Development of novel PIP4K inhibitors to treat p53-null cancer

开发新型 PIP4K 抑制剂来治疗 p53 缺失癌症

基本信息

批准号：
10260471
负责人：
YA HA
金额：
$ 33.88万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-10 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10260471
关键词：
Adult Animals Auxins Binding Biochemical Biological Biological Assay Biological Process CRISPR/Cas technology Cell Cycle Arrest Cell Line Cell Proliferation Cells Cellular Metabolic Process Chemicals Complex Crystallization Development Distant Energy Metabolism Family Family member Gene Expression Genetic Genetic study Homeostasis Hydrophobicity Hypersensitivity Insulin Knock-out Knockout Mice Li-Fraumeni Syndrome Link Lipids Malignant Neoplasms Metabolic Modification Molecular Conformation Muscle Muscle Cells Muscle Fibers Mutate Mutation Nuclear Pharmacology Phosphatidylinositol Phosphates Phosphatidylinositols Phosphorylation Phosphotransferases Play Proliferating Protein Kinase Refractory Regulation Roentgen Rays Role Route Second Messenger Systems Side Signal Transduction Specificity Stress Structure TP53 gene Testing Thinness Transgenic Animals analog base cancer cell design experimental study genetic approach high throughput screening human model inhibitor/antagonist insulin sensitivity kinase inhibitor male member mouse model mutant neoplastic cell novel novel strategies phosphatidylinositol 3-phosphate phosphatidylinositol 5-phosphate response small molecule inhibitor tumor tumorigenesis

项目摘要

A long-term objective of the project is to develop PI5P4Kα/β inhibitors as novel pharmacological agents to treat p53-null cancers. PI5P4Kβ and PI5P4Kβ are homologous lipid kinases that play important roles in regulating cell metabolism and proliferation. They catalyze the phosphorylation of PI(5)P to form PI(4,5)P2. Although this is not a major synthetic route for PI(4,5)P2, their activities eliminate PI(5)P, a stress-induced lipid second messenger. Transgenic animals with PI5P4Kβ knocked out are hypersensitive to insulin, and combined knockout with PI5P4Kα reduce spontaneous tumorigenesis in a mouse model of human Li-Fraumeni syndrome where tumor suppressor p53 is mutated in the germline. In preliminary studies several dihydropteridinone derivatives were identified from high throughput screening as weak inhibitors for PI5P4Kα. Initial syntheses, guided by X-ray crystallographic analysis of kinase inhibitor complexes, and exploiting a hydrophobic pocket unique to PI5P4Kα/β, have yielded compounds with 50-fold greater potency for both PI5P4Kα and PI5P4Kβ, and a high degree of selectivity against protein kinases. In specific aim 1, we will continue to modify the most potent inhibitor, based on a co-crystal structure of the compound with PI5P4Kβ, and focusing on a different region of the binding pocket that undergoes conformational change. We plan to solve the crystal structure of the inhibitor with PIKfyve, a distant member of the family, in order to design analogs that do not cross-inhibit it. In specific aim 2, we study how p53(+/+) and p53(-/-) cells respond to the chemical probe. In cultured myotubes, we found that lipid kinase inhibitor disrupted cell energy homeostasis, causing AMPK activation, which may explain enhanced insulin sensitivity observed in animal studies. The possibility that lipid kinase inhibition causes cell cycle arrest by disrupting energy homeostasis in proliferating p53-/- cancer cells will be examined. The synthetic lethal interaction between PI5P4Kα/β and p53 will be examined by both chemical biological and genetic approaches, and direct engagement of chemical probe with the lipid kinase within proliferating tumor cells will be studied by cellular thermal shift assay (CETSA).

该项目的长期目标是开发PI5P4Kα/β抑制剂作为治疗的新药理剂 p53-null癌症。 PI5P4Kβ和PI5P4Kβ是同源脂质激酶，在调节中起重要作用细胞代谢和增殖。它们催化Pi（5）P的磷酸化形成PI（4,5）P2。虽然这个不是PI（4,5）P2的主要合成途径，它们的活性消除了PI（5）P，是应力诱导的脂质第二信使。击倒PI5P4Kβ的转基因动物对胰岛素敏感，并结合用PI5P4Kα敲除在人li-fraumeni小鼠模型中自发性肿瘤发生综合征在种系中突变肿瘤抑制p53。在初步研究中从高吞吐量筛选中鉴定出二氢吡汀衍生物是PI5P4Kα的弱抑制剂。初始合成，以激酶抑制剂复合物的X射线晶体学分析为指导，并利用A PI5P4Kα/β独有的疏水口袋已经产生的化合物，两者都具有50倍的效力 PI5P4Kα和PI5P4Kβ，以及针对蛋白激酶的高度选择性。在特定目标1中，我们将继续根据PI5P4Kβ的化合物的共结晶来修饰最有效的抑制剂，并专注于经历构象变化的结合口袋的不同区域。我们计划用遥远的家族成员Pikfyve解决抑制剂的晶体结构，以设计不能交叉抑制它的类似物。在特定的目标2中，我们研究了p53（+/+）和p53（ - / - ）细胞如何响应化学探针。在培养的肌管中，我们发现脂质激酶抑制剂破坏了细胞能量稳态，引起AMPK激活，这可以解释在动物研究中观察到的增强的胰岛素敏感性。脂质激酶抑制作用的可能性通过破坏能量稳态而导致细胞周期停滞将检查p53 - / - 癌细胞。 PI5P4Kα/β和p53之间的合成致命相互作用将是通过化学生物学和遗传方法检查，并直接将化学探针与增殖肿瘤细胞中的脂质激酶将通过细胞热移测定法（CETSA）进行研究。