First molecules to reactivate RAS GTPase activity.
Aneta Wlodarczyk, Cezary Treda, Marcin Pacholczyk, Adrianna Rutkowska, Marta Wegierska, Amelia Kierasinska-Kalka, Katarzyna Wasiak, Damian Ciunowicz, Dagmara Grot, Pawel Glowacki, Ewelina Stoczynska-Fidelus, Piotr Rieske
Author Information
Aneta Wlodarczyk: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland. aneta.wlodarczyk@umed.lodz.pl.
Cezary Treda: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Marcin Pacholczyk: Department of Tumor Biology, Chair of Medical Biology, Medical University of Lodz, Zeligowskiego 7/9 St, Lodz, 90- 752, Poland.
Adrianna Rutkowska: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Marta Wegierska: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Amelia Kierasinska-Kalka: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Katarzyna Wasiak: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Damian Ciunowicz: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Dagmara Grot: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Pawel Glowacki: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Ewelina Stoczynska-Fidelus: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
Piotr Rieske: Department of Research and Development, Personather, LTD, Inwestycyjna 7 St, Konstantynow Lodzki, 95-050, Poland.
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
