Defining Stage-Specific Activity of Potent New Inhibitors of Cryptosporidium parvum Growth In Vitro.

摘要: ABSTRACT Cryptosporidium parvum and hominis have emerged as major enteric pathogens of infants in the developing world, addition to their known importance immunocompromised adults. Although there has been recent progress identifying new small molecules that inhibit sp. growth vitro or animal models, we lack information about mechanism action, potency across life cycle, cidal versus static activities. Here, explored four potent classes compounds include inhibitors likely target phosphatidylinositol 4 kinase (PI4K), phenylalanine-tRNA synthetase (PheRS), several with unknown mechanisms action. We utilized monoclonal antibodies gene expression probes for staging cycle development define timing when were active during grown vitro. These different targeted stages including blocked replication (PheRS inhibitors), prevented segmentation daughter cells thus egress (PI4K affected sexual-stage (a piperazine compound mechanism). Long-term cultivation C. epithelial cell monolayers derived from intestinal stem was used distinguish between activities based on ability parasites recover treatment. Collectively, these approaches should aid action designing vivo efficacy studies time-dependent concentrations needed achieve activity. IMPORTANCE Currently, nitazoxanide is only FDA-approved treatment cryptosporidiosis; unfortunately, it ineffective patients, varied immunocompetent individuals, not approved under 1 year age. Identifying cryptosporidiosis requires standardized quantifiable assays assessing potency, selectivity, activity, reversibility. provide protocols defining which are susceptible highly targets parasite. also utilize a newly developed long-term culture system monitoring reversibility means activity function concentration time valuable parameters establish conditions efficacious