Effectiveness of heparin in preventing thrombin generation and thrombin activity in patients undergoing coronary intervention

SummaryIn a thrombin generation test with continuous registration of thrombin activity in plasma we studied the ability of a variety of thrombin inhibitors of different type and mechanism of action to influence the activity of thrombin after activation of the coagulation system. Depending on the inhibitor, the peak of thrombin activity is delayed and/or reduced.By blocking the active site of generated thrombin inhibitors cause a concentration dependent reduction of the thrombin peak and inhibit feed-back reactions of thrombin resulting in a delay of thrombin generation. Highly potent synthetic active-site directed inhibitors (Ki ≤ 20 nM) reduce the thrombin activity formed in plasma after extrinsic or intrinsic activation with the same efficiency (IC50 0.1 - 0.6 μM) as hirudin. The delay and reduction of thrombin generation by inhibitors of the anion-binding exosite 1 of thrombin is only attributed to an inhibition of feed-back reactions of thrombin. For a 50% reduction of thrombin activity in plasma by this type of inhibitors relatively high concentrations were determined.

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The Effect of Procoagulative Phospholipids on the Synthetic Substrate Determined Thrombin Generation in Vitro

10.1055/s-0039-1680766 ◽

1977 ◽

Author(s):

F. Schulte ◽

O. Klug ◽

Ursula Roth

Keyword(s):

Thrombin Generation ◽

Chromogenic Substrate ◽

Chromogenic Substrates ◽

Platelet Factor ◽

Synthetic Substrate ◽

Thrombin Activity ◽

Vitro Effect ◽

Hydrolysis Of

The effect of procoagulative phospholipids (Procops) with platelet factor-3 like activity on the generation of thrombin in plasma can be determined in vitro with the aid of synthetic chromogenic substrates. Standard citrated plasma of two manufacturers and from different batches incubated with amounts of 2-10 meg Procops as 1:100 - 1:500 diluted Tachostyptan (micellar Procops in form of a pharmaceutical speciality) was activated. The generated amount of thrombin activity catalyses the hydrolysis of chromogenic substrate to a tripeptide and to p-nitroaniline which was measured kinetically with a spectrophotometer at 405 nm. At optimum concentrations of Procops, activities adequate to 80 (SD ± 7, VK 8. 8%) - 115 i. u. (SD ± 2. 5, VK 2.2%) thrombin per ml plasma were measured depending on the conditions of incubation and activation. Having made the basis of appropriate procedures for incubation and activation the in vitro effect of Procops on the thrombin generation in plasma is reproducibly measurable with the aid of chromogenic substrate.

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Peri-procedural thrombin generation predicts ischemic complications in primary percutaneous coronary intervention of ST-elevation myocardial infarction: a substudy of the ATOLL trial

European Heart Journal ◽

10.1093/eurheartj/eht310.4530 ◽

2013 ◽

Vol 34 (suppl 1) ◽

pp. 4530-4530

Author(s):

S. O'Connor ◽

A. Ankri ◽

M. Kerneis ◽

J. Abtan ◽

D. Brugier ◽

...

Keyword(s):

Myocardial Infarction ◽

Percutaneous Coronary Intervention ◽

Primary Percutaneous Coronary Intervention ◽

Thrombin Generation ◽

Coronary Intervention ◽

St Elevation Myocardial Infarction ◽

Percutaneous Coronary ◽

Ischemic Complications ◽

St Elevation

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EFFECT OF VARIOUS HEPARIN(OID)S ON HEPARIN COFACTOR II MEDIATED ANTI-THROMBIN ACTIVITY AND INHIBITION OF THROMBIN GENERATION IN VITRO

10.1055/s-0038-1644353 ◽

1987 ◽

Author(s):

T G van Dinther ◽

F Hol ◽

D G Meuleman

Keyword(s):

