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

项目4的转化目标是确定并将一种或两种铅引入临床试验 通过靶向NM 23-H2-DNA复合物抑制c-Myc转录的化合物。 本提案建立在上一个赠款期的进展基础上，并将进一步检验以下假设： NM 23-H2-DNA复合物在c-Myc的转录激活中起关键作用，而c-Myc是一个关键的 结肠癌的病因 本论文的主要目的是：（1）G-四链体-药物复合物的结构表征 在c-Myc启动子，（2）建立体外生化和细胞筛选，以确定小 抑制NM 23-H2与c-Myc沉默元件中的G-四链体结合的分子 启动子，（3）发现和优化G-四链体和NM 23-H2-相互作用的化合物， 辅助药物设计和基于结构和虚拟筛选方法，（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NMR将用于表征药物-G-四链体复合物， 分子模拟将用于识别新的先导化合物，荧光共振能量 转移（FRET）和双过滤器方法将用于鉴定体外生物化学命中。一 基于内切酶的高通量筛选已经用于识别命中，随后的测定将 在基因工程匹配的细胞系中进行，以提供原理证明， 行动如建议。基于结构的方法将用于帮助铅优化。以下 电极导线优化，将提交IND，临床开发计划将基于发现 具体目标1-4。免疫组织化学将用于指导患者选择以及I期和II期 将设计临床试验计划，以确定新药物是否有助于晚期 结肠直肠癌 本研究的长期目标是开发新型靶向药物治疗结直肠癌 癌

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.