Pentamidine: A Non-Peptide GPIIb/IIIa Antagonist – In Vitro Studies on Platelets from Humans and Other Species

Summary

In this paper we show that the non-peptide anti-parasite agent pentamidine is a broad spectrum anti-platelet agent with an IC₅₀ of 1.1 µM in ADP-induced platelet aggregation in human platelet rich plasma (PRP). It had similar activity when collagen, arachidonic acid, platelet activating factor, thrombin and epinephrine were used. It had no effect on platelet intracellular cAMP levels. It inhibited ¹²⁵I-fibrinogen, ¹²⁵I-fibronectin and ¹²⁵I-von Willebrand factor binding to ADP-activated fixed platelets with IC50 values of 160, 160 and 60 nM respectively. Pentamidine showed a high degree of species selectivity with slightly less activity in monkey and dog PRP and little activity in guinea pig, rabbit, rat and mouse PRP compared with human. This was similar to the other RGD analogues tested. This species specificity was shown to be dependent on the species of platelets and independent of the species of fibrinogen. Thus, pentamidine is a potent non-peptide inhibitor of fibrinogen binding to GPIIb/IIIa.

• REFERENCES

• 1 Trip MD, Cas VM, vanCapelle FJC, Vreeken J. Platelet hyperactivity and prognosis in survivors of myocardial infarction. N Engl J Med 1990; 322: 1549-1554
• 2 Henderson WG, Goldman S, Copeland JG, Moritz TE, Harker LA. Antiplatelet or anticoagulant therapy after coronary bypass surgery a meta-analysis of clinical trials. Ann Intern Med 1989; 111: 743-750
• 3 Stein B, Fuster V, Israel DM, Cohen M, Badimon L, Badimon J, Chesebro J. Platelet inhibitor agents in cardiovascular disease: an update. JACC 1989; 14(4): 813-836
• 4 RISC group Risk of myocardial infarction and death during treatment with low dose aspirin and intravenous heparin in men with unstable coronary artery disease. Lancet 1990; 336: 827-830
• 5 Coller B. Platelets and thrombolytic therapy. N Engl J Med 1990; 322: 33-42
• 6 Patrono C. Aspirin and human platelets: from clinical trials to acetylation of cyclooxygenase and back. TIPS 1989; 10: 453-458
• 7 Whittle BJR, Moncada S, Vane TR. Comparison of the effects of prostacyclin (PGI2), prostaglandin El and D2 on platelet aggregation in different species. Prostaglandins 1978; 16: 373-388
• 8 Topol EJ, Ellis SG, Califf RM, George BS, Stump DC, Bates ER, Nabel EG, Walton JA, Candela RJ, Lee KL, Kline EM, Pitt B. The thrombolysis and angioplasty in myocardial infarction (TAMI) 4 study group Combined tissue-type plasminogen activator and prostacyclin therapy for acute myocardial infarction. JACC 1989; 14(4): 877-884
• 9 Marguerie GA, Plow EF, Edington TS. Human platelets posses an inducible and saturable receptor specific for fibrinogen. J Biol Chem 1979; 254(12): 5357-5363
• 10 Hawiger J, Kloczewiak M, Bednarek MA, Timmons S. Platelet receptor recognition domains on the α chain of human fibrinogen: Structure-function analysis. Biochemistry 1989; 28: 2909-2914
• 11 Kloczewiak M, Timmons S, Bednarek MA, Sakon M, Hawiger J. Platelet receptor recognition domain on the γ-chain of human fibrinogen and its synthetic peptide analogues. Biochemistry 1989; 28: 2915-2919
• 12 Nachman RL, Leung LLK. Complex formation of platelet membrane glycoproteins IIb and IIa with fibrinogen. J Clin Invest 1982; 69: 263-269
• 13 Ruoslahti E. Integrins. J Clin Invest 1991; 87: 1-5
