Russian recombinant coagulation factors: the technological background and the results of clinical studies

The prolongation of survival and the improvement of quality of life in patients with hemophilia A and B are only possible if hemostatic disorders caused by coagulation factor VIII and IX deficiency are managed effectively. Recombinant coagulation factors are playing an ever-increasing role in the preventive care of affected patients. The development, production and use of domestic recombinant coagulation factors opened up new treatment opportunities and improved access to preventive care for hemophilia patients. The results of clinical studies on the efficacy and safety of the Russian recombinant factors showed that they had similar efficacy and safety compared to the plasma derived clotting factors.

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Inhibition of Factor XI by Myeloma Protein M.

10.1055/s-0039-1682822 ◽

1977 ◽

Author(s):

A. Rimon ◽

I. Raz ◽

M. Lahav

Keyword(s):

Coagulation Factor ◽

Coagulation Factors ◽

Normal Human Plasma ◽

Clotting Factors ◽

Factor Xi ◽

Myeloma Protein ◽

Myeloma Proteins ◽

Normal Human ◽

Time Test ◽

Hemostatic Disorders

Waldenstrom’s macroglobulinemia sometimes involves hemostatic disorders. It has been assumed that this may be due to interaction of the myeloma protein with soom blood coagulation factor(s). In the present investigation the sera of seven myeloma patients were studied for a possible interaction between the myeloma protein and clotting factors II, VIM, IX and XI.Normal human plasma was incubated for 30 mi η at 37°C with different dilutions of patients’ serum or of purified myeloma proteins. The ability of this mixture to restore the clotting times of various deficient plasmas was tested by the partial thromboplastin time test.Factor XI activity of normal plasma was substantially inhibited upon incubation with patient’s serum or with purified myeloma protein M. This effect seems to be specific for myeloma protein M since it was neither observed with myeloma proteins G or A, nor with normal IgM. Other coagulation factors investigated so far were not inhibited by myeloma protein M. It is concluded that this protein is specifically affine to factor XI resulting in hindering the latter’s activity.

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Demonstration of granulocytic proteases in plasma of patients with acute leukemia and septicemia with coagulation defects

Blood ◽

10.1182/blood.v49.2.219.bloodjournal492219 ◽

1977 ◽

Vol 49 (2) ◽

pp. 219-231 ◽

Cited By ~ 3

Author(s):

R Egbring ◽

W Schmidt ◽

G Fuchs ◽

K Havemann

Keyword(s):

Coagulation Factor ◽

Coagulation Factors ◽

Acute Myelocytic Leukemia ◽

Consumption Coagulopathy ◽

Clotting Factors ◽

Myelocytic Leukemia ◽

Coagulation Defects ◽

Alpha1 Antitrypsin ◽

Neutral Proteases

To show whether direct proteolysis of coagulation factors may play a role in patients with so-called consumption coagulopathy, granulocytic neutral proteases in the plasma of patients with acute myelocytic leukemia and septicemia were assayed by one- and two-dimensional Laurell electrophoresis. Complexes between serum alpha1-antitrypsin and elastase-like granulocytic protease could be demonstrated in those patients with acute myelocytic leukemia and septicemia who also had moderate or severe coagulation defects. Despite the presence of a high antiprotease potential, addition of the elastase-like enzyme to normal plasma resulted in coagulation defects in vitro comparable to those seen in the patients. These results and the ability of the elastase- like protease to destroy isolated clotting factors suggested that in certain types of coagulation factor deficiencies direct proteolysis rather than consumption of clotting factors due to disseminated intravascular coagulation may be operational.

