Drug that promotes hemostasis

Abstract

The invention relates to the use of prethrombin-1 as a drug.

Description

[0001] Medicines to promote hemostasis

[0002] The invention relates to a medicament for promoting hemostasis and for treating bleeding.

[0003] Hemostasis is the result of a cascade-like, proteolytic activation of inactive zymogens. The concerted interaction of activated coagulation factors with their cofactors results in the penultimate step of hemostasis, the activation of prothrombin to thrombin. When present in sufficient concentration, thrombin causes the fibrinogen present in the blood plasma to clot at the site of injury, where it, together with activated platelets, forms an insoluble fibrin matrix.

[0004] Thrombin formation is therefore a central element of hemostasis, with the generation of thrombin from prothrombin being a complex and tightly regulated enzymatic process. Other coagulation factors and cofactors, as well as platelets and certain endothelial factors, are required for the entire hemostatic process.

[0005] The enzymatic activation of prothrombin occurs through factor Xa, an enzyme produced by the activation of factor X. The activity of factor Xa is increased by several orders of magnitude when it forms the enzyme complex prothrombinase together with factor Va as a cofactor. Prothrombinase forms in the presence of calcium ions on phospholipid-containing membrane surfaces formed by activated platelets or the endothelium.

[0006] Factor Xa is produced by two different tenases: intrinsic and extrinsic tenase. Intrinsic tenase is characterized by the enzyme factor IXa and its cofactor VIIIa. Extrinsic tenase, in turn, represents the beginning of the coagulation cascade and consists of the enzyme factor VIIIa and tissue factor, a tissue factor that comes into contact with blood when the endothelium is injured. Factor Xa produced by extrinsic tenase is rapidly inhibited by TFPI. The tiny amounts of thrombin produced by the extrinsically produced factor Xa activate cofactors V and VIII as well as platelets at the site of the coagulation process (1). Factor V or a short form of factor V (factor V short) are cofactors of TFPI alongside protein S. Activation of factor V or factor V short by thrombin leads to a reduction in the activity of the cofactor and thus to a reduction in the inhibitory effect of TFPI (2).

[0007] Hemophilia A and B are pathological changes characterized by dysfunction of the components of intrinsic tenase. Dysfunction of intrinsic tenase leads to impaired hemostasis. These coagulation disorders are treated by substituting the missing coagulation factors. A common complication of this substitution treatment is the formation of alloantibodies against the substituted coagulation factor. These inhibitors make further substitution difficult or even render it completely ineffective. Circumventing these inhibitors represents a major challenge in hemophilia treatment. In the early 1970s, prothrombin complexes were developed as drugs for the treatment of bleeding from a variety of causes. These prothrombin complexes have also proven to be quite effective in inhibitor hemophilia.A further development was the use of activated prothrombin complexes, which increasingly replaced non-activated prothrombin complexes in the treatment of inhibitor hemophilia (3, 4). Factor Vila, as a component of extrinsic tenase, also emerged as a potential therapeutic option for inhibitor hemophilia.

[0008] Activated factors, especially factor Xa, in combination with prothrombin have been postulated as active components of activated prothrombin complex concentrates (5). Prothrombin has a relatively long half-life in the blood, which means that repeated administration of activated prothrombin complexes increases the prothrombin concentration to two to three times the normal value.

[0009] It was recognized early on that, in addition to the concentration of antithrombin, the concentration of prothrombin in the blood is the most important parameter for normal blood coagulation (6).

[0010] Highly purified prothrombin has been shown to have hemostatic activity in mouse models of hemophilia A and B (7). The prothrombin concentration in plasma is approximately 80–90 mg / L. The half-life is 48–70 hours. To significantly increase the prothrombin concentration in plasma, the administration of large amounts of prothrombin is required, but due to the long half-life, a long-lasting effect is also achievable. Nevertheless, the use of prothrombin as a drug was not pursued further. A clinical study evaluating the safety, toxicity, and pharmacodynamics of recombinant human prothrombin was terminated prematurely (8).

