Combined use of dihydroquinoline derivative and PPAR alpha agonist

Abstract

The invention provides a combined use of a dihydroquinoline derivative and a PPARalpha agonist, and particularly relates to an application of a specific dihydroquinoline derivative and a PPARalpha agonist in preparation of a product for preventing or treating fatty liver. It is found for the first time that the combination of the specific dihydroquinoline derivative and the PPARalpha agonist can significantly reduce the triglyceride level of the liver, so that alcohol-induced liver fatty degeneration is relieved, and the application has good clinical transformation prospects.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the combined use of a dihydroquinoline derivative and a PPARα agonist, particularly in the preparation of products for the prevention or treatment of fatty liver. Background Technology

[0002] Currently, there are no effective targeted therapies for fatty liver, such as alcoholic fatty liver disease (ALD), and clinical treatment mainly focuses on alcohol abstinence and supportive care. PPARα is a key nuclear receptor regulating fatty acid β-oxidation, but its nuclear transport in ALD is impaired, affecting transcriptional activity and leading to lipid accumulation. Existing PPARα agonists, such as fenofibrate, can partially improve lipid metabolism, but their effects depend on intact nuclear transport processes, making it difficult to overcome the treatment bottleneck caused by nuclear transport impairment. Summary of the Invention

[0003] To address the aforementioned problems, this invention utilizes a specific combination of dihydroquinoline derivatives and PPARα agonists to demonstrate therapeutic effects on fatty liver, particularly alcoholic fatty liver.

[0004] Specifically, this application discloses a combined use of a dihydroquinoline derivative and a PPARα agonist, characterized in that: in the preparation of products for the prevention or treatment of fatty liver, the structure of the dihydroquinoline derivative is as follows: , where R represents a C1-C4 alkyl group and X represents a halogen.

[0005] Furthermore, the fatty liver is selected from one of alcoholic fatty liver, non-alcoholic fatty liver, malnutrition-induced fatty liver, and drug-induced fatty liver.

[0006] Furthermore, the fatty liver mentioned is alcoholic fatty liver.

[0007] Furthermore, in the structural formula of the dihydroquinoline derivative, R is methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl.

[0008] Furthermore, in the structural formula of the dihydroquinoline derivative, X is F or Cl.

[0009] Furthermore, the structure of the dihydroquinoline derivative is as follows: (In this application, it is abbreviated as Fos).

[0010] Furthermore, the PPARα agonist is a fibrate or a non-fibrate drug.

[0011] Furthermore, the PPARα agonist is selected from one or more of fenofibrate (abbreviated as FB in this application), pemafibrate, gemfibrozil, sitagliptazakate, bezafibrate, ciprofibrate, clofibrate, aleglitazar, elafibrano (α / δ), and saroglitazar (α / γ).

[0012] Furthermore, the dosage of the dihydroquinoline derivative is 1-20 mg / kg / day, preferably 10 mg / kg / day.

[0013] Furthermore, the dosage of PPARα agonist is 20-80 mg / kg / day, preferably 20-50 mg / kg / day.

[0014] The products include pharmaceuticals.

[0015] This invention is the first to discover that dihydroquinoline derivatives significantly reduce liver triglyceride levels, thereby alleviating alcohol-induced hepatic steatosis, and has good prospects for clinical translation. Attached Figure Description

[0016] Figure 1 The plasma ALT levels in the Lieber-DeCarli alcoholic fatty liver model showed that liver damage was reduced after Fos treatment, and the combined use of PPARα agonists further reduced liver damage. Figure 2 In the Lieber-DeCarli alcoholic fatty liver model, liver H&E staining showed a decrease in hepatic lipid accumulation after Fos treatment, which was further reduced by the combined use of PPARα agonists. Figure 3 Oil Red O staining of the liver in the Lieber-DeCarli alcoholic fatty liver model showed a decrease in hepatic lipid accumulation after Fos treatment, which was further reduced by the combined use of PPARα agonists.

[0017] Figure 4 In the Lieber-DeCarli alcoholic fatty liver model, liver liposuction showed a decrease in hepatic triglyceride accumulation after Fos treatment, and the combined use of PPARα agonists further reduced it. Detailed Implementation

[0018] Example 1: Experimental methods and grouping: Eight-week-old female C57BL / 6J mice were divided into groups and modeled with a Lieber-DeCarli alcoholic liquid diet for 2 weeks. Then, they were given solvent (abbreviated as control), Fos (10 mg / kg / day, abbreviated as Fos), PPARα agonist fenofibrate (50 mg / kg / day), and Fos combination therapy (abbreviated as Fos+FB, with Fos dosages of 10 mg / kg / day and 40 mg / kg / day, respectively) for 2 weeks.

