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Thromb Haemost 1989; 61(01): 035-042
DOI: 10.1055/s-0038-1646523
Original Article
Schattauer GmbH Stuttgart

R 68 070: Thromboxane A2 Synthetase Inhibition and Thromboxane A2/Prostaglandin Endoperoxide Receptor Blockade Combined in One Molecule - I. Biochemical Profile In Vitro

F De Clerck
The Department of Haematology, Janssen Research Foundation, Beerse, Belgium
,
J Beetens
The Department of Haematology, Janssen Research Foundation, Beerse, Belgium
,
D de Chaffoy de Courcelles
1   The Department of Biochemistry, Janssen Research Foundation, Beerse, Belgium
,
E Freyne
2   The Department of Chemistry, Janssen Research Foundation, Beerse, Belgium
,
P A J Janssen
The Department of Haematology, Janssen Research Foundation, Beerse, Belgium
› Institutsangaben
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Publikationsverlauf

Received 25. April 1988

Accepted after revision 05. Oktober 1988

Publikationsdatum:
24. Juli 2018 (online)

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Summary

R 68 070 or (E)-5-[[[(3-pyridinyl)[3-(trifluoromethyl)phenyl]- methylen]amino]oxy] pentanoic acid (Janssen Research Foundation, Belgium) combines specific thromboxane A2 (TXA2) synthetase inhibition with TXA2/prostaglandin endoperoxide receptor blockade in one molecule.

In vitro, the compound specifically inhibits the production of TXB2 from [14C] arachidonic acid by washed human platelets (IC50 = 8.2 × 10-9 M) and by platelet microsomes (IC50 = 3.6 × 10-9 M), of MDA (IC50 = 1.91 × 10-8 M) and of TXB2 (IC50 = 1.47 × 10-8 M) by thrombin-coagulated human platelet-rich plasma (P.R.P.) and whole blood respectively and increases the levels of PGD2, PGE2, PGF2α and 6-keto-PGF1α. The activity of cyclo-oxygenase-, prostacyclin synthetase-, 5-, 12- and 15-lipoxygenase-enzymes are not affected. Additionally, R 68 070 inhibits human platelet aggregation in P.R.P. induced by U 46619 3 × 10-7 M to 2 × 10-6 M (IC50 = 2.08 × 10-6 M to 2.66 × 10-5 M), collagen 0.5 to 2 μg/ml (IC50 = 2.85 × 10-6 M to 4.81 × 10-5 M), arachidonic acid 7.5 × 10-4 M to 2 × 10- M (IC50 = 2.1 × 10-8 M to 3.3 × 10-8 M) and the U 46619 (1 × 10-7 M)-induced accumulation of [32P] phosphatidic acid (IC50 = 5.24 × 10-7 M) in washed human platelets. Collagen (0.75 μg/ml)-induced ATP release (IC50 = 4.1 × 10-6 M), ADP (1 to 2.5 × 10-6 M)-induced second wave aggregation (IC50 = 3.19 × 10-6 M) in P.R.P. as well as the collagen (1 μg/ml)-induced adhesion/aggregation reaction in human whole blood (IC50 = 1.02 × 10-5 M) are reduced as well by the compoun.

Primary platelet reactions induced by serotonin, ADP, PAF, or A 23187, platelet adenylate cyclase- and cAMP phosphodiesterase-activity, and platelet inhibitory activities of PGD2, PGI2, PGE2, PGE1 are not modified by R 68 070.

