New IMB16-4 Hot-Melt Extrusion Preparation Improved Oral Bioavailability and Enhanced Anti-Cholestatic Effect on Rats

Abstract

Background Cholestasis is challenging to treat due to lacked effective drugs. N-(3,4,5-trichlorophenyl)-2 (3-nitrobenzenesulfonamido) benzamide, abbreviated as IMB16-4, which may be effective for the treatment of cholestasis. However, its poor solubility and bioavailability seriously obstruct the research programs. Methods A hot-melt extrusion (HME) preparation was first applied to increase the bioavailability of IMB16-4, the oral bioavailability, anti-cholestatic effect and vitro cytotoxicity of IMB16-4 and IMB16-4-HME were evaluated. Meanwhile, the molecular docking and qRT-PCR were used to validate the mechanism behind. Results The oral bioavailability of IMB16-4-HME improved 65-fold compared with that of pure IMB16-4. Pharmacodynamics results demonstrated that IMB16-4-HME prominently decreased the serum levels of total bile acid (TBA) and alkaline phosphatase (ALP), but elevated the level of total bilirubin (TBIL) and direct bilirubin (DBIL). Histopathology revealed that IMB16-4-HME at lower dose exhibited stronger anti-cholestatic effect compared with pure IMB16-4. In addition, molecular docking demonstrated that IMB16-4 has great affinity with PPARα, and qRT-PCR results revealed that IMB16-4-HME significantly elevated the mRNA expression level of PPARα, but decreased the mRNA level of CYP7A1. Cytotoxicity assays demonstrated that the hepatotoxicity of IMB16-4-HME was absolutely attributed to IMB16-4, and the excipients of IMB16-4-HME may increase the drug load within HepG2 cells. Conclusion The HME preparation significantly increased the oral bioavailability and anti-cholestatic effect of pure IMB16-4, but caused liver injury at high dose, which require a dose balance between the curative effect and safety in the future research.

Effects of Long-Term High-Ergosterol Intake on the Cholesterol and Vitamin D Biosynthetic Pathways of Rats Fed a High-Fat and High-Sucrose Diet

“…42) Hence, in this study, we used the HFHS rat model to evaluate ergosterolmediated inhibition of dyslipidemia. Additionally, HFHS diets induce hepatic damage (indicated by increasing AST, ALT, ALP, and LDH levels at 2.6-to 23.0-fold), 26,27) cholestasis (indicated by increasing ALP, T-BIL, D-BIL, I-BIL, and TBA levels at 2.5to 23.7-fold), [27][28][29][30] hypercholesterolemia (indicated by increasing T-CHO, F-CHO, and LDL-C levels at 2.5-to 6.5-fold), 17) and an increased risk of cardiovascular disease…”

“…45) Furthermore, plasma ALP, T-BIL, D-BIL, and TBA levels were lower by 30%, 56%, 62%, and 43%, respectively, in the HFHS + ergosterol group than those in the HFHS group, suggesting that ergosterol might improve severe cholestasis. [28][29][30] This is because studies have a reduction in ALP, T-BIL, D-BIL, and TBA levels (31%, 51%, 34%, and 39%, respectively), which play a therapeutic role in resisting cholestasis. 26,27,42) Ergosterol, brassicasterol, and vitamin D2 were detected in the HFHS + ES group, indicating that the HFHS diet with long-term ergosterol intake facilitated the absorption ergosterol, its conversion to brassicasterol by DHCR7, [9][10][11][12] and ultimately its conversion to vitamin D2 by UV light.…”