Development of Zinc Metallochaperones as Mutant p53 Targeted Anti-cancer Drugs

锌金属伴侣作为突变 p53 靶向抗癌药物的开发

• 批准号: 9005386
• 负责人: Darren Richard Carpizo
• 金额: $ 37.99万
• 依托单位: RBHS -CANCER INSTITUTE OF NEW JERSEY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-15 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9005386
• 关键词: Affinity; Alleles; Amino Acids; Antineoplastic Agents; Apoptotic; Area; Binding; Biology; Biophysics; Cancer Patient; Cells; Chelating Agents; Clinic; Clinical; Complementary DNA; Complex; DNA; DNA Binding Domain; Data; Defect; Development; Dose; Drug Design; Drug Targeting; Feedback; Foundations; Frequencies; Genes; Genetic; Goals; Homeostasis; Hour; Human; In Vitro; Ionophores; Ions; Knowledge; Laboratories; Lead; Malignant Neoplasms; Measures; Mediating; Metalloproteins; Metals; Missense Mutation; Molecular Conformation; Mutate; Mutation; Outcome; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Post-Translational Protein Processing; Protein p53; Proteins; Public Health; Reactive Oxygen Species; Regulator Genes; Research; Research Personnel; Roentgen Rays; Specificity; Structure; TP53 gene; Therapeutic; Toxic effect; Validation; Zinc; Zinc deficiency; base; cancer cell; cancer type; design; drug development; functional restoration; in vivo; killings; member; monomer; mouse model; mutant; neoplastic cell; novel; novel therapeutics; programs; public health relevance; research study; restoration; small molecule; tumor; tumorigenesis

 DESCRIPTION (provided by applicant): Researchers have acknowledged for decades that the small-molecule restoration of wild type p53 function in tumors is one of the ultimate, yet elusive goals in cancer drug development. TP53 is the most commonly mutated gene in cancer and loss of its function is one of the central drivers of tumorigenesis. The majority of mutations are missense and generate a defective protein found at high levels in cancer cells. The p53-R175H is the most common missense mutant and is misfolded because the substitution impairs the protein's ability to bind zinc. We discovered a zinc chelating small-molecule (ZMC1) that can restore wild type structure/function to the p53-R175H by restoring zinc-binding and facilitating proper folding (so-called zinc metallochaperone (ZMC) mechanism). ZMC1 "reactivates" mutant p53 and selectively kills cancer cells by a p53 mediated apoptotic program both in vitro and in vivo. The pharmacologic restoration of a folding defect in a metalloprotein by metal ion delivery is unprecedented in drug development. We found that other zinc chelators with a similar affinity (Kd) for zinc can function as ZMCs and ZMCs can reactivate other mutants with impaired zinc-binding. Therefore, we hypothesize that ZMCs can be developed as effective mutant p53 targeted anti-cancer drugs. This will be investigated through the following specific aims: 1) Determine the impact of zinc homeostatic mechanisms on ZMC1 pharmacodynamics. Cells respond to the ZMC1 surge in zinc levels by normalizing zinc through homeostatic mechanisms. We hypothesize that these mechanisms regulate ZMC1 activity. We will explore this through mechanistic studies that measure the effect on ZMC1 activity of manipulating zinc regulatory genes in tumors cells. 2) Define the p53 mis-sense mutational spectrum that is amenable to ZMC reactivation. ZMC1 is proposed to reactivate the mutant class with impaired zinc binding; however the full spectrum of this class is unknown. We hypothesize that the mutants closest to the zinc-binding pocket are most likely to have impaired zinc binding. We will study the 15 most frequent mutants within 10Å of the zinc-binding pocket. This will define the potential patient poo for ZMCs. 3) Design a ZMC optimized for potency, toxicity, and efficacy in vivo. We have designed novel chemotypes that have ZMC activity and are more potent than ZMC1 in vitro. We will validate these results in vivo. Our team is composed of three laboratories that are leading this area of research. Their expertise is the following: 1) p53 biology (Darren Carpizo), Zinc/p53 folding biophysics (Stewart Loh), and zinc chelator drug design (David Augeri). Relevance to public health: The research performed in this proposal will provide the foundation for the development of a new class of anti-cancer drugs that target the most commonly mutated gene in human cancer. These drugs will have broad activity against all types of cancers.

 描述（由申请人提供）：几十年来，研究人员已经承认，肿瘤中野生型p53功能的小分子恢复是癌症药物开发的最终目标之一，但难以实现。TP 53是癌症中最常见的突变基因，其功能丧失是肿瘤发生的主要驱动因素之一。大多数突变是错义的，并在癌细胞中产生高水平的缺陷蛋白。p53-R175 H是最常见的错义突变体，并且是错误折叠的，因为取代损害了蛋白质结合锌的能力。我们发现了一种锌螯合小分子（ZMC 1），它可以通过恢复锌结合和促进正确折叠（所谓的锌金属伴侣（ZMC）机制）来恢复p53-R175 H的野生型结构/功能。ZMC 1在体外和体内通过p53介导的凋亡程序“重新激活”突变型p53并选择性地杀死癌细胞。通过金属离子递送的金属蛋白质中的折叠缺陷的药理学修复在药物开发中是前所未有的。我们发现，其他锌螯合剂具有类似的亲和力（Kd）的锌可以作为ZMCs和ZMCs的功能可以重新激活其他突变体与受损的锌结合。因此，我们假设ZMC可以被开发为有效的突变型p53靶向抗癌药物。这将通过以下具体目标进行研究：1）确定锌稳态机制对ZMC 1药效学的影响。细胞通过稳态机制使锌正常化来响应锌水平的ZMC 1激增。我们假设这些机制调节ZMC 1活性。我们将通过机制研究来探索这一点，这些研究测量了在肿瘤细胞中操纵锌调节基因对ZMC 1活性的影响。2)定义适合ZMC再激活的p53错义突变谱。ZMC 1被提议重新激活具有受损锌结合的突变体类别;然而，这类突变体的全部谱尚不清楚。我们推测，最接近锌结合口袋的突变体是最有可能有受损的锌结合。我们将研究15个最常见的突变体在10个锌结合口袋。这将定义ZMC的潜在患者粪便。3)设计针对体内效力、毒性和功效优化的ZMC。我们已经设计了具有ZMC活性并且在体外比ZMC 1更有效的新型化学型。我们将在体内验证这些结果。 我们的团队由三个实验室组成，这些实验室领导着这一研究领域。他们的专业知识如下：1）p53生物学（Darren Carpizo）、锌/p53折叠生物物理学（Stewart洛）和锌螯合剂药物设计（大卫·奥杰里）。与公共卫生的相关性：该提案中进行的研究将为开发一类新的抗癌药物提供基础，这些药物靶向人类癌症中最常见的突变基因。这些药物将对所有类型的癌症具有广泛的活性。