Inhibitors of VIM-2 by screening pharmacologically active and click-chemistry compound libraries.
VIM-2 is an Ambler class B metallo-beta-lactamase (MBL) capable of hydrolyzing a broad-spectrum of beta-lactam antibiotics. Although the discovery and development of MBL inhibitors continue to be an area of active research, an array of potent, small molecule inhibitors is yet to be fully characterized for VIM-2. In the presented research, a compound library screening approach was used to identify and characterize VIM-2 inhibitors from a library of pharmacologically active compounds as well as a focused 'click' chemistry library. The four most potent VIM-2 inhibitors resulting from a VIM-2 screen were characterized by kinetic studies in order to determine K(i) and mechanism of enzyme inhibition. As a result, two previously described pharmacologic agents, mitoxantrone (1,4-dihydroxy-5,8-bis([2-([2-hydroxyethyl]amino)ethyl]amino)-9,10-anthracenedione) and 4-chloromercuribenzoic acid (pCMB) were found to be active, the former as a non-competitive inhibitor (K(i)=K(i)(')=1.5+/-0.2microM) and the latter as a slowly reversible or irreversible inhibitor. Additionally, two novel sulfonyl-triazole analogs from the click library were identified as potent, competitive VIM-2 inhibitors: N-((4-((but-3-ynyloxy)methyl)-1H-1,2,3-triazol-5-yl)methyl)-4-iodobenzenesulfonamide (1, K(i)=0.41+/-0.03microM) and 4-iodo-N-((4-(methoxymethyl)-1H-1,2,3-triazol-5-yl)methyl)benzenesulfonamide (2, K(i)=1.4+/-0.10microM). Mitoxantrone and pCMB were also found to potentiate imipenem efficacy in MIC and synergy assays employing Escherichia coli. Taken together, all four compounds represent useful chemical probes to further investigate mechanisms of VIM-2 inhibition in biochemical and microbiology-based assays.
Minond
Dmitriy
D
Lead Identification, Translational Research Institute, The Scripps Research Institute, Scripps Florida, 130 Scripps Way #1A1, Jupiter, 33458 FL, USA.
Saldanha
S Adrian
SA
Subramaniam
Prem
P
Spaargaren
Michael
M
Spicer
Timothy
T
Fotsing
Joseph R
JR
Weide
Timo
T
Fokin
Valery V
VV
Sharpless
K Barry
KB
Galleni
Moreno
M
Bebrone
Carine
C
Lassaux
Patricia
P
Hodder
Peter
P
D000700:Analgesics / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D020134:Catalytic Domain; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology; D015378:Imipenem / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008942:Mitoxantrone / Q000494:pharmacology; D008958:Models, Molecular; D011485:Protein Binding; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases / Q000737:chemistry / Q000378:metabolism*; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
D000700:Analgesics; D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D065093:beta-Lactamase Inhibitors; D020245:p-Chloromercuribenzoic Acid; D015378:Imipenem; D008942:Mitoxantrone; C425108:beta-lactamase bla(vim-2); D001618:beta-Lactamases