TRAF2 and NCK-Interacting Kinase Inhibitors for Colorectal Cancer: In Vitro and Theoretical Validations

ACS Comb Sci. 2020 Nov 9;22(11):608-616. doi: 10.1021/acscombsci.0c00027. Epub 2020 Sep 29.


TRAF2 and NCK-interacting kinase (TNIK) is a critical factor in colorectal cancer (CRC) proliferation mediated by Wnt signaling. We attempted to identify efficient TNIK inhibitors using computational high-throughput virtual screening (HTVS) from various drug banks and databases. By performing/on performing e-pharmacophore screening and molecular docking methods, from ∼700 000 molecules, compounds LC_222150, LC_112060, and LC_64796 were identified as potential leads, through molecular dynamics (MD) simulations and density functional theory (DFT). These top 3 structures were commercially procured, and their inhibitory activity was assessed in vitro. Significant TNIK inhibition was observed, with an average IC50 of 18.33 ± 0.75 nM. In terms of anticancer activity, the observed average relative % activity (RPA) of 90.28 ± 1.04 for these compounds compared well with doxorubicin (86.75 ± 1.45) as a standard. Compounds LC_222150, LC_112060, and LC_64796, therefore, warrant further evaluation in vivo to assess their CRC therapeutic effects.

Keywords: anticancer; colorectal cancer (CRC); density functional theory (DFT); high-throughput virtual screening (HTVS); molecular docking.

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism*
  • Antineoplastic Agents / chemistry*
  • Antineoplastic Agents / pharmacology
  • Apoptosis / drug effects
  • Colorectal Neoplasms / drug therapy*
  • Density Functional Theory
  • Doxorubicin / pharmacology
  • Doxorubicin / standards
  • Drug Screening Assays, Antitumor
  • HCT116 Cells
  • High-Throughput Screening Assays
  • Humans
  • Molecular Docking Simulation
  • Molecular Dynamics Simulation
  • Oncogene Proteins / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein Kinase Inhibitors / chemistry*
  • Protein Kinase Inhibitors / pharmacology
  • Signal Transduction
  • Structure-Activity Relationship
  • TNF Receptor-Associated Factor 2 / metabolism*


  • Adaptor Proteins, Signal Transducing
  • Antineoplastic Agents
  • Nck protein
  • Oncogene Proteins
  • Protein Kinase Inhibitors
  • TNF Receptor-Associated Factor 2
  • Doxorubicin