The CERT antagonist HPA-12: first practical synthesis and individual binding evaluation of the four stereoisomers

Bioorg Med Chem. 2015 May 1;23(9):2004-9. doi: 10.1016/j.bmc.2015.03.019. Epub 2015 Mar 12.

Abstract

The first unified synthetic route to the four enantiopure HPA-12 stereoisomers in multi-gram scale is reported based on Crystallization-Induced Asymmetric Transformation (CIAT) technology. This preparative stereoselective synthesis allowed the unprecedented comparative evaluation of HPA-12 stereoisomers regarding their interaction with the CERT START domain. In vitro binding assay coupled to in silico docking approach indicate a possible interaction for the four derivatives. The first TR-FRET homogeneous-phase assay was developed to quantify their binding to the START domain, allowing complete determination of HPA-12 EC₅₀. Results indicate that not only the (1R,3S) lead to the strongest binding, but that both 1R and 3S stereocenters similarly contribute to extent of recognition This automated homogenous assay further opens up promising prospect for the identification of novel potential CERT antagonist by means of high throughput screening.

Keywords: CERT; CIAT; HPA-12; Sphingolipid; TR-FRET.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemical synthesis*
  • Amides / chemistry
  • Amides / pharmacology*
  • Binding Sites / drug effects
  • Dose-Response Relationship, Drug
  • Humans
  • Models, Molecular
  • Molecular Conformation
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Stereoisomerism
  • Structure-Activity Relationship

Substances

  • Amides
  • N-(3-hydroxy-1-hydroxymethyl-3-phenylpropyl)dodecanamide
  • CERT1 protein, human
  • Protein Serine-Threonine Kinases