This biochemical profile is compatible with a dual mechanism of action of R 68 070, namely TXA2 synthetase inhibition at low concentrations, plus additionally TXA2/prostaglandin endoperoxide receptor blockade at higher concentrations

Key words

R 68 070 - TXA2-synthetase - Receptor blockade - Man - Experimental models
 

  • References

  • 1 Needleman P, Moncada S, Bunting S, Vane JR, Hamberg M, Samuelsson B. Identification of an enzyme in platelet microsomes which generates thromboxane A2from prostaglandin endoperoxides. Nature. 1976; 261: 558-560
    PubMedGoogle Scholar
  • 2 Samuelsson B, Goldyne H, Granström E, Hamberg M, Hammarström S, Malmsten C. Prostaglandins and thromboxanes. Annu Rev Biochem 1978; 47: 997-1029
    CrossrefPubMedGoogle Scholar
  • 3 FitzGerald GA, Reilly IA G, Pedersen AK. The biochemical pharmacology of thromboxane synthetase inhibition in man. Circulation 1985; 72: 1194-1201
    CrossrefPubMedGoogle Scholar
  • 4 Vermylen J, Carreras LO, Van Schaeren J, De Freyn G, Machin SJ, Verstraete M. Thromboxane synthetase inhibition as anti-thrombotic strategy. Lancet 1981; 1: 1073-1075
    PubMedGoogle Scholar
  • 5 De Freyn G, Deckmyn H, Vermylen J. A thromboxane synthetase inhibitor reorients endoperoxide metabolism in whole blood towards prostacyclin and prostaglandin E2 . Thromb Res 1982; 26: 389-400
    CrossrefPubMedGoogle Scholar
  • 6 Gresele P, Deckmyn H, Huybrechts E, Vermylen J. Serum albumin enhances the impairment of platelet aggregation with thromboxane synthetase inhibition by increasing the formation of prostaglandin D2 . Biochem Pharmacol 1984; 33: 2083-2088
    CrossrefPubMedGoogle Scholar
  • 7 Bertelé V, Cerletti C, Schieppati A, Diminnio G, de Gaetano G. Inhibition of thromboxane synthetase does not necessarily prevent platelet aggregation. Lancet 1981; 1: 1057-1058
    PubMedGoogle Scholar
  • 8 Hornby EJ, Skidmore IF. Evidence that prostaglandin endoperoxides can induce platelet aggregation in the absence of thromboxane A2production. Biochem Pharmacol 1982; 31: 1153-1160
    CrossrefPubMedGoogle Scholar
  • 9 Bertelé V, Tomasiak M, Falanga A, Cerletti C, de Gaetano G. Aspirin inhibits platelet aggregation but not because it prevents thromboxane synthesis. Lancet 1982; 2: 775
    PubMedGoogle Scholar
  • 10 Rybicki JP, Le Breton GC. Prostaglandin H2directly lowers human platelet cAMP levels. Thromb Res 1983; 30: 407-417
    CrossrefPubMedGoogle Scholar
  • 11 Gresele P, Amout J, Deckmyn H, Vermylen J. Endogenous antiaggregatory prostaglandins can contribute to inhibition of hemostasis: a pharmacological study in vivo in humans. Adv Prostaglandin Thromboxane Leukotriene Res 1987; 17: 248-253
    PubMedGoogle Scholar
  • 12 Gresele P, Van Houtte E, Arnout J, Deckmyn H, Vermylen J. Thromboxane synthetase inhibition combined with thromboxane receptor blockade: a step forward in antithrombotic strategy. Thromb Haemostas 1984; 52: 364
    PubMedGoogle Scholar
  • 13 Bertelé V, de Gaetano G. Potentiation by dazoxiben, a thromboxane synthetase inhibitor, of platelet aggregation inhibitory activity of a thromboxane receptor antagonist and of prostacyclin. Eur J Pharmacol 1982; 85: 331-333
