Screening of a novel CRM1 targeting inhibitor and its effects on the proliferation and growth of extranodal NK/T cell lymphoma

LIU Meishuo; WANG Haina; LONG Feida; LENG Nan; YANG Yongliang

doi:10.11665/j.issn.1000-5048.20200108

Journal of China Pharmaceutical University > 2020 > 51(1): 44-51. > DOI: 10.11665/j.issn.1000-5048.20200108

LIU Meishuo, WANG Haina, LONG Feida, LENG Nan, YANG Yongliang. Screening of a novel CRM1 targeting inhibitor and its effects on the proliferation and growth of extranodal NK/T cell lymphoma[J]. Journal of China Pharmaceutical University, 2020, 51(1): 44-51. DOI: 10.11665/j.issn.1000-5048.20200108

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Screening of a novel CRM1 targeting inhibitor and its effects on the proliferation and growth of extranodal NK/T cell lymphoma

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The purpose of this study was to screen out the novel chromosome maintenance protein 1(CRM1)covalent targeting inhibitors by computer-assisted drug design(CADD), and to study their effects on the proliferation of extranodal nature killer/T cell lymphoma(ENKTL). A novel CRM1 inhibitor LFS-829 was designed based on the molecular structure of LFS-01 by means of ADME/T and covalent docking. The target binding of LFS-829 with CRM1 was analyzed by MALDI-TOF mass spectrometry. The effects of LFS-829 on the proliferation of extranodal NK/T cell lymphoma SNK6 and HANK-1 cells were detected by CCK-8. The cell morphology was observed by live cell workstation. Immunofluorescence experiments were used to analyze the effect of LFS-829 on nuclear export function of CRM1. The changes of NF-κB signaling pathway under different concentrations of LFS-829 were analyzed by Western blot, dual luciferase reporter gene assay and enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry, and the expression of proteins related to apoptosis pathway was detected by Western blot. Tests of peripheral blood mononuclear lymphocyte(PBMC)toxicity, platelet toxicity and mouse acute toxicity were done to make sure that it is not harmful to human. LFS-829 could bind covalently to the cysteine residue of the hydrophobic active pocket of CRM1. LFS-829 could selectively kill SNK6 and HANK-1 cells, with IC₅₀ of 366 nmol/L and 158 nmol/L in 72 h, respectively, and cell morphology was significantly changed. LFS-829 at 800 nmol/L significantly inhibited the nuclear export function of CRM1, promoted nuclear assembly of IκB-α, down-regulated the transcriptional activity of NF-κB signaling pathway, significantly up-regulated the expression of apoptotic pathway protein p53, cleaved Caspase 3 and cleaved Caspase 9 and induced apoptosis, with no obvious killing effect on PBMC or platelets. It did not cause substantial tissue damage to mice at the high dose of 300 mg/kg, which shows its great prospect of future application.

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[1]	Yamaguchi M,Suzuki R,Miyazaki K,et al.Improved prognosis of extranodal NK/T cell lymphoma,nasal type of nasal origin but not extranasal origin[J].Ann Hematol,2019,98(7):1647-1655.
[2]	Yong WB,Zheng W,Zhu J,et al.L-asparaginase in the treatment of refractory and relapsed extranodal NK/T-cell lymphoma,nasal type[J].Ann Hematol,2009,88(7):647-652.
[3]	Wen HJ,Ma HJ,Cai QC,et al.Recurrent ECSIT mutation encoding V140A triggers hyperinflammation and promotes hemophagocytic syndrome in extranodal NK/T cell lymphoma[J].Nat Med,2018,24(2):154-164.
[4]	Wu SJ,Zhang KR,Qin HQ,et al.Caffeic acid phenethyl ester(CAPE)revisited:covalent modulation of XPO1/CRM1 activities and implication for its mechanism of action[J].Chem Biol Drug Des,2017,89(5):655-662.
[5]	Niu MS,Wu SJ,Mao L,et al.CRM1 is a cellular target of curcumin:new insights for the myriad of biological effects of an ancient spice[J].Traffic,2013,14(10):1042-1052.
[6]	Mao L,Yang YL.Targeting the nuclear transport machinery by rational drug design[J].Curr Pharm Des,2013,19(12):2318-2325.
[7]	Wang HN,Wang FQ,Wu SJ,et al.Traditional herbal medicine-derived sulforaphene promotes mitophagic cell death in lymphoma cells through CRM1-mediated p62/SQSTM1 accumulation and AMPK activation[J].Chem Biol Interact,2018,281:11-23.
[8]	Liu Y,Zhao L,Li WT,et al.FIPSDock:a new molecular docking technique driven by fully informed swarm optimization algorithm[J].J Comput Chem,2013,34(1):67-75.
[9]	Sun QX,Carrasco YP,Hu YC,et al.Nuclear export inhibition through covalent conjugation and hydrolysis of Leptomycin B by CRM1[J].Proc Natl Acad Sci U S A,2013,110(4):1303-1308.
[10]	Tai YT,Landesman Y,Acharya C,et al.CRM1 inhibition induces tumor cell cytotoxicity and impairs osteoclastogenesis in multiple myeloma:molecular mechanisms and therapeutic implications[J].Leukemia,2014,28(1):155-165.
[11]	Kazim S,Malafa MP,Coppola D,et al.Selective nuclear export inhibitor KPT-330 enhances the antitumor activity of gemcitabine in human pancreatic cancer[J].Mol Cancer Ther,2015,14(7):1570-1581.
[12]	Kau TR,Schroeder F,Ramaswamy S,et al.A chemical genetic screen identifies inhibitors of regulated nuclear export of a Forkhead transcription factor in PTEN-deficient tumor cells[J].Cancer Cell,2003,4(6):463-476.
[13]	Wang X,Zhang JL,Kim HP,et al.Bcl-XL disrupts death-inducing signal complex formation in plasma membrane induced by hypoxia/reoxygenation[J].FASEB J,2004,18(15):1826-1833.
[14]	Yu Y,Wang DP,Kaosaard K,et al.C-Rel is an essential transcription factor for the development of acute graft-versus-host disease in mice[J].Eur J Immunol,2013,43(9):2327-2337.

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Screening of a novel CRM1 targeting inhibitor and its effects on the proliferation and growth of extranodal NK/T cell lymphoma

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