Adduct Formation of Delamanid with NAD in Mycobacteria

Mikayo Hayashi; Akihito Nishiyama; Ryuki Kitamoto; Yoshitaka Tateishi; Mayuko Osada-Oka; Yukiko Nishiuchi; Shaban A Kaboso; Xiuhao Chen; Mamoru Fujiwara; Yusuke Inoue; Yoshikazu Kawano; Masanori Kawasaki; Tohru Abe; Tsutomu Sato; Kentaro Kaneko; Kimiko Itoh; Sohkichi Matsumoto; Makoto Matsumoto

doi:10.1128/AAC.01755-19

Adduct Formation of Delamanid with NAD in Mycobacteria

Antimicrob Agents Chemother. 2020 Apr 21;64(5):e01755-19. doi: 10.1128/AAC.01755-19. Print 2020 Apr 21.

Authors

Mikayo Hayashi^#¹, Akihito Nishiyama^#², Ryuki Kitamoto¹, Yoshitaka Tateishi², Mayuko Osada-Oka^{2

3}, Yukiko Nishiuchi⁴, Shaban A Kaboso², Xiuhao Chen¹, Mamoru Fujiwara¹, Yusuke Inoue¹, Yoshikazu Kawano¹, Masanori Kawasaki¹, Tohru Abe⁵, Tsutomu Sato⁵, Kentaro Kaneko⁶, Kimiko Itoh⁶, Sohkichi Matsumoto^{7

8}, Makoto Matsumoto⁹

Affiliations

¹ Pharmaceutical Business Division, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan.
² Department of Bacteriology, Niigata University School of Medicine, Niigata, Japan.
³ Division of Applied Life Science, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto, Japan.
⁴ Toneyama Institute for Tuberculosis Research, Osaka City University Medical School, Osaka, Japan.
⁵ Department of Applied Biological Chemistry, Faculty of Agriculture and Graduate School of Science and Technology, Niigata University, Niigata, Japan.
⁶ Graduate School of Science and Technology, Niigata University, Niigata, Japan.
⁷ Department of Bacteriology, Niigata University School of Medicine, Niigata, Japan sohkichi@med.niigata-u.ac.jp Matsumoto.Makoto@otsuka.jp.
⁸ Laboratory of Tuberculosis, Institute of Tropical Disease, Universitas Airlangga, Mulyorejo, Surabaya, Indonesia.
⁹ Pharmaceutical Business Division, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan sohkichi@med.niigata-u.ac.jp Matsumoto.Makoto@otsuka.jp.

^# Contributed equally.

Abstract

Delamanid (DLM), a nitro-dihydroimidazooxazole derivative currently approved for pulmonary multidrug-resistant tuberculosis (TB) therapy, is a prodrug activated by mycobacterial 7,8-didemethyl-8-hydroxy 5-deazaflavin electron transfer coenzyme (F₄₂₀)-dependent nitroreductase (Ddn). Despite inhibiting the biosynthesis of a subclass of mycolic acids, the active DLM metabolite remained unknown. Comparative liquid chromatography-mass spectrometry (LC-MS) analysis of DLM metabolites revealed covalent binding of reduced DLM with a nicotinamide ring of NAD derivatives (oxidized form) in DLM-treated Mycobacterium tuberculosis var. Bacille de Calmette et Guérin. Isoniazid-resistant mutations in the type II NADH dehydrogenase gene (ndh) showed a higher intracellular NADH/NAD ratio and cross-resistance to DLM, which were restored by complementation of the mutants with wild-type ndh Our data demonstrated for the first time the adduct formation of reduced DLM with NAD in mycobacterial cells and its importance in the action of DLM.

Keywords: Mycobacterium; delamanid; drug; tuberculosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antitubercular Agents / pharmacology*
Chromatography, Liquid
Drug Resistance, Multiple, Bacterial / genetics
Isoniazid / pharmacology
Mass Spectrometry
Mycobacterium tuberculosis / drug effects*
Mycobacterium tuberculosis / genetics*
Mycolic Acids / metabolism
NAD / analysis
NADH Dehydrogenase / genetics
Nitroimidazoles / pharmacology*
Oxazoles / pharmacology*
Oxidation-Reduction
Polymorphism, Single Nucleotide / genetics
Tuberculosis, Multidrug-Resistant / drug therapy
Tuberculosis, Multidrug-Resistant / genetics*
Tuberculosis, Pulmonary / drug therapy*

Substances

Antitubercular Agents
Mycolic Acids
Nitroimidazoles
OPC-67683
Oxazoles
NAD
NADH dehydrogenase II
NADH Dehydrogenase
Isoniazid