The use of micelles to deliver potential hedgehog pathway inhibitor for the treatment of liver fibrosis

Abstract

Rationale: Hedgehog (Hh) pathway plays an essential role in liver fibrosis by promoting the proliferation of hepatic stellate cells (HSCs) by enhancing their metabolism via yes-associated protein 1 (YAP1). Despite the presence of several inhibitors, Hh signaling cannot be controlled exclusively due to their poor efficacy and the lack of a suitable delivery system to the injury site. Therefore, it is rationale to develop new potent Hh inhibitors and suitable delivery carriers.Methods: Based on the structure and activity of Hh inhibitor GDC-0449, we replaced its sulfonamide group with two methylpyridine-2yl at amide nitrogen to synthesize MDB5. We compared the Hh pathway inhibition and anti-fibrotic effect of MDB5 with GDC-0449 in vitro. Next, we developed MDB5 loaded micelles using our methoxy poly(ethylene glycol)-blockpoly(2-methyl-2-carboxyl-propylene carbonate-graft-dodecanol (PEG-PCC-g-DC) copolymer and characterized for physicochemical properties. We evaluated the therapeutic efficacy of MDB5 loaded micelles in common bile duct ligation (CBDL) induced liver fibrosis, mouse model. We also determined the intrahepatic distribution of fluorescently labeled micelles after MDB5 treatment.Results: Our results show that MDB5 was more potent in inhibiting Hh pathway components and HSC proliferation in vitro. We successfully developed MDB5 loaded micelles with particle size of 40 ± 10 nm and drug loading up to 10% w/w. MDB5 loaded micelles at the dose of 10 mg/kg were well tolerated by mice, without visible sign of toxicity. The serum enzyme activities elevated by CBDL was significantly decreased by MDB5 loaded micelles compared to GDC-0449 loaded micelles. MDB5 loaded micelles further decreased collagen deposition, HSC activation, and Hh activity and its target genes in the liver. MDB5 loaded micelles also prevented liver sinusoidal endothelial capillarization (LSEC) and therefore restored perfusion between blood and liver cells.Conclusions: Our study provides evidence that MDB5 was more potent in inhibiting Hh pathway in HSC-T6 cells and showed better hepatoprotection in CBDL mice compared to GDC-0449.

The Role of Autophagy and NLRP3 Inflammasome in Liver Fibrosis

“…It has been demonstrated that HSCs are Hh responsive and will be transformed into fibrogenic myofibroblasts (MFs). However, HSC activation can be inhibited by interfering Hh signaling [67,68]. It is well-known that excessive autophagy and NLRP3 inflammasome activation can induce autophagic cell death and pyroptosis, respectively [69].…”

When Viruses Cross Developmental Pathways

“…For instance, GANT-61 inhibited HH pathway members, GLI1 and GLI2 in HPV and KSHV associated tumors (Samarzija and Beard, 2012;Asha et al, 2020). Vismodegib, a SMO inhibitor that effectively terminates HH signaling, decreased liver fibrosis induced by HBV and HCV infection (Kumar et al, 2019). GSI repressed Notch signaling induced by KSHV, EBV, and HCV in associated diseases (Ito et al, 2011;Giunco et al, 2015;Jiang B.C.…”

Nanoapproaches to Modifying Epigenetics of Epithelial Mesenchymal Transition for Treatment of Pulmonary Fibrosis

“…These are summarized in Table 4 . Like with liposomes, the majority of these utilize APIs loaded into a lipophilic core, including pirfenidone (a drug used to treat pulmonary fibrosis) ( Singh et al, 2019 ), MDB5 (a novel form of Hedgehog (Hh) inhibitor GDC-0449) ( Kumar et al, 2019 ), salinomycin (an antibiotic with anti-cancer properties) ( Sousa et al, 2019 ), and doxorubicin (an anthracycline chemotherapeutic) ( Fang et al, 2014 ; Seifi-Najmi et al, 2016 ). Multiple formulations also make use of siRNA targeting genes along the EMT pathway ( Fang et al, 2014 ; Seifi-Najmi et al, 2016 ; Li et al, 2019 ; Suresh et al, 2019 ), including one directly conjugated to PEI to form a polyplex ( Ding et al, 2018 ; Wang Y. et al, 2019 ), as well as an inhibitor of CXCR4 ( Suresh et al, 2019 ), a chemokine receptor involved in the promotion of tumor migration and EMT ( Lin et al, 2018 ).…”

“…These all resulted in the inhibition of key EMT-biomarkers, including PAI-1, TGF-β, VEGF, Hh, Vimentin, and N-Cadherin. This led to decreased collagen deposition ( Wang Y. et al, 2019 ; Kumar et al, 2019 ; Li et al, 2019 ), cell migration and metastasis ( Fang et al, 2014 ; Seifi-Najmi et al, 2016 ; Sousa et al, 2019 ), and tumor growth ( Suresh et al, 2019 ). Polymeric NPs increased lung penetration and retention, provided stability by a polymer-siRNA conjugate, and facilitated codelivery further enhanced API efficacies.…”