CRUK Beatson Institute, Glasgow G61 1BD, United Kingdom.
Drug Discovery Program, CRUK Beatson Institute, Glasgow G61 1BD, United Kingdom.
J Med Chem. 2022 Feb 10;65(3):1898-1914. doi: 10.1021/acs.jmedchem.1c01265. Epub 2022 Feb 1.
RAS is a major anticancer drug target which requires membrane localization to activate downstream signal transduction. The direct inhibition of RAS has proven to be challenging. Here, we present a novel strategy for targeting RAS by stabilizing its interaction with the prenyl-binding protein PDE6D and disrupting its localization. Using rationally designed RAS point mutations, we were able to stabilize the RAS:PDE6D complex by increasing the affinity of RAS for PDE6D, which resulted in the redirection of RAS to the cytoplasm and the primary cilium and inhibition of oncogenic RAS/ERK signaling. We developed an SPR fragment screening and identified fragments that bind at the KRAS:PDE6D interface, as shown through cocrystal structures. Finally, we show that the stoichiometric ratios of KRAS:PDE6D vary in different cell lines, suggesting that the impact of this strategy might be cell-type-dependent. This study forms the foundation from which a potential anticancer small-molecule RAS:PDE6D complex stabilizer could be developed.
RAS 是一种主要的抗癌药物靶点，需要膜定位才能激活下游信号转导。直接抑制 RAS 已被证明具有挑战性。在这里，我们提出了一种通过稳定 RAS 与 prenyl-binding 蛋白 PDE6D 的相互作用并破坏其定位来靶向 RAS 的新策略。通过合理设计的 RAS 点突变，我们能够通过增加 RAS 与 PDE6D 的亲和力来稳定 RAS:PDE6D 复合物，从而导致 RAS 向细胞质和初级纤毛重新定位，并抑制致癌性 RAS/ERK 信号。我们开发了一种 SPR 片段筛选方法，并通过共晶结构显示鉴定出与 KRAS:PDE6D 界面结合的片段。最后，我们表明不同细胞系中 KRAS:PDE6D 的化学计量比不同，这表明该策略的影响可能取决于细胞类型。这项研究为开发潜在的抗癌小分子 RAS:PDE6D 复合物稳定剂奠定了基础。