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THE EFFECT OF THE VENOM OF THE ORIENTAL HORNET ON COAGULATION FACTORS

10.1055/s-0038-1644338 ◽

1987 ◽

Author(s):

A Kornberg ◽

S Kaufman ◽

L Silber ◽

J Ishay

Keyword(s):

Human Plasma ◽

Degradation Products ◽

Anticoagulant Activity ◽

Coagulation Factors ◽

Plasma Fibrinogen ◽

Thrombin Time ◽

Clotting Factors ◽

Viii And Ix

The extract from the venom sac of Vespa orientalis (VSE) inactivates exogenous and endogenous thromboplastin (Joshua and Ishay, Toxicon, 13:11-20,1975). The prolongation of both prothrombin time (PT) and recalcification time suggests inactivation of other factors. The aim of the present study is to investigate the effect of VSE on clotting factors. A lyophilized VSE with protein concentration of 5 mg/ml was used. Studies were performed in vitro with human plasma and in vivo in cats. Routine methods were employed for the assay of PT, activated tissue thromboplastin (APTT), thrombin time (TT), fibrinogen degradation products (FDP), fibrinogen and factors V,VII,VIII,IX,X. Human plasma was incubated with various concentrations of VSE (0,1,5,10,50,100 μg/ml) for 60 min and for various incubation times (0,5,15,30,+ 60,90,120 min) with 50 μg/ml VSE (n=8). 1 μg/ml VSE prolonged PT from 13.5 to 16 sec (p<0.05) and APTT from 62 to 180 sec. PT was maximal (17.7 sec) with 10 μg/ml and APTT (442 sec) with 50 μg/ml VSE. Factors V,VII,X decreased gradually from 94-105% to 11%,11% and 29% with 100 μg/ml VSE and VIII and IX to 1% even with 1 μg/ml VSE. After 5 min with constant concentration of VSE (50 μg/ml) PT was 14.9 sec (normal 13 sec) and APTT 165 sec (normal 54 sec). Both were maximal (17.5 and 298 sec) after 60 min. Factors VII and X decreased to 13% and 32% and VIII and IX to >1% after 60 min of incubation. Injection of 5 mg/kg VSE to cats (n=6-8) resulted in prolongation of PT from 9.4 to 11.2 sec and of APTT from 19.5 to 63 sec after 5 min. Both were maximal after 90 min (12.3 and 127 sec). Factors V,VII and X decreased from 100% to 7.6%, 13% and 37% and VIII and IX to 1% after 10 min. In all experiments TT and plasma fibrinogen were not affected and FDP were normal. Heating of VSE for 5 min at 80°C abolished completely the anticoagulant activity but dialysis for 24 hr at 4°C had no effect on it. The activity was eluted on Sephadex-25 both in void and post void volumes. The results show that VSE has a potent anticoagulant activity against various factors. Factors VIII and IX are markedly decreased. The effect on V, VII and X is moderate. Plasma fibrinogen is not affected. The nature and clinical significance of the anticoagulant activity merit further investigation.

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Hemostatic Levels of Coagulation Factors in Patients with Mild Prolongation of Prothrombin Time / INR: Implications for Transfusion Medicine Practice.

Blood ◽

10.1182/blood.v104.11.2728.2728 ◽

2004 ◽

Vol 104 (11) ◽

pp. 2728-2728

Author(s):

Karen Matevosyan ◽

Michael Cimo ◽

Christopher Madden ◽

Ravindra Sarode

Keyword(s):

