Conjugation, Identification and Bionomics Analysis of EGFR Targeting Drug TGF α-Saporin as a Bioactive Stent Coating Material

Abstract

Objective: To identify and testify the cytotoxicity of the EGFR targeting drug TGFa-SAP conjugated and synthesized with N-succinimidyl-3 (2-pyridyldithio) propionate on human hepatoma cell line BEL-7404 cells and proliferating vascular smooth muscle cells. Methods: Conjugation of saporin with TGFa was accomplished after derivatization of TGFa and saporin with N-succinimidyl-3 (2-pyridyldithio) proprionate and the purification of the conjugate was achieved through Eppendorf Centrifugal Filter. Cytotoxicity assays were measured by MTS assays. The value of Thymidine incorporation in BEL-7404 cells was measured by 3H-thymidine uptake. Results: Cytotoxicity assays testified that TGFa-SAP conjugate could remarkably inhibit the proliferation of human hepatoma cell line BEL-7404 cells in vitro. The value of thymidine incorporation of BEL-7404 cells in TGFa-SAP groups significantly decreased compared with the control group (P<0.05), and it had dose-dependence on TGFa-SAP’s concentration. But Saporin could not affect BEL-7404 cells even at higher level (10-5). TGFa-SAP conjugate had effective cytotoxicity on proliferating vascular smooth muscle cells, also. Conclusion: The results indicated that the conjugated EGFR targeting drug TGFa-SAP had effective cytotoxicity not only on BEL-7404 cells, but also on proliferating vascular smooth muscle cells, as a potential bioactive stent coating material.