Localization of Therapeutic Fab-CHP Conjugates to Sites of Denatured Collagen for the Treatment of Rheumatoid Arthritis.
Localization of Therapeutic Fab-CHP Conjugates to Sites of Denatured Collagen for the Treatment of Rheumatoid Arthritis.
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DOI：
10.1021/acs.bioconjchem.0c00324
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发表时间：
2020-08-19
影响因子：
4.7
通讯作者：
Owen, Shawn C.
中科院分区：
文献类型：
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作者：
Arlotta, Keith J.;San, Boi Hoa;Mu, Hong-Hua;Yu, S. Michael;Owen, Shawn C.
类风湿性关节炎（RA）是一种以滑膜慢性炎症和蛋白酶诱导的软骨降解为特征的自身免疫性疾病。目前RA的生物治疗主要通过中和促炎细胞因子TNFα可有效减轻症状;然而，持续、不加选择地过度抑制炎症因子可显著削弱宿主免疫系统，导致机会性感染并中断治疗。我们假设，通过与胶原杂交肽（CHP）结合，将抗TNF α治疗剂定位于关节炎关节处存在的变性胶原（dCol），将减少异位抗原结合并维持局部免疫抑制。我们分离了临床批准的抗TNF α治疗药物英夫利昔单抗（iFab）的抗原结合片段，并通过基于赖氨酸的结合与SMCC接头制备了iFab-CHP结合物。成功偶联后（经LC-MS证实），通过ELISA样试验表征iFab-CHP的结合亲和力，结果显示与英夫利西单抗的抗原结合相当，与CHP的dCol结合相当，并且具有同时结合dCol和TNFα的杂交能力。我们进一步证明了Fab-CHP在体内定位于高dCol区域，并在试点小鼠研究中通过组织学染色（番红-O和H&E）评估有希望的治疗功效。
Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation in synovial joints and protease-induced cartilage degradation. Current biologic treatments for RA can effectively reduce symptoms, primarily by neutralizing the proinflammatory cytokine TNFα; however, continued, indiscriminate overinhibition of inflammatory factors can significantly weaken the host immune system, leading to opportunistic infections and interrupting treatment. We hypothesize that localizing anti-TNFα therapeutics to denatured collagen (dCol) present at arthritic joints, via conjugation with collagen-hybridizing peptides (CHPs), will reduce off-site antigen binding and maintain local immunosuppression. We isolated the antigen-binding fragment of the clinically approved anti-TNFα therapeutic infliximab (iFab) and prepared iFab-CHP conjugates via lysine-based conjugation with an SMCC linker. After successful conjugation, confirmed by LC-MS, the binding affinity of iFab-CHP was characterized by ELISA-like assays, which showed comparable antigen binding relative to infliximab, comparable dCol binding relative to CHP, and the hybrid ability to bind both dCol and TNFα simultaneously. We further demonstrated localization of Fab-CHP to areas of high dCol in vivo and promising therapeutic efficacy, assessed by histological staining (Safranin-O and H&E), in a pilot mouse study.
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