Retrospective Analysis ◽

Coagulation Factor ◽

Coagulation Factors ◽

Median Number ◽

Clotting Factors ◽

Drastic Reduction ◽

Factor Viii Level ◽

Plasma Therapy ◽

Neurosurgical Patients ◽

Viii Level

Abstract OBJECTIVE: To investigate hemostatically important clotting factors in patients with mild to moderate prolongation of PT/INR, who generally receive prophylactic FFP transfusions, especially in neurosurgery, because there is scant data in the literature supporting prophylactic plasma therapy. METHODS: 36 plasma samples from 31 neurosurgical patients with PT/INR 12.5–15.0/1.3–1.5 were separated and frozen within 8 hours of collection; these were tested for coagulation factors II, VII and VIII levels, and PT/INR/PTT values were correlated with the factor levels. Retrospective analysis of all neurosurgery patients for whom FFP was ordered with similar PT/INR values was performed both during 5 months of study period and 5 months post-study period, when changes were implemented in FFP administration guidelines. The PT/INR/PTT values were studied 24 hours following the FFP requests in order to evaluate the effect of plasma infusion. Each group was divided in two subgroups: those who received FFP transfusion within 24 hours of the FFP requests, and those who did not. RESULTS: PT/INR/PTT and coagulation factor levels of the 36 samples are seen in Table 1. PT mean 13.78 INR mean 1.38 PTT mean 29.29 PT median 13.5 INR median 1.3 PTT median 29.5 PT range 10.7–18.6 INR range 0.9–1.9 PTT range 19.3–37.9 FII mean 68% FVII mean 53% FVIII mean 156% FII median 66% FVII median 51% FVIII median 126% FII range 34–107% FVII range 28–124% FVIII range 55–547% There was no correlation between PT/INR with FVII levels (p>0.05). All patients had FVII > 15–25%, level recommended as safe for surgery. FII was also hemostatically normal, whereas mean factor VIII level was elevated, that correlated with shortened PTT in many of these patients. Retrospective analysis revealed that during the study period there were a total of 99 FFP requests (69 patients). Of these 68 requests (46 patients) did and 31 (29 patients) did not receive FFP transfusions within the next 24 hours. Mean number of FFP units transfused was 3.03 (median 2, range 1–7). During the post study period a total of 15 requests (for 14 patients) were received; of these 2 did and 13 did not receive FFP transfusions within the next 24 hours. Mean and median number of FFP was 2. PT/INR/PTT at time of request and 24 hours post transfusion are presented in the Table 2 Study Period (99 FFP requests) Post Study Period (15 FFP requests) Transfused (68) Non-Transfused (31) Transfused (2) Non-Transfused (13) At FFP Request At 24 hrs At FFP Request At 24 hrs At FFP Request At 24 hrs At FFP Request At 24 hrs *PT;†INR;♠PTT 13.42* 12.1 11.9 11.29 14.1 12.8 12.22 11.54 1.3† 1.18 1.16 1.1 1.4 1.3 1.18 1.14 28.9♠ 27.98 27.65 27.43 26.6 27.1 27.4 26.99 CONCLUSION: In patients with a mild prolongation of PT/INR, plasma levels of FII and FVII were hemostatically normal, whereas FVIII levels were increased. After FFP transfusions, PT/INR and PTT values did not change suggesting that plasma therapy in these patients was not warranted. After the study there was a drastic reduction (85%) in FFP orders for prophylactic plasma transfusions.

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Demonstration of granulocytic proteases in plasma of patients with acute leukemia and septicemia with coagulation defects

Blood ◽

10.1182/blood.v49.2.219.219 ◽

1977 ◽

Vol 49 (2) ◽

pp. 219-231 ◽

Cited By ~ 164

Author(s):

R Egbring ◽

W Schmidt ◽

G Fuchs ◽

K Havemann

Keyword(s):

Coagulation Factor ◽

Coagulation Factors ◽

Acute Myelocytic Leukemia ◽

Consumption Coagulopathy ◽

Clotting Factors ◽

Myelocytic Leukemia ◽

Coagulation Defects ◽

Alpha1 Antitrypsin ◽

Neutral Proteases

Abstract To show whether direct proteolysis of coagulation factors may play a role in patients with so-called consumption coagulopathy, granulocytic neutral proteases in the plasma of patients with acute myelocytic leukemia and septicemia were assayed by one- and two-dimensional Laurell electrophoresis. Complexes between serum alpha1-antitrypsin and elastase-like granulocytic protease could be demonstrated in those patients with acute myelocytic leukemia and septicemia who also had moderate or severe coagulation defects. Despite the presence of a high antiprotease potential, addition of the elastase-like enzyme to normal plasma resulted in coagulation defects in vitro comparable to those seen in the patients. These results and the ability of the elastase- like protease to destroy isolated clotting factors suggested that in certain types of coagulation factor deficiencies direct proteolysis rather than consumption of clotting factors due to disseminated intravascular coagulation may be operational.

