Pharmacokinetics and Pharmacodynamics of a Low Molecular Weight Heparin (Enoxaparin) after Subcutaneous Injection, Comparison with Unfractionated Heparin – A Three Way Cross Over Study in Human Volunteers

Ana Victoria Bendetowicz; Suzette Béguin; H Caplain; H Coenraad Hemker

doi:10.1055/s-0038-1642435

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1994; 71(03): 305-313
DOI: 10.1055/s-0038-1642435

Original Article

Schattauer GmbH Stuttgart

Pharmacokinetics and Pharmacodynamics of a Low Molecular Weight Heparin (Enoxaparin) after Subcutaneous Injection, Comparison with Unfractionated Heparin – A Three Way Cross Over Study in Human Volunteers

Ana Victoria Bendetowicz

The Department of Biochemistry, Cardiovascular Research Institute Maastricht, University of Limburg, Maastricht, The Netherlands

,

Suzette Béguin

The Department of Biochemistry, Cardiovascular Research Institute Maastricht, University of Limburg, Maastricht, The Netherlands

,

H Caplain

¹Institut Aster, Paris, France

,

H Coenraad Hemker

The Department of Biochemistry, Cardiovascular Research Institute Maastricht, University of Limburg, Maastricht, The Netherlands

› Author Affiliations

Abstract

PDF Download

Summary

We determined, in volunteers, the plasma levels of heparin above and below the critical chainlength necessary for thrombin inhibition (ACLM and BCLM), from 1 to 24 h after subcutaneous injection of 5000IU unfractionated heparin (UFH), 40 mg enoxaparin and 1 mg/kg body weight of enoxaparin (LMWH) (n = 12 for each dose). The levels were calculated from the antithrombin- and anti-Xa activities using the specific activities of the materials injected. We also determined the course of thrombin- and of factor Xa generation after triggering the extrinsic system in the same samples. From the thrombin generation curves, we calculated the course of prothrombinase activity.

When the ACLM and BCLM plasma-levels are plotted against the inhibition of thrombin- and factor Xa generation, it appears that:

a) There is a unique dose response relationship between ACLM level and the inhibition of thrombin generation, independent of whether the ACLM is derived from UFH or LMWH. This relationship is not significantly altered by the BCLM appearing after LMWH injection.

b) There is a similar unique relationship between ACLM level and the inhibition of factor Xa generation, again independent of BCLM.

c) Inhibition of prothrombin activation hardly contributes to the overall effect on thrombin formation and is again independent of the source of ACLM.

d) ACLM levels were significantly higher after injection of LMWH than after UFH injection, even though the amounts of ACLM injected with the highest dose of LMWH were smaller than those administered in the UFH injection.

We conclude that the only functional difference between LMWH and UFH is the much higher bioavailability of the former. We surmise that, from the UFH injected, only the lower molecular weight species reach the circulation, i. e. a fraction similar to the ACLM injected with enoxaparin.

PDF (752 kb)

