Wijeratne Aruna, Xiao Junpeng, Reutter Christopher, Furness Kelly W, Leon Rebecca, Zia-Ebrahimi Mohammad, Cavitt Rachel N, Strelow John M, Van Horn Robert D, Peng Sheng-Bin, Barda David A, Engler Thomas A, Chalmers Michael J
Discovery Chemistry Research and Technologies and Oncology Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, United States.
ACS Med Chem Lett. 2018 May 21;9(6):557-562. doi: 10.1021/acsmedchemlett.8b00110. eCollection 2018 Jun 14.
The KRASG12C protein product is an attractive, yet challenging, target for small molecule inhibition. One option for therapeutic intervention is to design small molecule ligands capable of binding to and inactivating KRASG12C via formation of a covalent bond to the sulfhydryl group of cysteine 12. In order to better understand the cellular off-target interactions of , a covalent KRASG12C inhibitor, we have completed a series of complementary chemical proteomics experiments in H358 cells. A new thiol reactive probe (TRP) was designed and used to construct a cellular target occupancy assay for KRASG12C. In addition, the thiol reactive probes allowed us to profile potential off-target interactions of with over 3200 cysteine residues. In order to complement the TRP data we designed , an alkyne containing version of , to serve as bait in competitive chemical proteomics experiments. Herein, we describe and compare data from both the TRP and the click chemistry probe pull down experiments.
KRASG12C蛋白产物是小分子抑制的一个有吸引力但具有挑战性的靶点。治疗干预的一种选择是设计能够通过与半胱氨酸12的巯基形成共价键来结合并使KRASG12C失活的小分子配体。为了更好地理解共价KRASG12C抑制剂的细胞脱靶相互作用，我们在H358细胞中完成了一系列互补的化学蛋白质组学实验。设计了一种新的硫醇反应性探针（TRP），并用于构建KRASG12C的细胞靶点占有率测定。此外，硫醇反应性探针使我们能够分析与超过3200个半胱氨酸残基的潜在脱靶相互作用。为了补充TRP数据，我们设计了一种含炔烃的版本作为竞争性化学蛋白质组学实验的诱饵。在此，我们描述并比较了TRP和点击化学探针下拉实验的数据。
