Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**

Jamal El Bakali; Michal Blaszczyk; Joanna C. Evans; Jennifer A. Boland; William J. McCarthy; Imam Fathoni; Marcio V.B. Dias; Eachan O. Johnson; Anthony G. Coyne; Valerie Mizrahi; Tom L. Blundell; Chris Abell; Christina Spry

doi:10.1002/anie.202300221

Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**

Jamal El Bakali, Michal Blaszczyk, Joanna C. Evans, Jennifer A. Boland, William J. McCarthy, Imam Fathoni, Marcio V.B. Dias, Eachan O. Johnson, Anthony G. Coyne, Valerie Mizrahi, Tom L. Blundell, Chris Abell, Christina Spry^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

The coenzyme A (CoA) biosynthesis pathway has attracted attention as a potential target for much-needed novel antimicrobial drugs, including for the treatment of tuberculosis (TB), the lethal disease caused by Mycobacterium tuberculosis (Mtb). Seeking to identify inhibitors of Mtb phosphopantetheine adenylyltransferase (MtbPPAT), the enzyme that catalyses the penultimate step in CoA biosynthesis, we performed a fragment screen. In doing so, we discovered three series of fragments that occupy distinct regions of the MtbPPAT active site, presenting a unique opportunity for fragment linking. Here we show how, guided by X-ray crystal structures, we could link weakly-binding fragments to produce an active site binder with a K_D <20 μM and on-target anti-Mtb activity, as demonstrated using CRISPR interference. This study represents a big step toward validating MtbPPAT as a potential drug target and designing a MtbPPAT-targeting anti-TB drug.

Original language	English
Article number	e202300221
Journal	Angewandte Chemie - International Edition
Volume	62
Issue number	17
DOIs	https://doi.org/10.1002/anie.202300221
Publication status	Published - 17 Apr 2023

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El Bakali, J., Blaszczyk, M., Evans, J. C., Boland, J. A., McCarthy, W. J., Fathoni, I., Dias, M. V. B., Johnson, E. O., Coyne, A. G., Mizrahi, V., Blundell, T. L., Abell, C., & Spry, C. (2023). Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**. Angewandte Chemie - International Edition, 62(17), Article e202300221. https://doi.org/10.1002/anie.202300221

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title = "Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**",

abstract = "The coenzyme A (CoA) biosynthesis pathway has attracted attention as a potential target for much-needed novel antimicrobial drugs, including for the treatment of tuberculosis (TB), the lethal disease caused by Mycobacterium tuberculosis (Mtb). Seeking to identify inhibitors of Mtb phosphopantetheine adenylyltransferase (MtbPPAT), the enzyme that catalyses the penultimate step in CoA biosynthesis, we performed a fragment screen. In doing so, we discovered three series of fragments that occupy distinct regions of the MtbPPAT active site, presenting a unique opportunity for fragment linking. Here we show how, guided by X-ray crystal structures, we could link weakly-binding fragments to produce an active site binder with a KD <20 μM and on-target anti-Mtb activity, as demonstrated using CRISPR interference. This study represents a big step toward validating MtbPPAT as a potential drug target and designing a MtbPPAT-targeting anti-TB drug.",

keywords = "Coenzyme A, Drug Discovery, Enzymes, Fragment-Based, Tuberculosis",

author = "{El Bakali}, Jamal and Michal Blaszczyk and Evans, {Joanna C.} and Boland, {Jennifer A.} and McCarthy, {William J.} and Imam Fathoni and Dias, {Marcio V.B.} and Johnson, {Eachan O.} and Coyne, {Anthony G.} and Valerie Mizrahi and Blundell, {Tom L.} and Chris Abell and Christina Spry",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.",

year = "2023",

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doi = "10.1002/anie.202300221",

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El Bakali, J, Blaszczyk, M, Evans, JC, Boland, JA, McCarthy, WJ, Fathoni, I, Dias, MVB, Johnson, EO, Coyne, AG, Mizrahi, V, Blundell, TL, Abell, C & Spry, C 2023, 'Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**', Angewandte Chemie - International Edition, vol. 62, no. 17, e202300221. https://doi.org/10.1002/anie.202300221

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T1 - Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**

AU - El Bakali, Jamal

AU - Blaszczyk, Michal

AU - Evans, Joanna C.

AU - Boland, Jennifer A.

AU - McCarthy, William J.

AU - Fathoni, Imam

AU - Dias, Marcio V.B.

AU - Johnson, Eachan O.

AU - Coyne, Anthony G.

AU - Mizrahi, Valerie

AU - Blundell, Tom L.

AU - Abell, Chris

AU - Spry, Christina

PY - 2023/4/17

Y1 - 2023/4/17

N2 - The coenzyme A (CoA) biosynthesis pathway has attracted attention as a potential target for much-needed novel antimicrobial drugs, including for the treatment of tuberculosis (TB), the lethal disease caused by Mycobacterium tuberculosis (Mtb). Seeking to identify inhibitors of Mtb phosphopantetheine adenylyltransferase (MtbPPAT), the enzyme that catalyses the penultimate step in CoA biosynthesis, we performed a fragment screen. In doing so, we discovered three series of fragments that occupy distinct regions of the MtbPPAT active site, presenting a unique opportunity for fragment linking. Here we show how, guided by X-ray crystal structures, we could link weakly-binding fragments to produce an active site binder with a KD <20 μM and on-target anti-Mtb activity, as demonstrated using CRISPR interference. This study represents a big step toward validating MtbPPAT as a potential drug target and designing a MtbPPAT-targeting anti-TB drug.

AB - The coenzyme A (CoA) biosynthesis pathway has attracted attention as a potential target for much-needed novel antimicrobial drugs, including for the treatment of tuberculosis (TB), the lethal disease caused by Mycobacterium tuberculosis (Mtb). Seeking to identify inhibitors of Mtb phosphopantetheine adenylyltransferase (MtbPPAT), the enzyme that catalyses the penultimate step in CoA biosynthesis, we performed a fragment screen. In doing so, we discovered three series of fragments that occupy distinct regions of the MtbPPAT active site, presenting a unique opportunity for fragment linking. Here we show how, guided by X-ray crystal structures, we could link weakly-binding fragments to produce an active site binder with a KD <20 μM and on-target anti-Mtb activity, as demonstrated using CRISPR interference. This study represents a big step toward validating MtbPPAT as a potential drug target and designing a MtbPPAT-targeting anti-TB drug.

KW - Coenzyme A

KW - Drug Discovery

KW - Enzymes

KW - Fragment-Based

KW - Tuberculosis

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DO - 10.1002/anie.202300221

M3 - Article

SN - 1433-7851

VL - 62

JO - Angewandte Chemie - International Edition

JF - Angewandte Chemie - International Edition

IS - 17

M1 - e202300221

ER -

Chemical Validation of Mycobacterium tuberculosis Phosphopantetheine Adenylyltransferase Using Fragment Linking and CRISPR Interference**

Abstract

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