    CrossrefPubMedGoogle Scholar
  • 14 Rajtar G, Cerletti C, Castagnoli MN, Bertelé V, de Gaetano G. Prostaglandins and human platelet aggregation. Implications for the anti-aggregating activity of thromboxane-synthetase inhibitors Biochem Pharmacol 1985; 34: 307-310
    CrossrefPubMedGoogle Scholar
  • 15 FitzGerald DJ, Fragetta J, Fenelon LC, FitzGerald GA. Thromboxane synthetase inhibition and thromboxane/endoperoxide receptor antagonism in a chronic canine model of coronary thrombosis. Adv Prostaglandin Thromboxane Leukotriene Res 1987; 17: 496-500
    PubMedGoogle Scholar
  • 16 Beetens JR, Van Hoydonck AE, Herman AG. Stimulation of prostacyclin production by vitamin C in ram seminal vesicle microsomes: possible mode of action. Arch Int Pharmacodyn Ther 1985; 277: 56-65
    PubMedGoogle Scholar
  • 17 Beetens JR, Loots W, Somers Y, Coene MC, De Clerck F. Ketoconazole inhibits the biosynthesis of leukotrienes in vitro and in vivo. Biochem Pharmacol 1986; 35: 883-891
    CrossrefPubMedGoogle Scholar
  • 18 Takeguchi C, Kohno E, Sih CJ. Mechanism of prostaglandin biosynthesis I. Characterization and assay of bovine prostaglandin synthetase. Biochemistry 1971; 10: 2372-2376
    CrossrefPubMedGoogle Scholar
  • 19 De Clerck F, Van Nueten JM. Platelet-mediated vascular contractions. Inhibition of flunarizine, a calcium-entry blocker Biochem Pharmacol 1983; 32: 765-771
    CrossrefPubMedGoogle Scholar
  • 20 Beetens JR, Van Den Bossche R, Herman AG. Production of 6-oxo-PGF1αby aorta rings in the presence of ascorbic acid. Arch Int Pharmacodyn Ther 1982; 256: 151-152
    PubMedGoogle Scholar
  • 21 McMillan RM, MacIntyre DE, Gordon JC. Simple sensitive fluorimetric assay for malondialdehyde production by blood platelets. Thromb Res 1977; 11: 425-428
    CrossrefPubMedGoogle Scholar
  • 22 Patrono C. et al Low dose aspirin and inhibition of thromboxane B2production in healthy subjects. Thromb Res 1980; 17: 317-327
    CrossrefPubMedGoogle Scholar
  • 23 Salomon Y. Adenylate cyclase assay. Adv Cyclic Nucleotide Res 1979; 10: 35-55
    PubMedGoogle Scholar
  • 24 Bult H, Beetens J, Herman A. Blood levels of 6-oxo-prostaglandin Flαduring endotoxin-induced hypotension in rabbits. Eur J Pharmacol 1980; 63: 47-56
    CrossrefPubMedGoogle Scholar
  • 25 De Chaffoy de CourcellesD, Leysen JE, De Clerck F, Van Belle H, Janssen PA J. Evidence that phospholipid turnover is the signal transducing system coupled to serotonin-S2receptor sites. J Biol Chem 1985; 260: 7603-7608
    PubMedGoogle Scholar
  • 26 Mills DC B, Smith JB. The influence on platelet aggregation of drugs that affect the accumulation of adenosine 3’:5’-cyclic monophosphate in platelets. Biochem J 1971; 121: 185-196
    CrossrefPubMedGoogle Scholar
  • 27 De Clerck F, Xhonneux B, Tollenaere JP, Janssen PA J. Dependence of the antagonism at human platelet 5-HT2receptors by ketanserin on the reaction pH. Thromb Res 1985; 40: 581-596
    CrossrefPubMedGoogle Scholar
  • 28 De Clerck F, Van Gorp L, Beetens J, Reneman RS. Platelet- mediated vascular permeability in the rat: a predominant role for 5-hydroxytryptamine. Thromb Res 1985; 38: 321-339
    CrossrefPubMedGoogle Scholar
