start-ver=1.4
cd-journal=joma
no-vol=14
cd-vols=
no-issue=8
article-no=
start-page=e00484-26
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2026
dt-pub=20260804
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=High-throughput screening system for antimycobacterial compounds using in vitro infected cells: construction and application to compounds in the ?mura Natural Compound Library
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=The global increase in multidrug-resistant tuberculosis strains poses a major threat; consequently, there is an urgent need to identify and develop novel drugs. We have previously presented a simple fibroblast assay (SFA) screening method that enables the simultaneous assessment of antibacterial activity and cytotoxicity toward host cells. This method was developed based on our finding that live (but not dead) Mycobacterium tuberculosis bacilli exhibit cytotoxic activity against human lung fibroblasts, and that this cytotoxicity, which is proportional to the number of viable bacterial cells, is inhibited by known antimycobacterial drugs. While conventional methods to measure M. tuberculosis growth require 2?4 weeks to detect antibacterial activity; the SFA method enables detection within 2?3 days and can detect compounds that are active within host cells. Here, we applied high-throughput screening, based on assessment of cytotoxicity, to identify candidate antimycobacterial agents. In parallel, by comparing the direct antibacterial activity of compounds in liquid medium (using the broth dilution test) with that observed via SFA, we developed a new SFA-based screening system to select compounds with low cytotoxicity that are active outside or inside the host cells. Using this dual system, we screened 742 compounds from the ?mura Natural Compound Library (Kitasato University, Japan), with 62 hits. Of these, 19 were excluded owing to their toxicity, and six exhibited antibacterial activity only via SFA screening. Four promising antimycobacterial candidates with no cytotoxicity at the 10 ƒÊg/mL threshold?aridicin A, lankamycin, streptovaricin, and lariatin A?were identified. identified.This novel system supports the rapid identification of low-cytotoxicity antimycobacterial compounds that are active inside or outside the host cells.
en-copyright=
kn-copyright=

en-aut-name=TakiiTakemasa
en-aut-sei=Takii
en-aut-mei=Takemasa
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=

en-aut-name=KimishimaAoi
en-aut-sei=Kimishima
en-aut-mei=Aoi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=

en-aut-name=IwatsukiMasato
en-aut-sei=Iwatsuki
en-aut-mei=Masato
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=

en-aut-name=WatanabeYoshihiro
en-aut-sei=Watanabe
en-aut-mei=Yoshihiro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=

en-aut-name=OharaNaoya
en-aut-sei=Ohara
en-aut-mei=Naoya
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=

en-aut-name=AsamiYukihiro
en-aut-sei=Asami
en-aut-mei=Yukihiro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=

affil-num=1
en-affil=Department of Mycobacterium Reference and Research, The Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association
kn-affil=

affil-num=2
en-affil=?mura Satoshi Memorial Institute, Kitasato University
kn-affil=

affil-num=3
en-affil=?mura Satoshi Memorial Institute, Kitasato University
kn-affil=

affil-num=4
en-affil=?mura Satoshi Memorial Institute, Kitasato University
kn-affil=

affil-num=5
en-affil=Department of Oral Microbiology, Graduate School of Medicine, Density and Pharmaceutical Sciences, Okayama University
kn-affil=

affil-num=6
en-affil=?mura Satoshi Memorial Institute, Kitasato University
kn-affil=
en-keyword=Mycobacterium tuberculosis
kn-keyword=Mycobacterium tuberculosis
en-keyword=Mycobacterium avium
kn-keyword=Mycobacterium avium
en-keyword=Mycobacterium abscessus
kn-keyword=Mycobacterium abscessus
en-keyword=fibroblasts
kn-keyword=fibroblasts
en-keyword=the ?mura Natural Compound Library
kn-keyword=the ?mura Natural Compound Library
en-keyword=antibacterial activity
kn-keyword=antibacterial activity
END