S. Vertuani et al. / European Journal of Medicinal Chemistry 54 (2012) 202e209
209
decanted off and stored at ꢀ80 ꢁC before its employment for degra-
[20] V.L. Serebruany, S.R. Steinhubl, P.B. Berger, A.I. Malinin, D.L. Bhatt, E.J. Topol,
J. Am. Coll. Cardiol. 45 (2005) 246.
[21] T. Geisler, H. Langer, M. Wydymus, K. Göhring, C. Zürn, B. Bigalke, K. Stellos,
A.E. May, M. Gawaz, Eur. Heart J. 27 (2006) 2420.
[22] P.A. Gurbel, U.S. Tantry, Nat. Clin. Pract. Cardiovasc. Med. 3 (2006) 387.
[23] S. Heptinstall, J. Thromb. Haemost. 4 (2006) 539.
dation studies. The total protein concentration in the tissue homoge-
nate was determined using the Lowry procedure [60] (29.4 ꢂ 1.1
mg
protein/ L).
m
[24] D.J. Angiolillo, A. Fernandez-Ortiz, E. Bernardo, F. Alfonso, C. Macaya, T.A. Bass,
M.A. Costa, J. Am. Coll. Cardiol. 49 (2007) 1505.
[25] K.P. Bliden, J. DiChiara, U.S. Tantry, A.K. Bassi, S.K. Chaganti, P.A. Gurbel, J. Am.
Coll. Cardiol. 49 (2007) 657.
6.3. Stability studies in human plasma and rat liver homogenates
An aliquot of 15
L solution of each compounds (10ꢀ2 M in
m
[26] P. Buonamici, R. Marcucci, A. Migliorini, G.F. Gensini, A. Santini, R. Paniccia,
G. Moschi, A.M. Gori, R. Abbate, D. Antoniucci, J. Am. Coll. Cardiol. 49 (2007) 2312.
[27] J.T. Brandt, C.D. Payne, S.D. Wiviott, G. Weerakkody, N.A. Farid, D.S. Small,
J.A. Jakubowski, H. Naganuma, K.J. Winters, Am. Heart J. 153 (66) (2007) e9.
[28] T. Jernberg, C.D. Payne, K.J. Winters, C. Darstein, J.T. Brandt, J.A. Jakubowski,
H. Naganuma, A. Siegbahn, L. Wallentin, Eur. Heart J. 27 (2006) 1166.
[29] J.J.J. van Giezen, R.G. Humphries, Semin. Thromb. Hemost. 31 (2005) 195.
[30] B. Springthorpe, A. Bailey, P. Barton, T.N. Birkinshaw, R.V. Bonnert, R.C. Brown,
D. Chapman, J. Dixon, S.D. Guile, R.G. Humphries, S.F. Hunt, F. Ince, A.H. Ingall,
I.P. Kirk, P.D. Leeson, P. Leff, R.J. Lewis, B.P. Martin, D.F. McGinnity,
M.P. Mortimore, S.W. Paine, G. Pairaudeau, A. Patel, A.J. Rigby, R.J. Riley,
B.J. Teobald, W. Tomlinson, P.J. Webborn, P.A. Willis, Bioorg. Med. Chem. Lett.
17 (2007) 6013.
DMSO) was incubated (at a final concentration of 50 M) in rat liver
m
homogenate or human blood obtained from healthy subjects
(3 mL). Incubation of the aliquots was carried out at 37 ꢁC for
various periods up to 360 min. At different incubations times, an
aliquot of the solution (100
degradation was blocked by addition of ice water (100
mixing the solution, 50 L of SSA 10% and 50 L of internal standard
(50 M solution) were subsequently added. After centrifugation
(10 min at 14,000g), the supernatant was evaporated to dryness by
N2 flow. The residue was dissolved in 400 L of water and after
L) of supernatant
mL) was removed and compound
mL). After
m
m
m
m
[31] K. Balasubramaniam, G.N. Viswanathan, S.M. Marshall, A.G. Zaman, Cardiol.
Res. Pract. (2012) ID 909154.
[32] G. Cristalli, D.C. Mills, Biochem. J. 291 (1993) 875.
centrifugation (10 min at 19,800g) an aliquot (20
m
was injected into RP-HPLC.
[33] D.C. Mills, Blood 76 (1996) 835.
[34] N.J. Cusack, S.M.O. Hourani, J. Auton. Nerv. Syst. 81 (2000) 37.
[35] P. Crepaldi, B. Cacciari, M.C. Bonache, G. Spalluto, K. Varani, P.A. Borea,
I. Kügelgen, K. Hoffmann, M. Pugliano, C. Razzari, M. Cattaneo, Bioorg. Med.
Chem. 17 (2009) 4612.
