Identification of Small Molecules for Reactivation of p53 Cancer Mutants

鉴定用于 p53 癌症突变体再激活的小分子

• 批准号: 7617518
• 负责人: Peter Kaiser
• 金额: $ 14.62万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7617518
• 关键词: Amino Acids; Animal Model; Biochemical; Biological Assay; Chemicals; Compatible; Detection; Development; Disease regression; Fluorescence; Goals; Human; In Vitro; Lead; Length; Libraries; Malignant Neoplasms; Mammalian Cell; Methods; Molecular Conformation; Monitor; Mutate; Mutation; Patients; Preclinical Drug Evaluation; Protein p53; Public Health; Research; Resistance; Screening procedure; System; TP53 gene; Therapeutic; Thinking; Yeasts; base; cancer therapy; cell growth; functional restoration; high throughput screening; improved; in vitro Assay; mutant; research study; small molecule; thermostability; tissue/cell culture; tumor; tumor growth

DESCRIPTION (provided by applicant): Half of all human cancers carry mutations in the tumor suppressor p53. The majorities of p53 negative cancers do express full length, but mutated p53. Many of the p53 mutations found in cancers are single amino acid changes in the 200 amino acid core domain of p53. These mutations are thought to destabilize the correct p53 fold and thus lead to inactivation of the p53 tumor suppressor function. Our hypothesis is that small molecules exist that can stabilize the correct conformation in p53 cancer mutants. These molecules could therefore reactivate mutated forms of p53 in cancers. Experiments in animal models have demonstrated that restoring p53 function even in advanced tumors leads to tumor regression. Small molecules that can restore function to p53 cancer mutants are therefore predicted to significantly improve the outlook for patients carrying a p53 mutation. We propose to develop a screening platform compatible with high throughput small molecule screens (HTS) to identify compounds that reactivate p53 mutants that are frequently found in human cancers. The proposed drug screen relies on unique, cheap and very efficient yeast-based assays that allow us to screen compounds simultaneously against the 70 most commonly found p53 cancer mutants. We propose to implement fluorescence-based readouts for our yeast-based system to enable HTS. In addition we propose secondary yeast assays to confirm a specific effect of small molecules on p53 cancer mutants and to determine how many different p53 mutants are rescued by one specific small molecule. A biochemical strategy that looks at effects of the identified small molecules on thermostability of p53 is proposed to select molecules that directly improve structural integrity of p53. Lastly, adaptation of a tertiary screen is proposed that will evaluate rescue compounds in mammalian cells. The immediate goal of the proposal is to develop the framework to identify small molecules that restore function to p53 cancer mutants and use these molecules later as lead compounds to synthesize derivatives to increase their therapeutic activity. Restoring p53 function in tumors has been demonstrated to reverse tumor growth in animal models and is therefore an attractive strategy for cancer treatment. PUBLIC HEALTH RELEVANCE: About 50% of all human cancers carry mutations in the tumor suppressor p53. p53 negative cancers generally more resistant to current treatment strategies and alternative approaches are particularly necessary for effective treatment of these cancers. The objective of this proposal is to identify small molecules that restore function to p53 cancer mutants and to develop these molecules into therapeutics to restore p53 function in tumors.

描述（由申请人提供）：所有人类癌症的一半携带肿瘤抑制p53中的突变。大多数p53负癌确实表达了全长，但突变为p53。癌症中发现的许多p53突变是p53的200个氨基酸核心结构域中的单氨基酸变化。这些突变被认为破坏了正确的p53折叠，从而导致p53肿瘤抑制器功能失活。我们的假设是存在很小的分子可以稳定p53癌症突变体中正确构象。因此，这些分子可以重新激活癌症中p53的突变形式。动物模型中的实验表明，即使在晚期肿瘤中，恢复p53的功能也会导致肿瘤消退。因此，预计可以恢复p53癌症突变体功能的小分子可显着改善携带p53突变的患者的前景。我们建议开发与高吞吐量小分子筛选（HTS）兼容的筛选平台，以识别重新激活人类癌症经常发现的p53突变体的化合物。拟议的药物筛选依赖于独特，廉价且基于酵母菌的独特测定法，这使我们能够与最常见的p53癌症突变体同时筛选化合物。我们建议为基于酵母的系统实施基于荧光的读数以启用HTS。此外，我们建议辅助酵母分析确认小分子对p53癌症突变体的特定作用，并确定一个特定的小分子挽救了多少种不同的p53突变体。提出了一种生化策略，该策略研究了所鉴定的小分子对p53热稳定性的影响，以选择直接改善p53结构完整性的分子。最后，提出了三级筛查的适应，该筛查将评估哺乳动物细胞中的救援化合物。该提案的直接目标是开发框架，以鉴定恢复p53癌症突变体功能的小分子，并以后将这些分子用作铅化合物，以合成衍生物以增加其治疗活性。已证明恢复肿瘤中的p53功能可以在动物模型中逆转肿瘤的生长，因此是癌症治疗的有吸引力的策略。 公共卫生相关性：所有人类癌症中约有50％在肿瘤抑制p53中携带突变。 p53阴性癌症通常对当前治疗策略具有更大的抵抗力，而替代方法对于有效治疗这些癌症特别必要。该提案的目的是确定将功能恢复为p53癌症突变体的小分子，并将这些分子发展为治疗剂以恢复肿瘤中p53功能。