High‐throughput screen identifies MAS9 as a novel inhibitor of the C‐type lectin receptor‐2 (CLEC‐2)–podoplanin interaction

• DOI: 10.1111/bph.70036
• Publication Date: 2025-04-24T05:25:47Z
• Authors: Marcin A. Sowa; Yan Wu; Jan van Groningen; Ying Di; Helma van den Hurk; Johannes A. Eble; Jonathan M. Gibbins; Ángel García; Alice Y. Pollitt
• Keywords: Podoplanin; C-type lectin; Lymphangiogenesis

Abstract Background and Purpose The C‐type lectin‐like receptor‐2 (CLEC‐2) is a platelet receptor for the endogenous ligand podoplanin. This interaction contributes to several (patho)physiological processes, such as lymphangiogenesis, preservation of blood lymphatic vessel integrity organ development, tumour metastasis. Activation CLEC‐2 leads phosphorylation its cytoplasmic hemITAM domain initiates signalling cascade involving kinase Syk. aim this study was identify characterise novel small molecule inhibitor CLEC‐2. Experimental Approach An AlphaScreen‐based high‐throughput screening used CLEC‐2–podoplanin interaction. Binding site interactions were assessed using in silico modelling. Functional assays, including light transmission aggregometry, spreading evaluate effect on CLEC‐2‐mediated activation. Key Results A total 18,476 molecules screened resulting 14 candidates. Following secondary screening, one molecule, MAS9, taken forward further characterisation. binding sites MAS9 predicted share with ligands podoplanin rhodocytin. inhibited aggregation, signalling. also resulted fibrinogen binding. Conclusion Implications inhibits signalling, showing selectivity inhibition over GPVI. paves way future preclinical assays test potential therapeutic tool treat pathologies thromboinflammation cancer.