Discovery of novel wee1 inhibitors via structure-based virtual screening and biological evaluation
- 316 Downloads
Wee1 plays a critical role in the arrest of G2/M cell cycle for DNA repair before entering mitosis. Many cancer cells have been identified as overexpression of Wee1. In this research, pharmacophore modeling, molecular docking and molecular dynamics simulation approaches were constructed to identify novel potential Wee1 inhibitors. A compound 8 was found to have a novel skeleton against Wee1 with an IC50 value of 22.32 µM and a Ki value of 13.11 µM. Kinetic assays were employed to evaluate the compound 8 as a competitive inhibitor. Compound 8 was tested against A-549 tumor cell lines with IC50 value of 17.8 µM. To investigate the intermolecular interaction of Wee1 and compound 8, further molecular dynamics simulations were performed. It indicates that the binding mode of compound 8 and reference ligand is similar. The active core scaffold of compound 8 could represent a promising lead compound for studying Wee1 and be used for further structural optimization to design more potent Wee1 inhibitors.
KeywordsWee1 inhibitors Pharmacophore model Molecular docking Virtual screening Molecular dynamics simulation
This work was financially supported by CAS “Light of West China” Program (91 to Z.Z.), CAS Strategic biological resources service network (ZSTH-021), the Strategic Priority Research Program of the Chinese Academy of Sciences, Grant No. XDA12030206.
Compliance with ethical standards
Conflict of interest
The authors have declared no conflict of interest.
- 7.Mizuarai S, Yamanaka K, Itadani H, Arai T, Nishibata T, Hirai H, Kotani H (2009) Discovery of gene expression-based pharmacodynamic biomarker for a p53 context-specific anti-tumor drug Wee1 inhibitor. Mol Cancer 8(1):34. https://doi.org/10.1186/1476-4598-8-34 CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Van Linden AA, Baturin D, Ford JB, Fosmire SP, Gardner L, Korch C, Reigan P, Porter CC (2013) Inhibition of Wee1 sensitizes cancer cells to antimetabolite chemotherapeutics in vitro and in vivo, independent of p53 functionality. Mol Cancer Ther 12(12):2675–2684. https://doi.org/10.1158/1535-7163.MCT-13-0424 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Hashimoto O, Shinkawa M, Torimura T, Nakamura T, Selvendiran K, Sakamoto M, Koga H, Ueno T, Sata M (2006) Cell cycle regulation by the Wee1 inhibitor PD0166285, pyrido [2,3-d] pyimidine, in the B16 mouse melanoma cell line. BMC Cancer 6:292. https://doi.org/10.1186/1471-2407-6-292 CrossRefPubMedPubMedCentralGoogle Scholar
- 11.Bridges KA, Hirai H, Buser CA, Brooks C, Liu H, Buchholz TA, Molkentine JM, Mason KA, Meyn RE (2011) MK-1775, a novel Wee1 kinase inhibitor, radiosensitizes p53-defective human tumor cells. Clin Cancer Res 17(17):5638–5648. https://doi.org/10.1158/1078-0432.ccr-11-0650 CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Palmer BD, Thompson AM, Booth RJ, Dobrusin EM, Kraker AJ, Lee HH, Lunney EA, Mitchell LH, Ortwine DF, Smaill JB, Swan LM, Denny WA (2006) 4-Phenylpyrrolo[3,4-c]carbazole-1,3(2H,6H)-dione inhibitors of the checkpoint kinase Wee1. Structure-activity relationships for chromophore modification and phenyl ring substitution. J Med Chem 49(16):4896–4911. https://doi.org/10.1021/jm0512591 CrossRefPubMedGoogle Scholar
