Details

Autor(en) / Beteiligte

• Fernández, Delia I
• Troitiño, Sara
• Sobota, Vladimír
• Tullemans, Bibian M E
• Zou, Jinmi
• van den Hurk, Helma
• García, Ángel
• Honarnejad, Saman
• Kuijpers, Marijke J E
• Heemskerk, Johan W M

Titel

Ultra-high throughput-based screening for the discovery of antiplatelet drugs affecting receptor dependent calcium signaling dynamics

Ist Teil von

• Scientific reports, 2024-03, Vol.14 (1), p.6229-6229, Article 6229

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2024

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• Distinct platelet activation patterns are elicited by the tyrosine kinase-linked collagen receptor glycoprotein VI (GPVI) and the G-protein coupled protease-activated receptors (PAR1/4) for thrombin. This is reflected in the different platelet Ca responses induced by the GPVI agonist collagen-related peptide (CRP) and the PAR1/4 agonist thrombin. Using a 96 well-plate assay with human Calcium-6-loaded platelets and a panel of 22 pharmacological inhibitors, we assessed the cytosolic Ca signaling domains of these receptors and developed an automated Ca curve algorithm. The algorithm was used to evaluate an ultra-high throughput (UHT) based screening of 16,635 chemically diverse small molecules with orally active physicochemical properties for effects on platelets stimulated with CRP or thrombin. Stringent agonist-specific selection criteria resulted in the identification of 151 drug-like molecules, of which three hit compounds were further characterized. The dibenzyl formamide derivative ANO61 selectively modulated thrombin-induced Ca responses, whereas the aromatic sulfonyl imidazole AF299 and the phenothiazine ethopropazine affected CRP-induced responses. Platelet functional assays confirmed selectivity of these hits. Ethopropazine retained its inhibitory potential in the presence of plasma, and suppressed collagen-dependent thrombus buildup at arterial shear rate. In conclusion, targeting of platelet Ca signaling dynamics in a screening campaign has the potential of identifying novel platelet-inhibiting molecules.