Molecular Weight ◽

Thrombin Generation ◽

Factor Xa ◽

Weight Fraction ◽

Blood Platelet ◽

Low Molecular Weight ◽

Heparin Cofactor Ii ◽

Thrombin Activity ◽

At Iii

The effects of various heparin(oid)s, standard heparin VII (SH), dermatan sulphate (DS), a low molecular weight fraction of heparin (UMW-H), FragminR (FRA), Org 10172 = low molecular weight heparinoid, the fraction of Org 10172 with high affinity for AT-III (HA-10172) and the low affinity fraction (LA-10172) respectively were examined on in vitro thrombin generation and inactivation.Thrombin inactivation in the presence of either heparin cofactor II (HC-II) or anti-thrombin III (AT-III) was assessed with two newly developed assays using the purified cofactors, thrombin and chromogenic substrate S2238 on microtiterplates. Thrombin generation in the presence of HC-II and AT-III was studied using purified factor Xa, prothrombin and blood platelet lysate and the residual thrombin activity was assessed amidolytically.The inhibition of the compounds on thrombin activity are summarized in the tableThe following conclusions can be drawn:- SH, LMW-H, HA-10172 and FRA potentiate the AT-III mediated inactivation of Ha more strongly than the HC-II mediated inactivation.- DS and LA-10172 show the reverse pattern of inactivation, while Org 10172 potentiates both inactivaton pathways to a similar extent.Thrombin generation in the presence of HC-II is inhibited by mw-heparin(oid)s at approx. 2-5 times lower concentrations than the HC-II mediated thrombin inactivation, while the inhibiting effect of SH in both assays is comparable.AT-III mediated thrombin generation inhibition and AT-III mediated thrombin inactivation is comparable as well for SH, LMW-H and FRA. In contrast, Org 10172 and its subfractions are approx. 10 times more potent on AT-III mediated thrombin generation inhibition than on AT-III mediated thrombin inactivation.Org 10172 shows low anti-thrombin activity and this activity is mainly mediated via FC-II.

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The Recalcified Coagulation Activity

Clinical and Applied Thrombosis/Hemostasis ◽

10.1177/1076029607309228 ◽

2007 ◽

Vol 14 (4) ◽

pp. 447-453 ◽

Cited By ~ 7

Author(s):

Thomas W. Stief

Keyword(s):

Thrombin Generation ◽

Room Temperature ◽

Prothrombotic State ◽

Coagulation Activity ◽

Thrombin Activity ◽

Clinical Interest ◽

C 50

Thrombin activity generated after plasma recalcification is of analytical and clinical interest. Fifty microliters of citrated plasma was recalcified with 5 µL of 250 mM CaCl 2. After 0 to 90 minutes (37°C) 50 µl 2.5 M arginine, pH 8.6, was added. After 20 minutes, thrombin was chromogenically quantified. In normal recalcified plasma, the generated thrombin activity is about 0.1-0.2 IU/ml (37°C) when fibrin generation starts. Pooling of normal plasmas increases the generated thrombin activity about 3-fold. Plasmas of patients on heparin or coumarin generate about 10-fold less thrombin activity. Freezing of pooled plasma at −20°C and thawing at room temperature or 37°C increases thrombin generation approximately 1.5- or 2-fold, respectively. Only thrombin activities in the ascending part of the thrombin generation curve (RECA-t2/RECAt 1>1) are valid. So a prothrombotic state in blood or plasma can be diagnosed.

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Increased Thrombin Generation Capacity in Septic Patients with DIC.

Blood ◽

10.1182/blood.v108.11.4054.4054 ◽

2006 ◽

Vol 108 (11) ◽

pp. 4054-4054

Author(s):

Kazuo Kawasugi ◽

Ryousuke Shirasaki ◽

MIzuho Noguti ◽

Haruko Tashiro ◽

Moritaka Gotoh ◽

...

Keyword(s):

Thrombin Generation ◽

Bleeding Tendency ◽

Fibrin Deposition ◽

Activity Peak ◽

Thrombin Activity ◽

Healthy Female ◽

Generation Capacity ◽

Laboratory Changes ◽

Clinical Conditions ◽

Normal Controls

Abstract Variety clinical conditions may cause systemic activation of coagulation, ranging from insignificant laboratory changes to severe disseminated intravascular coagulation (DIC). DIC consists of widespread systemic activation of coagulation, resulting in diffuse fibrin deposition in small and midsize vessels. However, little is known about thrombin generation capacity in patients with DIC. To investigate the thrombin generation capacity, we measured thrombin generation in septic patients with DIC (n=20) and acute promyelocytic leukemia (APL)-induced DIC (n=5). Thrombin generation was determined by the Throbogram-Thrombinoscope assay (Thermo Electron Corporation, Netherlands). The analyzed TG parameters ware the peak of thrombin activity (Peak) and endogenous thrombin potential (ETP). The thrombin antithrombin complexes (TAT) levels were higher in both DIC patients as reported by others. In the septic patients with DIC, we found significant elevations in peak of thrombin activity and ETP as compared with normal controls (determined in 17 healthy males and 14 healthy female). However, the peak of thrombin activity and ETP levels were severely decreased by 60% n the APL patients with DIC. There was slightly correlation between the ETP and TAT levels in septic patients with DIC. Also, there was correlation between the ETP and bleeding tendency in APL patients with DIC. These results suggest that assess of thrombin generation capacity may be helpful in the making the diagnosis in septic patients with DIC. It appears that assess of ETP may contribute to evaluate bleeding tendency in APL patients with DIC. Sepsis with DIC APL with DIC Controls (n=31) *(P<0.05) significantly different controls Peak (nM) 325.3 ± 55.5* 88.2 ± 47.2* 281.2 ± 40.9 ETP (nM.min) 1 ± 326.1* 652.1 ± 274.5* 1469.2 ± 227.3 TAT (μg/ml) 28.4 ± 17.5* 22.5 ± 15.3* <3

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