- 13.Pokorny JL, Calligaris D, Gupta SK, Iyekegbe DO Jr, Mueller D, Bakken KK, Carlson BL, Schroeder MA, Evans DL, Lou Z, Decker PA, Eckel-Passow JE, Pucci V, Ma B, Shumway SD, Elmquist WF, Agar NY, Sarkaria JN (2015) The efficacy of the wee1 inhibitor MK-1775 combined with temozolomide is limited by heterogeneous distribution across the blood-brain barrier in glioblastoma. Clin Cancer Res 21(8):1916–1924. https://doi.org/10.1158/1078-0432.ccr-14-2588 CrossRefPubMedPubMedCentralGoogle Scholar
- 14.Rajeshkumar NV, De Oliveira E, Ottenhof N, Watters J, Brooks D, Demuth T, Shumway SD, Mizuarai S, Hirai H, Maitra A, Hidalgo M (2011) MK-1775, a potent Wee1 inhibitor, synergizes with gemcitabine to achieve tumor regressions, selectively in p53-deficient pancreatic cancer xenografts. Clin Cancer Res 17(9):2799–2806. https://doi.org/10.1158/1078-0432.ccr-10-2580 CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Smaill JB, Baker EN, Booth RJ, Bridges AJ, Dickson JM, Dobrusin EM, Ivanovic I, Kraker AJ, Lee HH, Lunney EA, Ortwine DF, Palmer BD, Quin J 3rd, Squire CJ, Thompson AM, Denny WA (2008) Synthesis and structure-activity relationships of N-6 substituted analogues of 9-hydroxy-4-phenylpyrrolo[3,4-c]carbazole-1,3(2H,6H)-diones as inhibitors of Wee1 and Chk1 checkpoint kinases. Eur J Med Chem 43(6):1276–1296. https://doi.org/10.1016/j.ejmech.2007.07.016 CrossRefPubMedGoogle Scholar
- 21.Smaill JB, Lee HH, Palmer BD, Thompson AM, Squire CJ, Baker EN, Booth RJ, Kraker A, Hook K, Denny WA (2008) Synthesis and structure-activity relationships of soluble 8-substituted 4-(2-chlorophenyl)-9-hydroxypyrrolo[3,4-c]carbazole-1,3(2H,6H)-diones as inhibitors of the Wee1 and Chk1 checkpoint kinases. Bioorg Med Chem Lett 18(3):929–933. https://doi.org/10.1016/j.bmcl.2007.12.046 CrossRefPubMedGoogle Scholar
- 24.Thangapandian S, John S, Sakkiah S, Lee KW (2011) Potential virtual lead identification in the discovery of renin inhibitors: application of ligand and structure-based pharmacophore modeling approaches. Eur J Med Chem 46(6):2469–2476. https://doi.org/10.1016/j.ejmech.2011.03.035 CrossRefPubMedGoogle Scholar
- 27.Lagorce D, Maupetit J, Baell J, Sperandio O, Tuffery P, Miteva MA, Galons H, Villoutreix BO (2011) The FAF-Drugs2 server: a multistep engine to prepare electronic chemical compound collections. Bioinformatics 27(14):2018–2020. https://doi.org/10.1093/bioinformatics/btr333 CrossRefPubMedGoogle Scholar
- 35.Barltrop JA, Owen TC, Cory AH, Cory JG (1991) 5-(3-carboxymethoxyphenyl)-2-(4,5-dimethylthiazolyl)-3-(4-sulfophenyl)tetrazolium, inner salt (MTS) and related analogs of 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) reducing to purple water-soluble formazans As cell-viability indicato. Bioorg Med Chem Lett 1(11):611–614CrossRefGoogle Scholar
- 36.Riss TL, Moravec RA (1992) Comparison of MTT, XTT, and a novel tetrazolium compound MTS for in vitro proliferation and chemosensitivity assays. Mol Biol Cell 3(1):184–190Google Scholar
- 37.Mir SE, De Witt Hamer PC, Krawczyk PM, Balaj L, Claes A, Niers JM, Van Tilborg AA, Zwinderman AH, Geerts D, Kaspers GJ, Peter Vandertop W, Cloos J, Tannous BA, Wesseling P, Aten JA, Noske DP, Van Noorden CJ, Wurdinger T (2010) In silico analysis of kinase expression identifies WEE1 as a gatekeeper against mitotic catastrophe in glioblastoma. Cancer cell 18(3):244–257. https://doi.org/10.1016/j.ccr.2010.08.011 CrossRefPubMedPubMedCentralGoogle Scholar
- 40.Masaki T, Shiratori Y, Rengifo W, Igarashi K, Yamagata M, Kurokohchi K, Uchida N, Miyauchi Y, Yoshiji H, Watanabe S, Omata M, Kuriyama S (2003) Cyclins and cyclin-dependent kinases: comparative study of hepatocellular carcinoma versus cirrhosis. Hepatology 37(3):534–543. https://doi.org/10.1053/jhep.2003.50112 CrossRefPubMedGoogle Scholar