Inhibitors of VIM-2 by screening pharmacologically active and click-chemistry compound libraries

Abstract

VIM-2 is an Ambler class B metallo-β-lactamase (MBL) capable of hydrolyzing a broad-spectrum of β-lactam antibiotics. Although the discovery and development of MBL inhibitors continues to be an area of active research, an array of potent, small molecule inhibitors has 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 Ki 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 (Ki = K′i = 1.5 ± 0.2 μM) 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, Ki = 0.41 ± 0.03 μM) and 4-iodo-N-((4-(methoxymethyl)-1H-1,2,3-triazol-5-yl)methyl)benzenesulfonamide (2, Ki = 1.4 ± 0.10 μM). Mitoxantrone and pCMB were also found to potentiate imipenem efficacy in MIC and synergy assays employing E. coli. Taken together, all four compounds represent useful chemical probes to further investigate mechanisms of VIM-2 inhibition in biochemical and microbiology-based assays.

Carbapenems: Past, Present, and Future

“…The biggest challenges are the metallo-␤-lactamases. However, some progress has been made in identifying potential inhibitors (cephalosporin-derived reverse hydroxamates and oximes, phthalic acid derivatives, mitoxantrone, 4-chloromercuribenzoic acid, sulfonyl-triazole analogs, and NH-1,2,3-triazole-based compounds) (66,84,138,202,248). Alternative inhibitors can include carbapenems with different stereochemistries.…”

Synthesis of N-unsubstituted 1,2,3-triazoles via a cascade including propargyl azides, allenyl azides, and triazafulvenes

“…On the other hand, such properties are highly welcome when NH-1,2,3-triazoles were synthesized as biologically active agents by the cascade method. [32][33][34][35][36][38][39][40][41][42]45 …”

“…Thus, the methods have already been applied to prepare some N-unsubstituted 1,2,3-triazoles. [32][33][34][35][36][37][38][39][40][41][42][43][44][45] Our previous studies were focused on the mechanisms to explain the formation of these heterocycles. [28][29][30][31] …”

An Automated Miniaturized Method to Perform and Analyze Antimicrobial Drug Synergy Assays

“…Traditional tube synergy testing requires a minimum of 36 tubes to test 25 combinations in a 1 mL volume to achieve reliable interpretation of synergy. 13,19 A 96-wpf synergy assay achieves 77 combinations per plate in 0.2 mL volume/well. 13 In contrast to both methods, the 384-wpf synergy test is able to obtain 260 test inhibitor combinations per plate using 0.06 mL/well.…”

“…Separate statistics are calculated for each side of the plate. BLI Left is the compound tested in columns 1-12 of the assay plate and BLI Right is the bacteria tested in columns [13][14][15][16][17][18][19][20][21][22][23] …”