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

J Chem Inf Model. 2024 Oct 14;64(19):7422-7431. doi: 10.1021/acs.jcim.4c00913. Epub 2024 Oct 3.

Abstract

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.

MeSH terms

  • Animals
  • Antineoplastic Agents* / chemistry
  • Antineoplastic Agents* / pharmacology
  • Apoptosis* / drug effects
  • Biological Assay
  • Cell Cycle Checkpoints / drug effects
  • Cell Line, Tumor
  • Cell Proliferation* / drug effects
  • Drug Discovery*
  • Drug Evaluation, Preclinical
  • Exportin 1 Protein*
  • Forkhead Box Protein O3 / metabolism
  • Humans
  • Karyopherins* / antagonists & inhibitors
  • Karyopherins* / metabolism
  • Molecular Docking Simulation
  • Receptors, Cytoplasmic and Nuclear* / antagonists & inhibitors
  • Receptors, Cytoplasmic and Nuclear* / chemistry
  • Receptors, Cytoplasmic and Nuclear* / metabolism
  • User-Computer Interface

Substances

  • Exportin 1 Protein
  • Karyopherins
  • Receptors, Cytoplasmic and Nuclear
  • Antineoplastic Agents
  • Forkhead Box Protein O3
  • FOXO3 protein, human