The antiproliferative and proapoptotic effects of cladosporols A and B are related to their different binding mode as PPARγ ligands

Biochem Pharmacol. 2016 May 15:108:22-35. doi: 10.1016/j.bcp.2016.03.007. Epub 2016 Mar 17.

Abstract

Cladosporols are secondary metabolites from Cladosporium tenuissimum characterized for their ability to control cell proliferation. We previously showed that cladosporol A inhibits proliferation of human colon cancer cells through a PPARγ-mediated modulation of gene expression. In this work, we investigated cladosporol B, an oxidate form of cladosporol A, and demonstrate that it is more efficient in inhibiting HT-29 cell proliferation due to a robust G0/G1-phase arrest and p21(waf1/cip1) overexpression. Cladosporol B acts as a PPARγ partial agonist with lower affinity and reduced transactivation potential in transient transfections as compared to the full agonists cladosporol A and rosiglitazone. Site-specific PPARγ mutants and surface plasmon resonance (SPR) experiments confirm these conclusions. Cladosporol B in addition displays a sustained proapoptotic activity also validated by p21(waf1/cip1) expression analysis in the presence of the selective PPARγ inhibitor GW9662. In the DMSO/H2O system, cladosporols A and B are unstable and convert to the ring-opened compounds 2A and 2B. Finally, docking experiments provide the structural basis for full and partial PPARγ agonism of 2A and 2B, respectively. In summary, we report here, for the first time, the structural characteristics of the binding of cladosporols, two natural molecules, to PPARγ. The binding of compound 2B is endowed with a lower transactivation potential, higher antiproliferative and proapoptotic activity than the two full agonists as compound 2A and rosiglitazone (RGZ).

Keywords: Apoptosis; Cladosporols; Colorectal cancer cells; Docking; Full and partial PPARγ agonists.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anilides / pharmacology
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Drug Partial Agonism
  • G1 Phase Cell Cycle Checkpoints
  • HEK293 Cells
  • Humans
  • Ligands
  • Molecular Docking Simulation
  • Naphthols / pharmacology*
  • Oxidative Stress
  • PPAR gamma / agonists*
  • PPAR gamma / antagonists & inhibitors
  • PPAR gamma / genetics
  • PPAR gamma / metabolism
  • Protein Binding
  • Protein Conformation
  • Stereoisomerism
  • Structure-Activity Relationship
  • Transcriptional Activation

Substances

  • 2-chloro-5-nitrobenzanilide
  • Anilides
  • Antineoplastic Agents
  • Cyclin-Dependent Kinase Inhibitor p21
  • Ligands
  • Naphthols
  • PPAR gamma
  • cladosporol A
  • cladosporol B