Uridine Triphosphate Thio Analogues Inhibit Platelet P2Y12Receptor and Aggregation.
Platelet P2Y12is an important adenosine diphosphate (ADP) receptor that is involved in agonist-induced platelet aggregation and is a valuable target for the development of anti-platelet drugs. Here we characterise the effects of thio analogues of uridine triphosphate (UTP) on ADP-induced platelet aggregation. Using human platelet-rich plasma, we demonstrate that UTP inhibits P2Y12but not P2Y₁ receptors and antagonises 10 µM ADP-induced platelet aggregation in a concentration-dependent manner with an IC50value of ~250 °µM. An eight-fold higher platelet inhibitory activity was observed with a 2-thio analogue of UTP (2S-UTP), with an IC50of 30 µM. The 4-thio analogue (4S-UTP) with an IC50of 7.5 µM was 33-fold more effective. A three-fold decrease in inhibitory activity, however, was observed by introducing an isobutyl group at the 4S- position. A complete loss of inhibition was observed with thio-modification of the γ phosphate of the sugar moiety, which yields an enzymatically stable analogue. The interaction of UTP analogues with P2Y12receptor was verified by P2Y12receptor binding and cyclic AMP (cAMP) assays. These novel data demonstrate for the first time that 2- and 4-thio analogues of UTP are potent P2Y12receptor antagonists that may be useful for therapeutic intervention.
Department of Cardiology/Angiology, University Hospital Giessen, 35392 Giessen, Germany. dursun.guenduez@med.uni-giessen.de.
Dursun
D
Gündüz
Department of Neurology, University Hospital Giessen, 35392 Giessen, Germany. Christian.Tanislav@neuro.med.uni-giessen.de.
Christian
C
Tanislav
Department of Cardiology/Angiology, Hannover Medical School, 30625 Hannover, Germany. sedding.daniel@mh-hannover.de.
Daniel
D
Sedding
Department of Cardiology/Angiology, University Hospital Marburg, 35043 Marburg, Germany. mariana.parahuleva@innere.med.uni-giessen.de.
Mariana
M
Parahuleva
Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, 35392 Giessen, Germany. sentot.santoso@med.uni-giessen.de.
Sentot
S
Santoso
Department of Cardiology/Angiology, University Hospital Giessen, 35392 Giessen, Germany. christian.troidl@med.uni-giessen.de.
Christian
C
Troidl
Department of Cardiology/Angiology, University Hospital Giessen, 35392 Giessen, Germany. christian.hamm@med.uni-giessen.de.
Christian W
CW
Hamm
Department of Cardiology/Angiology, University Hospital Giessen, 35392 Giessen, Germany. muhammad.aslam@med.uni-giessen.de.
Muhammad
M
Aslam
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000328:Adult; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism; D008875:Middle Aged; D015394:Molecular Structure; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D053657:Platelet-Rich Plasma; D058921:Purinergic P2Y Receptor Antagonists / Q000494:pharmacology*; D058925:Receptors, Purinergic P2Y12 / Q000378:metabolism*; D014544:Uridine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology*; D055815:Young Adult; D000098662:Vasodilator-Stimulated Phosphoprotein
D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D010750:Phosphoproteins; D010975:Platelet Aggregation Inhibitors; D058921:Purinergic P2Y Receptor Antagonists; D058925:Receptors, Purinergic P2Y12; D000098662:Vasodilator-Stimulated Phosphoprotein; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D014544:Uridine Triphosphate
2S-UTP; 4S-UTP; ADP; P2Y12 receptor; platelet aggregation