针对人血管性血友病因子 A3 结构域的嵌合单克隆抗体 MHCSZ-123 抑制恒河猴模型高剪切动脉血栓形成。

The chimeric monoclonal antibody MHCSZ-123 against human von Willebrand factor A3 domain inhibits high-shear arterial thrombosis in a Rhesus monkey model.

机构信息

The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, MOH Key Laboratory of Thrombosis and Hemostasis, Collaborative Innovation Center of Hematology, 188 Shizi Street, Suzhou, Jiangsu, 215006, China.

MOE Key Laboratory of Industrial Biotechnology, School of Biotechnology of Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu, 214122, China.

出版信息

J Hematol Oncol. 2017 May 19;10(1):111. doi: 10.1186/s13045-017-0475-2.

DOI:10.1186/s13045-017-0475-2

PMID:28526067

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5438484/

Abstract

BACKGROUND

SZ-123, a murine monoclonal antibody that targets the human von Willebrand factor (VWF) A3 domain and blocks the binding of collagen, is a powerful antithrombotic. In a Rhesus monkey model of thrombosis, SZ-123 had no side effects, such as bleeding or thrombocytopenia.

METHODS

The mouse/human chimeric version of SZ-123, MHCSZ-123, was developed and maintained inhibitory capacities in vitro and ex vivo after injection into monkeys. CHO-S cells were selected for stable expression of MHCSZ-123. Cell clones with high levels of MHCSZ-123 expression were screened with G418 then adapted to serum-free suspension culture. The antithrombotic effect of MHCSZ-123 on acute platelet-mediated thrombosis was studied in monkeys where thrombus formation was induced by injury and stenosis of the femoral artery, which allowed for cyclic flow reductions (CFRs). CFRs were measured in the femoral artery of anesthetized Rhesus monkeys before and after intravenous administration of MHCSZ-123. Ex vivo VWF binding to collagen, platelet aggregation, platelet counts, and template bleeding time were used as measurements of antithrombotic activity. In addition, plasma VWF and VWF occupancy were measured by ELISA.

RESULTS

Injection of 0.1, 0.3, and 0.6 mg/kg MHCSZ-123 significantly reduced CFRs by 29.4%, 57.9%, and 73.1%, respectively. When 0.3 and 0.6 mg/kg MHCSZ-123 were administered, 46.6%-65.8% inhibition of ristocetin-induced platelet aggregation was observed between 15 and 30 min after injection. We observed minimal effects on bleeding time, minimal blood loss, and no spontaneous bleeding or thrombocytopenia.

CONCLUSIONS

The VWF-A3 inhibitor MHCSZ-123 significantly reduced thrombosis in Rhesus monkeys and appeared to be safe and well tolerated.

摘要

背景

SZ-123 是一种靶向人血管性血友病因子（VWF）A3 结构域并阻断胶原蛋白结合的鼠源单克隆抗体，是一种强大的抗血栓药物。在恒河猴血栓模型中，SZ-123 无副作用，如出血或血小板减少。

方法

SZ-123 的鼠/人嵌合版本 MHCSZ-123 被开发出来，并在注射到猴子体内后保持体外和体外的抑制能力。CHO-S 细胞被选择用于稳定表达 MHCSZ-123。用 G418 筛选出高表达 MHCSZ-123 的细胞克隆，然后适应无血清悬浮培养。通过损伤和股动脉狭窄诱导血栓形成，允许循环流量减少（CFR），在接受麻醉的恒河猴中研究 MHCSZ-123 对急性血小板介导的血栓形成的抗血栓作用。在静脉注射 MHCSZ-123 前后测量股动脉的 CFR。VWF 与胶原蛋白的体外结合、血小板聚集、血小板计数和模板出血时间被用作抗血栓活性的测量。此外，通过 ELISA 测量血浆 VWF 和 VWF 占有率。

结果

注射 0.1、0.3 和 0.6mg/kg MHCSZ-123 分别使 CFR 降低 29.4%、57.9%和 73.1%。当给予 0.3 和 0.6mg/kg MHCSZ-123 时，在注射后 15 至 30 分钟观察到 46.6%-65.8%抑制瑞斯托菌素诱导的血小板聚集。我们观察到对出血时间的最小影响、最小的血液丢失、没有自发性出血或血小板减少。