Exploration of a Mutant p53 Reactivating Compound

描述（由申请人提供）：TP 53是人类癌症中最常见的突变基因，对于该基因，不存在有效的靶向抗癌药物。我们将NSC 319726（726）鉴定为突变型p53再活化化合物，以及突变型p53靶向药物开发的先导化合物[1]。先前的证据表明，726的作用是等位基因特异性的，通过恢复野生型结构和功能的第三个最常见的p53错义突变体（p53-R175 H）。这种效果取决于726的锌螯合和氧化还原调节性质。p53蛋白结合单个锌离子，这是p53结构和功能所必需的。p53-R175 H突变损害蛋白质结合锌，导致其错误折叠并失去转录功能。726突变p53再激活的机制尚不清楚。我们提供了初步的数据，描述了一种新的“双重”机制，其中726恢复野生型结构的p53-R175 H使用锌金属伴侣功能，以提供最佳浓度的锌，以促进正确的折叠。我们还证明，726的氧化还原调节特性的功能，以反式激活新确认的p53-R175，并诱导凋亡程序。我们还表明，这种锌金属伴侣功能适用于其他p53错义突变与受损的锌结合。在这个K 08职业发展奖中，Darren Carpizo博士和他的合作者计划从两个主要方向探索这种化合物。目的#1是剖析该化合物重新激活p53-R175突变蛋白和其他p53锌结合突变体的分子机制。目的#2是对该化合物进行临床前研究，以评估其作为突变型p53靶向抗癌药物的临床开发潜力。Carpizo博士是一名外科肿瘤学家，专门研究肝胆和胰腺癌，目前被任命为新泽西癌症研究所（CINJ）的外科助理教授，该研究所是NCI指定的综合癌症中心。他在洛杉矶的加州大学获得了正规的研究培训，在普通外科住院医师培训期间获得了分子和细胞生物学博士学位。Arnold Levine博士的实验室发现了p53肿瘤抑制因子，Joseph Bertino博士是国际公认的癌症发展治疗领域的专家，目前正在指导他。Carpizo博士的近期和长期职业目标是：1）建立一个独立的NIH研究项目资助的研究计划，重点是胰腺癌，2）成为一个专门的临床医生和科学家小组的领导者，专注于胰腺癌在NCI指定的综合癌症中心和3）在更广泛的外科肿瘤学学科中发展发展治疗学的利基。Carpizo博士和他的导师/合作导师设计的职业发展计划将通过三个主要组成部分实现这些目标：1）实验室研究：候选人将在Levine博士和Bertino博士的指导下执行上述研究计划。他将从几位合作者那里得到进一步的指导，其中包括Stewart洛博士，他是一位生物化学家，将用NSC 319726进行生物物理实验。Gaetano Montelione博士是一位NMR结构生物学家，他将尝试使用NMR技术研究药物-p53-R175蛋白质相互作用，Ken Olive博士是一位分子生物学家，他经营着一家由NIH资助的胰腺癌实验室，在胰腺癌转基因小鼠模型方面具有专业知识，该模型将用于测试NSC 319726。Carpizo博士还将接受Scott Lowe博士（纪念斯隆-凯特琳癌症中心）在小鼠癌症建模中使用RNAi技术的专业指导。2)转化临床试验经验：Carpizo博士将通过开展旨在研究Novartis Hedgehog抑制剂（LDE-225）在可手术切除胰腺癌患者中的作用机制的小型试点研究，获得转化临床试验经验。Carpizo博士是这项由诺华公司资助的临床试验的主要研究者。Bertino医生将指导他进行本试验。3)教学法：Carpizo博士将在冷泉港参加几门课程，包括1）先进的测序技术，2）癌症的机制和模型，3）胰腺癌研讨会。

DESCRIPTION (provided by applicant): TP53 is the most commonly mutated gene in human cancer for which no effective targeted anti-cancer drug exists. We identified NSC319726 (726) as a mutant p53 reactivating compound, and a lead compound for mutant p53 targeted drug development [1]. Prior evidence indicated that the effects of 726 were allele-specific, by restoring wildtype structure and function to the third most common p53 mis-sense mutant (p53-R175H). This effect was dependent on the zinc-chelating and redox modulating properties of 726. The p53 protein binds a single zinc ion, which is necessary for proper p53 structure and function. The p53-R175H mutation impairs the protein from binding zinc causing it to mis-fold and lose transcriptional function. The mechanism of 726 mutant p53 reactivation is unknown. We provide preliminary data that describe a novel "dual" mechanism where 726 restores wild-type structure to the p53-R175H using a zinc-metallochaperone function to provide an optimal concentration of zinc to facilitate proper folding. We also demonstrate that the redox modulating properties of 726 function to transactivate the newly conformed p53-R175 and induce an apoptotic program. We also demonstrate that this zinc-metallochaperone function applies to other p53 mis-sense mutants with impaired zinc binding. In this K08 Career Development Award, Dr Darren Carpizo and his collaborators plan to explore this compound in two major directions. Aim #1 is to dissect the molecular mechanism by which the compound reactivates the p53-R175 mutant protein and other p53 zinc-binding mutants. Aim #2 is conduct pre-clinical studies of the compound to evaluate its potential for clinical development as a mutant p53 targeted anti-cancer drug. Dr. Carpizo is a surgical oncologist specializing in hepatobiliary and pancreatic cancers currently appointed as an assistant professor of surgery at The Cancer Institute of New Jersey (CINJ), an NCI designated Comprehensive Cancer Center. He has obtained formalized training in research at the University of California at Los Angeles, earning a PhD in molecular and cell biology during his general surgical residency training. Dr. Arnold Levine whose laboratory discovered the p53 tumor suppressor and Dr. Joseph Bertino, an internationally recognized expert in the field of Developmental Therapeutics in cancer, are currently mentoring him. Dr. Carpizo's immediate and long term career goals are to 1) Build an independent NIH Research Project Grant supported research program focusing on pancreatic cancer, 2) Become a leader of a dedicated group of clinicians and scientists that focus on pancreatic cancer at an NCI designated Comprehensive Cancer Center and 3) To develop a niche of Developmental Therapeutics within the broader discipline of Surgical Oncology. The Career Development Plan designed by Dr. Carpizo and his Mentor/Co-Mentor will accomplish these goals with three major components: 1) Laboratory research: The candidate will execute the research plan outlined above under the mentorship of Drs. Levine and Bertino. He will receive further instruction from several collaborators including Dr. Stewart Loh, a biochemist who will conduct biophysical experiments with NSC319726. Dr. Gaetano Montelione, an NMR structural biologist who will attempt to use NMR techniques to study drug-p53-R175 protein interactions and Dr. Ken Olive, a molecular biologist who runs a NIH funded pancreatic cancer laboratory with expertise in the transgenic mouse model of pancreas cancer that will be used to test NSC319726. Dr. Carpizo will also receive specialized instruction in using RNAi technologies in mouse cancer modeling from Dr. Scott Lowe, (Memorial Sloan-Kettering cancer center). 2) Experience in Translational Clinical Trials: Dr. Carpizo will gain experience in translational clinical trials through the conduct of small pilot study designed to study the mechanism of action of the Novartis Hedgehog inhibitor (LDE-225) in patients with surgically resectable pancreas cancer. Dr. Carpizo is the principal investigator of this clinical trial that is being funded by Novartis. Dr. Bertino will mentor him through the conduct of this trial. 3) Didactics: Dr. Carpizo will take attend several courses at Cold Spring Harbor including 1) Advanced Sequencing Technologies, and 2) Mechanisms and Models of Cancer and 3) Workshop on Pancreatic Cancer.