[0019] Liver tissue samples were stained with hematoxylin and eosin (HE) and Oil Red O. The results showed that Fos significantly reduced liver lipid content. Liver liposuction also showed a significant reduction in triglycerides in the liver. Fos combined with the PPARα agonist fenofibrate further reduced liver lipid content.

[0020] Plasma was collected to detect alanine aminotransferase (ALT), a marker of liver damage. The results showed that Fos treatment significantly reduced ALT levels and improved liver damage; Fos combined with the PPARα agonist fenofibrate further reduced ALT and AST levels and improved liver damage.

[0021] The serum alanine aminotransferase (ALT) level in mice was measured using a fully automated biochemical analyzer. ALT levels were lower in the Fos (Fos) treatment group compared to the control group, and the decrease was more significant in the Fos+FB combination group, suggesting that this drug has a protective effect against alcoholic liver injury.

[0022] After paraffin sectioning, hematoxylin and eosin (HE) staining was performed, and hepatocyte structure and inflammatory changes were observed under a light microscope. In the control group, hepatocytes showed disordered arrangement and fatty degeneration; the Fos group showed improvement; and the combined Fos+FB group showed hepatocyte morphology closer to normal, indicating pathological improvement.

[0023] After freezing, sections were stained with Oil Red O to detect neutral lipid deposition. Significant lipid droplet deposition was observed in the control group, decreased in the Fos group, and significantly reduced in the Fos+FB group, suggesting that Fos can effectively reduce hepatic lipid deposition.

[0024] The level of triglycerides (TG) in liver tissue was quantitatively detected using a biochemical reagent kit, and the results were standardized to mg / g liver tissue. TG levels were significantly elevated in the control group; decreased in the Fos group; and showed the most significant decrease in the Fos+FB group, suggesting that Fos has a significant effect on improving hepatic lipid metabolism.

[0025] Figure 1 Serum alanine aminotransferase (ALT) level, test method: ALT activity (U / L) was detected by biochemical analyzer.

[0026] Labels: Control: control group, Fos: Fos monotherapy group, Fos+FB: Fos and fenofibrate combination group Elevated ALT levels in the control group indicated liver damage. ALT levels significantly decreased in the Fos group, with an even more significant decrease in the Fos+FB combination group, suggesting that Fos can improve alcoholic liver injury and has a synergistic effect with fenofibrate.

[0027] Figure 2 HE staining (structural changes in liver tissue), test method: paraffin sections, hematoxylin-eosin staining, observation under a microscope.

[0028] Control group: Disordered hepatocyte arrangement and significant fatty degeneration. Fos: Fos treatment group showed reduced fatty degeneration Fos+FB: Hepatocyte morphology in the combined treatment group was close to normal. Fos significantly improved alcohol-induced liver tissue pathological damage.

[0029] Figure 3 Oil Red O staining (lipid deposition detection): Test method: frozen sections, Oil Red O staining, detection of neutral fats.

[0030] In the control group, lipid droplet deposition was extensive; in the Fos group, deposition was reduced; and in the Fos+FB group, lipid droplets were minimal. These results indicate that Fos can effectively inhibit alcohol-induced lipid accumulation.

[0031] Figure 4 Liver triglyceride (TG) content, test method: quantitative detection using a kit, results expressed as mg / g liver. mg / g liver: triglyceride content per gram of liver tissue. In the control group, liver triglycerides (TG) were elevated; in the Fos group, they were decreased; and in the Fos+FB group, the decrease was most significant. These results were consistent with Oil Red O staining, further demonstrating that Fos reduces hepatic lipid accumulation.

[0032] In summary, the attached figures demonstrate from multiple perspectives—serological indicators, histological observation, lipid staining, and biochemical quantification—that Fos can significantly reduce alcohol-induced liver damage and lipid accumulation, and that it has a synergistic effect with fenofibrate.

[0033] Figure 1 (Serium ALT activity) Conditions: C57BL / 6J mice, alcohol feeding model (usually Lieber-DeCarli liquid diet or acute alcohol gavage), were divided into control group, Fos group, and Fos + fenofibrate (FB) combination group.

