Application of xanthine derivative in preparation of medicine for preventing and / or treating fatty liver disease related to metabolic dysfunction

Abstract

The invention provides application of a xanthine derivative in preparation of a medicine for preventing and / or treating fatty liver diseases related to metabolic dysfunction, and belongs to the technical field of medicine preparation. The invention relates to an application of a xanthine derivative 8-[(2, 3-dihydroxy propyl) sulfo]-3-methyl-7-[(4-methylphenyl) methyl]-1H-purine-2, 6-diketone in preparation of a medicine for preventing and / or treating fatty liver diseases related to metabolic dysfunction. The xanthine derivative has dual efficacy of improving liver cell insulin sensitivity and / or reducing liver cell lipid accumulation, and provides a new way for treatment of metabolic dysfunction related fatty liver diseases and preparation of related clinical drugs.

Description

Technical Field

[0001] This application belongs to the field of pharmaceutical preparation technology, specifically relating to the use of xanthine derivatives in the preparation of drugs for the prevention and / or treatment of fatty liver disease related to metabolic dysfunction. Background Technology

[0002] Metabolic dysfunction-associated fatty liver disease (MASLD, formerly known as NAFLD) is the most common chronic liver disease worldwide. Its disease spectrum includes simple hepatic steatosis (MASL), which can progress to metabolic-associated steatohepatitis (MASH), which is accompanied by hepatocellular damage, inflammation, and varying degrees of liver fibrosis, and may eventually develop into cirrhosis, liver failure, or even hepatocellular carcinoma.

[0003] Currently, there are no approved drugs worldwide for the treatment of MASLD. Standard clinical treatment mainly relies on lifestyle interventions (such as weight loss, exercise, and diet management), but due to poor long-term adherence, the overall treatment effect is limited. Summary of the Invention

[0004] The purpose of this invention is to provide the use of the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione in the preparation of a medicament for the prevention and / or treatment of fatty liver disease associated with metabolic dysfunction. The xanthine derivative has the dual effects of improving hepatocyte insulin sensitivity and lipid metabolism, and has excellent drug-like properties.

[0005] This invention provides the use of the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione in the preparation of medicaments for the prevention and / or treatment of fatty liver disease associated with metabolic dysfunction.

[0006] Preferably, the treatment of metabolic dysfunction-related fatty liver disease includes improving hepatocyte insulin sensitivity and / or reducing lipid accumulation in hepatocytes.

[0007] Preferably, the cell model of metabolic dysfunction-associated fatty liver disease is induced by free fatty acids.

[0008] Preferably, the free fatty acid includes palmitic acid.

[0009] Preferably, the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione comprises at least one of the following forms: a pharmaceutically acceptable salt form, a solvate, and a prodrug compound.

[0010] Preferably, the drug includes pharmaceutically acceptable excipients.

[0011] Preferably, the drug dosage form includes oral formulations and / or injectable formulations.

[0012] This invention provides a drug for treating fatty liver disease associated with metabolic dysfunction, the active ingredient of which includes the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione.

[0013] Preferably, the drug further includes pharmaceutically acceptable excipients.

[0014] Preferably, the dosage form of the drug includes at least one of granules, tablets, capsules, pills, and oral liquid preparations.

[0015] This invention provides the application of the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related fatty liver disease. Insulin sensitivity experiments showed that, compared with a blank control, the xanthine derivative significantly increased p-AKT and p-IR levels in mouse primary hepatocytes (MPHs) after insulin intervention. This indicates that the xanthine derivative can effectively enhance the insulin sensitivity of MPHs. Simultaneously, Oil Red O staining experiments showed that the xanthine derivative can effectively reduce palmitic acid-induced lipid accumulation in Huh7 cells and MPHs. Furthermore, cytotoxicity tests showed that the xanthine derivative has good drug safety. Therefore, the xanthine derivative exhibits excellent multiple effects in improving hepatocyte insulin sensitivity and lipid accumulation, and possesses excellent drug-like properties. This invention not only broadens the applications of the xanthine derivatives in pharmaceuticals, but also provides a new approach for the treatment of fatty liver disease related to metabolic dysfunction and the development of related drugs. Attached Figure Description

[0016] Figure 1 The results are for cytotoxicity assessment. Figure 2 To detect the effect of drugs on the phosphorylation levels of AKT and IR (p-AKT, p-IR) using Western blotting; Figure 3 The phosphorylation level of AKT at Ser473 by xanthine derivatives (p-AKT) S473 / AKT affects statistical results; Figure 4 p-IR, representing the phosphorylation level of xanthine derivatives at the Tyr1140 site. Y1140 / IR affects statistical results; Figure 5 The results are from Oil Red O staining. The scale bar for Huh7 cell images is 100 μm, and the scale bar for MPHs cell images is 50 μm. Detailed Implementation

[0017] This invention provides the use of the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione in the preparation of medicaments for the prevention and / or treatment of fatty liver disease associated with metabolic dysfunction.

