Multivalent Tranexamic Acid (TXA) and Benzamidine Derivatives for Serine Protease Inhibition

IF 4.9 Q1 CHEMISTRY, MEDICINAL

ACS Pharmacology and Translational Science Pub Date : 2025-05-14 DOI:10.1021/acsptsci.5c0003010.1021/acsptsci.5c00030

Tanmaye Nallan Chakravarthula, Rodrigo Santillan-Rodriguez, Ziqian Zeng, Abigail Hall, Andres Prieto Trujillo, Anushri Umesh and Nathan J. Alves*,

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引用次数: 0

Abstract

Blood coagulation and fibrinolysis pathways involve many serine proteases in a careful equilibrium. Disruption of this hemostatic balance can cause life-threatening thromboembolic and bleeding disorders that require therapeutic intervention. Heterobivalent molecules synthesized with both benzamidine (active site serine protease inhibitor) and tranexamic acid (TXA, kringle/lysine-site inhibitor) of increasing dPEG linker lengths (dPEG₄–dPEG₃₆) were synthesized and analyzed for plasmin, thrombin, and tissue plasminogen activator (tPA) inhibition using soluble enzymatic substrates. Linker lengths greater than the active and lysine binding site separation achieved improved inhibition with plasmin and tPA due to multivalent subsite binding effects. Despite TXA being a weak active site inhibitor, homomultivalent TXA (PAMAM⁸-TXA) demonstrated strong competitive plasmin inhibition (K_i = 2.5 ± 1.8 μM) due to the statistical rebinding effect. IC₅₀ values were also determined by assaying on physiologically relevant, fluorescently tagged, annular fibrin clots to capture the effect of kringle binding inhibition on fibrinolytic potential in the presence and absence of inhibitors.

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多价氨甲环酸（TXA）和苯脒衍生物对丝氨酸蛋白酶的抑制作用

凝血和纤溶途径涉及许多丝氨酸蛋白酶在一个小心的平衡。这种止血平衡的破坏可导致危及生命的血栓栓塞和出血性疾病，需要治疗干预。合成了由苄脒（活性位点丝氨酸蛋白酶抑制剂）和氨甲环酸（TXA， kringle/赖氨酸位点抑制剂）合成的增加dPEG连接长度的异二价分子（dPEG4-dPEG36），并利用可溶性酶底物对纤溶酶、凝血酶和组织纤溶酶原激活物（tPA）进行了抑制分析。由于多价亚位点结合效应，长度大于活性和赖氨酸结合位点分离的连接体对纤溶蛋白和tPA的抑制作用得到了改善。尽管TXA是一种弱活性位点抑制剂，但由于统计上的重结合效应，同价TXA （PAMAM8-TXA）表现出很强的竞争性纤溶酶抑制（Ki = 2.5±1.8 μM）。IC50值也通过分析生理相关的、荧光标记的环状纤维蛋白凝块来确定，以捕获kringle结合抑制在存在和不存在抑制剂的情况下对纤维蛋白溶解电位的影响。

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来源期刊

ACS Pharmacology and Translational Science Medicine-Pharmacology (medical)

CiteScore

10.00

自引率

3.30%

发文量

133

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