Wlodarczyk Aneta, Treda Cezary, Pacholczyk Marcin, Rutkowska Adrianna, Wegierska Marta, Kierasinska-Kalka Amelia, Wasiak Katarzyna, Ciunowicz Damian, Grot Dagmara, Glowacki Pawel, Stoczynska-Fidelus Ewelina, Rieske Piotr
Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Department of Tumor Biology, Chair of Medical Biology, Medical University of Lodz, Zeligowskiego 7/9 St, Lodz, 90- 752, Poland.
BMC Cancer. 2025 Jan 31;25(1):182. doi: 10.1186/s12885-025-13580-8.
Small-molecule compounds that even partially restore the GTPase activity of RAS can be used in anticancer therapy. Until now, attempts to obtain such compounds have failed. Compounds with this ability have been defined in our research.
The compounds were initially identified through virtual screening, and their optimal binding conformation in the RAS SW-II pocket was determined using the flexible docking technique. Efficacy was verified based on the IC50 determination, GTPase activity, as well as the AKT and ERK phospho WB assays.
The IC50 of the tested compounds was significantly lower against cells with the RAS mutation than against selected types of normal cells. The molecular mechanism of action of these compounds was proposed - minimization of the negative impact of the V12 sidechain on GTP hydrolysis of RAS. The work also indicates that the model of action of RAS mutants in cell lines is incomplete. The analysed cell line (SW-480) with RAS mutations does not always show increased ERK and AKT activity.
We have demonstrated molecules that partially restore the GTPase activity of RAS. Their mechanism of action is well explained based on current RAS mutant conformation and mechanistic models. These molecules inhibit the RAS-AKT pathway and show higher cytotoxicity against cancer cells with the RAS mutation (SW-480 cell line). However, SW-480 cells can switch into the subline proliferating independently of AKT phosphorylation and show partial resistance to the molecules described in this article.
即使能部分恢复RAS鸟苷三磷酸酶活性的小分子化合物也可用于抗癌治疗。到目前为止，获取此类化合物的尝试均告失败。我们的研究已确定了具有这种能力的化合物。
这些化合物最初通过虚拟筛选进行鉴定，并使用柔性对接技术确定它们在RAS SW-II口袋中的最佳结合构象。基于半数抑制浓度（IC50）测定、鸟苷三磷酸酶活性以及AKT和ERK磷酸化的蛋白质免疫印迹分析来验证疗效。
所测试化合物对具有RAS突变的细胞的IC50显著低于对选定类型正常细胞的IC50。提出了这些化合物的分子作用机制——将V12侧链对RAS鸟苷三磷酸水解的负面影响降至最低。这项研究还表明，RAS突变体在细胞系中的作用模式并不完整。分析的具有RAS突变的细胞系（SW-480）并不总是表现出ERK和AKT活性增加。
我们已证明了能部分恢复RAS鸟苷三磷酸酶活性的分子。基于当前RAS突变体构象和作用机制模型，很好地解释了它们的作用机制。这些分子抑制RAS-AKT途径，并且对具有RAS突变的癌细胞（SW-480细胞系）表现出更高的细胞毒性。然而，SW-480细胞可以转变为独立于AKT磷酸化而增殖的亚系，并对本文所述分子表现出部分抗性。
