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Thromb Haemost 2024; 124(03): 203-222
DOI: 10.1055/a-2211-5202
New Technologies, Diagnostic Tools and Drugs

Diphenyl-tetrazol-propanamide Derivatives Act as Dual-Specific Antagonists of Platelet CLEC-2 and Glycoprotein VI

Nobuo Watanabe 
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
2   Institute of Advanced Biosciences, Tokai University, Hiratsuka, Kanagawa, Japan
,
Yoshiko Shinozaki
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Sanae Ogiwara
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Riko Miyagasako
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Ayumi Sasaki
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Junko Kato
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Yusuke Suzuki
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Natsuko Fukunishi
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Yoshinori Okada
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Takeshi Saito
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Yumi Iida
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Misaki Higashiseto
3   Support Center for Medical Research and Education, Tokai University, Isehara, Kanagawa, Japan
,
Haruchika Masuda
4   Department of Physiology, Tokai University School of Medicine, Shimokasuya, Isehara, Kanagawa, Japan
,
Eiichiro Nagata
5   Department of Neurology, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Kazuhito Gotoh
6   Department of Laboratory Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Mari Amino
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Tomoatsu Tsuji
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Seiji Morita
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Yoshihide Nakagawa
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
,
Noriaki Hirayama
2   Institute of Advanced Biosciences, Tokai University, Hiratsuka, Kanagawa, Japan
7   The Institute of Medical Sciences, Tokai University, Isehara, Kanagawa, Japan
,
Sadaki Inokuchi 
1   Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
2   Institute of Advanced Biosciences, Tokai University, Hiratsuka, Kanagawa, Japan
› Author Affiliations
Funding This work was supported by grants from the Institute of Advanced Biosciences of Tokai University (S.I. and N.W.) and a Grant-in-Aid for Scientific Research Category C (23K07490 to N.W.) and AMED (A369-TS; to N.W.).
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Abstract

Background Platelet C-type lectin-like receptor 2 (CLEC-2) induces platelet activation and aggregation after clustering by its ligand podoplanin (PDPN). PDPN, which is not normally expressed in cells in contact with blood flow, is induced in inflammatory immune cells and some malignant tumor cells, thereby increasing the risk of venous thromboembolism (VTE) and tumor metastasis. Therefore, small-molecule compounds that can interfere with the PDPN–CLEC-2 axis have the potential to become selective antiplatelet agents.

Methods and Results Using molecular docking analysis of CLEC-2 and a PDPN–CLEC-2 binding-inhibition assay, we identified a group of diphenyl-tetrazol-propanamide derivatives as novel CLEC-2 inhibitors. A total of 12 hit compounds also inhibited PDPN-induced platelet aggregation in humans and mice. Unexpectedly, these compounds also fit the collagen-binding pocket of the glycoprotein VI molecule, thereby inhibiting collagen interaction. These compounds also inhibited collagen-induced platelet aggregation, and one compound ameliorated collagen-induced thrombocytopenia in mice. For clinical use, these compounds will require a degree of chemical modification to decrease albumin binding.

Conclusion Nonetheless, as dual activation of platelets by collagen and PDPN-positive cells is expected to occur after the rupture of atherosclerotic plaques, these dual antagonists could represent a promising pharmacophore, particularly for arterial thrombosis, in addition to VTE and metastasis.

Keywords

tumor metastasis - thrombosis - stroke - myocardial infarction

Authors' Contribution

N.H. and S.I. conceived and planned the project. N.W., Y.S., S.O., and R.M. conducted the mouse experiments. N.W., A.S., Y.I., M.H., E.N., M.A., K.G., and Y.O. conducted in vitro experiments. J.K., Y.S., and N.F. conducted the electron microscopic analysis. N.H. conducted the molecular docking analysis. T.S. conducted the mass-spectrometry analysis. S.I. organized the project, and N.W., H.M., T.T., S.M., and Y.N. analyzed the data and wrote the manuscript. N.W. and S.I. edited the final manuscript. All authors have read and approved the final manuscript.


Supplementary Material



Publication History

Received: 09 March 2023

Accepted: 12 November 2023

Accepted Manuscript online:
15 November 2023

Article published online:
29 December 2023

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