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Abstract
Collagen prolyl 4-hydroxylase 1 (C-P4H1) is an α-ketoglutarate (α-KG)-dependent dioxygenase that catalyzes 4-hydroxylation of proline on collagen. C-P4H1-induced prolyl hydroxylation is required for proper collagen deposition and cancer metastasis. Therefore, targeting C-P4H1 is considered a potential therapeutic strategy for collagen-related cancer progression and metastasis. However, no C-P4H1 inhibitors are available for clinical testing, and the high content assay is currently not available for C-P4H1 inhibitor screening. In the present study, we developed a high-throughput screening assay by quantifying succinate, a byproduct of C-P4H-catalyzed hydroxylation. C-P4H1 is the major isoform of collagen prolyl 4-hydroxylases (CP4Hs) that contributes the majority prolyl 4-hydroxylase activity. Using C-P4H1 tetramer purified from the eukaryotic expression system, we showed that the Succinate-GloTM Hydroxylase assay was more sensitive for measuring C-P4H1 activity compared with the hydroxyproline colorimetric assay. Next, we performed high-throughput screening with the FDA-approved drug library and identified several new C-P4H1 inhibitors, including Silodosin and Ticlopidine. Silodosin and Ticlopidine inhibited C-P4H1 activity in a dose-dependent manner and suppressed collagen secretion and tumor invasion in 3D tissue culture. These C-P4H1 inhibitors provide new agents to test clinical potential of targeting C-P4H1 in suppressing cancer progression and metastasis.
Document Type
Article
Publication Date
9-10-2020
Digital Object Identifier (DOI)
https://doi.org/10.3390/ijms21186613
Funding Information
This study was supported by funding from NIH (1R01CA207772, and 1R01CA215095 to R.X.).
Repository Citation
Wang, Shike; Lee, Kuo-Hao; Araujo, Nathália Victoria; Zhan, Chang-Guo; Rangnekar, Vivek M.; and Xu, Ren, "Develop a High-Throughput Screening Method to Identify C-P4H1 (Collagen Prolyl 4-Hydroxylase 1) Inhibitors from FDA-Approved Chemicals" (2020). Pharmaceutical Sciences Faculty Publications. 150.
https://uknowledge.uky.edu/ps_facpub/150
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Cancer Biology Commons, Oncology Commons, Pharmacology Commons, Pharmacy and Pharmaceutical Sciences Commons, Radiation Medicine Commons, Toxicology Commons
Notes/Citation Information
Published in International Journal of Molecular Sciences, v. 21, issue 18, 6613.
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).