[0011] Prothrombin is a vitamin K-dependent proenzyme and possesses a complex structure characterized by an N-terminal Gla domain and the kringle 1 domain (fragment 1), the kringle 2 domain (fragment 2), and the actual enzyme domain. To form thrombin, prothrombin must be cleaved at two sites: R271 and R320. Cleavage at site R320 results in the formation of the enzymatically active meizothrombin. Subsequent cleavage at site R271 results in the release of thrombin. If the R271 cleavage site of prothrombin is cleaved first, the enzymatically inactive prethrombin-2 is formed, which can subsequently be converted into thrombin by cleavage at site R320.

[0012] In the presence of cofactor Va, the R320 cleavage site is preferentially cleaved, yielding meizothrombin as an intermediate. In the absence of factor Va, however, the R271 cleavage site is preferentially cleaved, yielding primarily prethrombin-2 (9).

[0013] The majority of thrombin required for blood coagulation is formed by the prothrombinase complex, consisting of factor Xa and its cofactor Va, on phospholipid-containing surfaces in a concerted mechanism (10). However, thrombin also plays a crucial role in the activation of platelets and the formation of the cofactors Va and VIIIa, which are important for the two tenases.

[0014] Prethrombin-1 is an enzymatically inactive cleavage product formed by cleavage of fragment 1 from prothrombin by thrombin in a feedback reaction (11, 12). Both prethrombin-1 and prethrombin-2 lack the Gla domain required for membrane binding and are therefore not efficiently activated in vitro by prothrombinase bound to phospholipid vesicles (13). In the absence of factor Va or phospholipids, free factor Xa activates prothrombin and prethrombin-1 at rates similar to those of thrombin.

[0015] When prethrombin-1 is generated on activated platelet surfaces, it rapidly activates to thrombin (14). Fragment 2, present in prethrombin-1, contains a sequence that binds factor Va (15). Prethrombin-2 lacks this sequence, which is located on kringle 2.

[0016] McDuffie et al. (16) used specific immunoassays to determine the concentrations of prothrombin, thrombin, and prothrombin fragments in the plasma of normal individuals and patients with suspected disseminated intravascular coagulation (DIC). From their results, they concluded that measurements of prethrombin-1 levels are not useful for the diagnosis or treatment management of DIC.

[0017] Owen et al. (17) investigated the catabolism of prothrombin and its fragments in normal dogs. To determine whether prethrombin-1 might play a role as a degradation product of prothrombin, they infused dogs with radiolabeled prethrombin-1. Based on the half-lives and plasma concentrations of prethrombin-1 determined in normal individuals, they found that prothrombin is not catabolized via the release of prethrombin-1.

[0018] Lanchantin et al. (18) recognized early on that the action of thrombin on prothrombin results in the loss of prothrombin activity in a coagulation test performed with prothrombin-deficient plasma. The two cleavage products resulting from the action of thrombin on prothrombin were eventually named prethrombin-1 and fragment 1. Prethrombin-1 is enzymatically inactive.

[0019] Heldebrandt et al. (19) have reported that prothrombin and prethrombin-1 (intermediate 1) can be activated to thrombin by factor Xa at approximately the same rate in a dilute aqueous buffer. Due to the missing fragment 1, prethrombin-1 cannot bind to phospholipid membranes and can therefore only be metabolized by prothrombinase at a much lower rate than prothrombin. This is also the reason why prethrombin-1 has hardly any activity in the coagulation test with prothrombin-deficient plasma. The test method is based on a coagulometric coagulation test in which all coagulation factors except prothrombin are present in the plasma. The activity of prothrombin is measured by adding calcium and thromboplastin. Prothrombin is the limiting factor and the clotting time is inversely proportional to the concentration of prothrombin. Seegers et al. (20) and Baker et al.(21) have shown that prethrombin-1 can be converted into thrombin only with difficulty when phospholipids, factor Xa, factor V and calcium ions are used as procoagulants.