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The Role of GRP and MGP in the Development of Non-Hemorrhagic VKCFD1 Phenotypes

International Journal of Molecular Sciences ◽

10.3390/ijms23020798 ◽

2022 ◽

Vol 23 (2) ◽

pp. 798

Author(s):

Suvoshree Ghosh ◽

Johannes Oldenburg ◽

Katrin J. Czogalla-Nitsche

Keyword(s):

Vitamin K ◽

Treatment Strategies ◽

Coagulation Factor ◽

Matrix Gla Protein ◽

Clotting Factors ◽

Factor Deficiency ◽

Coagulation Factor Deficiency ◽

New Treatment ◽

Deficiency Type

Vitamin K dependent coagulation factor deficiency type 1 (VKCFD1) is a rare hereditary bleeding disorder caused by mutations in γ-Glutamyl carboxylase (GGCX) gene. The GGCX enzyme catalyzes the γ-carboxylation of 15 different vitamin K dependent (VKD) proteins, which have function in blood coagulation, calcification, and cell signaling. Therefore, in addition to bleedings, some VKCFD1 patients develop diverse non-hemorrhagic phenotypes such as skin hyper-laxity, skeletal dysmorphologies, and/or cardiac defects. Recent studies showed that GGCX mutations differentially effect γ-carboxylation of VKD proteins, where clotting factors are sufficiently γ-carboxylated, but not certain non-hemostatic VKD proteins. This could be one reason for the development of diverse phenotypes. The major manifestation of non-hemorrhagic phenotypes in VKCFD1 patients are mineralization defects. Therefore, the mechanism of regulation of calcification by specific VKD proteins as matrix Gla protein (MGP) and Gla-rich protein (GRP) in physiological and pathological conditions is of high interest. This will also help to understand the patho-mechanism of VKCFD1 phenotypes and to deduce new treatment strategies. In the present review article, we have summarized the recent findings on the function of GRP and MGP and how these proteins influence the development of non-hemorrhagic phenotypes in VKCFD1 patients.

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Effect of Neutral Proteases of Human Granulocytes on Isolated Clotting-Factors

10.1055/s-0039-1689546 ◽

1975 ◽

Author(s):

W. Schmidt ◽

R. Egbring ◽

K. Havemann ◽

H. Beeser

Keyword(s):

Coagulation Factor ◽

Coagulation Factors ◽

Factor Vii ◽

Factor V ◽

Clotting Factors ◽

Human Granulocytes ◽

Rapid Destruction ◽

Weak Activity ◽

Moderate Effect ◽

Antigen Antibody

To examine whether direct proteolysis of coagulation factors may play a role in patients with so called consumption coagulopathy, an elastase-like and a chymotrypsin-like neutral protease isolated from human granulocytes were investigated for their influence on several purified clotting factors. The elastaselike protease induced a rapid destruction of fibrinogen, factors II, VIII, XII and XIII activity, whereas a moderate effect on factor V and VII activity was observed. The chymotrypsin-like enzyme showed a rapid inactivation of factor VIII, moderate effect on factor VII and XIII and only a weak activity against fibrinogen, factor II and XIII. Incubation of factor V with both enzymes leads to a transitory activation. In spite of the presence of a high antiprotease potential in plasma, addition of the elastase-like enzyme to normal plasma resulted in an activation of several coagulation factors. As it has been shown that the proteases are activity released from granulocytes in presence of antigen-antibody complexes, endotoxin and polynucleotides, the results given above together with the appearence of granulocytic proteases in the plasma of patients with acute leucemia and septicemia suggest that in certain types of coagulation factor deficiencies direct proteolysis rather than consumption of clotting factors due to dissiminated intravascular coagulation may be operational.

