Pharmacodynamic And Pharmacokinetic Study Of Anti TNF-α×anti ED-B Bispecific Antibody α EBTA

Rheumatoid arthritis is an autoimmune disease featuring arthrosynovitis and also a highly disabling disease which can lead to irreversible joint destruction, joint malformation and joint dysfunction. Tumor necrosis factor-alpha, TNF-a is involved in synovial tissues hyperplasia of RA,and also autoimmune responses、inflammation and many other pathophysiological processes. It is located in vertex of cytokine regulating networks and acts as pro-inflammatory factor playing a crucial role in RA pathogenesis. It has been proved that the antagonist of TNF-a has been to be an effective therapeutic agent in RA, nevertheless, its disadvantage lies in undesirable success rate of targeting and large number of side-effects.Extra-domain B of fibronectin, ED-B FN has demonstrated specific expression in RA patients’arthrosynovitis. As a consequence, the antibody with effects of both TNF-αand ED-B FN which is constructed by connecting the targeting vector which is formed by ED-B FN monoclonal antibody or antibody fragment to TNF-a monoclonal antibody or antibody fragment is in a position to reduce or remove its immunosuppressive action to normal structures and joints in targeted therapy, improving pharmaceutical targeting and safety.By virtue of pichia pastoris constructed by genetic engineering, concentration of bifunctional antibody in the supernatant was3μg/mL, after nickel ion affinity chromatography, concentration is0.3mg/mL.The study has performed the secretory expression bispecific antibodyaEBTA and conducted pharmaceutical analysis in vitro; carried out an investigation into AIA model mouse pharmacokinetics and pharmacodynamics. The results have shown thataEBTA is able to withstand cytotoxic effect of TNF-a;aEBTA has manifested specificity aggregation in AIA mouse arthritis with distinctive difference in terms of density compared to other organs (liver, kidney, spleen and lungs); that pharmacology experiment has indicated bispecific antibodyaEBTA has agreeable therapeutic effect on AIA mouse. To sum up, bispecific antibodyaEBTA has taken on admirable therapeutic effect in targeting and treating AIA mouse arthritis, laying a sound foundation for future application and development of related medicines.