A "Click Chemistry Platform" for the Rapid Synthesis of Bispecific Molecules for Inducing Protein Degradation
A "Click Chemistry Platform" for the Rapid Synthesis of Bispecific Molecules for Inducing Protein Degradation
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DOI：
10.1021/acs.jmedchem.6b01781
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发表时间：
2018-01-25
影响因子：
7.3
通讯作者：
Ceet, Victor J.
中科院分区：
文献类型：
--
作者：
Wurz, Ryan P.;Dellamaggiore, Ken;Ceet, Victor J.
靶向嵌合体的蛋白水解是含有靶蛋白粘合剂的双特异性分子和连接器连接的泛素连接酶粘合剂。通过将泛素连接酶募集到靶蛋白中，protac可以促进靶蛋白的泛素化和蛋白酶体降解。有效的Protac的产生取决于蛋白质/连接酶配对对，链接位点，接头长度和接头组成的性质，所有这些都难以以系统的方式解决。本文中，我们描述了一种合成Protacs的“点击化学”方法。我们证明了这种方法使用溴结构域和外部域-4（BRD4）配体JQ-1（3）和靶向Cereblon（CRBN）和von Hippel-Lindau（VHL）蛋白的连接酶结合剂的实用性。使用Alphascreen接近测定法来确定Protac形成三元酶-protac-target蛋白复合物的能力和MSD测定法，以测量由Protac促进的靶蛋白的细胞降解。
Proteolysis targeting chimeras (PROTACs) are bispecific molecules containing a target protein binder and an ubiquitin ligase binder connected by a linker. By recruiting an ubiquitin ligase to a target protein, PROTACs promote ubiquitination and proteasomal degradation of the target protein. The generation of effective PROTACs depends on the nature of the protein/ligase ligand pair, linkage site, linker length, and linker composition, all of which have been difficult to address in a systematic way. Herein, we describe a "click chemistry" approach for the synthesis of PROTACs. We demonstrate the utility of this approach with the bromodomain and extraterminal domain-4 (BRD4) ligand JQ-1 (3) and ligase binders targeting cereblon (CRBN) and Von Hippel-Lindau (VHL) proteins. An AlphaScreen proximity assay was used to determine the ability of PROTACs to form the ternary ligase-PROTAC-target protein complex and a MSD assay to measure cellular degradation of the target protein promoted by PROTACs.