Department of Pharmaceutical Chemistry, the, Cardiovascular Research Institute, Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, 94158, San Francisco, California, United States.
Angew Chem Int Ed Engl. 2024 Nov 18;63(47):e202412843. doi: 10.1002/anie.202412843. Epub 2024 Oct 15.
Sulfur fluoride exchange (SuFEx) and phosphorus fluoride exchange (PFEx) click chemistries are advancing research across multiple disciplines. By genetically incorporating latent bioreactive unnatural amino acids (Uaas), these chemistries have been integrated into proteins, enabling precise covalent linkages with biological macromolecules and paving the way for new applications. However, their suboptimal reaction rates in proteins limit effectiveness, and traditional catalytic methods for small molecules are often incompatible with biological systems or in vivo applications. We demonstrated that introducing an arginine adjacent to the latent bioreactive Uaa significantly boosts SuFEx and PFEx reaction rates between proteins. This method is effective across various Uaas, target residues, and protein environments. Notably, it also enables efficient SuFEx reactions in acidic conditions, common in certain cellular compartments and tumor microenvironments, which typically hinder SuFEx reactions. Furthermore, we developed the first covalent cell engager that substantially enhances natural killer cell activation through improved covalent interaction facilitated by arginine. These findings provide mechanistic insights and offer a biocompatible strategy to harness these robust chemistries for advancing biological research and developing new biotherapeutics.
硫氟交换（SuFEx）和磷氟交换（PFEx）点击化学正在多个学科领域推动研究进展。通过将潜在的生物反应性非天然氨基酸（Uaas）基因整合到蛋白质中，可以实现与生物大分子的精确共价连接，并为新的应用铺平道路。然而，这些化学反应在蛋白质中的反应速率不理想，限制了其效果，而传统的小分子催化方法通常与生物系统或体内应用不兼容。我们证明，在潜在的生物反应性 Uaas 旁边引入精氨酸可以显著提高蛋白质之间的 SuFEx 和 PFEx 反应速率。该方法在各种 Uaas、靶标残基和蛋白质环境中均有效。值得注意的是，它还可以在酸性条件下实现高效的 SuFEx 反应，酸性条件在某些细胞区室和肿瘤微环境中很常见，通常会阻碍 SuFEx 反应。此外，我们开发了第一个共价细胞衔接物，通过精氨酸促进的改进共价相互作用，显著增强了自然杀伤细胞的激活。这些发现提供了机制见解，并提供了一种生物相容性策略，以利用这些强大的化学方法来推进生物研究和开发新的生物疗法。
