Exploration of a Mutant p53 Reactivating Compound

突变型 p53 重新激活化合物的探索

• 批准号: 8883424
• 负责人: Darren Richard Carpizo
• 金额: $ 15.9万
• 依托单位: RBHS -CANCER INSTITUTE OF NEW JERSEY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-16 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8883424
• 关键词: Acetylcysteine; Address; Affinity; Alleles; Antineoplastic Agents; Apoptosis; Apoptotic; Area; Binding; Buffers; California; Cancer Institute of New Jersey; Cancer Model; Cellular biology; Chelating Agents; Chemistry; Clinic; Clinical; Clinical Trials; Comprehensive Cancer Center; Computer Simulation; Copper; DNA Binding Domain; Data; Development; Development Plans; Developmental Therapeutics Program; Diamide; Discipline; Doctor of Philosophy; Dose; Drug Kinetics; Drug Targeting; Educational workshop; Enrollment; Environment; Erinaceidae; Family; Funding; Future; Genes; Genetically Engineered Mouse; Goals; Health; Human; Instruction; Ions; Iron; K-Series Research Career Programs; Knowledge; Laboratories; Laboratory Research; Lead; Left; Los Angeles; MDM2 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Memorial Sloan-Kettering Cancer Center; Mentors; Mentorship; Metals; Methodology; Modeling; Molecular; Molecular Biology; Mus; Mutate; Mutation; Normal Cell; Olives - dietary; Operative Surgical Procedures; Outcome; Oxidation-Reduction; Patients; Pharmaceutical Preparations; Pharmacodynamics; Pilot Projects; Post-Translational Protein Processing; Principal Investigator; Property; Protein p53; Proteins; RNA Interference; Reducing Agents; Research; Research Design; Research Project Grants; Research Support; Resectable; Residencies; Running; Scientist; Source; Structural Biologist; Structure; Surgical Oncologist; Surgical Oncology; TP53 gene; Techniques; Technology; Testing; Therapeutic; Thiosemicarbazones; Training; Transgenic Mice; Tumor Suppressor Proteins; United States; United States National Institutes of Health; Universities; Xenograft procedure; Zinc; anti-cancer therapeutic; cancer cell; career; career development; chelation; design; drug development; experience; hepatobiliary cancer; in vivo; inhibitor/antagonist; killings; mouse model; mutant; novel; pre-clinical; preclinical study; professor; programs; protein folding; research study; restoration; screening; translational clinical trial; tumor growth; zinc-binding protein

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.

描述（由申请人提供）：TP53是人类癌症中最常见的突变基因，目前尚无有效的靶向抗癌药物。我们发现NSC319726（726）是突变型p53再激活化合物，也是突变型p53靶向药物开发[1]的先导化合物。先前的证据表明，726的作用是等位基因特异性的，通过恢复野生型的结构和功能到第三个最常见的p53错义突变体（p53- r175h）。这种效果取决于726的锌螯合和氧化还原调节特性。p53蛋白结合一个锌离子，这是p53正常结构和功能所必需的。p53-R175H突变破坏了蛋白质与锌的结合，导致其错误折叠并失去转录功能。726突变体p53再激活的机制尚不清楚。我们提供的初步数据描述了一种新的“双重”机制，其中726利用锌-金属伴侣蛋白功能提供最佳的锌浓度来促进适当的折叠，从而恢复p53-R175H的野生型结构。我们还证明了726的氧化还原调节特性可以反激活新形成的p53-R175并诱导凋亡程序。我们也证明这种锌-金属伴侣的功能适用于其他锌结合受损的p53错义突变体。在本次K08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。目标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博士是一名专攻肝胆和胰腺癌的外科肿瘤学家，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。他在加州大学洛杉矶分校（University of California at Los Angeles）获得了正式的研究培训，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。目前，他的导师是发现p53肿瘤抑制因子的实验室的Arnold Levine博士和癌症发育治疗领域国际公认的专家Joseph Bertino博士。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项目，重点关注胰腺癌；2)在NCI指定的综合癌症中心成为专注于胰腺癌的临床医生和科学家团队的领导者；3)在外科肿瘤学的更广泛学科中发展发育治疗学。由Dr. Carpizo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组成部分来实现这些目标：1)实验室研究：候选人将在Dr. Carpizo的指导下执行上述研究计划。Levine和Bertino。他将接受几位合作者的进一步指导，其中包括生物化学家Stewart Loh博士，他将用NSC319726进行生物物理实验。Gaetano Montelione博士是核磁共振结构生物学家，他将尝试使用核磁共振技术研究药物-p53- r175蛋白质的相互作用；Ken Olive博士是分子生物学家，他管理着一个由美国国立卫生研究院资助的胰腺癌实验室，专门研究胰腺癌的转基因小鼠模型，该模型将用于测试NSC319726。Carpizo博士还将接受Scott Lowe博士（Memorial Sloan-Kettering癌症中心）的专门指导，在小鼠癌症建模中使用RNAi技术。2)转化临床试验经验：Carpizo博士将通过开展小型试点研究获得转化临床试验经验，该研究旨在研究诺华Hedgehog抑制剂（LDE-225）在手术切除的胰腺癌患者中的作用机制。Carpizo博士是这项临床试验的首席研究员，该试验由诺华公司资助。伯蒂诺医生会指导他进行试验。3)教学：Dr. Carpizo将参加冷泉港的几门课程，包括：1)先进测序技术，2)癌症机制和模型，3)胰腺癌研讨会。