TY - JOUR
T1 - Galloylated polyphenols represent a new class of antithrombotic agents with broad activity against thiol isomerases
AU - Yang, Moua
AU - Hancco Zirena, Ivan
AU - Kennedy, Quinn P.
AU - Patel, Anika
AU - Merrill-Skoloff, Glenn
AU - Sack, Kelsey D.
AU - Fulcidor, Emmy
AU - Scartelli, Christina
AU - Guo, Shihui
AU - Bekendam, Roelof H.
AU - Owegie, Osamede C.
AU - Xie, Huanzhang
AU - Ghiran, Ionita C.
AU - Levy, Oren
AU - Lin, Lin
AU - Flaumenhaft, Robert
N1 - Publisher Copyright:
© 2025 The Authors
PY - 2025/1/1
Y1 - 2025/1/1
N2 - Background: Both protein disulfide isomerase (PDI) and SARS-CoV-2 main protease (Mpro) 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 Mpro. Objectives: Our goal was to identify phytochemical inhibitors of SARS-CoV-2 Mpro 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 Mpro. 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 Mpro and viral replication.
AB - Background: Both protein disulfide isomerase (PDI) and SARS-CoV-2 main protease (Mpro) 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 Mpro. Objectives: Our goal was to identify phytochemical inhibitors of SARS-CoV-2 Mpro 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 Mpro. 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 Mpro and viral replication.
KW - galloylated polyphenol
KW - protein disulfide isomerase
KW - thiol isomerase
KW - thrombosis
UR - http://www.scopus.com/inward/record.url?scp=86000492623&partnerID=8YFLogxK
U2 - 10.1016/j.jtha.2025.01.021
DO - 10.1016/j.jtha.2025.01.021
M3 - Article
C2 - 39952360
AN - SCOPUS:86000492623
SN - 1538-7933
JO - Journal of Thrombosis and Haemostasis
JF - Journal of Thrombosis and Haemostasis
ER -