  • 29 Feinman RD, Lubowsky J, Charo I, Zabinski M. The lumiaggregometer: a new instrument for simultaneous measurement of secretion and aggregation by platelets. J Lab Clin Med 1977; 90: 125-129
    PubMedGoogle Scholar
  • 30 De Clerck F, Somers Y, Van Gorp L. Platelet-vessel wall interactions in haemostasis: implication of 5-hydroxytryptamine. Agents Actions 1984; 15: 627-635
    CrossrefPubMedGoogle Scholar
  • 31 Gresele P, Arnout J, Janssens W, Deckmyn H, Lemmens J, Vermylen J. BM 13177, a selective blocker of platelet and vessel wall thromboxane receptors, is active in man. Lancet 1984; : 991-994
    PubMedGoogle Scholar
  • 32 Brittain RT, Boutal L, Carter MC, Coleman RA, Collington EW, Geisow HP, Hallett P, Hornby EJ, Humphrey PP A, Jack D, Kennedy I, Lumley P, McCabe PJ, Skidmore IF, Thomas M, Wallis CJ. AH 23848 a thromboxane receptor-blocking drug that can clarify the pathophysiologic role of thromboxane A2 . Circulation 1985; 72: 1208-1218
    CrossrefPubMedGoogle Scholar
  • 33 Smith JB, Jubil W. OKY-1581 a selective inhibitor of thromboxane synthesis in vivo and in vitro. Prostaglandins 1981; 22: 353-363
    CrossrefPubMedGoogle Scholar
  • 34 Ruppert D, Weithman KU. HL 725, an extremely potent inhibitor of platelet phosphodiesterase and induced platelet aggregation in vitro. Life Sci 1982; 31: 2037-2043
    CrossrefPubMedGoogle Scholar
  • 35 Coleman RA, Humphrey PP A, Kennedy I, Levy GP, Lumley P. U 46619, a selective thromboxane A2-like agonist. Br J Pharmacol 1980; 68: 1278
    PubMedGoogle Scholar
  • 36 Coleman RA, Humphrey PP A, Kennedy I, Levy GP, Lumley P. Comparison of the actions of U 46619, a prostaglandin H2-analogue, with those of prostaglandin H2and thromboxane A2on isolated smooth muscle preparations. Br J Pharmacol 1981; 73: 773-778
    CrossrefPubMedGoogle Scholar
  • 37 Draper NR, Smith M. Applied Regression Analysis. Wiley & Sons; New York: 1981
    Google Scholar
  • 38 Pedersen EK, Watson ML, FitzGerald GA. Inhibition of thromboxane biosynthesis in serum; limitations of the measurements of immunoreactive 6-keto-PGF1α . Thromb Res 1983; 33: 99-103
    PubMedGoogle Scholar
  • 39 Carey F, Haworth D. Thromboxane synthetase inhibition: implications for prostaglandin endoperoxide metabolism. Prostaglandins 1986; 31: 47-59
    CrossrefPubMedGoogle Scholar
  • 40 Mehta J, Mehta P, Lawson DL, Ostrowski N, Brignon L. Influence of selective thromboxane synthetase blocker CGS-13080 on thromboxane and prostacyclin biosynthesis in whole blood: evidence for synthesis of prostacyclin by leukocytes from platelet-derived endoperoxides. J Lab Clin Med 1985; 106: 246-252
    PubMedGoogle Scholar
  • 41 Uotila P, Matintalo M. Inhibition of thromboxane synthetase by OKY-1581 stimulates the formation of PGE2, PGF2α, PGD2 and 6-keto-PGF1αin human platelets. Prostaglandins Leukotrienes Med 1984; 14: 41-46
    CrossrefPubMedGoogle Scholar
  • 42 Ambler J, Butler KD, Ku EC, Maguire ED, Smith JR, Wallis RB. CGS 12970: a novel, long acting thromboxane synthetase inhibitor. Br J Pharmacol 1985; 86: 497-504
    CrossrefPubMedGoogle Scholar
