title: The CERT antagonist HPA-12: first practical synthesis and individual binding evaluation of the four stereoisomers.
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.
authors.affiliation: SPCMIB, UMR-CNRS 5068, Université Paul Sabatier-Toulouse III, 118, route de Narbonne, F-31062 Toulouse Cedex 9, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Santos
authors.affiliation: Unité de Service et de Recherche CNRS-Pierre Fabre n°3388 ETaC, CRDPF, 3 avenue H. Curien, 31035 Toulouse Cedex 01, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Fleury
authors.affiliation: SPCMIB, UMR-CNRS 5068, Université Paul Sabatier-Toulouse III, 118, route de Narbonne, F-31062 Toulouse Cedex 9, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Rodriguez
authors.affiliation: Department of Organic Chemistry, Slovak University of Technology, Radlinského 9, 81237 Bratislava, Slovakia.
authors.forename: Jozef
authors.initials: J
authors.lastname: Markus
authors.affiliation: Department of Organic Chemistry, Slovak University of Technology, Radlinského 9, 81237 Bratislava, Slovakia. Electronic address: dusan.berkes@stuba.sk.
authors.forename: Dušan
authors.initials: D
authors.lastname: Berkeš
authors.affiliation: URCOM, EA 3221, INC3M CNRS FR-3038, UFR Sciences & Techniques, University of Le Havre, 25, rue Philippe Lebon, B.P. 540, F-76058 Le Havre Cedex, France.
authors.forename: Adam
authors.initials: A
authors.lastname: Daïch
authors.affiliation: Unité de Service et de Recherche CNRS-Pierre Fabre n°3388 ETaC, CRDPF, 3 avenue H. Curien, 31035 Toulouse Cedex 01, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Ausseil
authors.affiliation: SPCMIB, UMR-CNRS 5068, Université Paul Sabatier-Toulouse III, 118, route de Narbonne, F-31062 Toulouse Cedex 9, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Baudoin-Dehoux
authors.affiliation: SPCMIB, UMR-CNRS 5068, Université Paul Sabatier-Toulouse III, 118, route de Narbonne, F-31062 Toulouse Cedex 9, France. Electronic address: ballereau@chimie.ups-tlse.fr.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Ballereau
authors.affiliation: SPCMIB, UMR-CNRS 5068, Université Paul Sabatier-Toulouse III, 118, route de Narbonne, F-31062 Toulouse Cedex 9, France. Electronic address: genisson@chimie.ups-tlse.fr.
authors.forename: Yves
authors.initials: Y
authors.lastname: Génisson
mesh_terms: D000577:Amides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008958:Models, Molecular; D008968:Molecular Conformation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: D000577:Amides; C441643:N-(3-hydroxy-1-hydroxymethyl-3-phenylpropyl)dodecanamide; C119090:CERT1 protein, human; D017346:Protein Serine-Threonine Kinases
keywords: CERT; CIAT; HPA-12; Sphingolipid; TR-FRET