[36] B. Xu, A. Stephens, G. Kirschenheuter, A.F. Greslin, X. Cheng, J. Sennelo,
M. Cattaneo, M.L. Zighetti, A. Chen, S.A. Kim, H.S. Kim, N. Bischofberger,
G. Cook, K.A. Jacobson, J. Med. Chem. 45 (2002) 5694.
[37] J.G. Douglass, R.I. Patel, B.R. Yerxa, S.S. Shaver, P.S. Watson, K. Bednarski,
R. Plourde, C.C. Redick, K. Brubaker, A.C. Jones, J.L. Boyer, J. Med. Chem. 51
(2008) 1007.
[38] J.G. Douglass, J.B. deCamp, E.H. Fulcher, W. Jones, S. Mahanty, A. Morgan,
D. Smirnov, A.C. Jones, J.L. Boyer, P.S. Watson, Bioorg. Med. Chem. Lett. 18
(2008) 2167.
Acknowledgements
Authors wish to thank Alberto Casolari and Elisa Durini for
careful NMR experiments. This work was supported by Ministry of
Education, University and Research (grant FIRB RBNE03YA3L_008).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
[39] S. Manfredini, P.G. Baraldi, R. Bazzanini, M. Marangoni, D. Simoni, J. Balzarini,
E. De Clercq, J. Med. Chem. 38 (1995) 199.
[40] S. Manfredini, P.G. Baraldi, E. Durini, S. Vertuani, J. Balzarini, E. De Clercq,
A. Karlsson, V. Buzzoni, L. Thelander, J. Med. Chem. 42 (1999) 3243.
[41] S. Manfredini, P.G. Baraldi, R. Bazzanini, E. Durini, S. Vertuani, A. Pani,
T. Marceddu, F. Demontis, L. Vargiu, P. La Colla, Nucleosides Nucleotides
Nucleic Acids 19 (2000) 705.
[42] S. Manfredini, N. Solaroli, A. Angusti, F. Nalin, E. Durini, S. Vertuani, S. Pricl,
M. Ferrone, S. Spadari, F. Focher, A. Verri, E. De Clercq, J. Balzarini, Antivir.
Chem. Chemother. 14 (2003) 183.
[43] S. Manfredini, A. Angusti, C.A. Veronese, E. Durini, S. Vertuani, F. Nalin,
N. Solaroli, S. Pricl, M. Ferrone, M. Mura, M.A. Piano, B. Poddesu, A. Cadeddu,
P. La Colla, R. Loddo, Pure Appl. Chem. 76 (2004) 1007.
[44] A. Angusti, S. Manfredini, E. Durini, N. Ciliberti, S. Vertuani, N. Solaroli, S. Pricl,
M. Ferrone, M. Fermeglia, R. Loddo, B. Secci, A. Visioli, T. Sanna, G. Collu,
M. Pezzullo, P. La Colla, Chem. Pharm. Bull. (Tokyo) 56 (2008) 423.
[45] Y. Chen, S. Morera, C. Pasti, A. Angusti, N. Solaroli, M. Veron, J. Janin,
S. Manfredini, D. Deville-Bonne, Med. Chem. 1 (2005) 529.
[46] N. Kohyama, T. Katashima, Y. Yamamoto, Synthesis 17 (2004) 2799.
[47] G.S. Buenger, V. Nair, Synthesis 10 (1990) 962.
References
[1] G. Hollopeter, H.M. Jantzen, D. Vincent, G. Li, L. England, V. Ramakrishnan,
R.B. Yang, P. Nurden, A. Nurden, D. Julius, P.B. Conley, Nature 409 (2001) 202.
[2] E. ten Boekel, P. Bartels, Pathophysiol. Haemost. Thromb. 32 (2002) 137.
[3] J. Jin, J.L. Daniel, S.P. Kunapuli, J. Biol. Med. 273 (1998) 2030.
[4] G.E. Jarvis, R.G. Humphries, M.J. Robertson, P. Leff, Br. J. Pharmacol. 129 (2000)
275.
[5] M.P. Abbracchio, C. Verderio, Novartis Found. Symp. 276 (2006) 91.
[6] C. Léon, C. Vial, C. Gachet, P. Ohlmann, B. Hechler, J.P. Cazenave, A. Lecchi,
M. Cattaneo, Thromb. Haemost. 81 (1999) 775.
[7] E.A. Barnard, J. Simon, Trends Pharmacol. Sci. 22 (2001) 388.
[8] F. Marteau, E. Le Poul, D. Communi, C. Labouret, P. Savi, J.M. Boeynaems,
N.S. Gonzalez, Mol. Pharmacol. 64 (2003) 104.
