1887

Abstract

A high-throughput screening (HTS) assay was developed for identifying compounds with inhibitory effect on , one of the key regulators positively controlling pathogenesis. An inhibitory effect on was expected to lead to attenuation in the secretion of the major pathogenicity factors of , cholera toxin and toxin co-regulated pilus. The plasmid construct pAKSB was developed with a kanamycin resistance (Km) gene under the control of the -like promoter for conferring a Km phenotype under -expressing conditions. The HTS assay was performed to identify compounds with inhibitory effect on the growth of O139 MO10 carrying the construct pAKSB in growth medium containing Km (30 g ml), but not in its absence. Of 20 338 compounds screened, six compounds were identified to inhibit the pAKSB-induced Km phenotype and these compounds caused transcriptional inhibition of in O139 strain MO10 as well as variant O1 El Tor strain NM06-058. Of the three most active substances, compound 53760866 showed lowest half-maximal cytotoxicity in a eukaryotic cell viability assay and was characterized further. Compound 53760866 caused reduction in cholera toxin secretion and expression of TcpA in vitro. The virulence attenuation corroborated well in a suckling mouse model , which showed reduction of colonization by NM06-058 when co-administered with 53760866. The screening method and the compounds may lead to new preventive strategies for cholera by reducing the pathogenicity of .

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2016-07-01
2020-09-28
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