Howell Rebecca A, Wang Shikun, Khambete Mihir, McDonald David M, Spiegel David A
Department of Chemistry, Yale University, 225 Prospect Street, New Haven, Connecticut 06520, United States.
J Am Chem Soc. 2024 Jun 10. doi: 10.1021/jacs.3c13320.
Targeted protein degradation (TPD) has emerged as an effective therapeutic strategy for a wide range of diseases; however, the blood-brain barrier (BBB) limits access of degraders into the central nervous system (CNS). Here, we present a new class of bifunctional small molecules, called TransMoDEs (cytosis-inducing lecular egraders of xtracellular proteins), capable of both (1) removal of target protein via lysosomal proteolysis and (2) transcytosis of protein targets across brain endothelial cells. TransMoDEs are derived from Angiopep-2, a peptide motif previously employed as a covalent tag to facilitate receptor-mediated transcytosis across the BBB. We demonstrate that TransMoDEs containing either a biotin or chloroalkane ligand can trigger endocytosis of streptavidin or HaloTag protein, respectively. Interestingly, although low-density lipoprotein receptor-related protein 1 (LRP1) has been reported as the primary receptor for Angiopep-2, TransMoDE-mediated target uptake does not rely exclusively on this pathway. Furthermore, TransMoDE-mediated endocytosis of streptavidin in a bEnd.3 BBB model occurs in a clathrin-mediated mechanism and results in both lysosomal localization and transcytosis of the target protein. This study demonstrates that TransMoDEs can recruit, transcytose, and degrade proteins of interest in cells relevant to the CNS, supporting their further development for the removal of pathogenic neuroproteins.
靶向蛋白质降解（TPD）已成为治疗多种疾病的有效策略；然而，血脑屏障（BBB）限制了降解剂进入中枢神经系统（CNS）。在此，我们提出了一类新型双功能小分子，称为跨细胞转运降解剂（TransMoDEs，胞外蛋白的胞吞诱导分子降解剂），其能够（1）通过溶酶体蛋白水解作用去除靶蛋白，以及（2）使蛋白靶标跨脑内皮细胞进行转胞吞作用。TransMoDEs衍生自血管活性肠肽-2（Angiopep-2），这是一种先前用作共价标签以促进受体介导的跨血脑屏障转胞吞作用的肽基序。我们证明，含有生物素或氯代烷烃配体的TransMoDEs分别可以触发抗生物素蛋白或卤代标签蛋白的内吞作用。有趣的是，尽管低密度脂蛋白受体相关蛋白1（LRP1）已被报道为Angiopep-2的主要受体，但TransMoDE介导的靶标摄取并不完全依赖于该途径。此外，在bEnd.3血脑屏障模型中，TransMoDE介导的抗生物素蛋白内吞作用以网格蛋白介导的机制发生，并导致靶蛋白的溶酶体定位和转胞吞作用。这项研究表明，TransMoDEs可以在与中枢神经系统相关的细胞中募集、转胞吞和降解感兴趣的蛋白质，支持它们在去除致病性神经蛋白方面的进一步开发。
