Chlorobutanol, a Preservative of Desmopressin, Inhibits Human Platelet Aggregation and Release In Vitro

Summary

Therapeutic preparations of desmopressin for parenteral use contain the preservative chlorobutanol (5 mg/ml). We show here that chlorobutanol is a potent inhibitor of platelet aggregation and release. It exhibited a significant inhibitory activity toward several aggregation inducers in a concentration- and time-dependent manner. Thromboxane B₂ formation, ATP release, and elevation of cytosolic free calcium caused by collagen, ADP, epinephrine, arachidonic acid and thrombin respectively were markedly inhibited by chlorobutanol. Chlorobutanol had no effect on elastase- treated platelets and its antiplatelet effect could be reversed. It is concluded that the antiplatelet effect of chlorobutanol is mainly due to its inhibition on the arachidonic acid pathway but it is unlikely to have a nonspecitic toxic effect. This antiplatelet effect of chlorobutanol suggests that desmopressin, when administered for improving hemostasis, should not contain chlorobutanol as a preservative.

• References

• 1 Mannucci PM, Remuzzi G, Pusineri I, Lombardi R, Valsecchi C, Mecca G, Zimmerrnan TS. Deamino-S-arginine vasopressin shortens the bleeding time in uremia. N Engl J Med 1983; 308: 8-12
• 2 Watson A J, Keogh JA. 1-deamino-8-D-arginine vasopressin (DDAVP): a potential new treatment for the bleeding diathesis of acute renal failure. Pharmatherapeutica 1984; 3: 618-622
• 3 Canavese C, Salomone M, Pacitti A, Angiarotti G, Calitri V. Reduced response of uraemic bleeding time to repeated doses of desmopressin (letter). Lancet 1985; 8433: 867-868
• 4 Hermsmeyer K, Aprigliano O. Effects of chlorobutanol and bradykinin on myocardial excitation. Am J Physiol 1976; 230: 306-10
• 5 Sequeira SJ, McKenna TJ. Chlorbutol, a new inhibitor of aldosterone biosynthesis identified during examination of heparin effect on aldosterone production. J CIin Endocrinol Metab 1986; 63: 780-784
• 6 Mustard JE, Perry DW, Ardlie NG, Packham MA. preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204
• 7 Born GV R. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962; 194: 927-9
• 8 Deluca M, McElroy WD. d-Luciferase and Luciferin. In: Methods in Enzymology Colowich SP, Kaplan NO. eds Academic Press 1978; Vol. LVII, 3
• 9 Kornecki E, Ehrlich YH, De Mars DD, Lenox R. Exposure of fibrinogen receptors in human platelets by surface proteolysis with elastase. J Clin Invest 1986; 77: 750-756
• 10 Erne P, Bolli P, Burgisser E, Buhler FR. Correlation of platelet calcium with blood pressure: effect of antihypertensive therapy. N Engl J Med 1984; 310: 1084-8
• 11 Grynkiewicz G, Peonie M, Tsien Ry. A new generation of ca²⁺ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440-50
• 12 Tsien RX, Pozzan T, Rink TJ. Calcium homeostasis in intact Iymphocytes: cytoplasmic free calcium monitored with a new, intracellular trapped fluorescent indicator. J cell Biol 1982; 94: 325-334
• 13 Hamberg M, Svensson J, Wakabayshi T, Samuelsson B. Isolation and structure of two prostaglandin endoperoxides that cause platelet aggregation. Proc Natl Acad Sci USA 1974 72: 345-349
• 14 Hamberg M, Svensson J, Samuelsson B. Thromboxanes - a new group of biologically active compounds derived from prostaglandin endoperoxides. Proc Natl Acad Sci USA 1975; 72: 2994-2998
• 15 Plow EE, Marguerie GA. Participation of ADP in the binding of fibrinogen to thrombin-stimulated platelets. Blood 1980; 56: 553-555
• 16 Nachman RL, Leung LL K. Complex formation of platelet membrane glycoproteins IIb and IIIa with fibrinogen. J Clin Invest 1982; 69: 263-269
• 17 McEver RR, Bennett EM, Martin NM. Identification of two structurally and functionally distinct sites on human platelet membrane glycoprotein IIFIIIa using monoclonal antibodies. J Biol Chem 1983; 258: 5269-5275
• 18 Greenberg JP, Packham MA, Guccione MA, Harfenist EJ, Orr JI, Kinlough-Rathbone RL, Perry DW, Mustard JE. The effect of pretreatment of human or rabbit platelets with chymotrypsin on their responses to human fibrinogen and aggregating agents. Blood 1979; 54: 753-765
• 19 Niewiarowski S, Budzynski AZ, Morinelli TA, Budzynski TM, Stewart GJ. Exposure of fibrinogen receptor on human platelets by proteolytic enzymes. J Biol Chem 1981; 256: 917-925
• 20 Feinstein MB, Halenda SP, Zavoico GB. Calcium and platelet function. In: Calcium and Cell Physiology Marme D. ed Springer Verlag; New York: 1985: 345-376
• 21 Daniel JL, Selak MA, Purdon AD, Salganicoff L. Methods for the study of the role of calcium in platelet function. In: Methods for Studing Platelets and Megakaryocytes Colman Rw. ed Alan R. Liss, Inc.; New York: 1987: 185-215
• 22 Feinstein MB, Zavoico GB. Calcium and cyclic AMP. In: The Platelets Longenecker GL. ed Academic Press, Inc; 1985: 234-269
• 23 Hallam TJ, Rink TJ. Insights into platelet function gained with fluorescent. Ca²⁺ indicators. In: Platelets in Biology urd Pathology III Maclntyre DE. ed Elsevier; 1987: 353-70
• 24 Ditullio NW, Kinter LB, Storer BL. Pharmacological characterization of vasopressin aggregation response of human platelets. Thromb Res 1985; 40: 425-432
• 25 Vicente Y, Alberca I, Mannucci PM. Reduced effect of exercise and DDAVP on factor VIII-von Willebrand factor and plasminogen activator after sequential application of both the stimuli. Thromb Haemostas 1984; 51: 129-130
• 26 Holmberg L, Nilsson IM. AHF related protein in clinical praxis. Scand J Haematol 1974; 12: 221-231
• 27 Tung C, Graham GG, Wade DN, Williams KM. The pharmacokinetics of chlorbutol in man. Biopharmacol Drug Dispos 1982; 3: 371-378