[0018] In this invention, the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione has the CAS number 303969-88-2 and the molecular formula C 17 H 20 N4O4S has a molecular weight of 376.43 and its structural formula is shown in Formula I.

[0019] Formula I.

[0020] In this embodiment of the invention, the xanthine derivative was purchased from Lifechemicals, catalog number F0373-1257.

[0021] In this invention, the xanthine derivative preferably comprises at least one of the following forms: a pharmaceutically acceptable salt form, a solvate, and a prodrug compound.

[0022] In this invention, the treatment of metabolic dysfunction-associated fatty liver disease (MASLD) preferably includes improving hepatocyte insulin sensitivity and / or reducing lipid accumulation in hepatocytes. The cell model of MASLD is preferably induced by free fatty acids. The free fatty acids preferably include palmitic acid. The treatment concentration of palmitic acid is preferably 0.3 mM. The improvement in hepatocyte insulin sensitivity is characterized by detecting the phosphorylation levels (p-AKT, p-IR) of the key insulin signaling pathway protein AKT at Ser473 and the key protein IR at Tyr1140. Insulin resistance is the "first hit" and a persistent driver of MASLD. The xanthine derivative of this invention improves liver metabolic disorders at their source by directly enhancing hepatocyte sensitivity to insulin, which goes beyond the superficial effects of simply "protecting the liver" or "lowering lipids."

[0023] In this invention, the drug preferably includes pharmaceutically acceptable excipients. This invention does not impose any particular limitation on the type of excipients; common excipients well known in the art can be used, such as fillers, binders, wetting agents, disintegrants, lubricants, solubilizers, emulsifiers, preservatives, flow aids, coating materials, flavoring agents, osmotic pressure regulators, and pH adjusters. The drug dosage form preferably includes oral and / or injectable formulations. The xanthine derivative meets the first requirement of Lipinski's "five rules," indicating good oral absorption potential. The number of hydrogen bond donors (HBD=3) and hydrogen bond acceptors (HBA=8) are as follows: the number of hydrogen bond donors fully complies with the "five rules" (HBD≤5), and although the number of hydrogen bond acceptors slightly exceeds the upper limit (HBA≤10), considering that its structure contains an essential, highly hydrophilic dihydroxypropylthiol side chain, this value is within an acceptable optimization range and does not negatively affect the overall properties of the compound. The lipid-water partition coefficient (calculated LogP = 0.95) is particularly outstanding, falling within the ideal range (0-3) for drug-like molecules. This indicates that the compound achieves an excellent balance between hydrophilicity and lipophilicity; moderate lipophilicity (contributed by the p-methylbenzyl group at position 7) facilitates its penetration through cell membranes and reaching its intracellular target site; sufficient hydrophilicity (mainly contributed by the 2,3-dihydroxypropylthio group at position 8) ensures adequate solubility in aqueous media (such as blood and cytoplasm), avoiding absorption problems caused by low solubility. In summary, the xanthine derivative fully conforms to the classic drug-likeness rules, laying a solid physicochemical foundation for its development as an oral drug.

[0024] This invention provides a drug for treating fatty liver disease associated with metabolic dysfunction, the active ingredient of which includes the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione.

[0025] In this invention, the drug preferably further includes pharmaceutically acceptable excipients. This invention does not impose any particular limitation on the type of excipients; any excipients well-known in the art can be used. The dosage form of the drug preferably includes at least one of granules, tablets, capsules, pills, and oral liquid preparations. This invention does not impose any particular limitation on the preparation method of the drug; any preparation method well-known in the art can be used. The mass percentage of xanthine derivatives in the drug is 1%~99%, which can be 10%~800%, 15%~70%, 20%~60%, or 40%.

[0026] The following examples illustrate the application of the xanthine derivatives provided by the present invention in the preparation of medicaments for the prevention and / or treatment of fatty liver disease related to metabolic dysfunction. However, these examples should not be construed as limiting the scope of protection of the present invention.

[0027] Example 1 Cytotoxicity assessment (MTS assay) MTS experiments were conducted to assess the cellular safety of the compounds.

