Design and synthesis of tag-free photoprobes for the identification of the molecular target for CCG-1423, a novel inhibitor of the Rho/MKL1/SRF signaling pathway

Jessica L. Bell, Andrew J. Haak, Susan M. Wade, Yihan Sun, Richard R. Neubig, Scott D. Larsen

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

CCG-1423 and related analogues represent a new class of inhibitors of Rho/MKL1/SRF-mediated gene transcription, a pathway that has been implicated in both cancer and fibrosis. The molecular target for these compounds is unknown. To facilitate its identification, a series of tag-free photoaffinity probes was designed and synthesized, each one containing a photoactivatable group and an acetylenic end group for subsequent attachment to a fluorescent tag using click chemistry. All were confirmed to maintain biological activity in a cell-based assay for inhibition of SRE-Luc expression. The functional activity of the most potent probe 24 was further confirmed in an assay for PC-3 cell migration. Photolysis of 24 in intact PC-3 cells followed by cell lysis, click ligation of a fluorescent dye, and gel electrophoresis revealed specific labeling of a single 24 kDa band that could be blocked with an active competitor. Future work will focus on identifying the labeled protein(s).

Original languageEnglish (US)
Pages (from-to)966-973
Number of pages8
JournalBeilstein Journal of Organic Chemistry
Volume9
DOIs
StatePublished - May 21 2013

Keywords

  • CCG-1423
  • Click ligation
  • Photoaffinity labeling
  • Rho pathway inhibitor
  • Tag-free photoprobe
  • Target identification

ASJC Scopus subject areas

  • Organic Chemistry

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