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：
10427407
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/10427407
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 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 抑制剂作为新型药物来治疗 p53 缺失的癌症中 PI5P4Kβ 和 PI5P4Kβ 是同源脂质激酶，在调节中发挥重要作用。它们催化 PI(5)P 磷酸化形成 PI(4,5)P2。不是 PI(4,5)P2 的主要合成途径，它们的活性消除了 PI(5)P（一种应激诱导的脂质） PI5P4Kβ 被敲除的转基因动物对胰岛素过敏，并且联合起来。 PI5P4Kα 敲除可减少人类 Li-Fraumeni 小鼠模型中的自发肿瘤发生在一些初步研究中，肿瘤抑制基因 p53 在种系中发生突变。通过高通量筛选，二氢蝶啶酮衍生物被鉴定为 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 PI5P4Kα/β 独特的疏水口袋，产生的化合物对两者的效力均高 50 倍 PI5P4Kα 和 PI5P4Kβ，以及针对蛋白激酶的高度选择性在具体目标 1 中，我们将。基于化合物与 PI5P4Kβ 的共晶结构，继续修改最有效的抑制剂，并关注经历构象变化的结合袋的不同区域。使用该家族的远亲成员 PIKfyve 解析抑制剂的晶体结构，以便设计在特定目标 2 中，我们研究 p53(+/+) 和 p53(-/-) 细胞如何响应。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， AMPK 激活，这可以解释动物研究中观察到的胰岛素敏感性增强的原因。脂质激酶抑制可能通过破坏增殖过程中的能量稳态而导致细胞周期停滞将检查 p53-/- 癌细胞 PI5P4Kα/β 和 p53 之间的合成致死相互作用。通过化学生物学和遗传学方法进行检查，并通过化学探针直接接触将通过细胞热位移测定（CETSA）研究增殖肿瘤细胞内的脂质激酶。