Study Of Targeting Inhibitors Of CRM1 With Therapeutic Potential For Extranodal NK/T-cell Lymphoma

Chromosome maintenance protein 1(CRM1)is a nuclear exporter responsible for the active transport of proteins in cells.Studies have shown that abnormal activation of CRM1 plays an important role in tumorigenesis,anti-apoptosis and other aspect,and has become a potential target for the development of anti-tumor drugs.Extranodal NK/T cell lymphoma(ENKTL)is a rare and highly malignant non-Hodgkin's lymphoma.The pathogenic mechanism of ENKTL has been unclear.At present,although there are some treatment methods that can play a role,the prognosis is poor,and the treatment effect is not good.In this study,a novel CRM1 inhibitor LFS-829 was designed based on the molecular structure of LFS-01 by computer-assisted drug design(CADD)consisiting: ADME/T,covalent docking and molecular dynamics simulation.The target binding of LFS-829 with CRM1 was analyzed by MALDI-TOF mass spectrometry and SPR assay.The extranodal NK/T cell lymphoma cells were used as experimental subjects to explore the therapeutic potential and mechanism of the effect of LFS-829 on the proliferation of ENKTL.The safety of small molecule inhibitors to the host was evaluated by peripheral blood mononuclear cell(PBMC)toxicity test,platelet toxicity test and animal experiment.It provided a novel targeted therapy strategy for the treatment of ENKTL.The main conclusions are shown as follows:(1)A novel CRM1 inhibitor LFS-829 was designed based on the molecular structure of LFS-01 by computer-assisted drug design.It was demonstrated that LFS-829 was targeted to CRM1 by covalent binding to the active amino acid CYS of the CRM1 hydrophobic cleft by the means of covalent docking and molecular dynamics simulation.(2)X-ray crystal diffraction technique,mass spectrometry analysis and surface plasmon resonance(SPR)analysis coNFirmed that LFS-829 covalently binds to the active amino acid CYS of the CRM1 hydrophobic cleft to target CRM1.The affinity constant KD between small molecule compound LFS-829 and CRM1 is approximately 26.95 nM.293 T cells was used as the research object,the half-inhibitory concentration of LFS-829 on CRM1 nuclear export function was about 158.1 nM.(3)LFS-829 can effectively inhibit the cell proliferation activity of ENKTL cell lines such as SNK6 and HANK-1,and the half-inhibitory concentration(IC50)for 72 h is 366 nM and 158 nM,respectively.In SNK6 cells,LFS-829 inhibits CRM1-mediated nuclear export,allowing I?B? to accumulate in the nucleus,thereby inhibiting the NF-?B signal transduction pathway down-regulating the expression of anti-apoptotic proteins and promoting tumor cell apoptosis.In the PBMC toxicity test,the platelet toxicity test,and the animal's acute toxicity test,LFS-829 showed great safety to the host.