新型凝血酶抑制肽Ultravariegin的PEG修饰以延长体内循环时间与增强抗血栓作用

PEGylation of New Thrombin Inhibitor Peptide Ultravariegin for Prolonged In Vivo Circulation and Enhanced Antithrombotic Effects

DOI 原文链接

用sci-hub下载

MOLECULAR PHARMACEUTICS

影响因子:4.5

分区:医学2区 / 药学2区医学：研究与实验3区

发表日期:2024 Dec 02

作者: Xia Song, Yuting Wen, Aaron Wei Liang Li, Jingling Zhu, Cho Yeow Koh, R Manjunatha Kini, Mark Yan Yee Chan, Jun Li

DOI: 10.1021/acs.molpharmaceut.4c00769

摘要

抗凝治疗常用于预防和治疗心血管疾病、脑血管疾病及癌症患者的动脉和静脉血栓。静脉血栓是继急性冠脉疾病和中风之后的第三大心血管死亡原因。亟需开发具有良好效果且出血风险低的有效抗凝药物。Variegin及其变体是一类新型抗凝肽，在临床前研究中表现出良好的前景。Variegin及其最优变体Ultravariegin（UV）能更有效抑制血栓形成，同时引起的出血较传统药物如肝素和比伐卢定少。然而，UV的短半衰期限制了其临床应用。PEG化是一种将聚乙二醇（PEG）链共价连接到肽或药物上的方法，可能通过延长其在体内的循环时间来改善UV的效果。本研究采用Maleimide-PEG5k和10k对UV进行PEG修饰。体外和体外外周血浆中的动物模型（大鼠和兔子）实验显示，PEG修饰对UV的抗凝活性影响微乎其微。在大鼠和兔子体内实验中，PEG修饰的UV表现出更长的半衰期和更强的抗凝效果，尤其是在皮下注射时。PEG修饰显著延长了兔子体内UV的半衰期，持续抗凝作用可达4天。这说明增加UV的分子大小并用PEG包覆可以减少肾脏清除，延长循环时间。PEG修饰UV的半衰期和抗凝活性得到改善，使其成为更优的抗凝治疗候选药物。

Abstract

Anticoagulant therapy is commonly used to prevent and treat arterial and venous blood clots in patients with cardiovascular disease, cerebrovascular disease, and cancer. Venous blood clots are the third leading cause of cardiovascular death following acute coronary artery disease and stroke. There is a significant need for effective anticoagulant therapy with minimal risk of bleeding. Variegin and its variants are a new type of antithrombin peptide that has shown promising results in preclinical studies. Variegin and its best variant, ultravariegin (UV), can more effectively inhibit blood clot formation while causing less bleeding than traditional medications such as heparin and bivalirudin. However, the short lifespan of UV remains a limitation for its use in clinical settings. PEGylation, a method of conjugating poly(ethylene glycol) (PEG) chains to peptides or drugs, may help improve the effectiveness of UV by extending its circulation time in the body. In this study, UV was PEGylated using maleimide-PEG5k and 10k. The impact of PEGylation on the antithrombin activity of UV was assessed in vitro and ex vivo in rat and rabbit plasma, showing minimal effects on the efficacy. In vivo studies in rats and rabbits revealed that PEGylated UV had a longer half-life and greater anticoagulant effects than unmodified UV did, especially when it was administered subcutaneously. PEGylation significantly extended the half-life of UV in rabbits, resulting in sustained anticoagulant effects for up to 4 days. This demonstrated that increasing the size of UV and shielding it with PEG could reduce clearance by the kidneys and prolong its circulation time. The improved half-life and antithrombin activity of PEGylated UV make it a more favorable choice for anticoagulant therapy.