title: Fragment-Based Approaches to the Development of Mycobacterium tuberculosis CYP121 Inhibitors.
abstract: The essential enzyme CYP121 is a target for drug development against antibiotic resistant strains of Mycobacterium tuberculosis. A triazol-1-yl phenol fragment 1 was identified to bind to CYP121 using a cascade of biophysical assays. Synthetic merging and optimization of 1 produced a 100-fold improvement in binding affinity, yielding lead compound 2 (KD = 15 μM). Deconstruction of 2 into its component retrofragments allowed the group efficiency of structural motifs to be assessed, the identification of more LE scaffolds for optimization and highlighted binding affinity hotspots. Structure-guided addition of a metal-binding pharmacophore onto LE retrofragment scaffolds produced low nanomolar (KD = 15 nM) CYP121 ligands. Elaboration of these compounds to target binding hotspots in the distal active site afforded compounds with excellent selectivity against human drug-metabolizing P450s. Analysis of the factors governing ligand potency and selectivity using X-ray crystallography, UV-vis spectroscopy, and native mass spectrometry provides insight for subsequent drug development.
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Madeline E
authors.initials: ME
authors.lastname: Kavanagh
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Anthony G
authors.initials: AG
authors.lastname: Coyne
authors.affiliation: Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester , 131 Princess Street, Manchester M1 7DN, U.K.
authors.forename: Kirsty J
authors.initials: KJ
authors.lastname: McLean
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Guy G
authors.initials: GG
authors.lastname: James
authors.affiliation: Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester , 131 Princess Street, Manchester M1 7DN, U.K.
authors.forename: Colin W
authors.initials: CW
authors.lastname: Levy
authors.affiliation: Laboratory of Mycobacterial Metabolism and Antibiotic Research, Francis Crick Institute, The Mill Hill Laboratory , London NW7 1AA, U.K.|School of Pharmaceutical Sciences, São Paulo State University (UNESP) , 4801-902 Araraquara, SP, Brazil.
authors.forename: Leonardo B
authors.initials: LB
authors.lastname: Marino
authors.affiliation: Laboratory of Mycobacterial Metabolism and Antibiotic Research, Francis Crick Institute, The Mill Hill Laboratory , London NW7 1AA, U.K.
authors.forename: Luiz Pedro S
authors.initials: LP
authors.lastname: de Carvalho
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Daniel S H
authors.initials: DS
authors.lastname: Chan
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Sean A
authors.initials: SA
authors.lastname: Hudson
authors.affiliation: Department of Biochemistry, University of Cambridge , 80 Tennis Court Road, Cambridge CB2 1GA U.K.
authors.forename: Sachin
authors.initials: S
authors.lastname: Surade
authors.affiliation: Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester , 131 Princess Street, Manchester M1 7DN, U.K.
authors.forename: David
authors.initials: D
authors.lastname: Leys
authors.affiliation: Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester , 131 Princess Street, Manchester M1 7DN, U.K.
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Munro
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Chris
authors.initials: C
authors.lastname: Abell
mesh_terms: D001426:Bacterial Proteins / Q000037:antagonists & inhibitors*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008024:Ligands; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000201:enzymology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014376:Tuberculosis / Q000188:drug therapy* / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D008024:Ligands; C464825:cytochrome P-450 CYP121; D003577:Cytochrome P-450 Enzyme System