[9] M. Cattaneo, C. Gachet, Arterioscler. Thromb. Vasc. Biol. 19 (1999) 2281.
[10] P. Savi, J.M. Pereillio, M.F. Uzabiaga, J. Combalbert, C. Picard, J.P. Maffrand,
M. Pascal, J.M. Herbert, Thromb. Haemost. 84 (2000) 891.
[11] J.M. Boeynaems, H. van Giezen, P. Savi, J.M. Herbert, Curr. Opin. Invest. Drugs 6
(2005) 275.
[12] S. Yusuf, F. Zhao, S.R. Mehta, S. Chrolavicius, G. Tognoni, K.K. Fox, N. Engl. J.
Med. 345 (2001) 494.
[13] S.R. Steinhubl, P.B. Berger, J.T. Mann III, E.T. Fry, A. DeLago, C. Wilmer,
E.J. Topol, JAMA 288 (2002) 2411 (published correction appears in JAMA
289:987 (2003)).
[14] M.S. Sabatine, C.P. Cannon, C.M. Gibson, J.L. López-Sendón, G. Montalescot,
P. Theroux, M.J. Claeys, F. Cools, K.A. Hill, A.M. Skene, C.H. McCabe,
E. Braunwald, N. Engl. J. Med. 352 (2005) 1179.
[48] A. Lanver, H.G. Schmalz, Molecules 10 (2005) 508.
[49] M.J. Robins, P.W. Hatfield, Can. J. Chem. 60 (1982) 547.
[50] C.J. Marasco Jr., D.L. Kramer, J. Miller, C.W. Porter, C.J. Bacchi, D. Rattendi,
L. Kucera, N. Iyer, R. Bernacki, P. Pera, J.R. Sufrin, J. Med. Chem. 45 (2002)
5112.
[51] S.P. Vincent, S. Grenier, C. Mioskowski, C. Salesse, L. Lebeau, Bioorg. Med.
Chem. Lett. 11 (2001) 1185.
[52] B. Pavan, A. Dalpiaz, N. Ciliberti, C. Biondi, S. Manfredini, S. Vertuani, Mole-
cules 13 (2008) 1035.
N.M. Nielsen, H. Bundgaard, J. Med. Chem. 32 (1989) 727.
[53]
[54] P. Masson, M.T. Froment, P.L. Fortier, J.E. Visicchio, C.F. Bartels, O. Lockridge,
Biochim. Biophys. Acta 1387 (1998) 41.
[15] M.S. Sabatine, C.P. Cannon, C.M. Gibson, J.L. López-Sendón, G. Montalescot,
P. Theroux, B.S. Lewis, S.A. Murphy, C.H. McCabe, E. Braunwald, JAMA 294
(2005) 1224.
[16] J.L. Anderson, C.D. Adams, E.M. Antman, C.R. Bridges, R.M. Califf, D.E. Casey Jr.,
W.E. Chavey 2nd, F.M. Fesmire, J.S. Hochman, T.N. Levin, A.M. Lincoff,
E.D. Peterson, P. Theroux, N.K. Wenger, R.S. Wright, Circulation 116 (2007) 803.
[17] J.P. Bassand, C.W. Hamm, D. Ardissino, E. Boersma, A. Budaj, F. Fernández-
Avilés, K.A. Fox, D. Hasdai, E.M. Ohman, L. Wallentin, W. Wijns, Eur. Heart J. 28
(2007) 1598.
[55] A. Dalpiaz, B. Cacciari, C.B. Vicentini, F. Bortolotti, G. Spalluto, S. Federico,
B. Pavan, F. Vincenzi, P.A. Borea, K. Varani, Mol. Pharm. 9 (2012) 591.
[56] A. Hampton, J. Am. Chem. Soc. 83 (1961) 3640.
[57] Y. Xu, H. Jin, Z. Yang, L. Zhang, L. Zhang, Tetrahedron 65 (2009) 5228.
[58] J. Howarth, P. Kenny, S. McDonnell, A. O’Connor, Bioorg. Med. Chem. Lett. 13
(2003) 2693.
[59] S. Dionisotti, S. Ferrara, C. Molta, C. Zocchi, E. Ongini, J. Pharmacol. Exp. Ther.
298 (1996) 726.
[60] O.H. Lowry, N.J. Rosebrough, A.L. Farr, R.J. Randall, J. Biol. Chem. 193 (1951)
265.
[18] P.A. Gurbel, K.P. Bliden, Am. J. Cardiol. 91 (2003) 1123.
[19] S. Matetzky, B. Shenkman, V. Guetta, M. Shechter, R. Bienart, I. Goldenberg,
I. Novikov, H. Pres, N. Savion, D. Varon, H. Hod, Circulation 109 (2004) 3171.