatty liver disease, alleviating the symptoms of fatty liver disease, curing fatty liver disease, and the like.

[0063] As specific examples of fatty liver disease, examples include NAFLD, NASH, metabolic associated fatty liver disease (MAFLD), and fatty liver. Although not particularly limited, the therapeutic agent of the present embodiment can be preferably used for the treatment of NAFLD and / or NASH.

[0064] The therapeutic agent of the present embodiment, when treating fatty liver disease, can improve one or more symptoms selected from the group consisting of abnormal liver function (e.g., an increase in ALT and / or AST in the blood plasma), abnormal lipid metabolism (e.g., an increase in total cholesterol in the blood plasma), liver steatosis (e.g., an increase in triglyceride in the liver), liver fibrosis (e.g., an increase in the area ratio of picrosirius positivity), and hepatitis (e.g., an increase in the number of hCLS).

[0065] Examples

[0066] Hereinafter, the present application will be described in more detail using examples, but the technical scope of the present application is not limited thereto.

[0067] In the present embodiment, the effect of the test drug was evaluated using a high-fat food·high-fructose-induced NASH model (Fast Food Diet model: FFD model). For the FFD model, see BMC Gastroenterol. 2020; 20: 210. and Am J Physiol Gastrointest Liver Physiol. 2013 Oct 1; 305(7): G483-95.

[0068] <Administered substance>

[0069] • 0.5% (w / v) methylcellulose solution (hereinafter also referred to as 0.5% MC.)

[0070] • Test drug

[0071] The test drug was prepared by adding a hydrochloride salt of the compound represented by the above formula (1) (hereinafter referred to as "Compound 1") to 0.5% MC and performing ultrasonic disruption.

[0072] <Experimental animals>

[0073] Six-week-old C57BL / 6JJms Slc male mice were supplied with standard feed (D09100304, research diet) or disease-inducing FFD feed (D09100310N, research diet) for 20 weeks. Thereafter, the mice were grouped based on three items, i.e., alanine aminotransferase (ALT) values in plasma, body weight, and blood glucose values at the time of fasting, as shown in Table 1.

[0074] <Administration method>

[0075] Each group was subjected to forced oral administration of the administration substance for 18 weeks in accordance with the usage and amount shown in Table 1.

[0076] [Table 1]

[0077] Table 1

[0078]

[0079] * Morning: 9 o'clock ± 1 hour, afternoon: 17 o'clock ± 1 hour,

[0080] * Groups 3 to 5 are also expressed as "Compound 1 (10) group", "Compound 1 (30) group", and "Compound 1 (60) group", respectively.

[0081] <Evaluation method>

[0082] ALT was measured by blood collection from the tail vein at 12 weeks of administration. At 18 weeks of administration, autopsy was performed, and aspartate aminotransferase (AST) in plasma, total cholesterol (Tcho) in plasma, and triglyceride (TG) in the liver were measured. In addition, the area rate of picrosirius and the number of Hepatic crown-like structure (hCLS) were measured by liver analysis.

[0083] <Statistical analysis>

[0084] Statistical analysis was performed using GraphPad Prism 6 (GraphPad Software, Inc.). For the normal group and the control (vehicle) group, a t-test or a Mann-Whitney's test was performed. For the control group and the compound 1 group, a Dunnett-type multiple comparison test or a Dunn's-type multiple comparison test was performed. Either test was selected as appropriate depending on the distribution of each parameter. The significance level for all tests was set to p = 0.05.

[0085] ALT and AST in Plasma

[0086] Figure 1A ALT in plasma at 12 weeks of administration is shown. Figure 1B ALT in plasma at 18 weeks of administration is shown. Figure 1C AST in plasma at 18 weeks of administration is shown. The control group showed a statistically significant increase in ALT and AST in plasma relative to the normal group. In addition, the compound 1 (60) group showed a statistically significant inhibitory effect on the increase in ALT and AST in plasma relative to the control group. It was confirmed from this result that compound 1 improved liver dysfunction.

[0087] Tcho in Plasma

[0088] Figure 2 The value of Tcho in plasma at 18 weeks of administration is shown. The control group showed a statistically significant increase in Tcho in plasma relative to the normal group. In addition, the compound 1 (60) group showed a statistically significant inhibitory effect on the increase in Tcho in plasma relative to the control group. It was confirmed from this result that compound 1 improved lipid metabolism.