References

References
1 Johnson EA, Kirkwood TBL, Striding Y, Perez-Requejo JL, Ingram GIC, Bangham DR, Brozovic M. Four heparin preparations: anti-Xa potentiating effect of heparin after subcutaneous injection. Thromb Haemost 1976; 35: 586-91
2 Yin ET, Wessler S, Stoll PJ. Biological properties of the naturally occurring inhibitor to activated factor X. J Biol Chem 1971; 246: 3703-11
3 Hemker HC, Béguin S, Bendetowicz AV, Wielders S. The determination of the levels of unfractionated heparin and low molecular weight heparins in plasma. Their effect on thrombin mediated feedback reactions in vivo. Preliminary results on samples after subcutaneous injection. Haemostasis 1991; 21 (04) 258-72
4 Bendetowicz AV, Pacaud E, Béguin S, Uzan A, Hemker HC. On the relationship between molecular mass and anticoagulant activity of a low-molecular weight heparin (enoxaparin). Thromb Haemost 1992; 67: 556-62
5 Béguin S, Wielders S, Lormeau JC, Hemker HC. The mode of action of CY 216 and CY 222 in plasma. Thromb Haemost 1992; 67: 33-41
6 Béguin S, Kessels H, Dol F, Hemker HC. The consumption of Antithrombin III during coagulation, its consequences for the calculation of prothrombinase activity and the standardisation of heparin activity. Thromb Haemost 1992; 68: 136-12
7 Hemker HC, Willems GM, Béguin S. A computer assisted method to obtain the prothrombin activation velocity in whole plasma independent of thrombin decay processes. Thromb Haemostas 1986; 56: 9-17
8 Béguin S, Lindhout T, Hemker HC. The mode of action of heparin in plasma. Thromb Haemost 1988; 60: 457-62
9 Owren PA, Aas K. The control of dicumarol therapy and the quantitative determination of prothrombin and proconvertin. Scand J Clin Lab Invest 1951; 3: 201-18
10 Josso F, Prou-Wartelle O. Exploration de 1/hemostase In: Techniques en Hematologie. Alagille D. edts. Flammarion; Paris: 1972: 101-8
11 Lindhout T, Govers-Riemslag JWP, der Waart PVan, Hemker HC, Rosing J. Factor Va - Factor Xa interaction. Effect of phospholipid vesicles of different composition. Biochemistry 1982; 21: 5494-502
12 Pieters J, Lindhout T, Hemker HC. In situ generated thrombin is the only enzyme that effectively activates factor VIII and factor V in thromboplastin activated plasma. Blood 1989; 1021-4
13 Schoen P, Hemker HC, Lindhout T. Ratios of anti-factor Xa to anti-thrombin activities in recalcified human plasma. Brit J Haemat 1992; 81: 255-62
14 Hemker HC, Wielders S, Kessels H, Béguin S. Continuous registration of thrombin generation in plasma, its use for the determination of the thrombin potential. Thromb Haemost 1993; 70: 617-24
15 Béguin S, Mardiguian J, Lindhout T, Hemker HC. The Mode of Action of Low Molecular Weight Heparin Preparation (PK 10169) and Two of its Major Components on Thrombin Generation in Plasma. Thromb Haemost 1989; 61 (01) 30-4
16 Hirsh J, Levine MN. Low molecular weight heparin. Blood 1992; 79: 1-17
17 Danishefsky I, Eiber HB. Studies on the metabolism of heparin. Arch Biochem Biophys 1959; 85: 53-61
18 Shanberge JN, Gruhl M, Kitani T, Ambegaonkar S, Kambayashi J, Nakagawa M, Lenter D. Fractionated tritium labelled heparin studies in vitro and in vivo. Thromb Res 1978; 13: 767-83
19 Dawes J, Pepper DS. Catabolism of low dose heparin in man. Thromb Res 1979; 14: 845-60
20 Dawes J, Prowse CV, Pepper DSHeparin. LMW heparin and SP54 concentrations after subcutaneous injection. Thromb Res 1986; 44: 683-93
21 Boneu B, Caranobe C, Sie P. Pharmacokinetics of heparin and low molecular weight heparin. Balliere’s Clinical Haematology 1990; 3: 531-44
22 Frydman AM, Bara L, Roux YLe, Woler M, Chauliac F, Samama MM. The antithrombotic activity and pharmacokinetics of enoxaparine, a low molecular weight heparin, in humans given single subcutaneous doses of 20 to 80 mg. J Clin Pharm 1988; 28: 609-18
23 Thomas DP, Merton RE. A low molecular weight heparin compared with unfractionated heparin. Thromb Res 1982; 28: 343-50
24 Bratt G, Tornebohm E, Lockner D, Bergstrom K. A human pharmacological study comparing conventional heparin and a low molecular weight heparin fragment. Thromb Haemost 1985; 53: 208-11
25 Bratt G, Tornebohm W, Widlund L, Lockner D. Low molecular weight heparin (KABI2165, Fragmin): pharmacokinetics after intravenous and subcutaneous administration in human volunteers. Thromb Res 1986; 42: 613-9
26 Dawes J, Bara L, Roux YLe, Woler M, Chauliac F, Samama MM. The anti thrombotic activity and concentration of a low-molecular-weight-heparin (PK 10169) and unfractionated heparin after intravenous and subcutaneous administration. Haemostasis 1986; 16: 116-22
27 Matzsch T, Bergqvist D, Hedner U, Gstergaard P. Effects of an enzymatically depolymerized heparin as compared with conventional heparin in healthy volunteers. Thrombosis and Haemostasis 1987; 17: 97-101
28 Dryjski M, Schneider DE, Mojaverian P, Be-Sheng Kuo, Bjomsson TD. Investigations on plasma activity of low molecular weight heparin after intravenous and oral administrations. British J Clin Pharm 1989; 28: 188-92
29 Barrowcliffe TW, Johnson EA, Thomas DP. Low molecular weight heparin. John Wiley & Sons; Chichester: etc 1992
30 Hemker HC. The mode of action of heparin in plasma. In: Thrombosis and Haemostasis Verstraete M, Vermylen J, Lijnen HR, Amout J. Eds. I. S. T. H. Leuven University Press; 1987: 17-36
31 Barrowcliffe TW, Havercroft SJ, Kemball-Cook G, Lindahl U. The effect of Ca²⁺, phospholipid and factor V on the anti (Factor Xa) activity of heparin and its high-affinity oligosaccharides. Biochem J 1987; 243: 31-7
32 Ellis V, Scully MF, Kakkar VV. The acceleration of the inhibition of plate let prothrombinase complex by heparin. Biochemical Journal 1986; 233: 161-5
33 Schoen P, Lindhout T, Willems G, Hemker HC. Antithrombin III Dependent Anti-Prothrombinase Activity of Heparins of Various Molecular Weight. J Biol Chem 1989; 264 (17) 10002-7
34 Sandset P, Abilgaard U, Larsen AL. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI). Thromb Res 1988; 50: 803-13
35 Lindahl AK, Abilgaard U, Larsen ML, Aamodt LM, Nordfang P, Beck TC. Extrinsic pathway inhibitor (EPI) and the post-heparin anticoagulant effect in tissue thromboplastin induced coagulation. Thromb Res 1991; Suppl XIV: 39-48
36 Béguin S, Lindhout T, Hemker HC. The effect of trace amounts of thromboplastin on the generation of thrombin in platelet rich plasma, its inhibition by heparin. Thrombosis and Haemostasis 1989; 61: 25-9