[0034] Figure 2 (Hematologic staining, histological changes in the liver) Conditions: Same as above, mouse liver tissue was fixed in 4% paraformaldehyde and embedded in paraffin.

[0035] Methods: Sections (4 μm) were stained with hematoxylin and eosin (H&E) and observed under an optical microscope to examine hepatocyte arrangement, ballooning degeneration, and inflammatory infiltration.

[0036] Figure 3 (Oil Red O staining, lipid droplet deposition) Conditions: Same as above, fresh liver tissue was taken and frozen sections were prepared (7 μm).

[0037] Methods: Neutral lipids were detected by Oil Red O staining, cell nuclei were counterstained with hematoxylin, and the distribution of red lipid droplets was observed under an optical microscope.

[0038] Figure 4 (Liver triglyceride (TG) content) Conditions: Same as above, liver tissue homogenate.

[0039] Methods: The colorimetric method using a commercial reagent kit was used for detection, and the results were standardized to mg TG / g liver.

[0040] Explanation of symbols in the diagram p > 0.05 is considered to be statistically insignificant; p < 0.05 is considered to be statistically significant.

[0041] Figure 1 (Serum alanine aminotransferase (ALT) level) In mice fed with alcohol, serum ALT levels were significantly elevated in the control group, indicating hepatocellular damage. Fos monotherapy significantly reduced ALT levels, and the reduction was even more significant when combined with fenofibrate, indicating that Fos can alleviate alcohol-induced liver injury and exhibit a synergistic protective effect when used in combination with other drugs.

[0042] Figure 2 (HE staining, liver histology) HE staining results showed that hepatocytes in the control group mice exhibited disordered arrangement and significant fatty degeneration. Pathological damage was reduced in the Fos-treated group, while the Fos combined with fenofibrate group showed more normal hepatocyte structure, significantly improved lipid droplet deposition and inflammatory infiltration, further demonstrating its protective effect at the histological level.

[0043] Figure 3 (Oil Red O staining, liver lipid droplet deposition) Oil Red O staining results showed that red lipid droplets were widely deposited in hepatocytes of the control group. Fos monotherapy reduced the number and size of lipid droplets, while Fos combined with fenofibrate significantly cleared lipid droplet deposits, suggesting that the drug can effectively improve alcohol-induced intrahepatic lipid accumulation and has a complementary effect with lipid metabolism regulating drugs.

[0044] Figure 4 (Liver triglyceride (TG) content) Liver tissue analysis showed that TG levels were significantly elevated in the control group. Fos treatment significantly reduced liver TG levels, with the combined effect of fenofibrate being the most pronounced, approaching normal levels. This indicates that Fos can restore hepatic lipid balance by promoting fatty acid β-oxidation, and that combined treatment further enhances its lipid-lowering effect.

[0045] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The combined use of a dihydroquinoline derivative and a PPARα agonist, characterized in that: For use in the preparation of products for the prevention or treatment of fatty liver, the structure of the dihydroquinoline derivative is as follows: , where R represents a C1-C4 alkyl group and X represents a halogen.

2. The use according to claim 1, characterized in that: The fatty liver is selected from one of the following: alcoholic fatty liver, non-alcoholic fatty liver, malnutrition-induced fatty liver, and drug-induced fatty liver.

3. The use according to claim 1, characterized in that: The fatty liver mentioned is alcoholic fatty liver.

4. The use according to claim 1, characterized in that: In the structural formula of the dihydroquinoline derivative, R is methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl.

5. The use according to claim 1, characterized in that: In the structural formula of the dihydroquinoline derivative, X is either F or Cl.

6. The use according to claim 1, characterized in that: The structure of the dihydroquinoline derivative is as follows: .

7. The use according to claim 1, characterized in that: The PPARα agonist is a fibrate or a non-fibrate drug.

8. The use according to claim 7, characterized in that: The PPARα agonist is selected from one or more of the following: fenofibrate, gemfibrozil, sitagliptazakate, bezafibrate, ciprofibrate, clofibrate, aleglitazar, elafibranor, and saroglitazar.

9. The use according to any one of claims 1-8, characterized in that: The dosage of dihydroquinoline derivatives is 1-20 mg / kg / day.

10. The use according to any one of claims 7-8, characterized in that: The dosage of PPARα agonists is 20-80 mg / kg / day.

Images