Novel aptamer to von Willebrand factor A1 domain (TAGX-0004) shows total inhibition of thrombus formation superior to ARC1779 and comparable to caplacizumab.
Novel aptamer to von Willebrand factor A1 domain (TAGX-0004) shows total inhibition of thrombus formation superior to ARC1779 and comparable to caplacizumab.
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DOI：
10.3324/haematol.2019.235549
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发表时间：
2020-11-01
期刊：
影响因子：
10.1
通讯作者：
Matsumoto M
中科院分区：
文献类型：
--
作者：
Sakai K;Someya T;Harada K;Yagi H;Matsui T;Matsumoto M
血管性血友病因子（VWF）是一种血液糖蛋白，通过其A1结构域与血小板糖蛋白Ib之间的相互作用在血小板血栓形成中起重要作用。ARC 1779是一种VWF A1结构域的适体，在获得性血栓性血小板减少性紫癜（aTTP）的临床试验中进行了评估。随后，caplacizumab（一种抗VWF A1结构域纳米抗体）在欧洲和美国获批用于aTTP。我们最近开发了一种新的DNA适体，TAGX-0004，VWF A1结构域，它包含一个人工基地，并表现出高亲和力的VWF。为了比较这三种药物对VWF A1的影响，分析了它们在静态条件下抑制瑞斯托霉素或肉毒杆菌素诱导的血小板聚集的能力，并在微芯片流动室系统中研究了在高剪切应力下抑制血栓形成的能力。在两项试验中，TAGX-0004显示出比ARC 1779更强的抑制作用，并且与caplacizumab具有相当的抑制作用。使用VWF A1结构域的丙氨酸扫描诱变进行表面等离子体共振，分析TAGX-0004和ARC 1779的结合位点。电泳迁移率变动分析表明，R1395和R1399在A1结构域结合到两个适体。R1287、K1362和R1392促进ARC 1779结合，F1366对TAGX-0004结合至关重要。caplacizumab结合位点的表面等离子体共振分析鉴定了VWF A1结构域中的5个氨基酸（K1362、R1392、R1395、R1399和K1406）。这些结果表明，TAGX-0004在体外具有比caplacizumab更好的药理学特性，可能同样有希望用于aTTP治疗。
Von Willebrand factor (VWF) is a blood glycoprotein that plays an important role in platelet thrombus formation through interaction between its A1 domain and platelet glycoprotein Ib. ARC1779, an aptamer to the VWF A1 domain, was evaluated in a clinical trial for acquired thrombotic thrombocytopenic purpura (aTTP). Subsequently, caplacizumab, an anti-VWF A1 domain nanobody, was approved for aTTP in Europe and the USA. We recently developed a novel DNA aptamer, TAGX-0004, to the VWF A1 domain; it contains an artificial base and demonstrates high affinity for VWF. To compare the effects of these three agents on VWF A1, their ability to inhibit ristocetin- or botrocetin-induced platelet aggregation under static conditions was analyzed, and the inhibition of thrombus formation under high shear stress was investigated in a microchip flow chamber system. In both assays, TAGX-0004 showed stronger inhibition than ARC1779, and had comparable inhibitory effects to caplacizumab. The binding sites of TAGX-0004 and ARC1779 were analyzed with surface plasmon resonance performed using alanine scanning mutagenesis of the VWF A1 domain. An electrophoretic mobility shift assay showed that R1395 and R1399 in the A1 domain bound to both aptamers. R1287, K1362, and R1392 contributed to ARC1779 binding, and F1366 was essential for TAGX-0004 binding. Surface plasmon resonance analysis of the binding sites of caplacizumab identified five amino acids in the VWF A1 domain (K1362, R1392, R1395, R1399, and K1406). These results suggest that TAGX-0004 possesses better pharmacological properties than caplacizumab in vitro and might be similarly promising for aTTP treatment.
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DOI：
10.1016/j.str.2009.09.011
发表时间：
2009-11-11
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Huang RH;Fremont DH;Diener JL;Schaub RG;Sadler JE
通讯作者：
Sadler JE
影响因子：
158.5
作者：
Scully, M.;Cataland, S. R.;Zeldin, R. K.
通讯作者：
Zeldin, R. K.
影响因子：
37.8
作者：
Gilbert, James C.;DeFeo-Fraulini, Tia;Schaub, Robert G.
通讯作者：
Schaub, Robert G.
影响因子：
158.5
作者：
Gragoudas, ES;Adamis, AP;Guyer, DR
通讯作者：
Guyer, DR
影响因子：
3.3
作者：
Bath, PMW;Blann, A;Butterworth, RJ
通讯作者：
Butterworth, RJ