[0028] Human hepatocellular carcinoma Huh7 cells were seeded at standard density in 96-well plates. After the cells adhered, the medium was replaced with fresh medium containing either 10 μM xanthine derivative (8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione) or an equal volume of DMSO (control solvent), and cultured for 24 h, 48 h, and 72 h. At each time point, MTS reagent was added to each well, and after incubation for 1 h, the absorbance was measured at 490 nm using a microplate reader.

[0029] See results Figure 1 Compared with the solvent control group, the 10 μM purine derivative treatment group did not cause a significant decrease in absorbance in Huh7 cells, indicating that the compound had no significant toxic effect on cell viability at this concentration, providing a safe concentration basis for subsequent pharmacodynamic experiments.

[0030] Example 2 Insulin sensitivity test To verify the efficacy of purine derivatives in improving insulin sensitivity, the study was conducted in mouse primary hepatocytes (MPHs).

[0031] Model establishment and drug administration: Primary mouse hepatocytes (MPHs) were seeded at an appropriate density in 12-well plates. After cell adhesion, the medium was replaced with fresh medium containing 10 μM purine derivatives or DMSO (solvent control) and treated for 24 h. Subsequently, the cells were starved for 6 h in serum-free medium and treated with insulin (MPHs 10 nM insulin) for 0, 5, and 15 min. The phosphorylation levels (p-AKT, p-IR) of key insulin signaling proteins AKT and IR were detected by Western blotting. A blank control of primary mouse hepatocytes (MPHs) was also set up.

[0032] See results Figures 2-4 Compared with the control group, the phosphorylation level of AKT (p-AKT) in cells treated with purine derivatives was significantly higher. S473 / AKT) and the phosphorylation level of IR (p-IR) Y1140 Both insulin sensitivity (IR) and insulin sensitivity (PHS) were significantly increased. This indicates that purine derivatives can effectively enhance the insulin sensitivity of primary mouse hepatocytes (MPHs).

[0033] Example 3 Oil Red O staining experiment To verify the efficacy of purine derivatives in inhibiting lipid accumulation in hepatocytes, an Oil Red O staining experiment was performed.

[0034] A hepatic steatosis model was established by stimulating Huh7 cells and mouse primary hepatocytes (MPHs) with free fatty acids (0.3 mM palmitic acid PA) for 24 h. Simultaneously, 10 μM purine derivatives were added for a co-intervention for 24 h, serving as the experimental group, while the model group used DMSO as a control. After the intervention, the culture medium was discarded, and the cells were fixed with 4% paraformaldehyde, followed by Oil Red O staining to label neutral lipid droplets. After staining, the cells were observed and photographed under an optical microscope, or quantitative absorbance was measured after dissolving the dye in isopropanol. BSA-treated control groups for both cell types were also included.

[0035] See results Figure 5 In the control group, numerous bright red lipid droplets appeared in the cytoplasm of cells. However, in cells treated with purine derivatives, both the number and size of these red lipid droplets were significantly reduced, indicating a significant decrease in lipid content. This suggests that purine derivatives can effectively reduce lipid accumulation in hepatocytes.

[0036] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione in the preparation of drugs for the prevention and / or treatment of fatty liver disease associated with metabolic dysfunction.

2. The application according to claim 1, characterized in that, The treatment of metabolic dysfunction-related fatty liver disease includes improving hepatocyte insulin sensitivity and / or reducing lipid accumulation in hepatocytes.

3. The application according to claim 1, characterized in that, The cell model of metabolic dysfunction-associated fatty liver disease was induced by free fatty acids.

4. The application according to claim 3, characterized in that, The free fatty acids include palmitic acid.

5. The application according to claim 1, characterized in that, The xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione comprises at least one of the following forms: a pharmaceutically acceptable salt form, a solvate, and a prodrug compound.

6. The application according to claim 1, characterized in that, The drug includes pharmaceutically acceptable excipients.

7. The application according to any one of claims 1 to 6, characterized in that, The drug dosage forms include oral formulations and / or injectable formulations.

8. A drug for treating fatty liver disease associated with metabolic dysfunction, characterized in that, The active ingredients include the xanthine derivative 8-[(2,3-dihydroxypropyl)thio]-3-methyl-7-[(4-methylphenyl)methyl]-1H-purine-2,6-dione.

9. The drug according to claim 8, characterized in that, It also includes pharmaceutically acceptable excipients.

10. The medicament for treating metabolic dysfunction-related fatty liver disease according to claim 8 or 9, characterized in that, The dosage form of the drug includes at least one of granules, tablets, capsules, pills, and oral liquid preparations.

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