将近端反应弹头基因编码到蛋白质粘合剂/药物中,已成为共价结合蛋白质靶标实现广泛应用的有效策略。为了扩大在生理条件下靶向不同天然残留物的反应范围,迫切需要开发一种具有优异稳定性和广泛反应性的基因编码反应弹头。
该文中,研究人员报道了含环氧酪氨酸(EPOY)的基因编码,用于开发共价蛋白药物。研究表明,当EPOY结合到纳米体(KN035)中时,可以与PD-L1蛋白相同位置的不同侧链(突变)交联。值得注意的是,首次实现了能够共价和位点特异性靶向10个不同亲核残基的单一基因编码反应弹头。这将在很大程度上扩大共价弹头的范围,并启发小分子药物和蛋白质药物共价弹头的发展。
此外,将EPOY结合到设计的锚蛋白重复蛋白(DarpinK13)中,以产生KRAS的共价结合物。这种共价KRAS结合物有可能基于所有致癌KRAS突变体之间的结构相似性,实现KRAS的泛共价靶向,同时通过与KRAS特异性残基(H95和E107)的共价相互作用避免与NRAS/HRAS的脱靶结合。研究认为,共价靶向H95将是未来开发共价泛KRAS抑制剂的一种很有前途的策略。
附:英文原文
Title: Genetically Encoded Epoxide Warhead for Precise and Versatile Covalent Targeting of Proteins
Author: Jinpeng Zhang, Xia Wang, Qingjun Huang, Jinsong Ye, Jie Wang
Issue&Volume: May 31, 2024
Abstract: Genetically encoding a proximal reactive warhead into the protein binder/drug has emerged as an efficient strategy for covalently binding to protein targets, enabling broad applications. To expand the reactivity scope for targeting the diverse natural residues under physiological conditions, the development of a genetically encoded reactive warhead with excellent stability and broad reactivity is highly desired. Herein, we reported the genetic encoding of epoxide-containing tyrosine (EPOY) for developing covalent protein drugs. Our study demonstrates that EPOY, when incorporated into a nanobody (KN035), can cross-link with different side chains (mutations) at the same position of PD-L1 protein. Significantly, a single genetically encoded reactive warhead that is capable of covalent and site-specific targeting to 10 different nucleophilic residues was achieved for the first time. This would largely expand the scope of covalent warhead and inspire the development of covalent warheads for both small-molecule drugs and protein drugs. Furthermore, we incorporate the EPOY into a designed ankyrin repeat protein (DarpinK13) to create the covalent binders of KRAS. This covalent KRAS binder holds the potential to achieve pan-covalent targeting of KRAS based on the structural similarity among all oncogenic KRAS mutants while avoiding off-target binding to NRAS/HRAS through a covalent interaction with KRAS-specific residues (H95 and E107). We envision that covalently targeting to H95 will be a promising strategy for the development of covalent pan-KRAS inhibitors in the future.
DOI: 10.1021/jacs.4c03974
Source: https://pubs.acs.org/doi/abs/10.1021/jacs.4c03974
JACS:《美国化学会志》,创刊于1879年。隶属于美国化学会,最新IF:16.383
官方网址:https://pubs.acs.org/journal/jacsat
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