Dvořáková Petra, Bušek Petr, Knedlík Tomáš, Schimer Jiří, Etrych Tomáš, Kostka Libor, Stollinová Šromová Lucie, Šubr Vladimír, Šácha Pavel, Šedo Aleksi, Konvalinka Jan
Institute of Organic Chemistry and Biochemistry of The Czech Academy of Sciences , Flemingovo nám 2, 16610 Prague 6, Czech Republic.
Department of Cell Biology, Faculty of Science, Charles University , Viničná 7, 12843 Prague 2, Czech Republic.
J Med Chem. 2017 Oct 26;60(20):8385-8393. doi: 10.1021/acs.jmedchem.7b00767. Epub 2017 Oct 16.
Proteases are directly involved in cancer pathogenesis. Expression of fibroblast activation protein (FAP) is upregulated in stromal fibroblasts in more than 90% of epithelial cancers and is associated with tumor progression. FAP expression is minimal or absent in most normal adult tissues, suggesting its promise as a target for the diagnosis or treatment of various cancers. Here, we report preparation of a polymer conjugate (an iBody) containing a FAP-specific inhibitor as the targeting ligand. The iBody inhibits both human and mouse FAP with low nanomolar inhibition constants but does not inhibit close FAP homologues dipeptidyl peptidase IV, dipeptidyl peptidase 9, and prolyl oligopeptidase. We demonstrate the applicability of this iBody for the isolation of FAP from cell lysates and blood serum as well as for its detection by ELISA, Western blot, flow cytometry, and confocal microscopy. Our results show the iBody is a useful tool for FAP targeting in vitro and potentially also for specific anticancer drug delivery.
蛋白酶直接参与癌症发病机制。在超过90%的上皮癌中，成纤维细胞激活蛋白（FAP）在基质成纤维细胞中的表达上调，且与肿瘤进展相关。在大多数正常成人组织中，FAP表达极少或不存在，这表明它有望成为各种癌症诊断或治疗的靶点。在此，我们报告了一种聚合物偶联物（一种iBody）的制备，该偶联物含有FAP特异性抑制剂作为靶向配体。该iBody以低纳摩尔抑制常数抑制人和小鼠的FAP，但不抑制密切相关的FAP同源物二肽基肽酶IV、二肽基肽酶9和脯氨酰寡肽酶。我们证明了这种iBody可用于从细胞裂解物和血清中分离FAP，以及通过酶联免疫吸附测定、蛋白质免疫印迹、流式细胞术和共聚焦显微镜对其进行检测。我们的结果表明，该iBody是一种在体外靶向FAP的有用工具，也可能用于特定抗癌药物的递送。
