RESUMO
Metallo-ß-lactamases (MBLs) enable bacterial resistance to almost all classes of ß-lactam antibiotics. We report studies on enethiol containing MBL inhibitors, which were prepared by rhodanine hydrolysis. The enethiols inhibit MBLs from different subclasses. Crystallographic analyses reveal that the enethiol sulphur displaces the di-Zn(II) ion bridging 'hydrolytic' water. In some, but not all, cases biophysical analyses provide evidence that rhodanine/enethiol inhibition involves formation of a ternary MBL enethiol rhodanine complex. The results demonstrate how low molecular weight active site Zn(II) chelating compounds can inhibit a range of clinically relevant MBLs and provide additional evidence for the potential of rhodanines to be hydrolysed to potent inhibitors of MBL protein fold and, maybe, other metallo-enzymes, perhaps contributing to the complex biological effects of rhodanines. The results imply that any medicinal chemistry studies employing rhodanines (and related scaffolds) as inhibitors should as a matter of course include testing of their hydrolysis products.
Assuntos
Rodanina/química , Compostos de Sulfidrila/química , Inibidores de beta-Lactamases/síntese química , beta-Lactamases/química , Enedi-Inos/química , Concentração Inibidora 50 , Estrutura Molecular , Rodanina/síntese química , Rodanina/farmacologia , Relação Estrutura-Atividade , Compostos de Sulfidrila/farmacologia , Inibidores de beta-Lactamases/farmacologia , beta-Lactamases/efeitos dos fármacosRESUMO
Resistance to ß-lactam antibiotics mediated by metallo-ß-lactamases (MBLs) is a growing problem. We describe the use of protein-observe 19 F-NMR (PrOFâ NMR) to study the dynamics of the São Paulo MBL (SPM-1) from ß-lactam-resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, which flank the di-ZnII active site, were selectively 19 F-labeled using 3-bromo-1,1,1-trifluoroacetone. The PrOFâ NMR results reveal roles for the mobile α3 and L3 regions in the binding of both inhibitors and hydrolyzed ß-lactam products to SPM-1. These results have implications for the mechanisms and inhibition of MBLs by ß-lactams and non-ß-lactams and illustrate the utility of PrOFâ NMR for efficiently analyzing metal chelation, identifying new binding modes, and studying protein binding from a mixture of equilibrating isomers.
Assuntos
Imagem por Ressonância Magnética de Flúor-19 , Inibidores de beta-Lactamases/farmacologia , beta-Lactamases/metabolismo , Sítios de Ligação/efeitos dos fármacos , Modelos Moleculares , Conformação Molecular , Inibidores de beta-Lactamases/síntese química , Inibidores de beta-Lactamases/químicaRESUMO
Acetamido derivatives of the naturally antibacterial non-ß-lactam lactivicin (LTV) have improved activity against their penicillin binding protein targets and reduced hydrolysis by ß-lactamases, but penetration into Gram-negative bacteria is still relatively poor. Here we report that modification of the LTV lactone with a catechol-type siderophore increases potency 1,000-fold against Stenotrophomonas maltophilia, a species renowned for its insusceptibility to antimicrobials. The MIC90 of modified lactone compound 17 (LTV17) against a global collection of extensively drug-resistant clinical S. maltophilia isolates was 0.063 µg · ml(-1) Sideromimic modification does not reduce the ability of LTVs to induce production of the L1 and L2 ß-lactamases in S. maltophilia and does not reduce the rate at which LTVs are hydrolyzed by L1 or L2. We conclude, therefore, that lactivicin modification with a siderophore known to be preferentially used by S. maltophilia substantially increases penetration via siderophore uptake. LTV17 has the potential to be developed as a novel antimicrobial for treatment of infections by S. maltophilia More generally, our work shows that sideromimic modification in a species-targeted manner might prove useful for the development of narrow-spectrum antimicrobials that have reduced collateral effects.
Assuntos
Antibacterianos/farmacologia , Peptídeos/farmacologia , Stenotrophomonas maltophilia/efeitos dos fármacos , Antibacterianos/química , Farmacorresistência Bacteriana , Testes de Sensibilidade Microbiana , Peptídeos/química , Peptídeos Cíclicos , Relação Estrutura-AtividadeRESUMO
Two cats, two paths: two novel domino reactions starting from 6-hydroxy-2-alkyl-2-alkynylcyclohexanones have been discovered. While redox-neutral platinum catalysis gives rise to furans through a sequence of cyclization, 1,2-shift, and Grob fragmentation, oxidative copper catalysis provides an entry to bicyclic 2,3-dihydrofurans. Upon cyclization and oxidation, an unusual benzilic acid rearrangement can take place in this case.
Assuntos
Furanos/síntese química , Compostos Organometálicos/química , Elementos de Transição/química , Catálise , Ciclização , Furanos/química , Estrutura Molecular , OxirreduçãoRESUMO
The use of ß-lactam antibiotics is compromised by resistance, which is provided by ß-lactamases belonging to both metallo (MBL)- and serine (SBL)-ß-lactamase subfamilies. The rhodanines are one of very few compound classes that inhibit penicillin-binding proteins (PBPs), SBLs and, as recently reported, MBLs. Here, we describe crystallographic analyses of the mechanism of inhibition of the clinically relevant VIM-2 MBL by a rhodanine, which reveal that the rhodanine ring undergoes hydrolysis to give a thioenolate. The thioenolate is found to bind via di-zinc chelation, mimicking the binding of intermediates in ß-lactam hydrolysis. Crystallization of VIM-2 in the presence of the intact rhodanine led to observation of a ternary complex of MBL, a thioenolate fragment and rhodanine. The crystallographic observations are supported by kinetic and biophysical studies, including (19)F NMR analyses, which reveal the rhodanine-derived thioenolate to be a potent broad-spectrum MBL inhibitor and a lead structure for the development of new types of clinically useful MBL inhibitors.