A new strategy for hit generation: Novel in cellulo active inhibitors of CYP121A1 from Mycobacterium tuberculosis via a combined X-ray crystallographic and phenotypic screening approach (XP screen)

Eur J Med Chem. 2022 Feb 15;230:114105. doi: 10.1016/j.ejmech.2022.114105.

Martyn Frederickson ¹, Irwin R Selvam ², Dimitrios Evangelopoulos ³, Kirsty J McLean ², Mona M Katariya ¹, Richard B Tunnicliffe ², Bethany Campbell ¹, Madeline E Kavanagh ¹, Sitthivut Charoensutthivarakul ¹, Richard T Blankley ⁴, Colin W Levy ⁵, Luiz Pedro S de Carvalho ³, David Leys ², Andrew W Munro ⁶, Anthony G Coyne ⁷, Chris Abell ¹

Affiliations

1 Yusuf Hamied Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, United Kingdom.
2 Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, United Kingdom.
3 Mycobacterium Metabolism and Antibiotic Research Laboratory, Francis Crick Institute, 1 Midland Road, London, NW1 1AT, United Kingdom.
4 Agilent Technologies U.K. Ltd, 5500 Lakeside, Cheadle Royal, Cheshire, SK8 3GR, United Kingdom.
5 Manchester Protein Structure Facility (MPSF), Manchester Institute of Biotechnology, University of Manchester, Manchester, M1 7DN, United Kingdom.
6 Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, United Kingdom. Electronic address: andrew.munro@manchester.ac.uk.
7 Yusuf Hamied Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, United Kingdom. Electronic address: agc40@cam.ac.uk.

PMID: 35065413 DOI: 10.1016/j.ejmech.2022.114105

Abstract

There is a pressing need for new drugs against tuberculosis (TB) to combat the growing resistance to current antituberculars. Herein a novel strategy is described for hit generation against promising TB targets involving X-ray crystallographic screening in combination with phenotypic screening. This combined approach (XP Screen) affords both a validation of target engagement as well as determination of in cellulo activity. The utility of this method is illustrated by way of an XP Screen against CYP121A1, a Cytochrome P450 enzyme from Mycobacterium tuberculosis (Mtb) championed as a validated drug discovery target. A focused screening set was synthesized and tested by such means, with several members of the set showing promising activity against Mtb strain H37Rv. One compound was observed as an X-ray hit against CYP121A1 and showed improved activity against Mtb strain H37Rv under multiple assay conditions (pan-assay activity). Data obtained during X-ray crystallographic screening were utilized in a structure-based campaign to design a limited number of analogues (less than twenty), many of which also showed pan-assay activity against Mtb strain H37Rv. These included the benzo[b][1,4]oxazine derivative (MIC₉₀ 6.25 μM), a novel hit compound suitable as a starting point for a more involved hit to lead candidate medicinal chemistry campaign.

Keywords

CYP121; Drug discovery; Mycobacterium tuberculosis; Tuberculosis; X-ray crystallography.

Products

Cat. No.

Product Name

Description

Target

Research Area
HY-144325

CYP121A1-IN-1

CYP121A1 Inhibitor

Cytochrome P450

Infection

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