Structure-based design and biological evaluation of novel mTOR inhibitors as potential anti-cervical agents

doi:10.5246/jcps.2020.09.056

Abstract

Abstract:

The mammalian target of rapamycin (mTOR) is a critical component of the PI3K-AKT signaling pathway. It is highly activated in cervical cancer, which continues to pose an important clinical challenge with an urgent need for new and improved therapeutic approaches. Herein, we describe the structure-based drug discovery and biological evaluation of a series of mTOR kinase inhibitors as potential anti-cervical cancer agents. The results of enzymatic activity assays supported C3 as a potential mTOR inhibitor, which exhibited high inhibitory activity with an IC₅₀ of 1.57 μM. Molecular docking and dynamics simulation were conducted to predict the binding patterns, suggesting relationships between structure and activity. The anti-proliferative assay against diverse cancer cell lines was displayed subsequently, revealing that C3 exhibited significant proliferation inhibition against cervical cancer cell HeLa (IC₅₀ = 0.38 μM) compared with other cell lines. Moreover, C3 could effectively reduce the expression of phospho-ribosomal S6 protein (p-S6) in HeLa cells in a dose-dependent manner. Noteworthily, mTOR signaling and other cellular pathways might contribute to the significant effect of C3 against cervical cancer simultaneously. These data indicated that C3 represented a good lead molecule for further development as a therapeutic agent for cervical cancer treatment.

Key words: Structure-based drug discovery, mTOR inhibitors, Molecular modeling, Cellular pathway, Cervical cancer

CLC Number:

R916.1

Supporting:

Peili Jiao, Yiyan Li, Xing Wu, Yuxi Wang, Beibei Mao, Hongwei Jin, Lihe Zhang, Liangren Zhang, Zhenming Liu. Structure-based design and biological evaluation of novel mTOR inhibitors as potential anti-cervical agents[J]. Journal of Chinese Pharmaceutical Sciences, 2020, 29(9): 603-616.

References

[1] Laplante, M.; Sabatini, D.M. mTOR signaling in growth control and disease. Cell. 2012, 149, 274-293.

[2] Keith, C.T.; Schreiber, S.L. PIK-related kinases: DNA repair, recombination, and cell cycle checkpoints. Science. 1995, 270, 50-51.

[3] Guertin, D.A.; Sabatini, D.M. Defining the role of mTOR in cancer. Cancer Cell. 2007, 12, 9-22.

[4] Marosvári, D.; Nagy, N.; Kriston, C.; Deák, B.; Hajdu, M.; Bödör, C.; Csala, I.; Bagó, A.G.; Szállási, Z.; Sebestyén, A.; Reiniger, L. Discrepancy between low levels of mTOR activity and high levels of P-S6 in primary central nervous system lymphoma may be explained by PAS domain-containing serine/threonine-protein kinase-mediated phosphorylation. J. Neuropathol. Exp. Neurol. 2018, 77, 268-273.

[5] Tavares, M.K.; Dos Reis, S.; Platt, N.; Heinrich, I.A.; Wolin, I.A.V.; Leal, R.B.; Kaster, M.P.; Rodrigues, A.L.S.; Freitas, A.E. Agmatine potentiates neuroprotective effects of subthreshold concentrations of ketamine via mTOR/S6 kinase signaling pathway. Neurochem. Int. 2018, 118, 275-285.

[6] Bauriedel, G.; Jabs, A.; Kraemer, S.; Nickenig, G.; Skowasch, D. Neointimal expression of rapamycin receptor FK506-binding protein FKBP12: postinjury animal and human in-stent restenosis tissue characteristics. J. Vasc. Res. 2008, 45, 173-178.

[7] Yang, H.J.; Rudge, D.G.; Koos, J.D.; Vaidialingam, B.; Yang, H.J.; Pavletich, N.P. mTOR kinase structure, mechanism and regulation. Nature. 2013, 497, 217-223.

[8] Choi, J.; Chen, J.; Schreiber, S.L.; Clardy, J. Structure of the FKBP12-rapamycin complex interacting with the binding domain of human FRAP. Science. 1996, 273, 239-242.

[9] Li, S.Y.; Liang, Y.; Wu, M.L.; Wang, X.J.; Fu, H.X.; Chen, Y.H.; Wang, Z.G. The novel mTOR inhibitor CCI-779 (temsirolimus) induces antiproliferative effects through inhibition of mTOR in Bel-7402 liver cancer cells. Cancer Cell Int. 2013, 13, 30.

[10] Liu, H.Y.; Zang, C.B.; Schefe, J.H.; Schwarzlose-Schwarck, S.; Regierer, A.C.; Elstner, E.; Schulz, C.O.; Scholz, C.; Possinger, K.; Eucker, J. The mTOR inhibitor RAD001 sensitizes tumor cells to the cytotoxic effect of carboplatin in breast cancer in vitro. Anticancer Res. 2011, 31, 2713-2722.

[11] Rivera, V.M.; Squillace, R.M.; Miller, D.; Berk, L.; Wardwell, S.D.; Ning, Y.Y.; Pollock, R.; Narasimhan, N.I.; Iuliucci, J.D.; Wang, F.; Clackson, T. Ridaforolimus (AP23573; MK-8669), a potent mTOR inhibitor, has broad antitumor activity and can be optimally administered using intermittent dosing regimens. Mol. Cancer Ther. 2011, 10, 1059-1071.

