Exploration of a Mutant p53 Reactivating Compound

描述(申请人提供)：TP53是人类癌症中最常见的突变基因，目前还没有有效的靶向抗癌药物。我们鉴定了NSC319726(726)为突变型P53再激活化合物和突变型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，第三常见的p53错义突变体(P53-R175H)。这一效应依赖于726的锌螯合和氧化还原调节特性。P53蛋白结合单一的锌离子，这是正常的P53结构和功能所必需的。P53-R175H突变损害了蛋白质与锌的结合，导致其错误折叠并失去转录功能。726突变型P53重新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使用锌-金属分配伴侣功能将野生型结构恢复到p53-R175H，以提供最佳的锌浓度，以促进适当的折叠。我们还证明了726的氧化还原调节特性能够反式激活新整合的p53-R175并诱导细胞凋亡。我们还证明这种锌-金属配位体功能适用于其他锌结合受损的p53错义突变体。在K08职业发展奖中，达伦·卡皮索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分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正在进行该化合物的临床前研究，以评估其作为突变型p53靶向抗癌药物的临床开发潜力。卡皮波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，CINJ是NCI指定的综合癌症中心。他在加州大学洛杉矶分校接受了正式的研究培训，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。阿诺德·莱文博士和约瑟夫·贝尔蒂诺博士目前正在指导他。莱文博士的实验室发现了p53肿瘤抑制基因，约瑟夫·贝尔蒂诺博士是国际公认的癌症开发治疗专家。卡皮波博士的近期和长期职业目标是：1)建立一个独立的、由美国国立卫生研究院资助的、专注于胰腺癌的研究项目；2)成为美国国立卫生研究院指定的综合癌症中心专门研究胰腺癌的临床医生和科学家团队的领导者；3)在更广泛的外科肿瘤学学科中发展开发治疗学的利基市场。卡皮索博士和他的导师/共同导师设计的职业发展计划将通过三个主要部分实现这些目标：1)实验室研究：应聘者将在Levine博士和Bertino博士的指导下执行上面概述的研究计划。他将从几位合作者那里得到进一步的指导，其中包括将用NSC319726进行生物物理实验的生物化学家陆恭蕙博士。将尝试使用核磁共振技术研究药物与P53-R175蛋白质相互作用的核磁共振结构生物学家Gaetano Montelione博士和将用于测试NSC319726的NIH资助的胰腺癌实验室的分子生物学家Ken Olive博士，他在胰腺癌转基因小鼠模型方面拥有专业知识。卡皮佐博士还将接受斯隆-凯特琳纪念癌症中心Scott Lowe博士的专业指导，指导他如何在老鼠癌症模型中使用RNAi技术。2)转化性临床试验经验：卡皮波博士将通过开展旨在研究诺华Hedgehog抑制剂(LDE-225)在可手术切除的胰腺癌患者中的作用机制的小型先导性研究，获得转化性临床试验方面的经验。卡皮佐博士是这项由诺华公司资助的临床试验的首席研究员。贝尔蒂诺博士将在这项试验的过程中指导他。3)教学：卡皮波博士将在冷泉港参加几个课程，包括1)先进测序技术，2)癌症机制和模型，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.