Galloylated polyphenols represent a new class of antithrombotic agents with broad activity against thiol isomerases

IF 5 2区医学 Q1 HEMATOLOGY Journal of Thrombosis and Haemostasis Pub Date : 2025-06-01 Epub Date: 2025-02-12 DOI:10.1016/j.jtha.2025.01.021

Moua Yang , Ivan Hancco Zirena , Quinn P. Kennedy , Anika Patel , Glenn Merrill-Skoloff , Kelsey D. Sack , Emmy Fulcidor , Christina Scartelli , Shihui Guo , Roelof H. Bekendam , Osamede C. Owegie , Huanzhang Xie , Ionita C. Ghiran , Oren Levy , Lin Lin , Robert Flaumenhaft

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Abstract

Background

Both protein disulfide isomerase (PDI) and SARS-CoV-2 main protease (M^pro) are reliant on active-site cysteines stabilized by adjacent amino acids. We reasoned that redox-active compounds might interfere with both enzymes by acting in the vicinity of these reactive sites thus interfering with viral replication and thrombus formation. Our previous screen of 1019 flavonoids identified several compounds that inhibit SARS-CoV-2 M^pro.

Objectives

Our goal was to identify phytochemical inhibitors of SARS-CoV-2 M^pro that block thiol isomerases and are antithrombotic.

Methods

PDI, ERp57, ERp5, ERp46, isolated domains of PDI, and PDI mutants were used to evaluate the effects of galloylated polyphenols and their analogs on thiol isomerase reductase activity. Laser-injury and ferric chloride models of thrombus formation and a tail snip assay were used to assess the effects on thrombosis and hemostasis.

Results

Pinocembrin 7-O-(3''-galloyl-4'',6''-(S)-hexahydroxydiphenoyl)-β-D-glucose (PGHG) inhibited both PDI and SARS-CoV-2 M^pro. Evaluation of isolated PDI fragments and active-site cysteine mutants showed that PGHG acts at the catalytic domains. Structure-function studies showed that PGHG interacts with histidines within the Cys53-Gly54-His55-Cys56 motifs of PDI. PGHG was equally active against other thiol isomerases, including ERp57, ERp5, ERp72, and ERp46. Screening numerous galloylated polyphenols demonstrated a class effect on thiol isomerase inhibition. Structure-activity relationships indicated that the galloyl moieties within large galloylated polyphenols were important for their inhibitory activity. PGHG and punicalagin were antithrombotic in murine models of thrombus formation.

Conclusions

Galloylated polyphenols represent a large class of antithrombotic compounds with broad activity against thiol isomerases. Many of these compounds also inhibit SARS-CoV-2 M^pro and viral replication.

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没食子酸多酚是一类新的抗血栓药物，对巯基异构酶具有广泛的活性。

背景：蛋白二硫异构酶（PDI）和SARS-CoV-2主蛋白酶（Mpro）都依赖于邻近氨基酸稳定的活性位点半胱氨酸。我们推断，氧化还原活性化合物可能通过作用于这些活性位点附近而干扰这两种酶，从而干扰病毒复制和血栓形成。我们之前对1019类黄酮的筛选发现了几种抑制SARS-CoV-2 Mpro的化合物。目的：我们的目标是鉴定阻断硫醇异构酶并具有抗血栓作用的SARS-CoV-2 Mpro的植物化学抑制剂。方法：采用PDI、ERp57、ERp5、ERp46、PDI分离结构域和PDI突变体，评价没食子酸多酚及其类似物对巯基异构酶还原酶活性的影响。采用激光损伤和FeCl3血栓形成模型及尾巴剪法评估其对血栓形成和止血的影响。结果：7-O-（3′-没食子酰-4′，6′-(S)-六羟基二酚）- β - d -葡萄糖（PGHG）对PDI和SARS-CoV-2 Mpro均有抑制作用。对分离的PDI片段和活性位点半胱氨酸突变体的评价表明，PGHG在催化结构域起作用。结构-功能研究表明PGHG与PDI的CGHC基序中的组氨酸相互作用。PGHG对包括ERp57、ERp5、ERp72和ERp46在内的其他硫醇异构酶也具有同样的活性。筛选多种没食子酸多酚显示出对巯基异构酶的抑制作用。构效关系表明，大型没食子酰化多酚中的没食子酰部分对其抑制活性起重要作用。PGHG和punicalagin在小鼠血栓形成模型中具有抗血栓作用。结论：没食子酸多酚是一类抗血栓化合物，对巯基异构酶具有广泛的活性。其中许多化合物还能抑制SARS-CoV-2 Mpro和病毒复制。

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来源期刊

Journal of Thrombosis and Haemostasis 医学-外周血管病

CiteScore

24.30

自引率

3.80%

发文量

321

审稿时长

1 months

期刊介绍： The Journal of Thrombosis and Haemostasis (JTH) serves as the official journal of the International Society on Thrombosis and Haemostasis. It is dedicated to advancing science related to thrombosis, bleeding disorders, and vascular biology through the dissemination and exchange of information and ideas within the global research community. Types of Publications: The journal publishes a variety of content, including: Original research reports State-of-the-art reviews Brief reports Case reports Invited commentaries on publications in the Journal Forum articles Correspondence Announcements Scope of Contributions: Editors invite contributions from both fundamental and clinical domains. These include: Basic manuscripts on blood coagulation and fibrinolysis Studies on proteins and reactions related to thrombosis and haemostasis Research on blood platelets and their interactions with other biological systems, such as the vessel wall, blood cells, and invading organisms Clinical manuscripts covering various topics including venous thrombosis, arterial disease, hemophilia, bleeding disorders, and platelet diseases Clinical manuscripts may encompass etiology, diagnostics, prognosis, prevention, and treatment strategies.