Anti-cancer Effect And Mechanism Of GZD2202 As A Novel Small Molecular Trk Inhibitor Against Neuroblastoma In Vitro And In Vivo

Background and objective:The tropomyocin receptor kinase?Trk?family is a kind of neurotrophin receptor,which includes Trk A,TrkB,and TrkC proteins.The binding of neurotrophins to Trk induces its dimerization,phosphorylation and activation of Trk and the downstream signaling pathways via PI3K/Akt/mTOR?PLC?and Ras/Raf/MEK/ERK,which induces cell growth,proliferation,invasion,metastasis,angiogenesis and drug resistence.Here we mainly investigated the anti-tumor effect of GZD2202 as a novel Trk inhibitor on neuroblastoma in vitro and in vivo.Methods:Firstly,we studied the kinase inhibitory effect of GZD2202 by Z'-LYTE^TMM kinase assay in vitro,and constructed TrkB stable expression cell line SH-SY5Y-TrkB.Then the inhibitory effect of GZD2202 on BDNF mediated cell proliferation,migration and invasion was evaluated using CCK-8,Wound healing and Transwell assay,respectively.We also made investigation of the underlying mechanism by Western Blot.Furthermore,we studied the anticancer effect of GZD2202 on SH-SY5Y-TrkB xenograft,and evaluated its pro-apoptosis effect on tumor tissues.Results:The results demonstrated that GZD2202 shows selective kinase inhibitory effect on TrkB,with an kinase inhibitory IC₅₀0 of 34.97±19.18 nM,and GZD2202 could dose-dependently inhibit BDNF mediated SH-SY5Y-TrkB cell proliferation,migration,invasion,phosphorylation of the downstream signaling pathways and upregulation of MMP2/9.It was also shown that GZD2202 exhibited about 36.1%growth inhibiton in xenograft mouse with SH-SY5Y-TrkB neuroblastoma cells,TUNEL assays in the tumor tissue showed that GZD2202 induced apoptosis in SH-SY5Y-TrkB xenograft model.Conclusions:These data collectively showed that GZD2202 as a novel Trk inhibitor,demonstrated promising anticancer effect on neuroblastoma in vitro and in vivo,and is potential to be a candidate drug for clinically TrkB high expression neuroblastoma patients.