Discovery of Potent Covalent CRM1 Inhibitors Via a Customized Structure-Based Virtual Screening Pipeline and Bioassays

• DOI: 10.1021/acs.jcim.4c00913
• Publication Date: 2024-10-03T20:29:18Z
• Authors: He Liu; Chao Shen; Haonan Li; Tingjun Hou; Yongliang Yang
• Keywords: Forkhead Box Protein O3; Drug Evaluation, Preclinical; Receptors, Cytoplasmic and Nuclear; Exportin 1 Protein; Apoptosis; Antineoplastic Agents; Cell Cycle Checkpoints; Karyopherins; Molecular Docking Simulation; User-Computer Interface; Cell Line, Tumor; Drug Discovery; Humans; Animals; Biological Assay; Cell Proliferation

CRM1 (chromosomal region maintenance 1, also referred to as exportin 1 or XPO1) plays a crucial role in maintaining the appropriate nuclear levels of tumor suppressor proteins (TSPs), growth regulatory proteins (GRPs), and antiapoptotic proteins, thereby contributing significantly to their anticancer effects. Dysregulation of CRM1-mediated nuclear transport, observed in a range of cancers such as colon cancer as well as autoimmune diseases, highlights its significance in various disease processes. In this paper, we employed a customized structure-based virtual screening campaign to search for novel covalent CRM1 inhibitors and purchased 50 potentially active compounds for in vitro bioassays. Among these candidates, AN-988 displayed a notably higher binding affinity (KD = 615 nM) toward CRM1, as determined by the biolayer interferometry (BLI) assay. Furthermore, AN-988 exhibited a strong suppression of colorectal cancer cell proliferation and remarkable anti-inflammatory effects. Notably, AN-988 induced cell apoptosis and cell cycle arrest in a time- and dose-dependent manner by effectively inhibiting the translocation of FOXO3a from the nucleus to the cytosol, thereby preserving the activity of FOXO3a. Collectively, our study identified AN-988 as a promising CRM1 inhibitor, underscoring its potential as a preclinical colon cancer therapy candidate.