抗TNF-α抗ED-B双特异性抗体αEBTA的药效学和药代动力学研究
发布时间：2019-03-07 10:22
【摘要】：类风湿性关节炎(rheumatoid arthritis,RA)是一种以关节滑膜炎症为特征的自身免疫性疾病,致残性强,可导致不可修复的关节畸形、破坏及功能丧失。肿瘤坏死因子-α(TNF-α,Tumor Necrosis Factor-alpha)在RA病变的滑膜组织增生、自身免疫反应和炎症等生理病理过程中起着重要作用,处在细胞因子调节网络的顶点,是RA发病机制中处于关键位置的促炎性因子。TNF-α的拮抗剂已被证明是类风湿关节炎的有效治疗剂,但缺点是靶向性低,副作用多。 含额外结构域B的纤维连接蛋白(Extra-domain B of fibronectin, ED-B FN)在RA患者的关节炎滑膜中呈现特异性表达。因此,以ED-B FN单克隆抗体或抗体片段作为靶向载体,与TNF-α单克隆抗体或抗体片段连接,构建抗TNF-α抗ED-B FN的双特异性抗体(Bispecific antibody)αEBTA,在RA的靶向治疗中能降低或消除其对正常组织和关节的免疫抑制作用,能有效提高药物的靶向性和安全性。 本研究通过由基因工程方法构建的毕赤酵母菌,分泌表达双抗aEBTA,上清中双抗的浓度为3μg／mL,经过镍离子亲和层析后,浓度达到了0.3mg/mL。对其进行了体外药效分析,AIA(佐剂诱导型)模型小鼠药代动力学和药效学研究。并由结果分析出双抗aEBTA能对抗TNF-a的细胞毒作用；双抗在类风湿关节模型(AIA)小鼠炎症关节中呈现特异性聚集,浓度比在其他器官(肝、肾、脾、肺)有显著差异；药效学实验表明双抗对AIA小鼠有较好的治疗效果。 结论：双抗aEBTA对AIA小鼠炎症关节有靶向分布和较好的治疗效果,为以后相关药物的应用和开发打下了很好的基础。
[Abstract]:Rheumatoid arthritis (rheumatoid arthritis,RA) is an autoimmune disease characterized by synovial inflammation. It is highly disabled and can lead to irreparable deformities, destruction and loss of function. Tumor necrosis factor-伪 (TNF- 伪, Tumor Necrosis Factor-alpha) plays an important role in the physiological and pathological process of RA disease, such as synovial tissue proliferation, autoimmunity and inflammation, and is at the apex of cytokine regulation network. The antagonist of TNF- 伪 has been proved to be an effective therapeutic agent for rheumatoid arthritis (RA), but its disadvantage is low targeting and many side effects. Fibronectin (Extra-domain B of fibronectin, ED-B FN) containing extra domain B was specifically expressed in synovium of RA patients with arthritis. Therefore, using ED-B FN monoclonal antibody or antibody fragment as target vector, the bispecific antibody (Bispecific antibody) 伪 EBTA, against TNF- 伪 anti ED-B FN was constructed by ligating with (Bispecific antibody) 伪 monoclonal antibody or antibody fragment. The immunosuppressive effect on normal tissues and joints can be reduced or eliminated in the targeted therapy of RA, and the targeting and safety of the drug can be improved effectively. In this study, Pichia pastoris was constructed by genetic engineering method. The concentration of double antibody in the supernatant of double antibody aEBTA, was 3 渭 g / mL, and the concentration was 0.3 mg / mol 路L-1 after Ni ~ (2 +) affinity chromatography. Pharmacodynamics and pharmacodynamics of, AIA (adjuvant-induced) mice were studied in vitro. The results showed that biantibody aEBTA could antagonize the cytotoxic effect of TNF-a, and the biantibody showed specific aggregation in inflammatory joints of rheumatoid joint model (AIA) mice, and the concentration was significantly different compared with other organs (liver, kidney, spleen, lung). Pharmacodynamic experiments showed that the double antibody had a good therapeutic effect on AIA mice. Conclusion: biantibody aEBTA has targeted distribution and good therapeutic effect on inflammatory joints in AIA mice, which lays a good foundation for the application and development of related drugs in the future.
【学位授予单位】：湖北大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
本文编号：2436027
[Abstract]:Rheumatoid arthritis (rheumatoid arthritis,RA) is an autoimmune disease characterized by synovial inflammation. It is highly disabled and can lead to irreparable deformities, destruction and loss of function. Tumor necrosis factor-伪 (TNF- 伪, Tumor Necrosis Factor-alpha) plays an important role in the physiological and pathological process of RA disease, such as synovial tissue proliferation, autoimmunity and inflammation, and is at the apex of cytokine regulation network. The antagonist of TNF- 伪 has been proved to be an effective therapeutic agent for rheumatoid arthritis (RA), but its disadvantage is low targeting and many side effects. Fibronectin (Extra-domain B of fibronectin, ED-B FN) containing extra domain B was specifically expressed in synovium of RA patients with arthritis. Therefore, using ED-B FN monoclonal antibody or antibody fragment as target vector, the bispecific antibody (Bispecific antibody) 伪 EBTA, against TNF- 伪 anti ED-B FN was constructed by ligating with (Bispecific antibody) 伪 monoclonal antibody or antibody fragment. The immunosuppressive effect on normal tissues and joints can be reduced or eliminated in the targeted therapy of RA, and the targeting and safety of the drug can be improved effectively. In this study, Pichia pastoris was constructed by genetic engineering method. The concentration of double antibody in the supernatant of double antibody aEBTA, was 3 渭 g / mL, and the concentration was 0.3 mg / mol 路L-1 after Ni ~ (2 +) affinity chromatography. Pharmacodynamics and pharmacodynamics of, AIA (adjuvant-induced) mice were studied in vitro. The results showed that biantibody aEBTA could antagonize the cytotoxic effect of TNF-a, and the biantibody showed specific aggregation in inflammatory joints of rheumatoid joint model (AIA) mice, and the concentration was significantly different compared with other organs (liver, kidney, spleen, lung). Pharmacodynamic experiments showed that the double antibody had a good therapeutic effect on AIA mice. Conclusion: biantibody aEBTA has targeted distribution and good therapeutic effect on inflammatory joints in AIA mice, which lays a good foundation for the application and development of related drugs in the future.
【学位授予单位】：湖北大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
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相关期刊论文 前2条
1 郑立运;方先珍;;酵母表达系统最新研究进展[J];郑州牧业工程高等专科学校学报;2005年04期
2 张欣;张春明;;双特异性抗体及其在肿瘤治疗中的应用[J];生命的化学;2007年02期
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