title: Synergistic Inhibition of Both P2Y1 and P2Y12 Adenosine Diphosphate Receptors As Novel Approach to Rapidly Attenuate Platelet-Mediated Thrombosis.
abstract: OBJECTIVE
  Unlike currently approved adenosine diphosphate receptor antagonists, the new diadenosine tetraphosphate derivative GLS-409 targets not only P2Y12 but also the second human platelet adenosine diphosphate receptor P2Y1 and may, therefore, be a promising antiplatelet drug candidate. The current study is the first to investigate the in vivo antithrombotic effects of GLS-409.
  
  
  RESULTS
  We studied (1) the in vivo effects of GLS-409 on agonist-stimulated platelet aggregation in anesthetized rats, (2) the antithrombotic activity of GLS-409 and the associated effect on the bleeding time in a canine model of platelet-mediated coronary artery thrombosis, and (3) the inhibition of agonist-stimulated platelet aggregation by GLS-409 versus selective P2Y1 and P2Y12 inhibition in vitro in samples from healthy human subjects before and 2 hours after aspirin intake. In vivo treatment with GLS-409 significantly inhibited adenosine diphosphate- and collagen-stimulated platelet aggregation in rats. Further, GLS-409 attenuated cyclic flow variation, that is, platelet-mediated thrombosis, in vivo in our canine model of unstable angina. The improvement in coronary patency was accompanied by a nonsignificant 30% increase in bleeding time. Of note, GLS-409 exerted its effects without affecting rat and canine hemodynamics. Finally, in vitro treatment with GLS-409 showed effects similar to that of cangrelor and the combination of cangrelor with the selective P2Y1 inhibitor MRS 2179 on agonist-stimulated platelet aggregation in human platelet-rich plasma and whole blood before and 2 hours after aspirin intake.
  
  
  CONCLUSIONS
  Synergistic inhibition of both P2Y1 and P2Y12 adenosine diphosphate receptors by GLS-409 immediately attenuates platelet-mediated thrombosis and effectively blocks agonist-stimulated platelet aggregation irrespective of concomitant aspirin therapy.
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.). thomas.gremmel@meduniwien.ac.at.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gremmel
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Ivan B
authors.initials: IB
authors.lastname: Yanachkov
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Milka I
authors.initials: MI
authors.lastname: Yanachkova
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: George E
authors.initials: GE
authors.lastname: Wright
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Joseph
authors.initials: J
authors.lastname: Wider
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Vishnu V R
authors.initials: VV
authors.lastname: Undyala
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Alan D
authors.initials: AD
authors.lastname: Michelson
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Frelinger
authors.affiliation: From the Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (T.G., A.D.M., A.L.F.); Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria (T.G.); GLSynthesis Inc, Worcester, MA (I.B.Y., M.I.Y., G.E.W.); and Cardiovascular Research Institute and Departments of Physiology and Emergency Medicine, Wayne State University School of Medicine, Detroit, MI (J.W., V.V.R.U., K.P.).
authors.forename: Karin
authors.initials: K
authors.lastname: Przyklenk
mesh_terms: D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000328:Adult; D000818:Animals; D001241:Aspirin / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D003328:Coronary Thrombosis / Q000097:blood / Q000188:drug therapy*; D015226:Dinucleoside Phosphates / Q000494:pharmacology*; D004195:Disease Models, Animal; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D005260:Female; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D008297:Male; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D010979:Platelet Function Tests; D058921:Purinergic P2Y Receptor Antagonists / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D058472:Receptors, Purinergic P2Y1 / Q000097:blood / Q000187:drug effects*; D058925:Receptors, Purinergic P2Y12 / Q000097:blood / Q000187:drug effects*; D013997:Time Factors; D055815:Young Adult
chemical_list: D015226:Dinucleoside Phosphates; D005343:Fibrinolytic Agents; C111914:N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate; C546382:P2RY1 protein, human; C548576:P2RY12 protein, human; C546384:P2ry1 protein, rat; C553058:P2ry12 protein, rat; D010975:Platelet Aggregation Inhibitors; D058921:Purinergic P2Y Receptor Antagonists; D058472:Receptors, Purinergic P2Y1; D058925:Receptors, Purinergic P2Y12; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; C117446:cangrelor; D001241:Aspirin
keywords: P2Y1; P2Y12; adenosine diphosphate; antiplatelet therapy; aspirin