Computational and in vitro targeting of HUVECs by ARA-Linker-TGFαL3 through VEGFR2

Angiogenesis blockade represents a therapeutic strategy to inhibit the growth of the tumour and its progression and metastasis. Targeting the vascular endothelial growth factor receptor 2 (VEGFR2) is among the approaches approved as anti-cancer drugs. In this study, we examined ARA-linker-TGF alpha L3 and TGF alpha L3 binding affinity to VEGFR2. Next, the cytotoxicity effect of ARA-linker-TGF alpha L3 on human umbilical vein endothelial cells was evaluated. The 100 ns molecular dynamic simulation was employed to estimate the stability of the ligand-receptor interaction and the free energy decomposition was performed by molecular mechanics generalised Born surface area method. The MTT test evaluated the proliferation inhibition of ARA-linker-TGF alpha L3 and arazyme on HUVECs cells. Also, the change in VEGFR2 gene expression was investigated after treatment with recombinant proteins. Analysis of the trajectory using the root-mean-square deviation and fluctuation, radius of gyration, H-bond calculation, and binding free energy showed that ARA-linker-TGF alpha L3 formed the more stable complex with the D2 and D3 domains of VEGFR2 than TGF alpha L3. Meanwhile, ARA-linker-TGF alpha L3 revealed higher cytotoxicity and inhibition of VEGFR2 expression than arazyme. The results of the current study suggest the anti-angiogenesis effects of ARA-linker-TGF alpha L3 that can be useful in cancer therapy.