Abstract
The G protein-coupled receptor (GPCR) smoothened (SMO) is a key signaling component of the sonic hedgehog (Hh) pathway and a clinically validated target for cancer treatment. The FDA-approved SMO inhibitors GDC-0449/Vismodegib and LDE225/Sonidegib demonstrated clinical antitumor efficacy. Nevertheless, relatively high percentage of treated patients would eventually develop acquired cross resistance to both drugs. Here, based on published structure and activity of GDC-0449 inhibitor class, we replaced its amide core with benzimidazole which retained bulk of the SMO-targeting activity as measured in our Hh/SMO/Gli1-reporter system. Synthesis and screening of multiple series of benzimidazole derivatives identified HH-1, HH-13, and HH-20 with potent target suppression (IC50: <0.1 μmol/L) in the reporter assays. In NIH3T3 cells stimulated with a secreted Hh (SHH), these inhibitors dose dependently reduced mRNA and protein expression of the endogenous pathway components PTCH-1, Gli1, and cyclin D1 resulting in growth inhibition via G0/G1 arrest. Mechanistically, the SMO-targeted growth inhibition involved downregulation of mTOR signaling inputs and readouts consistent with diminished mTORC1/mTORC2 functions and apoptosis. In mice, as with GDC-0449, orally administered HH inhibitors blocked paracrine activation of stromal Hh pathway in Calu-6 tumor microenvironment and attenuated growth of PTCH+/−/P53−/− medulloblastoma allograft tumors. Furthermore, HH-13 and HH-20 potently targeted the drug-resistant smoothened SMO-D473H (IC50: <0.2 μmol/L) compared to the poor inhibition by GDC-0449 (IC50: >60 μmol/L). These results identify HH-13 and HH-20 as potent inhibitors capable of targeting naïve and drug-resistant Hh/SMO-driven cancers. The current leads may be optimized to improve pharmaceutical property for potential development of new therapy for treatment of Hh pathway-driven cancers.
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Acknowledgements
This work was supported by a startup grant from Fudan University (EZF301002) and National Natural Science Foundation of China (81273367). We thank Fudan University School of Pharmacy Animal Facility and Instrument Center for excellent technical support.
Author contributions
Conception and design: H.L., K.Y. Development of methodology: H.Z., Q.L., K.Y. Acquisition of data: H.Z., Q.L., X.Z. Analysis and interpretation of data: H.Z., Q.L., X.Z. Writing, review, and/or revision of the manuscript: Q.L., H.L., K.Y. Administrative, technical, or material support: H.L., K.Y. Study supervision: K.Y.
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Li, Qr., Zhao, H., Zhang, Xs. et al. Novel-smoothened inhibitors for therapeutic targeting of naïve and drug-resistant hedgehog pathway-driven cancers. Acta Pharmacol Sin 40, 257–267 (2019). https://doi.org/10.1038/s41401-018-0019-5
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DOI: https://doi.org/10.1038/s41401-018-0019-5
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