[0020] Seegers et al. (22, 23) described a previously unknown procoagulant, which they termed protein M. This protein accelerated the formation of thrombin in a five-component system consisting of a thrombin zymogen, factor Xa, factor V, phospholipids, and calcium ions.

[0021] Topical application of thrombin for hemostasis is widely used clinically (24, 25).

[0022] Disorders of the coagulation system can lead to severe impairment of hemostasis, although direct administration of thrombin is not a treatment option in hemorrhagic diatheses because of the high risk of fatal systemic coagulation.

[0023] It is therefore of great interest to be able to provide treatment options with suitable drugs that normalize prolonged bleeding times, particularly in hemophilia A or B in the presence of inhibitors.

[0024] Currently available treatment options primarily include activated prothrombin complexes (FEIBA®, Takeda) or activated factor VII (NovoSeven®, Novo Nordisk). Activated factor VII has a very short half-life and is therefore not suitable for prophylactic use. Activated prothrombin complexes have a complex composition, with zymogens and traces of activated forms of the procoagulant coagulation factors II, VII, IX, and X, anticoagulant factors such as protein C and TFPI, and low amounts of the cofactors FV, ​​FVIII, and protein S present in a balanced ratio. (26)

[0025] The present invention aims to provide a medicament for promoting hemostasis and treating bleeding, which contains a molecularly well-defined active ingredient and has a composition that is easy to formulate. Furthermore, the medicament is intended to represent an alternative to currently common hemostasis-promoting medicaments, such as activated prothrombin complexes and activated factor VIa. Description of the invention:

[0026] This object is achieved with a medicament containing prethrombin-1 as the active ingredient. The invention thus consists in the use of prethrombin-1 as a medicament or in the use of prethrombin-1 for the production of a medicament.

[0027] A preferred embodiment of the invention consists in the specific use of prethrombin-1 to promote hemostasis or in the use of prethrombin-1 for the preparation of a medicament for promoting hemostasis.

[0028] Further preferred embodiments of the invention are:

[0029] Prethrombin-1 for use in the treatment of bleeding or the use of prethrombin-1 for the manufacture of a medicament for the treatment of bleeding;

[0030] Prethrombin-1 for use in the treatment of bleeding in patients with coagulation disorders or the use of prethrombin-1 for the manufacture of a medicament for the treatment of bleeding in patients with coagulation disorders; Prethrombin-1 for use in the treatment of bleeding resulting from trauma or internal injury or the use of prethrombin-1 for the manufacture of a medicament for the treatment of bleeding resulting from trauma or internal injury;

[0031] Prethrombin-1 for use in the treatment of breakthrough bleeding during substitution treatments with coagulation factors, factor VIII-mimicking therapies or gene therapies or the use of prethrombin-1 for the manufacture of a medicament for the treatment of breakthrough bleeding during substitution treatments with coagulation factors, factor VIII-mimicking therapies or gene therapies; and

[0032] Prethrombin-1 for use in reversing anticoagulation by direct-acting, orally administered anticoagulants or the use of prethrombin-1 for the manufacture of a medicament for reversing anticoagulation by direct-acting, orally administered anticoagulants.

[0033] The invention further encompasses a method of treating patients to promote hemostasis, to treat bleeding, to treat coagulation disorders, to treat bleeding resulting from trauma or internal injury, to treat breakthrough bleeding under replacement therapies with coagulation factors, factor VIII-mimicking therapies or gene therapies, and to reverse anticoagulation by direct-acting, orally administered anticoagulants, which methods comprise administering to a patient an effective amount of prethrombin-1.

[0034] As described in the example below, prethrombin-1 can be readily prepared from prothrombin by cleavage with thrombin followed by chromatographic purification. Prethrombin-1 can be formulated with a suitable pharmaceutical vehicle, sterilized by filtration, and aseptically filled. Freeze-drying is a preferred pharmaceutical preparation.

