The humanized platelet glycoprotein VI Fab inhibitor EMA601 protects from arterial thrombosis and ischaemic stroke in mice.

IF 8.9 2区环境科学与生态学 Q1 ENGINEERING, ENVIRONMENTAL

Environmental Science & Technology Letters Environ. Pub Date : 2024-11-14 DOI:10.1093/eurheartj/ehae482

Stefano Navarro, Ivan Talucci, Vanessa Göb, Stefanie Hartmann, Sarah Beck, Valerie Orth, Guido Stoll, Hans M Maric, David Stegner, Bernhard Nieswandt

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引用次数: 0

Abstract

Background and aims: Glycoprotein VI (GPVI) is a platelet collagen/fibrin(ogen) receptor and an emerging pharmacological target for the treatment of thrombotic and thrombo-inflammatory diseases, notably ischaemic stroke. A first anti-human GPVI (hGPVI) antibody Fab-fragment (ACT017/glenzocimab, KD: 4.1 nM) recently passed a clinical phase 1b/2a study in patients with acute ischaemic stroke and was found to be well tolerated, safe, and potentially beneficial. In this study, a novel humanized anti-GPVI antibody Fab-fragment (EMA601; KD: 0.195 nM) was developed that inhibits hGPVI function with very high potency in vitro and in vivo.

Methods: Fab-fragments of the mouse anti-hGPVI IgG Emf6.1 were tested for functional GPVI inhibition in human platelets and in hGPVI expressing (hGP6tg/tg) mouse platelets. The in vivo effect of Emf6.1Fab was assessed in a tail bleeding assay, an arterial thrombosis model and the transient middle cerebral artery occlusion (tMCAO) model of ischaemic stroke. Using complementary-determining region grafting, a humanized version of Emf6.1Fab (EMA601) was generated. Emf6.1Fab/EMA601 interaction with hGPVI was mapped in array format and kinetics and quantified by bio-layer interferometry.

Results: Emf6.1Fab (KD: 0.427 nM) blocked GPVI function in human and hGP6tg/tg mouse platelets in multiple assays in vitro at concentrations ≥5 µg/mL. Emf6.1Fab (4 mg/kg)-treated hGP6tg/tg mice showed potent hGPVI inhibition ex vivo and were profoundly protected from arterial thrombosis as well as from cerebral infarct growth after tMCAO, whereas tail-bleeding times remained unaffected. Emf6.1Fab binds to a so far undescribed membrane proximal epitope in GPVI. The humanized variant EMA601 displayed further increased affinity for hGPVI (KD: 0.195 nM) and fully inhibited the receptor at 0.5 µg/mL, corresponding to a >50-fold potency compared with ACT017.

Conclusions: EMA601 is a conceptually novel and promising anti-platelet agent to efficiently prevent or treat arterial thrombosis and thrombo-inflammatory pathologies in humans at risk.

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人源化血小板糖蛋白 VI Fab 抑制剂 EMA601 可防止小鼠动脉血栓形成和缺血性中风。

背景和目的：糖蛋白Ⅵ（GPVI）是血小板胶原蛋白/纤维蛋白（ogen）受体，也是治疗血栓性和血栓炎症性疾病（尤其是缺血性中风）的新兴药物靶点。首个抗人 GPVI（hGPVI）抗体 Fab 片段（ACT017/glenzocimab，KD：4.1 nM）最近通过了急性缺血性中风患者的 1b/2a 期临床研究，结果表明该抗体耐受性良好、安全且可能有益。本研究开发了一种新型人源化抗 GPVI 抗体 Fab 片段（EMA601；KD：0.195 nM），可在体外和体内以极高的效力抑制 hGPVI 功能：方法：测试了小鼠抗 hGPVI IgG Emf6.1 的 Fab 片段对人类血小板和表达 hGPVI 的（hGP6tg/tg）小鼠血小板中 GPVI 功能的抑制作用。在尾部出血试验、动脉血栓形成模型和缺血性中风的短暂性大脑中动脉闭塞（tMCAO）模型中评估了 Emf6.1Fab 的体内效应。通过互补决定区移植，生成了人源化版本的 Emf6.1Fab（EMA601）。以阵列形式绘制了 Emf6.1Fab/EMA601 与 hGPVI 的相互作用图，并通过生物层干涉测量法对动力学进行了量化：结果：Emf6.1Fab（KD：0.427 nM）在浓度≥5 µg/mL的多种体外试验中阻断了人类和 hGP6tg/tg 小鼠血小板中 GPVI 的功能。经 Emf6.1Fab（4 毫克/千克）处理的 hGP6tg/tg 小鼠在体外表现出了强效的 hGPVI 抑制作用，并在 tMCAO 后对动脉血栓形成和脑梗塞生长起到了极大的保护作用，而尾部出血时间则不受影响。Emf6.1Fab与GPVI中迄今尚未描述的膜近端表位结合。人源化变体 EMA601 对 hGPVI 的亲和力进一步提高（KD：0.195 nM），在 0.5 µg/mL 浓度下完全抑制受体，与 ACT017 相比药效提高了 50 倍以上：EMA601是一种概念新颖、前景广阔的抗血小板药物，可有效预防或治疗高危人群的动脉血栓形成和血栓炎症病变。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Environmental Science & Technology Letters Environ. ENGINEERING, ENVIRONMENTALENVIRONMENTAL SC-ENVIRONMENTAL SCIENCES

CiteScore

17.90

自引率

3.70%

发文量

163

期刊介绍： Environmental Science & Technology Letters serves as an international forum for brief communications on experimental or theoretical results of exceptional timeliness in all aspects of environmental science, both pure and applied. Published as soon as accepted, these communications are summarized in monthly issues. Additionally, the journal features short reviews on emerging topics in environmental science and technology.