Subscribe to RSS
DOI: 10.1160/TH11-06-0400
BF061, a novel antiplatelet and antithrombotic agent targeting P2Y12 receptor and phosphodiesterase
Financial support: This work was supported in part by National Natural Science Foundation of China [No.30772564, 30973529], National Drug Innovative Program from Ministry of Science and Technology of China [2009ZX09301–011], Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning to ZD, and a grant from National Institutes of Health National Heart, Lung and Blood Institute [grant HL60683] to SPK. KY is a recipient of Hui-Chun Chin and Tsung-Dao Lee Chinese Undergraduate Research Endowment (CURE).Publication History
Received:
13 June 2011
Accepted after major revision:
31 August 2011
Publication Date:
27 November 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/201106/lookinside/thumbnails/10-1160-th11-06-0400-1.jpg)
Summary
The addition of phosphodiesterase (PDE) inhibitors has been reported to potentiate the antithrombotic effects of P2Y12 antagonists without increasing bleeding risk. In this study, we report that a potent antiplatelet agent, 2-ethylthio-6-phenethylaminoadenosine (BF061), inhibits platelet activation and thrombosis via P2Y12 antagonism and PDE inhibition. We explored the antiplatelet mechanism of BF061 by measuring cAMP, cGMP levels, PDE activity, and the interaction between ADP and P2Y12 using atomic force microscopy. The antithrombotic effect of BF061 was evaluated in mice using intravital microscopy in FeCl3-induced mesenteric and laser-induced cremasteric arterial thrombosis models. BF061 robustly inhibited platelet aggregation and ATP release induced by multiple platelet agonists via P2Y12 antagonism and PDE inhibition. Interestingly, despite being structurally similar to BF061, P2Y12 receptor antagonist AR-C69931MX had no effect on human platelet PDE. In FeCl3-induced mesenteric arterial thrombosis model, BF061 effectively prevented thrombus formation similarly to clopidogrel; it also reduced thrombus volume in laser-injured cremaster arteriole model. In contrast, BF061 induced dramatically less bleeding at an antithrombotic dose compared to clopidogrel. In summary, we developed a novel antiplatelet and antithrombotic agent targeting both P2Y12 and PDE. Given the prevalence of combined antiplatelet therapy in clinical practice, an antiplatelet agent bearing dual activities may have therapeutic advantage as a potential antithrombotic drug.
* These authors contributed equally to this work.
-
References
- 1 Michelson AD. P2Y12 antagonism: promises and challenges. Arterioscler Thromb Vasc Biol 2008; 28: 33-38.
- 2 Chen KY, Rha SW, Li YJ. et al. Triple versus dual antiplatelet therapy in patients with acute ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention. Circulation 2009; 119: 3207-3214.
- 3 Lee SW, Park SW, Kim YH. et al. A Randomized, Double-Blind, Multicenter Comparison Study of Triple Antiplatelet Therapy With Dual Antiplatelet Therapy to Reduce Restenosis After Drug-Eluting Stent Implantation in Long Coronary Lesions Results From the DECLARE-LONG II (Drug-Eluting Stenting Followed by Cilostazol Treatment Reduces Late Restenosis in Patients with Long Coronary Lesions) Trial. J Am Coll Cardiol 2011; 57: 1264-1270.
- 4 Suh JW, Lee SP, Park KW. et al. Multicenter randomized trial evaluating the efficacy of cilostazol on ischemic vascular complications after drug-eluting stent implantation for coronary heart disease: Results of the CILON-T (Influence of CILostazol-based triple antiplatelet therapy ON ischemic complication after drug-eluting stenT implantation) trial. J Am Coll Cardiol 2011; 57: 280-289.
- 5 Lee SW, Park SW, Hong MK. et al. Triple versus dual antiplatelet therapy after coronary stenting: impact on stent thrombosis. J Am Coll Cardiol 2005; 46: 1833-1837.
- 6 Lee K, Kim JY, Yoo BS. et al. Cilostazol augments the inhibition of platelet aggregation in clopidogrel low-responders. J Thromb Haemost 2010; 8: 2577-2579.
- 7 Shankar H, Murugappan S, Kim S. et al. Role of G protein-gated inwardly rectifying potassium channels in P2Y12 receptor-mediated platelet functional responses. Blood 2004; 104: 1335-1343.
- 8 Ding Z, Bynagari YS, Mada SR. et al. Studies on the role of the extracellular cysteines and oligomeric structures of the P2Y receptor when interacting with antagonists. J Thromb Haemost 2009; 7: 232-234.
- 9 Zhang S, Hu L, Du H. et al. BF0801, a novel adenine derivative, inhibits platelet activation via phosphodiesterase inhibition and P2Y12 antagonism. Thromb Haemost 2010; 104: 845-857.
