title: Biochemical investigations of the mechanism of action of small molecules ZL006 and IC87201 as potential inhibitors of the nNOS-PDZ/PSD-95-PDZ interactions.
abstract: ZL006 and IC87201 have been presented as efficient inhibitors of the nNOS/PSD-95 protein-protein interaction and shown great promise in cellular experiments and animal models of ischemic stroke and pain. Here, we investigate the proposed mechanism of action of ZL006 and IC87201 using biochemical and biophysical methods, such as fluorescence polarization (FP), isothermal titration calorimetry (ITC), and (1)H-(15)N HSQC NMR. Our data show that under the applied in vitro conditions, ZL006 and IC87201 do not interact with the PDZ domains of nNOS or PSD-95, nor inhibit the nNOS-PDZ/PSD-95-PDZ interface by interacting with the β-finger of nNOS-PDZ. Our findings have implications for further medicinal chemistry efforts of ZL006, IC87201 and analogues, and challenge the general and widespread view on their mechanism of action.
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
authors.forename: Anders
authors.initials: A
authors.lastname: Bach
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
authors.forename: Søren W
authors.initials: SW
authors.lastname: Pedersen
authors.affiliation: NMR Centre for Structural Biology, University of Liverpool, UK L69 7ZB Liverpool.
authors.forename: Liam A
authors.initials: LA
authors.lastname: Dorr
authors.affiliation: Clermont Université, ENSCCF, CNRS UMR 6296, Institut de Chimie de Clermont-Ferrand, BP10187, F-63174 Aubière, France.
authors.forename: Gary
authors.initials: G
authors.lastname: Vallon
authors.affiliation: Clermont Université, ENSCCF, CNRS UMR 6296, Institut de Chimie de Clermont-Ferrand, BP10187, F-63174 Aubière, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Ripoche
authors.affiliation: Clermont Université, ENSCCF, CNRS UMR 6296, Institut de Chimie de Clermont-Ferrand, BP10187, F-63174 Aubière, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Ducki
authors.affiliation: NMR Centre for Structural Biology, University of Liverpool, UK L69 7ZB Liverpool.
authors.forename: Lu-Yun
authors.initials: LY
authors.lastname: Lian
mesh_terms: D000636:Aminosalicylic Acids / Q000494:pharmacology*; D001596:Benzylamines / Q000494:pharmacology*; D002151:Calorimetry; D002733:Chlorophenols / Q000494:pharmacology*; D000076122:Disks Large Homolog 4 Protein; D005454:Fluorescence Polarization; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors*; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D052248:Nitric Oxide Synthase Type I / Q000037:antagonists & inhibitors*; D054731:PDZ Domains / Q000187:drug effects*; D014230:Triazoles / Q000494:pharmacology*
chemical_list: C546030:2-((1H-benzo(d)(1,2,3)triazol-5-ylamino)methyl)-4,6-dichlorophenol; D000636:Aminosalicylic Acids; D001596:Benzylamines; D002733:Chlorophenols; C501444:DLG4 protein, human; D000076122:Disks Large Homolog 4 Protein; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D014230:Triazoles; C000607694:ZL006 compound; D052248:Nitric Oxide Synthase Type I