蛋白酶降解靶向嵌合物（PROTAC）是一种具有巨大潜力的异双功能分子，能够克服传统抑制剂的局限性。然而，它的固有缺点越来越显现，如大分子重量引起的细胞渗透性差。因此，在生物正交反应和分子片段的基础上提出了一种胞内自组装策略，以克服固有缺陷，形成一种新型自组装的PROTACs。设计和合成了两种包含四氮唑和诺伯烯作为生物正交基团的前体物质，它们可以在细胞中连接形成新型的PROTACs。幸运的是，超快速HRMS和HPLC测定表明，在活体U87细胞中检测到了由生物正交反应驱动的自组装PROTACs。生物评价表明，前体分子LN-1能以浓度依赖的方式降解PDGFR-β蛋白，而癌细胞与另一前体分子TzB共同处理时，效果更佳。我们的发现验证了自组装策略在未来开发新型PROTACs中的可行性。Copyright © 2023 Elsevier Inc. All rights reserved.

Proteolysis targeting chimera (PROTAC) is a heterobifunctional molecule with enormous potential for its ability to overcome the limitations of traditional inhibitors. However, its inherent disadvantages have been increasingly revealed, such as poor cell permeability caused by large molecule weight. Herein, to overcome the inherent shortcomings, intracellular self-assembly was proposed based on bioorthogonal reaction and molecular fragments, affording a novel type of self-assembled PROTACs. Two types of precursors incorporated with tetrazine and norbornene as bioorthogonal groups were designed and synthesized, and they could subsequently be conjugated in cells to generate novel PROTACs. Fortunately, ultrafast HRMS and HPLC assays indicated that self-assembled PROTACs driven by the bio-orthogonal reaction were detected in living U87 cells. Biological evaluation suggested that the precursor molecule LN-1 could degrade PDGFR-β protein in a concentration-dependent manner, while cancer cells were co-treated with another precursor molecule, TzB. Our findings verified the feasibility of a self-assembly strategy in future development of novel PROTACs.Copyright © 2023 Elsevier Inc. All rights reserved.