STAT3を分解するPROTAC/SNIPERの分子設計、合成と活性評価

降解STAT3的PROTAC/SNIPER的分子设计、合成及活性评价

基本信息

批准号：
22KF0090
负责人：
内藤幹彦
金额：
$ 1.47万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2023
资助国家：
日本
起止时间：
2023-03-08 至 2024-03-31
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22KF0090/
关键词：
PROTAC STAT3 SNIPER protein degradation

项目摘要

Proteolysis targeting chimera (PROTAC) is a technology that uses chimeric chemicals to bind to a protein of interest (POI) and an E3 ligase so as to induce POI degradation through the ubiquitin-proteasome system. In this project, we intended to develop novel PROTAC degraders targeting STAT3.First, we employed sulfonamide derivatives that are believed to bind to the SH2 domain of STAT3 as small molecule-warheads and synthesized PROTACs by conjugating to thalidomide (a cereblon E3 ligase binder) via alkyl linkers. The molecular weights of the resulting PROTACs are approximately 900. Then, the STAT3 degradation activity of these PROTACs were tested along with their parental STAT3 binders in MCF7 and SU-DHL-1 cells. The western blot analysis indicated that PS314 (PROTAC, using PS313 as a warhead) showed a significant activity to reduce STAT3 protein level in both cell lines, while PS312 (PROTAC, using PS311 as a warhead) did not reduce the STAT3 protein level.We also developed oligonucleotide-warheaded PROTAC molecules against STAT3. A decoy oligonucleotide to which STAT3 binds was conjugated to three different E3 ligands to be LCL-STAT3 decoy, POM-STAT3 decoy and VH-STAT3 decoy. Tested in MCF7 cells, POM-STAT3 decoy was found to be more able to degrade STAT3 than VH-STAT3 decoy, and LCL-STAT3 decoy showed no degradation activity.

蛋白水解靶向嵌合体（PROTAC）是一种利用嵌合化学物质与目的蛋白（POI）和E3连接酶结合，通过泛素-蛋白酶体系统诱导POI降解的技术。在这个项目中，我们打算开发针对 STAT3 的新型 PROTAC 降解剂。首先，我们采用磺酰胺衍生物作为小分子弹头，据信可以与 STAT3 的 SH2 结构域结合，并通过与沙利度胺（一种 cereblon E3 连接酶结合剂）缀合来合成 PROTAC。通过烷基接头。所得 PROTAC 的分子量约为 900。然后，在 MCF7 和 SU-DHL-1 细胞中测试这些 PROTAC 的 STAT3 降解活性及其亲本 STAT3 结合物。蛋白质印迹分析表明，PS314（PROTAC，使用PS313作为弹头）在两种细胞系中均表现出显着降低STAT3蛋白水平的活性，而PS312（PROTAC，使用PS311作为弹头）没有降低STAT3蛋白水平。还开发了针对 STAT3 的寡核苷酸弹头 PROTAC 分子。 STAT3结合的诱饵寡核苷酸与三种不同的E3配体缀合，成为LCL-STAT3诱饵、POM-STAT3诱饵和VH-STAT3诱饵。在 MCF7 细胞中进行测试，发现 POM-STAT3 诱饵比 VH-STAT3 诱饵更能够降解 STAT3，而 LCL-STAT3 诱饵则没有表现出降解活性。