新型TNF-α特异性affibody分子对川崎病冠脉内皮损伤的靶向治疗作用及机制研究

基于单抗的靶向治疗已在炎症性疾病治疗中取得了重大的研究进展，但仍存在毒副作用大、价格昂贵等问题限制了其广泛应用。亲和体(affibody)分子具备以高亲和力与靶分子结合的特性，同时因易于制备、穿透性强和免疫原性低等诸多优点，使其在炎症性疾病治疗方面比抗体更具优势。我们在前期研究中通过噬菌体展示技术，成功筛选出与TNFα高亲和力和特异性结合的affibody分子，且体外实验已初步探明其具有很好的中和毒力效果。本研究计划将白蛋白结合结构域ABD融合入优选的二价TNFα结合亲合体，设计为双特异性融合蛋白（ZTAT），以期能够提高靶向特性和半衰期获得更好的体内治疗效果。在评估确认了ZTAT蛋白的靶向结合能力、中和活力、免疫原性和药代学等特征后，进一步在川崎病体内外模型中调查ZTAT在川崎病血管损伤中的治疗作用及其可能的作用机制。为发展基于TNFα亲和体的炎症性疾病靶向治疗提供新途径，极具应用前景。

Targeted therapy based on monoclonal antibodies has made significant research progress in the treatment of inflammatory diseases, but there are still problems such as severe toxic side effects and high prices that limit its wide application.Affibody molecules, the next generation of antibodies, are small and highly robust proteins engineered to bind to specific target proteins with high affinity, and are therefore suitable for development as potential biopharmaceutical drugs for therapeutic applications with low immunogenicity and cost-efficient production.In our previous research, using phage display technology, we successfully screened and obtained several TNFα specific affibody molecules with high affinity and have preliminarily proved that ZTNF-α affibodies could well neutralize the TNFα induced cytotoxicity in vitro. This research aim to integrate the albumin binding domain ABD into the preferred bivalent TNFα binding affibody molecules (hereinafter denoted ZTAT) and designed as a bispecific fusion protein. After determining the binding ability, TNFα neutralizing capacity, immunogenicity and pharmacokinetics of the engineered ZTAT protein,we further investigate the therapeutic effect and potential mechanism of action of ZTAT protein in vitro and in vivo. We believe that TNFα-binding affibody molecules have great potential for utilisation and development as agents for targeted therapy of inflammatory disease .