[0035] For the use of prethrombin-1 to produce a medicament from prothrombin, various methods for virus inactivation or virus elimination may be used. Examples of these methods are known to those skilled in the art.

[0036] As an alternative to obtaining prethrombin-1 from plasma, it can also be produced using suitable molecular biological methods, particularly recombinant DNA technology. An additional, special embodiment of the present invention thus consists in using recombinant prethrombin-1 instead of native prethrombin-1 for the above-mentioned indications.

[0037] Prothrombin complex concentrates cannot be produced using recombinant DNA technology. Their composition is too complex. Prothrombin, as the primary procoagulant factor, must be balanced by sufficient amounts of anticoagulant components. Since these have a shorter half-life than prothrombin, an excess of prothrombin and associated undesirable thrombogenic effects can occur. Prethrombin-1 is defined by the protein sequence resulting from the cleavage of prothrombin fragment 1 at position R155.

[0038] The dosage of prethrombin-1 depends on the cause and severity of the bleeding, but is preferably 100 units / kg body weight, where one unit of prethrombin-1 is defined as the amount that molecularly corresponds to one unit of prothrombin. Lower doses can also promote hemostasis. Higher doses of up to 300 units / kg can be effective if the bleeding cannot be controlled with standard doses. Plasma levels can be further increased by repeated administration of prethrombin-1.

[0039] Prethrombin-1 has been shown to have only a low propensity to cause adverse thromboembolic events.

[0040] The efficacy of prethrombin-1 as a hemostatic drug is demonstrated below in an FVIII inhibitor mouse model. The inventors used this model to investigate the promotion of hemostasis by prethrombin-1. Surprisingly, prethrombin-1 was found to possess excellent hemostatic effects in this model, as demonstrated in the following example.

[0041] Prethrombin-2 has no hemostatic effect in vivo.

[0042] The invention is therefore based, among other things, on the finding that prethrombin-1 promotes hemostasis in vivo.

[0043] Prethrombin-1 can also be used in particular for the following bleeding conditions that are difficult to control:

[0044] • Severe acute bleeding in patients with coagulation disorders

[0045] • Acute life-threatening bleeding as a result of trauma or internal injuries

[0046] • Reversal of anticoagulation during therapy with direct-acting oral anticoagulants (NOACs) due to life-threatening or uncontrolled bleeding. NOACs (new oral anticoagulants) are anticoagulant and antithrombotic agents. The anticoagulant effect is based on the direct inhibition of blood clotting factors. • Other critical bleeding situations in which activated or non-activated prothrombin complex concentrates or recombinant factor VIa are used.

[0047] “Severe” bleeding refers to bleeding that cannot be stopped using conventional methods.

[0048] Prethrombin-1 should be effective when administered both intravenously and subcutaneously.

[0049] The invention is described in more detail in the following examples.

[0050] Example 1: Production of prethrombin-1

[0051] 1 ml of a prothrombin concentrate with an activity of 425 U prothrombin / ml was mixed with 50 U thrombin. The mixture was incubated for approximately 24 h at room temperature. After incubation, the mixture was diluted to 10 ml with the buffer solution 10 mM citrate, 137 mM NaCl, pH 7.0 and applied to 15 ml Heparin Sepharose FF. Prethrombin-1 was isolated from the fraction of the 2nd UV 280 nm (18 ml) signal peak of an isocratic elution with the buffer solution 15 mM citrate, 150 mM NaCl, pH 7.0, concentrated to 5 ml over a 5 kDa UF membrane, diluted to 10 ml with distilled water, and applied to 10 ml AIEX CaptoQ ImpRes. Prethrombin-1 was isolated from the fraction of the second UV 280 nm signal peak (22 ml) of an isocratic elution with the buffer solution 15 mM citrate, 150 mM NaCl, pH 7.0. The prethrombin-1 fraction was again concentrated to 1 ml using a 5 kDa UF membrane. The resulting prethrombin-1 was adjusted to a protein concentration of 5.0 mg / ml.

