Department of Pharmacology & Healthy Aging Research Center, Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan City, Taiwan, Republic of China.
Department of Neurology, Chang Gung Memorial Hospital, Taoyuan City, Taiwan, Republic of China.
Thromb Haemost. 2018 Jan;118(1):90-102. doi: 10.1160/TH17-05-0330. Epub 2018 Jan 5.
Without conjunctive administration of an anticoagulant, endothelial injury-induced thrombosis is resistant to thrombolysis and prone to re-thrombosis. We hypothesized that co-delivery of recombinant tissue plasminogen activator (rtPA) with annexin V-containing anticoagulants that specifically target the injured endothelium may passivate the thrombogenic elements of the vascular injury site and enhance rtPA-induced thrombolysis. In this study, the effects of conjunctive administration of Kinexins (Kunitz inhibitor-annexin V fusion proteins) with rtPA on thrombolysis were determined and . Thromboelastometry showed that both TAP-A (tick anticoagulant peptide-annexin V fusion protein; an inhibitor of factor Xa [FXa] and prothrombinase) and A-6L15 (annexin V-6L15 fusion protein; an inhibitor of tissue factor/FVIIa) exerted concentration-dependent (10-100 nM) effects on clot formation, with TAP-A being several folds more potent than A-6L15 in whole blood. Combination of TAP-A or A-6L15 with rtPA (1 μg/mL) led to decrease in lysis index, suggesting conjunctive enhancement of thrombolysis by combined use of rtPA with TAP-A or A-6L15. In a rat cremaster muscle preparation subjected to photochemical injury, conjunctive administration of rtPA and TAP-A significantly restored tissue perfusion to 56%, which is approximately two fold of that by rtPA or TAP-A alone. Near-infrared fluorescence images demonstrated local retention of a fluorescent A-6L15-S288 at the injury site, suggesting a targeting effect of the fusion protein. Pharmacokinetic analysis showed that I-labelled TAP-A and A-6L15 had initial distribution half-lives (T) of approximately 6 minutes and elimination half-lives (T) of approximately 2.3 hours. In conclusion, Kinexins were potentially useful adjunctive agents with rtPA thrombolytic therapy especially for thrombosis induced by endothelial injury.
没有联合抗凝治疗，内皮损伤诱导的血栓对溶栓治疗有抵抗力，并且容易再次形成血栓。我们假设，将含有 Annexin V 的抗凝剂与组织型纤溶酶原激活剂（rtPA）共同给药，该抗凝剂专门针对受损的内皮细胞，可能使血管损伤部位的血栓形成因素失活，并增强 rtPA 诱导的溶栓作用。在这项研究中，确定了与 rtPA 共同给药 Kinexins（Kunitz 抑制剂- Annexin V 融合蛋白）对溶栓的影响。血栓弹性描记术表明，TAP-A（蜱抗凝肽- Annexin V 融合蛋白；因子 Xa [FXa] 和凝血酶原酶的抑制剂）和 A-6L15（ Annexin V-6L15 融合蛋白；组织因子/FVIIa 的抑制剂）均以浓度依赖性方式（10-100 nM）影响血栓形成，TAP-A 在全血中的作用比 A-6L15 强几倍。TAP-A 或 A-6L15 与 rtPA（1 μg/mL）联合使用会降低溶解指数，这表明通过将 rtPA 与 TAP-A 或 A-6L15 联合使用可增强溶栓作用。在光化学损伤的大鼠提睾肌制备中，rtPA 和 TAP-A 的联合给药可将组织灌注恢复到 56%，这大约是 rtPA 或 TAP-A 单独给药的两倍。近红外荧光图像显示，荧光 A-6L15-S288 局部保留在损伤部位，表明融合蛋白具有靶向作用。药代动力学分析表明，I 标记的 TAP-A 和 A-6L15 具有约 6 分钟的初始分布半衰期（T）和约 2.3 小时的消除半衰期（T）。总之，Kinexins 可能是 rtPA 溶栓治疗的有用辅助剂，尤其是对于内皮损伤引起的血栓形成。