  • 43 Terashita Z, Imura Y, Tanabe M, Kawazoe K, Nishikawa K, Kato K, Terao S. CV-4151 - a potent, selective thromboxane A2synthetase inhibitor. Thromb Res 1986; 41: 223-237
    CrossrefPubMedGoogle Scholar
  • 44 Hiraku S, Taniguchi K, Wakitani K, Omawari N, Kira H, Miyamoto T, Okegawa T, Kawasaki A, Ujiie A. Pharmacological studies on the TXA2synthetase inhibitor (E)-3-[p-(1H-imidazol-1-ylmethyl)phenyl]- 2-propenoic acid (OKY-046). Jpn J Pharmacol 1986; 41: 393-401
    CrossrefPubMedGoogle Scholar
  • 45 Beetens JR, Claeys M, Herman AG. Antioxidants increase the formation of 6-oxo-PGF1α by ram seminal vesicle microsomes. Biochem Pharmacol 1981; 30: 2811-2851
    CrossrefPubMedGoogle Scholar
  • 46 Flower RJ, Cheung HS, Cushman DW. Quantitative determination of prostaglandins and malondialdehyde formed by the arachidonate oxygenase (prostaglandin synthetase) system of bovine seminal vesicle. Prostaglandins 1973; 4: 325-341
    CrossrefPubMedGoogle Scholar
  • 47 De Clerck F, Van Nueten JM. Platelet-mediated vascular contractions: inhibition of the serotonergic component by ketanserin. Thromb Res 1982; 27: 713-727
    CrossrefPubMedGoogle Scholar
  • 48 Patscheke H, Stegmeier K. BM 13.177 is a selective antagonist of prostaglandin H2and thromboxane A2in human platelets. IRCS Med Sci 1984; 12: 9-10
    PubMedGoogle Scholar
  • 49 Ogletree ML, Harris DN, Greenberg R, Haslanger MF, Nakane M. Pharmacological actions of SQ 29548, a novel selective thromboxane antagonist. J Pharmacol Exp Ther 1985; 234: 435-441
    PubMedGoogle Scholar
  • 50 Armstrong RA, Jones RL, Wilson NH. Ligand binding to thromboxane receptors on human platelets: correlation with biological activity. Br J Pharmacol 1983; 79: 953-964
    CrossrefPubMedGoogle Scholar
  • 51 De Clerck F, David JL. Pharmacological Control of platelet and red blood cell function in the microcirculation. J Cardiovasc Pharmacol 1981; 3: 1388-1412
    CrossrefPubMedGoogle Scholar
  • 52 Best LC, Holland TK, Jones PB B, Russell RG G. The interrelationship between thromboxane biosynthesis, aggregation and 5-hydroxytryptamine secretion in human platelets in vitro. Thromb Haemostas 1980; 43: 38-40
    PubMedGoogle Scholar
  • 53 Carter AJ, Heptinstall S. Platelet aggregation in whole blood: the role of thromboxane A2and adenosine diphosphate. Thromb Haemostas 1985; 54: 612-616
    PubMedGoogle Scholar
  • 54 Heptinstall S, Bevan J, Cockbill SR, Hanley SP, Parry MJ. Effects of a selective inhibitor of thromboxane synthetase on human blood platelet behaviour. Thromb Res 1980; 20: 219-230
    CrossrefPubMedGoogle Scholar
  • 55 Heptinstall S, Bientz N, Cockbill SR, Hanley SP, Peacock I. Different effects of thromboxane synthetase inhibitors on platelets from different individuals. Lancet 1982; 2: 1156
    PubMedGoogle Scholar
  • 56 Sills T, Heptinstall S. Effects of a thromboxane synthetase inhibitor and a cAMP phosphodiesterase inhibitor, singly and in combination, on platelet behaviour. Thromb Haemostas 1986; 55: 305-308
    PubMedGoogle Scholar
  • 57 Parise LV, Venton DL, Lebreton GC. Arachidonic acid-induced platelet aggregation is mediated by a thromboxane A2/prostaglandin H2receptor interaction. J Pharmacol Exp Ther 1984; 228: 240-244
    PubMedGoogle Scholar