[0052] The resulting band-pure prethrombin-1 was formulated with a suitable pharmaceutically acceptable vehicle and formulated into a drug using conventional methods. The resulting prethrombin-1 exhibited only approximately 1% of the activity of prothrombin in the prothrombin assay using prothrombin-deficient plasma. With a complex of factor Xa and its cofactor Va, one unit of the resulting prethrombin-1 could produce approximately 200 NIH units of thrombin. Example 2: Determination of the hemostatic effect in an FVIII inhibitor mouse model

[0053] FVB mice were anesthetized and treated with an FVIII antibody via the tail vein in such a way that they exhibited an increased tendency to bleed after tail incision. After application of the test substance via the femoral vein, the tip of the tail was amputated with a scalpel at a distance of 3 mm. Blood loss was determined gravimetrically. Bleeding time was defined as the time until the bleeding stopped. After the bleeding stopped, slight rebleeding could occur. The test duration was 30 minutes in all cases.

[0054] The results obtained are clearly presented in the following tables, where the abbreviation "AK" stands for the antibody against Factor VIII.

[0055]

[0056] * ... minor bleeding

[0057] The tables show the following:

[0058] The placebo group, in which five animals received isotonic saline solution, showed bleeding times > 30 min, which were associated with blood losses of 424.8 to 823.6 mg.

[0059] When prethrombin-1 was administered at a dose of 100 U / kg body weight, the bleeding time was reduced in four animals to 6 min, 9 min, 2 min, or 1 min 30 sec. Only in one animal was the bleeding time greater than 30 min, with very slight bleeding. Blood loss was significantly reduced in all animals, with no measurable blood loss in three animals, 3.7 mg in one animal, and 10.1 mg in another.

[0060] When the prethrombin-1 dose was increased to 300 mg / kg body weight, bleeding times were 5 min 10 sec, >30 min, 4 min 30 sec, 1 min 30 sec, and 1 min 40 sec. Here, too, the bleeding was extremely mild compared to the placebo group. In five animals, no blood loss was measurable at all, and in one animal, the loss was 51.4 mg.

[0061] The experiments show that both at a prethrombin-1 dose of 100 mg / kg body weight and at a dose of 300 mg / kg body weight, the bleeding characteristics were significantly reduced compared to the placebo group and, above all, blood loss was significantly reduced compared to the placebo group.

[0062] Example 3: Thrombogenic potential of prethrombin-1 in a modified Wessler test The Wessler test has been used for many years to determine the thrombogenicity of various substances in an in vivo model (27, 28).

[0063] New Zealand white rabbits (2.5–3.5 kg body weight, Charles River, Germany) were used. The animals were anesthetized, and venous access was established via an ear vein. The contralateral common jugular vein was then dissected, and the animals were placed into a temporary hemophilic state with a factor VI I antibody.

[0064] After intravenous administration of the test substance, a 1.5 cm long vein segment was ligated. After a waiting period of 20 minutes, the ligated vein segment was removed and incised in an isotonic sodium citrate buffer solution. The luminal vein surface was macroscopically inspected, and thrombi, if present, were removed and weighed. The following scale was used to evaluate thrombi:

[0065] 0 no thrombus

[0066] 1 small thrombi weighing < 2 mg

[0067] 2 one or more thrombi

[0068] 3 a thrombus completely filling the vein

[0069] The results show that prethrombin-1 is not thrombogenic in the Wessler test, even at high doses.

[0070] Example 4: Efficacy of prethrombin-1 in reversing the anticoagulation of oral Xa inhibitors

[0071] The efficacy of prethrombin-1 in reversing the anticoagulation of oral Xa inhibitors was demonstrated in a rivaroxaban rabbit model. Anesthetized New Zealand White rabbits were treated with the Factor Xa inhibitor rivaroxaban to achieve anticoagulation. The increased bleeding tendency associated with anticoagulation was determined by claw clipping. After intravenous administration of prethrombin-1, blood loss was measured again after claw clipping.

