Plasmodium falciparum Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation.

Authors

Kirthana M. V. Sindhe, Department of Medicine, University of California, San Francisco
Wesley Wu, Department of Biochemistry and Biophysics, University of California, San FranciscoFollow
Jenny Legac, Department of Medicine, University of California, San FranciscoFollow
Yong-Kang Zhang, Anacor Pharmaceuticals, Inc.Follow
Eric E. Easom, Anacor Pharmaceuticals, Inc.
Roland A. Cooper, Dominican University of CaliforniaFollow
Jacob J. Plattner, Anacor Pharmaceuticals, Inc., Palo Alto, CaliforniaFollow
Yvonne R. Freund, Anacor Pharmaceuticals, Inc.Follow
Joseph L. DeRisi, University of California, San Francisco
Philip J. Rosenthal, University of California, San FranciscoFollow

Department

Natural Sciences and Mathematics

Document Type

Article

Source

mBio

Publication Date

1-28-2020

ISSN

2150-7511

Volume

11

Issue

1

First Page

e02640-19

Abstract

New antimalarial drugs are needed. The benzoxaborole AN13762 showed excellent activity against cultured Plasmodium falciparum, against fresh Ugandan P. falciparum isolates, and in murine malaria models. To gain mechanistic insights, we selected in vitro for P. falciparum isolates resistant to AN13762. In all of 11 independent selections with 100 to 200 nM AN13762, the 50% inhibitory concentration (IC50) increased from 18–118 nM to 180–890 nM, and whole-genome sequencing of resistant parasites demonstrated mutations in prodrug activation and resistance esterase (PfPARE). The introduction of PfPARE mutations led to a similar level of resistance, and recombinant PfPARE hydrolyzed AN13762 to the benzoxaborole AN10248, which has activity similar to that of AN13762 but for which selection of resistance was not readily achieved. Parasites further selected with micromolar concentrations of AN13762 developed higher-level resistance (IC50, 1.9 to 5.0 μM), and sequencing revealed additional mutations in any of 5 genes, 4 of which were associated with ubiquitination/sumoylation enzyme cascades; the introduction of one of these mutations, in SUMO-activating enzyme subunit 2, led to a similar level of resistance. The other gene mutated in highly resistant parasites encodes the P. falciparum cleavage and specificity factor homolog PfCPSF3, previously identified as the antimalarial target of another benzoxaborole. Parasites selected for resistance to AN13762 were cross-resistant with a close analog, AN13956, but not with standard antimalarials, AN10248, or other benzoxaboroles known to have different P. falciparum targets. Thus, AN13762 appears to have a novel mechanism of antimalarial action and multiple mechanisms of resistance, including loss of function of PfPARE preventing activation to AN10248, followed by alterations in ubiquitination/sumoylation pathways or PfCPSF3.

PubMed ID

31992618

Rights

Copyright © 2020 Sindhe et al.

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.