Novel Targets of Chemotherapeutic Drugs that Trap Protein on DNA

 描述(由申请人提供)：一些非常有效的化疗药物，通过稳定作为正常细胞功能所必需的中间体形成的共价蛋白质-DNA加合物，用于临床功能。尽管这些药物的有效性得到了证明，但还没有努力系统地确定这类药物的所有目标。我们的建议描述了一种新的方法，将药物治疗中严格浓缩共价蛋白质加合物与无偏见的蛋白质组学相结合，以识别和发现药物靶点。使用这种方法，我们已经确认了拓扑替康和依托泊苷的已知靶点，并且我们已经确定了这些药物的新靶点，从而确立了这种方法作为发现工具的能力。我们建议应用这种方法来确定六种通过这种机制发挥作用的化疗药物的靶点：依托泊苷、阿霉素、拓扑替康、地西他滨(5-aza-DC)、氮胞苷(5-aza-C)和奥拉帕利，并验证这些靶点。这些实验的成功将确定当前使用的药物的新靶点，并建立确定药物作用机制的一般方法，从而使未来能够识别新的药物靶点对。

 DESCRIPTION (provided by applicant): Some very potent chemotherapeutics used in the clinic function by stabilizing covalent protein-DNA adducts formed as obligatory intermediates in normal cellular function. Despite the demonstrated effectiveness of these drugs, there has been no effort to systematically identify all targets of drugs in this class. Our proposal describes a novel approach that combines stringent enrichment of covalent protein adducts upon drug treatment with unbiased proteomics to identify and discover drug targets. Using this approach, we have confirmed known targets of topotecan and etoposide, and we have identified novel targets of these drugs, thereby establishing the power of this approach as a discovery tool. We propose to apply this approach to identify the targets of six chemotherapeutic drugs that function by this mechanism: etoposide, doxorubicin, topotecan, decitabine (5-aza-dC), azacitidine (5-aza-C) and olaparib, and to validate these targets. Success in these experiments will identify new targets of drugs in current use, and establish a general approach for determining mechanisms of drug action to enable future identification of novel drug-target pairs.