Identification of Small Molecule Inhibitors That Block the Toxoplasma gondii Rhoptry Kinase ROP18

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• 作者：Catherine Simpson ; Nathaniel G. Jones ; Emily A. Hull-Ryde ; Dmitri Kireev ; Michael Stashko ; Keliang Tang ; James W. Janetka ; Scott A. Wildman ; William J. Zuercher ; Matthieu Schapira ; Raymond Hui ; William Janzen ; L. David Sibley
• 刊名：ACS Infectious Diseases
• 出版年：2016
• 出版时间：March 11, 2016
• 年：2016
• 卷：2
• 期：3
• 页码：194-206
• 全文大小：743K
• ISSN：2373-8227

文摘

The protozoan parasite Toxoplasma gondii secretes a family of serine-threonine protein kinases into its host cell in order to disrupt signaling and alter immune responses. One prominent secretory effector is the rhoptry protein 18 (ROP18), a serine-threonine kinase that phosphorylates immunity related GTPases (IRGs) and hence blocks interferon gamma-mediated responses in rodent cells. Previous genetic studies show that ROP18 is a major virulence component of T. gondii strains from North and South America. Here, we implemented a high throughput screen to identify small molecule inhibitors of ROP18 in vitro and subsequently validated their specificity within infected cells. Although ROP18 was not susceptible to many kinase-directed inhibitors that affect mammalian kinases, the screen identified several sub-micromolar inhibitors that belong to three chemical scaffolds: oxindoles, 6-azaquinazolines, and pyrazolopyridines. Treatment of interferon γ-activated cells with one of these inhibitors enhanced immunity related GTPase recruitment to wild type parasites, recapitulating the defect of Δrop18 mutant parasites, consistent with targeting ROP18 within infected cells. These compounds provide useful starting points for chemical biology experiments or as leads for therapeutic interventions designed to reduce parasite virulence.