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Intracellular maturation of the γ-carboxyglutamic acid (Gla) region in prothrombin coincides with release of the propeptide

Biochemical Journal ◽

10.1042/bj2910723 ◽

1993 ◽

Vol 291 (3) ◽

pp. 723-727 ◽

Cited By ~ 13

Author(s):

R Wallin ◽

C Stanton ◽

S M Hutson

Keyword(s):

Conformational Change ◽

Vitamin K ◽

Secretory Pathway ◽

Coagulation Factor ◽

Coagulation Factors ◽

Coagulation System ◽

Clotting Factor ◽

Clotting Factors ◽

Post Translational Modifications ◽

Carboxyglutamic Acid

Vitamin K-dependent coagulation factors undergo several post-translational modifications before the proteins are secreted into the blood as functional zymogens of the coagulation system. The modifications include Asn-linked glycosylation, Asn/Asp hydroxylation, removal of a signal peptide for translocation of the polypeptide into the endoplasmic reticulum and removal of a propeptide which, when attached to the intracellular coagulation factor precursor, directs the protein for vitamin K-dependent gamma-carboxylation. gamma-Carboxylation of targeted Glu residues results in formation of Ca(2+)-binding gamma-carboxyglutamic acid (Gla) residues. Ca2+ binding by these residues induces a conformational change in the protein which is a necessary event for optimal activation or activity of the clotting factor in blood. In the present study we have monitored the intracellular prothrombin precursor in the secretory pathway of liver cells to determine the effect that the propeptide has on Ca(2+)-dependent folding of the protein. The data provide evidence that the Ca(2+)-induced conformational change required for activation of prothrombin coincides with release of the propeptide in the trans-Golgi apparatus of the liver cell and elucidates an important function for the endoproteinase furin in biosynthesis of vitamin K-dependent clotting factors.

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Increased Activity of Vitamin K-Dependent Clotting Factors in Human Hyperlipoproteinaemia – Association with Cholesterol and Triglyceride Levels

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651852 ◽

1977 ◽

Vol 38 (02) ◽

pp. 0465-0474 ◽

Cited By ~ 46

Author(s):

M Constantino ◽

C Merskey ◽

D. J Kudzma ◽

M. B Zucker

Keyword(s):

Vitamin K ◽

Plasma Lipids ◽

Coagulation Factors ◽

Clotting Factor ◽

Factor X ◽

Clotting Factors ◽

Blood Coagulation Factors ◽

Cholesterol Levels ◽

Type V ◽

Increased Activity

SummaryLevels of blood coagulation factors, cholesterol and triglyceride were measured in human plasma. Prothrombin was significantly elevated in type Ha hyperlipidaemia; prothrombin and factors VII, IX and X in type lib; and prothrombin and factors VII and IX in type V. Multiple regression analysis showed significant correlation between the levels of these plasma lipids and the vitamin K-dependent clotting factors (prothrombin, factors VII, IX and X). Higher cholesterol levels were associated with higher levels of prothrombin and factor X while higher triglyceride levels were associated with higher levels of these as well as factors VII and IX. Prothrombin showed a significant cholesterol-triglyceride interaction in that higher cholesterol levels were associated with higher prothrombin levels at all levels of triglyceride, with the most marked effects in subjects with higher triglyceride levels. Higher prothrombin levels were noted in subjects with high or moderately elevated (but not low) cholesterol levels. Ultracentrifugation of plasma in a density of 1.21 showed activity for prothrombin and factors VII and X only in the lipoprotein-free subnatant fraction. Thus, a true increase in clotting factor protein was probably present. The significance of the correlation between levels of vitamin K-dependent clotting factors and plasma lipids remains to be determined.

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