IN VITRO ANTIVIRAL SCREEN FOR RESPIRATORY VIRUSES

本合同的总体目标是提供资源， 新化学物质对成员的临床前体外评价 呼吸道病毒--甲型流感，B型流感， 副流感病毒3型，呼吸道合胞病毒，麻疹，和 腺病毒5型。 在最初的研究中，实验化合物将 使用CPE抑制试验筛选， 观察和中性红吸收后的比色变化。 将通过目视检查处理的细胞来确定初始细胞毒性。 细胞和中性红测定。 选择性指数为10或更高的物质将被 使用病毒产率降低测定证实。 细胞毒性 确认实验将使用Coulter测定的活细胞计数 柜台 这些验证性测试旨在选择 具有在动物模型和临床中进一步评价的潜力 审判 将对活性物质进行进一步检测，以确定 细胞毒性水平（即，DNA、RNA和蛋白质合成抑制） 以及作用机制（即，药物添加时间，代谢产物 挑战）。 当结果证明，额外的研究，如测试 与其他化合物组合用于毒性和抗病毒的物质 功效和杀病毒研究。

The overall objective of this contract is to provide a resource for preclinical in vitro evaluation of new chemical substances against members of the respiratory virus group -- influenza type A, influenza type B, parainfluenza virus type 3, respiratory syncytial virus, measles, and adenovirus type 5. In the initial studies, experimental compounds will be screened using the CPE inhibition assay determined both by microscopic observation and by colorimetric changes following neutral red uptake. Initial cytotoxicity will be determined by visual inspection of treated cells and by the neutral red assay. Substances that have a selectivity index of ten or greater will be confirmed using the virus yield reduction assay. Cytotoxicity in confirming experiments will use viable cell count determined by Coulter counter. These confirmatory tests are designed to select substances that have potential for further evaluation in animal models and clinical trials. Further testing of active substances will be performed to determine the level of cell toxicity (i.e., DNA, RNA, and protein synthesis inhibition) and the mechanism of action (i.e., time of drug addition, metabolite challenge). When results warrant, additional studies such as testing the substances in combination with other compounds for toxicity and antiviral efficacy, and virucidal studies.