[0089] TG in Liver

[0090] The liver sample was homogenized with a phosphate buffer, methanol and chloroform were added, and after extraction overnight, further chloroform and distilled water were added. The mixture was centrifuged and the chloroform layer was collected, and the solvent was dried. The residue was dissolved with isopropyl alcohol / Triton X-100, which was used as a measurement sample, and measured using Triglyceride E-Test Wako (Fuji Film Wako Pure Chemical Industries, Ltd.).

[0091] Figure 3 The value of TG in liver at 18 weeks of administration is shown. The control group showed a statistically significant increase in TG in liver relative to the normal group. In addition, the compound 1 (30) group and the compound 1 (60) group showed a statistically significant inhibitory effect on the increase in TG in liver relative to the control group. It was confirmed from this result that compound 1 had an inhibitory effect on the fattening of the liver.

[0092] <Centaur positive area rate>

[0093] The excised livers were fixed with a 10% neutral buffered formalin solution, paraffin-embedded according to the conventional method, and prepared into centaur-stained specimens. Using the centaur-stained specimens, the positive area rate was calculated. The positive area rate of the normal group was averaged as the background value.

[0094] Figure 4 The centaur positive area rate of the liver at the 18th week of administration is shown. The control group showed a statistically significant increase in the centaur positive area rate of the liver relative to the normal group. In addition, the compound 1 (60) group showed a statistically significant increase in the inhibition of the centaur positive area rate of the liver relative to the control group. From this result, it was confirmed that compound 1 has an anti-fibrosis effect on the liver.

[0095] <Number of hCLS>

[0096] A CD68 immunostained specimen (rabbit anti-CD68 polyclonal antibody [ab125212], abcam) was prepared, and the hCLS was counted.

[0097] Figure 5 The number of CD68-positive hCLS based on immunostaining of the liver at the 18th week of administration is shown. The control group showed a statistically significant increase in the number of hCLS of the liver relative to the normal group. In addition, the compound 1 (10) group and the compound 1 (60) group showed a statistically significant increase in the inhibition of the number of hCLS of the liver relative to the control group. From this result, it was confirmed that compound 1 has an anti-inflammatory effect on the liver.

[0098] <Pathological analysis>

[0099] The results of the observation of the pathological tissues of the liver are shown in Table 2. In the pathological analysis, it was also confirmed that compound 1 has an improving effect on the fattening and fibrosis of the liver.

[0100] [Table 2]

[0101] Table 2 Pathological tissue observation results

[0102]

[0103] Note) -: negative / normal range, ±: slight, +: moderate, ++: significant

[0104] Regarding the fibrosis evaluation, individuals with nodules were excluded

[0105] <Overall evaluation>

[0106] Since Compound 1 inhibits the increase in ALT / AST / Tcho in plasma and TG in the liver, it significantly improves the indices related to fatty liver disease (liver escape enzyme and lipid metabolism). Further, Compound 1 also significantly inhibits the percentage of the area positive to picrosirius and the number of hCLS, improving the indices of fibrosis and inflammation of the liver. The results confirming these improvement effects are also shown in the results of pathological tissue observation. From the above results, the efficacy of Compound 1 in the NASH model can be confirmed.

Claims

1. Use of a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof in the manufacture of a therapeutic drug for fatty liver disease, [Chemical Formula 1] ， wherein the fatty liver disease is metabolic-associated fatty liver disease (MAFLD).

2. Use according to claim 1, wherein, the pharmaceutically acceptable salt is a hydrochloride.

3. Use according to claim 1, wherein, the MAFLD is accompanied by fibrosis of the liver.

4. The use according to claim 1, wherein, the MAFLD is accompanied by inflammation of the liver.

5. Use of a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof in the manufacture of a therapeutic drug for fatty liver disease, [Chemical Formula 1] ， wherein, the fatty liver disease is nonalcoholic steatohepatitis (NASH).

6. Use according to claim 5, wherein, the pharmaceutically acceptable salt is a hydrochloride.

7. Use according to claim 5, wherein, the NASH is accompanied by fibrosis of the liver.

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