- 10 Berlot C. Expression and functional analysis of G protein alpha subunits in mammalian cells. Boca Raton: CRC Press. 1999
- 11 Ma T, Zhu X. Interleukin-6 increases the levels of cyclic GMP and nitrite in rat hippocampal slices. Eur J Pharmacol 1997; 321: 343-347.
- 12 Spoto G, Whitehead E, Ferraro A. et al. A reverse-phase HPLC method for cAMP phosphodiesterase activity. Anal Biochem 1991; 196: 207-210.
- 13 Tang HF, Song YH, Chen JC. et al. Upregulation of phosphodiesterase-4 in the lung of allergic rats. Am J Respir Crit Care Med 2005; 171: 823-828.
- 14 Ding Z, Kim S, Kunapuli SP. Identification of a Potent Inverse Agonist at a Constitutively Active Mutant of Human P2Y12 Receptor. Mol Pharmacol 2006; 69: 338-345.
- 15 Horber JK, Miles MJ. Scanning probe evolution in biology. Science 2003; 302: 1002-1005.
- 16 Hinterdorfer P, Dufrene YF. Detection and localization of single molecular recognition events using atomic force microscopy. Nat Methods 2006; 3: 347-355.
- 17 Banno F, Chauhan AK, Kokame K. et al. The distal carboxyl-terminal domains of ADAMTS13 are required for regulation of in vivo thrombus formation. Blood 2009; 113: 5323-5329.
- 18 Chauhan AK, Motto DG, Lamb CB. et al. Systemic antithrombotic effects of ADAMTS13. J Exp Med 2006; 203: 767-776.
- 19 Frenette PS, Johnson RC, Hynes RO. et al. Platelets roll on stimulated endothelium in vivo: an interaction mediated by endothelial P-selectin. Proc Natl Acad Sci USA 1995; 92: 7450-7454.
- 20 Angelillo-Scherrer A, Burnier L, Flores N. et al. Role of Gas6 receptors in platelet signaling during thrombus stabilization and implications for antithrombotic therapy. J Clin Invest 2005; 115: 237-246.
- 21 Stolla M, Stefanini L, Roden RC. et al. The kinetics of alphaIIbbeta3 activation determines the size and stability of thrombi in mice: implications for antiplatelet therapy. Blood 2011; 117: 1005-1013.
- 22 Paul BZ, Jin J, Kunapuli SP. Molecular mechanism of thromboxane A(2)-induced platelet aggregation. Essential role for p2t(ac) and alpha(2a) receptors. J Biol Chem 1999; 274: 29108-29114.
- 23 Zhao L, Bath P, Heptinstall S. Effects of combining three different antiplatelet agents on platelets and leukocytes in whole blood in vitro. Br J Pharmacol 2001; 134: 353-358.
- 24 Orlowski E, Chand R, Yip J. et al. A platelet tetraspanin superfamily member, CD151, is required for regulation of thrombus growth and stability in vivo. J Thromb Haemost 2009; 7: 2074-2084.
- 25 Lenain N, Freund M, Leon C. et al. Inhibition of localized thrombosis in P2Y1-deficient mice and rodents treated with MRS2179, a P2Y1 receptor antagonist. J Thromb Haemost 2003; 1: 1144-1149.
- 26 Hechler B, Nonne C, Roh EJ. et al. MRS2500 [2-Iodo-N6-methyl-(N)-methanocarba-2'-deoxyadenosine-3',5'-bisphosphate], a Potent, Selective, and Stable Antagonist of the Platelet P2Y1 Receptor with Strong Antithrombotic Activity in Mice. J Pharmacol Exp Ther 2006; 316: 556-563.
- 27 Lee SW, Park SW, Yun SC. et al. Triple antiplatelet therapy reduces ischemic events after drug-eluting stent implantation: Drug-Eluting stenting followed by Cilostazol treatment REduces Adverse Serious cardiac Events (DECREASE registry). Am Heart J 2010; 159: 284-291.
- 28 Han Y, Li Y, Wang S. et al. Cilostazol in addition to aspirin and clopidogrel improves long-term outcomes after percutaneous coronary intervention in patients with acute coronary syndromes: a randomized, controlled study. Am Heart J 2009; 157: 733-739.
- 29 Andre P, Deguzman F, Haberstock-Debic H. et al. Thienopyridines, but Not Eli-nogrel, Result in Off-Target Effects at the Vessel Wall That Contribute to Bleeding. J Pharmacol Exp Ther 2011; epub ahead of print.
- 30 Sibbing D, Steinhubl SR, Schulz S. et al. Platelet aggregation and its association with stent thrombosis and bleeding in clopidogrel-treated patients: initial evidence of a therapeutic window. J Am Coll Cardiol 2010; 56: 317-318.
- 31 Ferreiro JL, Sibbing D, Angiolillo DJ. Platelet function testing and risk of bleeding complications. Thromb Haemost 2010; 103: 1128-1135.