Characterizing Compounds Targeting Colorectal Cancer Derived From Monastrol Using High-Through Screening of an Extensive Combinatorial Library

BackgroundCancer remains a critical global health concern. Among its various forms, colorectal cancer (CRC) stands out due to its high prevalence and mortality rates, emphasizing the urgent need for novel therapeutic agents to enhance treatment efficacy and prolong patient survival. Monastrol, an antimitotic compound known to bind kinesin Eg5, is employed in some cancer therapies. Recent studies have revealed that monastrol also interacts with fascin, a protein implicated in tumor aggressiveness and metastasis, thereby disrupting microtubule dynamics and actin bundling, ultimately impairing cell migration. MethodsIn this work, we developed a workflow to identify fascin-binding compounds based on a monastrol-derived pharmacophore model, integrating in silico predictions with in vitro validation. We performed ligand-based virtual screening using a pharmacophore model constructed from monastrol, applied to a high-throughput screening (HTS) library of 1.6 million compounds. The top-ranking candidates from the virtual screening were subsequently subjected to physicochemical characterization and cellular assays. ResultsTwo compounds (designated Z118298144 and Z17544625) were identified that exhibited strong binding to fascin and inhibited actin bundling in physicochemical assays. Furthermore, cellular experiments demonstrated that both compounds reduced proliferation and impaired migration of CRC cells at micromolar concentrations. ConclusionsWe established an optimized pipeline combining virtual screening with experimental validation to efficiently identify fascin inhibitors. Using this approach, we discovered two promising compounds with anticancer activity in CRC cell cultures. Moreover, the protocol has been successfully adapted for application to additional cancer-related targets, expanding its potential utility in drug discovery. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/667829v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@dc470eorg.highwire.dtl.DTLVardef@1bd0759org.highwire.dtl.DTLVardef@12d755dorg.highwire.dtl.DTLVardef@1742d45_HPS_FORMAT_FIGEXP M_FIG C_FIG