DRUG TARGETING OF G-QUADRAPLEX-NM23-H2 COMPLEX IN THE c-MYC PROMOTER

项目4的翻译目标是确定并将一两个领先优势引入临床试验 通过靶向NM23-H2-DNA复合体抑制c-Myc转录的化合物。 这项提议建立在上一次赠款期间的进展基础上，并将进一步检验以下假设： NM23-H2-DNA复合体在c-Myc转录激活中起关键作用，而c-Myc是 结肠癌的致病因素。 该提案的具体目标是(1)G-四链药物络合物的结构表征 在c-Myc启动子，(2)建立体外生化和细胞为基础的筛选，以鉴定小 抑制NM23-H2与c-Myc沉默元件中G-四链结合的分子 启动子，(3)利用计算机发现和优化G-四链体和NM23-H2-相互作用化合物- 辅助药物设计和基于结构和虚拟筛选的方法，(4)体内评价和 随后的临床前开发，以及(5)提交I期临床试验和整体临床试验的IND 发展计划。高场核磁共振将用于表征药物-G-四链复合体， 分子模拟将被用于识别新的先导化合物，以及荧光共振能量 将使用转移(FRET)和双滤过法来鉴定体外生化结果。一个 基于荧光素酶的高通量筛选已经被用于识别命中，后续的分析将 在匹配的基因工程细胞系中进行，以提供原理证据，即 行动的结果是按照提议的。将使用基于结构的方法来协助进行领导优化。跟随 领导优化，将提交IND，临床开发计划将基于发现 从具体目标1到4。免疫组织化学将用于指导患者的选择和I、II期 将设计临床试验计划，以确定新药物是否有助于晚期糖尿病患者 结直肠癌。 这项研究的长期目标是开发治疗结直肠癌的新型靶向药物。 癌症。

The translational goal in Project 4 is to identify and bring forward into clinical trials one or two lead compounds that will suppress c-Myc transcription by targeting the NM23-H2-DNA complex. This proposal builds upon progress from the last grant period and will further test hypothesis that the NM23-H2-DNA complex is critically involved in transcriptional activation of c-Myc and c-Myc is a critical factor in colon cancer. The specific aims of the proposal are (1) structural characterization of the G-quadruplex-drug complexes in the c-Myc promoter, (2) to establish in vitro biochemical and cell-based screens to identify small molecules that inhibit binding of NM23-H2 to the G-quadruplex in the silencer element of the c-Myc promoter, (3) to discover and optimize G-quadruplex- and NM23-H2-interactive compounds using computer- aided drug design and structure-based and virtual-screening approaches, (4) In vivo evaluation and subsequent preclinical development, and (5) to file an IND for Phase I clinical trial and overall clinical development program. High-field NMR will be used to characterize the drug-G-quadruplex complex, molecular modeling will be used to identify new lead compounds, and fluorescence resonance energy transfer (FRET) and double-filter methods will be used for identification of in vitro biochemical hits. A luciferase-based high-throughput screen has already been used to identify hits, and subsequent assays will be carried out in matched cell lines genetically engineered to provide proof of principle that the mechanism of action is as proposed. Structure-based approaches will be used to aid in lead optimization. Following lead optimization, an IND will be filed and the clinical development program will be based upon discoveries from specific aims 1-4. Immunohistochemistry will be used to guide patient selection and phase I and II clinical trial plans will be designed to determine whether the new agents will help patients with advanced colorectal cancer. The long-term goal of this research is to develop novel target-directed drugs for treatment of colorectal cancer.