[0072] The results obtained are summarized in the table below.

[0073] The rivaroxaban dose was increased from 7 to 70 mg / kg body weight. Since a sufficient bleeding tendency was achieved at a dose of 70 mg / kg body weight, a further dose increase was not necessary. After intravenous administration of prethrombin-1, no blood loss was measurable in 10 animals. Blood loss was measurable in two animals, but this was less than after anticoagulation.

[0074] From these measurements, it can be concluded that prethrombin-1 can reverse the pro-bleeding effect of oral Xa inhibitors when reversal of anticoagulation is necessary due to life-threatening or uncontrolled bleeding. References:

[0075] (1) Orfeo T, Butenas S, Brummel-Ziedins KE, Mann KG. The tissue factor requirement in blood coagulation. J Biol Chem. 2005; 280(52):42887-96.

[0076] (2) Petrillo T, Ayombil F, Van't Veer C, Camire RM. Regulation of factor V and factor V-short by TFPIa: Relationship between B-domain proteolysis and binding. J Biol Chem. 2021 Jan-Jun; 296:100234.

[0077] (3) Eibl H, Schwarz O and Eisinger F. Patent AT 350726 (1976) Process for the preparation of a blood coagulation-promoting preparation from human blood plasma

[0078] (4) Eibl H, Schwarz O and Eisinger F. Patent AT 350726 USP 4,160,025 (1979) Method of producing a blood-coagulation-promoting preparation from human blood plasma.

[0079] (5) Turecek P. and Schwarz HP. (2013) "Factor eight inhibitor bypassing activity" in Production of Plasma Proteins for Therapeutic use, published by John Wiley & Sons inc.

[0080] (6) Butenas S, van't Veer C, Mann KG "Normal" thrombin generation. Blood. 1999 Oct l;94(7):2169-78.

[0081] (7) Hansson KM, Lindblom A, Elg M, Lövgren A. Recombinant human prothrombin (MEDI8111) prevents bleeding in haemophilia A and B mice. Haemophilia. 2016 May;22(3):453-61.

[0082] (8) To Assess Safety, Tolerability and Pharmacodynamics of Intravenous MEDI8111 After Single Ascending Doses. - Study Results - ClinicalTrials.gov (9) Stojanovski BM, Di Cera E. Role of sequence and position of the cleavage sites in prothrombin activation. J Biol Chem. 2021 Aug; 297(2):100955.

[0083] (10) Mann KG, Elion J, Butkowski RJ, Downing M, Nesheim ME. Prothrombin. Methods Enzymol. 1981;80 Pt C:286-302.

[0084] (11) Haynes LM, Bouchard BA, Tracy PB, Mann KG. Prothrombin activation by platelet- associated prothrombinase proceeds through the prethrombin-2 pathway via a concerted mechanism. J Biol Chem. 2012 Nov 9;287(46):38647-55.

[0085] (12) Chen Z, Pelc LA, Di Cera E. Crystal structure of prethrombin-1. Proc Natl Acad Sei U S A. 2010 Nov 9;107(45):19278-83.

[0086] (13) Bukys MA, Orban T, Kim PY, Nesheim ME, Kalafatis M. The interaction of fragment 1 of prothrombin with the membrane surface is a prerequisite for optimum expression of factor Va cofactor activity within prothrombinase. Thromb Haemost. 2008 Mar;99(3):511-22

[0087] (14) Ayombil F, Woodet al. Prethrombin-1, the Gla-domainless prothrombin intermediate, is activated efficiently to thrombin by prothrombinase assembled on the activated platelet surface. July 2011. Conference: International Society of Thrombosis and Haemostasis. Japan Volume: Journal of Thrombosis and Haemostasis 9, 352-35

[0088] (15) Friedmann AP, Koutychenko A, Wu C, Fredenburgh JC, Weitz JI, Gross PL, Xu P, Ni F, Kim PY. Identification and characterization of a factor Va-binding site on human prothrombin fragment 2. Sei Rep. 2019 Feb 21;9(1):2436.