• 14 Coller BS, Folts JD, Smith SR, Scudder LE, Lordan R. Abolition of in vivo platelet thrombus formation in primates with monoclonal antibodies to the platelet GPIIb/IIIa receptor. Circulation 1989; 80: 1766-1774
• 15 Yasuda T, Gold HK, Leinbach RC, Saito T, Guerrero JL, Jang I, Holt R, Fallon JT, Collen D. Lysis of plasminogen activator-resistant platelet rich coronary artery thrombus with combined bolus injection of recombinant tissue type plasminogen activator and anti-platelet GPIIb/IIIa antibody. JACC 1990; 16(7): 1728-1735
• 16 Gan ZR, Gould RJ, Jacobs JW, Friedman PA, Polokoff MA. Echistatin: a potent platelet aggregation inhibitor from the venom of the viper Echis carinatus . J Biol Chem 1988; 263(36): 19827-19832
• 17 Mohri H, Hashimoto K, Noguchi T, Matsuzaki M, Ohkubo T, Ohba M, Kumagai H. The effects of cycloGRGDSPA on platelet aggregation (abstr, Japanese). Blood Vessel 1989; 20 (5) 431
• 18 Nichols A, Vasko J, Koster P, Smith J, Barone F, Nelson A, Stadel J, Powers P, Rhodes G, Miller-Stein C, Boppana V, Bennet D, Berry D, Romoff T, Calvo R, Ali F, Sorenson E, Samanen J. SK+F 106760, a novel GPIIIb/IIIa antagonist: antithrombotic activity and potentiation of streptokinase-mediated thrombolysis (abst). Eur J Pharm 1990; 183: 2019
• 19 Hodohara K, Fujiyama Y, Hosada S, Yasunaga K. Effect of amidinonaphtol derivatives on the cellular interaction with adhesive proteins. Blood Vessel 1989; 20: 553-559
• 20 Harfenist EJ, Packham MA, Mustard JF. Effects of the cell adhesion peptide Arg-Gly-Asp-Ser, on responses of washed platelets from humans, rabbits and rats. Blood 1988; 71: 132-136
• 21 Offord SJ, Shatynski DA, Moore JB, Falotico R, Tobia AJ. Arg-Gly-Asp-Ser (RGDS) does not inhibit mouse platelet aggregation or protect mice against arachidonic acid-induced lethality. Pharmacologist 1989; 31 (3) 163
• 22 Nicholson NS, Panzer-Knodle SG, Salyers AK, Taite BB, King LW, Miyano M, Gorczynski RJ, Williams MH, Zupec ME, Tjoeng FS, Adams SP, Feigen LP. Antiplatelet and antithrombotic effects of platelet glycoprotein IIb/IIIa (GPIIb/IIIa) inhibition by arginine-glycine-aspartic acid-serine (RGDS) and arginine-glycine-aspartic acid (RGD) (o-me) Y (SC-46749). J Pharm Exp Ther 1991; 256: 876-882
• 23 Beviglia L, Rossi C, Calabrese R, Soszka T, Niewiarowski S, Poggi A. Potent anti-thrombotic effect of albolabrin, a disintegrin with RGD sequence isolated from viper venom (abst). Thromb Res Suppl 1991; XIII: 40
• 24 Cadroy Y, Houghton RA, Hanson SR. RGDV peptide selectivity inhibits platelet-dependent thrombus formation in vivo-studies using a baboon model. J Clin Invest 1989; 84: 939-944
• 25 Geratz JD, Tidwell RR, Brinkhous KM, Mohammad SF, Dann O, Loewe H. Specific inhibition of platelet agglutination and aggregation by aromatic amidino compounds. Thromb Haemostas 1978; 39: 411-425
• 26 Goa K, Campoli-Richards DM. Pentamidine Isethionate: A review of its antiprotozoal activity, pharmacokinetic properties and therapeutic use in Pneumocystis carinii pneumonia. Drugs 1987; 33: 242-258
• 27 Anderson R, Boedecker M, Ma M, Goldstein JC. Adverse reactions associated with pentamidine isothionate in AIDS patients: Recommendations for monitoring therapy. Drug Intell Clin Pharm. Drug Intell Clin Pharm 1986; 20: 862-868
• 28 Golden JA, Chernoff D, Hollander H, Feigel D, Conte JE. Prevention of Pneumocystis carinii pneumonia by inhaled pentamidine. Lancet 1989; i: 654-657
• 29 Kekwick A, Mackay ME, Nance MH, Record BR. The purification of human fibrinogen. Biochemistry 1955; 60: 671-683