[12] Chresta, C.M.; Davies, B.R.; Hickson, I.; Harding, T.; Cosulich, S.; Critchlow, S.E.; Vincent, J.P.; Ellston, R.; Jones, D.; Sini, P.; James, D.; Howard, Z.; Dudley, P.; Hughes, G.; Smith, L.; Maguire, S.; Hummersone, M.; Malagu, K.; Menear, K.; Jenkins, R.; Jacobsen, M.; Smith, G.C.M.; Guichard, S.; Pass, M. AZD8055 is a potent, selective, and orally bioavailable ATP-competitive mammalian target of rapamycin kinase inhibitor with in vitro and in vivo antitumor activity. Cancer Res. 2010, 70, 288-298.

[13] Tian, T.; Li, X.Y.; Zhang, J.H. mTOR signaling in cancer and mTOR inhibitors in solid tumor targeting therapy. Int. J. Mol. Sci. 2019, 20, 755.

[14] Rodrik-Outmezguine, V.S.; Okaniwa, M.; Yao, Z.; Novotny, C.J.; McWhirter, C.; Banaji, A.; Won, H.; Wong, W.; Berger, M.; de Stanchina, E.; Barratt, D.G.; Cosulich, S.; Klinowska, T.; Rosen, N.; Shokat, K.M. Overcoming mTOR resistance mutations with a new-generation mTOR inhibitor. Nature. 2016, 534, 272-276.

[15] Bray, F.; Ferlay, J.; Soerjomataram, I.; Siegel, R.L.; Torre, L.A.; Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 2018, 68, 394-424.

[16] Yuan, L.Y.; Qin, X.M.; Li, L.; Zhou, J.T.; Zhou, M.; Li, X.X.; Xu, Y.; Cheng, L.; Xing, H. Overexpression of LINC00037 represses cervical cancer progression by activating mTOR signaling pathway. J. Cell Physiol. 2019, 234, 13353-13360.

[17] Klempner, S.J.; Myers, A.P.; Cantley, L.C. What a tangled web we weave: emerging resistance mechanisms to inhibition of the phosphoinositide 3-kinase pathway. Cancer Discov. 2013, 3, 1345-1354.

[18] Xia, J.; Jin, H.W.; Liu, Z.M.; Zhang, L.R.; Wang, X.S. An unbiased method to build benchmarking sets for ligand-based virtual screening and its application to GPCRs. J. Chem. Inf. Model. 2014, 54, 1433-1450.

[19] Chiem, K.; Jani, S.; Fuentes, B.; Lin, D.L.; Rasche, M.E.; Tolmasky, M.E. Identification of an inhibitor of the aminoglycoside 6'-N-acetyltransferase type ib [AAC(6')-ib] by glide molecular docking. Medchemcomm. 2016, 7, 184-189.

[20] Xia, J.; Hu, H.B.; Xue, W.J.; Wang, X.S.; Wu, S. The discovery of novel HDAC3 inhibitors via virtual screening and in vitro bioassay. J. Enzyme. Inhib. Med. Chem. 2018, 33, 525-535.

[21] Sattarinezhad, E.; Bordbar, A.K.; Fani, N. Virtual screening of Piperine analogs as Survivin inhibitors and their molecular interaction analysis by using consensus docking, MD simulation, MMPB/GBSA and alanine scanning techniques. J. Biomol. Struct. Dyn. 2017, 35, 1824-1832.

[22] Śledź, P.; Caflisch, A. Protein structure-based drug design: from docking to molecular dynamics. Curr. Opin. Struct. Biol. 2018, 48, 93-102.

[23] Ward, R.A.; Bethel, P.; Cook, C.; Davies, E.; Debreczeni, J.E.; Fairley, G.; Feron, L.; Flemington, V.; Graham, M.A.; Greenwood, R.; Griffin, N.; Hanson, L.; Hopcroft, P.; Howard, T.D.; Hudson, J.; James, M.; Jones, C.D.; Jones, C.R.; Lamont, S.; Lewis, R.; Lindsay, N.; Roberts, K.; Simpson, I.; St-Gallay, S.; Swallow, S.; Tang, J.; Tonge, M.; Wang, Z.H.; Zhai, B.C. Structure-guided discovery of potent and selective inhibitors of ERK1/2 from a modestly active and promiscuous chemical start point. J. Med. Chem. 2017, 60, 3438-3450.

[24] Aliwaini, S.; Awadallah, A.M.; Morjan, R.Y.; Ghunaim, M.; Alqaddi, H.; Abuhamad, A.Y.; Awadallah, E.A.; Abughefra, Y.M. Novel imidazo[1,2-a]pyridine inhibits AKT/mTOR pathway and induces cell cycle arrest and apoptosis in melanoma and cervical cancer cells. Oncol. Lett. 2019, 18, 830-837.

[25] Vichai, V.; Kirtikara, K. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat. Protoc. 2006, 1, 1112-1116.

[26] Wang, X.M.; Xin, M.H.; Xu, J.; Kang, B.R.; Li, Y.; Lu, S.M.; Zhang, S.Q. Synthesis and antitumor activities evaluation of m-(4-morpholinoquinazolin-2-yl)benzamides in vitro and in vivo. Eur. J. Med. Chem. 2015, 96, 382-395.