Drug Discovery Chemistry Platform Unit, RIKEN Center for Sustainable Resource Science, Saitama, Japan.
Social Cooperation Program of Targeted Protein Degradation, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
Methods Enzymol. 2023;681:41-60. doi: 10.1016/bs.mie.2022.09.001. Epub 2022 Nov 18.
Specific and nongenetic IAP-dependent Protein Erasers (SNIPERs) are a kind of PROTACs recruiting IAP ubiquitin ligases to induce degradation of target proteins. We have developed a series of SNIPERs against BCR-ABL oncogenic kinases by employing kinase inhibitors as target ligands. Some of these SNIPERs show potent activities to degrade BCR-ABL protein and inhibit CML cell growth. However, since SNIPERs also inhibit kinase activity, it takes some ingenuity to show that degradation of BCR-ABL plays a significant role on growth inhibitory activity. Here we describe protocols for synthesizing SNIPERs against BCR-ABL oncogenic kinase that contain kinase inhibitors as target ligands, and methods for evaluating the growth inhibitory activity against cancer cells, especially focusing on a method to discriminate the significance of protein degradation from that of kinase inhibition.
特异性和非遗传的 IAP 依赖性蛋白清除剂（SNIPERs）是一类通过招募 IAP 泛素连接酶来诱导靶蛋白降解的 PROTACs。我们通过使用激酶抑制剂作为靶配体，开发了一系列针对 BCR-ABL 致癌激酶的 SNIPERs。其中一些 SNIPERs 显示出有效降解 BCR-ABL 蛋白和抑制 CML 细胞生长的活性。然而，由于 SNIPERs 也抑制激酶活性，因此需要一些巧妙的方法来证明 BCR-ABL 的降解在生长抑制活性中起着重要作用。在这里，我们描述了合成针对 BCR-ABL 致癌激酶的 SNIPERs 的方案，这些 SNIPERs 包含作为靶配体的激酶抑制剂，以及评估针对癌细胞的生长抑制活性的方法，特别是侧重于区分蛋白降解和激酶抑制的意义的方法。