• 30 Fay PJ, Chavin SI, Schroeder D, Young FE, Marder VJ. Purification and characterization of a highly purified factor VIII consisting of a single type of polypeptide chain. Proc Natl Acad Sci (USA) 1982; 79: 7200-7204
• 31 Greenwood FC, Hunter WM, Glover JS. The preparation of ^13II-labelled human growth hormone of high specific radioactivity. Biochem J 1963; 89: 114-123
• 32 Tuszynski G, Knight L, Piperno J. Removal of free radioiodide from solutions of radiolabelled protein by a rapid centrifugation technique. J Labeled Compounds and Radiopharmaceuticals 1981; XVIII(1–2): 206-207
• 33 Plow E, Marguerie G. Inhibition of fibrinogen binding to human platelets by the tetrapeptide glycyl-L-prolyl-L-arginyl-L-proline. Proc Natl Acad Sci (USA) 1982; 79: 3711-3715
• 34 Klimm J, Kloczewiak M, Linden J. Comparison of the inhibitory activity of free and albumin bound thromboxane receptor antagonist BM 13505 on U46619 induced platelet aggregation. Thromb Haemostas 1989; 62: 191
• 35 Motoyama Y, Sakata Y, Seki J, Asada T, Namikawa Y, Horiai H, Ono T. The effects of TFC-612, a 7-thia prostaglandin E1 derivative, on platelet aggregation. Thromb Res 1991; 62: 471-480
• 36 O’Brien J, Etherington M, Salmon G. DN9693, a phosphodiesterase inhibitor compared with prostaglandin El and prostacyclin in four platelet function tests. Thromb Res 1988; 50: 237-241
• 37 Lee H, Paton R, Ruan C, Caen J. The in vitro effect of ticlopidine on fibrinogen and factor VIII binding to human platelets. Thromb Haemostas 1981; 46(3): 590-592
• 38 Geratz J, Whitmore A. Diamidino-α,ω-diphenoxylalkanes. Structure activity relationships for the inhibition of thrombin, pancreatic Kallikrein and trypsin. J Med Chem 1973; 16(9): 970-975
• 39 Turini P, Kurooka S, Steer M, Corbascio A, Singer T. The action of phenylmethylsulfonylfluoride on human acetylcholin esterase, chymotrypsin and trypsin. J Pharm Exp Ther 1969; 167: 098-104
• 40 Kroll M, Schafer A. Biochemical mechanisms of platelet activation. Blood 1989; 74: 1181-1195
• 41 Allain J, Cooper H, Wagner R, Brinkhous K. Platelets fixed with paraformaldehyde: a new reagent for assay of von Willebrand factor and platelet aggregation factor. J Lab Clin Med 1975; 85: 318-328
• 42 Plow E, Marguerie G, Ginsberg M. Fibrinogen, fibrinogen receptors and the peptides that inhibit these interactions. Biochem Pharm 1987; 36: 4035-4040
• 43 Plow EF, Marguerie GA, Ginsberg MH. Fibronectin binding to thrombin-stimulated platelets: Evidence for fibrin(ogen) independent and dependent pathways. Blood 1985; 66(1): 26-32
• 44 Verhallen PFJ, Barth M. Species comparison of anti-aggregatory properties of three fibrinogen receptor antagonists: RGDS, echistatin and the carboxyterminal duodecapeptide of the fibrinogen gamma-chain (abst). Thromb Haemostas 1990; 65 (6) 1144
• 45 Pottratz S, Martin II W. Role of fibronectin in Pneumocystis carinii attachment to cultured lung cells. J Clin Invest 1990; 85: 351-356
• 46 Pottratz S, Martin II W. Mechanism of Pneumocystis carinii attachment to cultured rat alveolar macrophages. J Clin Invest 1990; 86: 1678-1683
• 47 Osei K, Falko J, Nelso K, Stephens R. Diabetogenic effect of pentamidine. In vitro and in vivo studies in a patient with a malignant insulinoma. Am J Med 1984; 77: 41-46
• 48 Miller R, Semple S. Bronchial bleeding with nebulised pentamidine. Lancet 1988; ii: 1488