[0089] (16) McDuffie FC, Giffin C, Niedringhaus R, Mann KG, Owen CA Jr, Bowie EJ, Peterson J, Clark G, Hunder GG. Prothrombin, thrombin, and prothrombin fragments in plasma of normal individuals and of patients with laboratory evidence of disseminated intravascular coagulation. Thromb Res. 1979;16(5-6):759-73. (17) Owen CA Jr, Mann KG, McDuffie FC. The turnover in normal dogs of prothrombin and its fragments; effect of induced intravascular coagulation. Thromb Haemost. 1979 Aug 31;42(2):548-55.

[0090] (18) Lanchantin GF, Friedmann JA, Hart DW. On the occurrence of polymorphic human prothrombin. Electrophoretic and chromatographic alterations of the molecule due to the action of thrombin. Biol Chem. 1968 Feb 10;243(3):476-86.

[0091] (19) Heldebrant CM, Butkowski RJ, Bajaj SP, Mann KG The activation of prothrombin. II. Partial reactions, physical and chemical characterization of the intermediates of activation. J Biol Chem. 1973 Oct 25;248(20):7149-63.

[0092] (20) Seegers WH, Hassouna HI, Novoa E. Immunological aspects of some vitamin K- dependent factors and preparation of depleted plasmas. Thromb Res. 1977 Nov;ll(5):633-42.

[0093] (21) Baker WJ, Seegers WH. The conversion of prethrombin to thrombin. Thromb Diath Haemorrh. 1967 Feb 28;17(l-2):205-13.

[0094] (22) Seegers WH, Ghosh A. Activation of prothrombin and factor X: function of previously unrecognized plasma protein. Thromb Res. 1980 Jan 1-15;17(1-2):71- 81.

[0095] (23) Seegers WH, Hassouna HI, Novoa E. Immunological aspects of some vitamin K- dependent factors and preparation of depleted plasmas. Thromb Res. 1977 Nov;ll(5):633-42.

[0096] (24) Meehan WP et al. Recombinant human thrombin: Demonstration of efficacy in models of surgical bleedings. Journal of surgical research (2004-10-01) 323

[0097] (25) Stroethmann M EP 0068 048 A2 (26) Varadi K, Tangada S, Loeschberger M, Montsch P, Schrenk G, Ewenstein B, Turecek PL. Pro- and anticoagulant factors facilitate thrombin generation and balance the haemostatic response to FEIBA® in prophylactic therapy. Haemophilia. 2016 Jul;22(4):615-24.

[0098] (27) S. Wessler, S.M. Reimer, M.C. Sheps Biologic assay of a thrombosis-inducing activity in human serum J Appl Physiol, 14 (1959), pp. 943-946

[0099] (28) A.R. Giles, M. Johnston, H. Hoogendoorn, M. Blajchman, J. Hirsh The thrombogenicity of prothrombin complex concentrates: I. The relationship between in vitro characteristics and in vivo thrombogenicity in rabbits Thromb Res, 17 (1980), pp. 353-366

Claims

Patent claims:

1. Prethrombin-1 for use as a medicinal product.

2. Prethrombin 1 for specific use in promoting hemostasis.

3. Prethrombin-1 for use according to claim 2 in the treatment of bleeding.

4. Prethrombin-1 for use according to claim 3 in the treatment of bleeding in patients with coagulation disorders.

5. Prethrombin-1 for use according to claim 3 in the treatment of bleeding resulting from trauma or internal injury.

6. Prethrombin-1 for use according to claim 3 in the treatment of breakthrough bleeding during replacement treatments with coagulation factors, factor VIII-mimicking therapies or gene therapies.

7. Prethrombin-1 for use in the reversal of anticoagulation by direct-acting, orally administered anticoagulants.

8. Recombinant prethrombin-1 for use according to any one of claims 1 to 7.

Images