MXPA99006754A - A stabilized mixture comprising fibrinogen - Google Patents
A stabilized mixture comprising fibrinogenInfo
- Publication number
- MXPA99006754A MXPA99006754A MXPA/A/1999/006754A MX9906754A MXPA99006754A MX PA99006754 A MXPA99006754 A MX PA99006754A MX 9906754 A MX9906754 A MX 9906754A MX PA99006754 A MXPA99006754 A MX PA99006754A
- Authority
- MX
- Mexico
- Prior art keywords
- arginine
- fibrinogen
- solution
- component
- lysine
- Prior art date
Links
- 108010049003 Fibrinogen Proteins 0.000 title claims abstract description 36
- 102000008946 Fibrinogen Human genes 0.000 title claims abstract description 36
- 229940012952 Fibrinogen Drugs 0.000 title claims abstract description 36
- 229940019698 Fibrinogen containing hemostatics Drugs 0.000 title claims abstract description 36
- 239000000203 mixture Substances 0.000 title claims abstract description 28
- GYDJEQRTZSCIOI-LJGSYFOKSA-N (1r,4r)-4-(aminomethyl)cyclohexane-1-carboxylic acid Chemical compound NC[C@H]1CC[C@H](C(O)=O)CC1 GYDJEQRTZSCIOI-LJGSYFOKSA-N 0.000 claims abstract description 28
- 229960000401 tranexamic acid Drugs 0.000 claims abstract description 28
- 239000004475 Arginine Substances 0.000 claims abstract description 24
- 229960003121 arginine Drugs 0.000 claims abstract description 23
- 239000004472 Lysine Substances 0.000 claims abstract description 19
- 229960003646 lysine Drugs 0.000 claims abstract description 18
- 239000011780 sodium chloride Substances 0.000 claims abstract description 4
- 210000002381 Plasma Anatomy 0.000 claims abstract description 3
- 150000003839 salts Chemical class 0.000 claims abstract description 3
- 239000000126 substance Substances 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims abstract 8
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 claims description 11
- 239000003106 tissue adhesive Substances 0.000 claims description 11
- 230000000694 effects Effects 0.000 claims description 10
- 229960003589 Arginine hydrochloride Drugs 0.000 claims description 9
- 108010054218 Factor VIII Proteins 0.000 claims description 8
- 102000001690 Factor VIII Human genes 0.000 claims description 8
- 229960000301 Factor VIII Drugs 0.000 claims description 8
- 230000002797 proteolythic Effects 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 7
- 102100019017 VWF Human genes 0.000 claims description 7
- 108010047303 von Willebrand Factor Proteins 0.000 claims description 7
- 229960001134 von Willebrand factor Drugs 0.000 claims description 7
- 229940088598 Enzyme Drugs 0.000 claims description 5
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 claims description 5
- GYDJEQRTZSCIOI-UHFFFAOYSA-N AMCHA Chemical compound NCC1CCC(C(O)=O)CC1 GYDJEQRTZSCIOI-UHFFFAOYSA-N 0.000 claims description 4
- 229950003499 FIBRIN Drugs 0.000 claims description 4
- 102100008658 FN1 Human genes 0.000 claims description 4
- 108010071289 Factor XIII Proteins 0.000 claims description 4
- 108010073385 Fibrin Proteins 0.000 claims description 4
- 102000009123 Fibrin Human genes 0.000 claims description 4
- 108010067306 Fibronectins Proteins 0.000 claims description 4
- 229960003766 Thrombin (Human) Drugs 0.000 claims description 4
- 229940012444 factor XIII Drugs 0.000 claims description 4
- 230000037320 fibronectin Effects 0.000 claims description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 4
- 102100009661 VTN Human genes 0.000 claims description 3
- 108010031318 Vitronectin Proteins 0.000 claims description 3
- UIIMBOGNXHQVGW-UHFFFAOYSA-M buffer Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 3
- 239000004471 Glycine Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000010188 recombinant method Methods 0.000 claims description 2
- 239000007853 buffer solution Substances 0.000 claims 2
- BVHLGVCQOALMSV-JEDNCBNOSA-N L-lysine hydrochloride Chemical compound Cl.NCCCC[C@H](N)C(O)=O BVHLGVCQOALMSV-JEDNCBNOSA-N 0.000 claims 1
- 229960005337 Lysine Hydrochloride Drugs 0.000 claims 1
- 239000012460 protein solution Substances 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 abstract description 5
- 108090000623 proteins and genes Proteins 0.000 abstract description 5
- 239000008366 buffered solution Substances 0.000 abstract 1
- 239000000306 component Substances 0.000 description 32
- 108010080379 Fibrin Tissue Adhesive Proteins 0.000 description 5
- 108091005771 Peptidases Proteins 0.000 description 5
- 102000035443 Peptidases Human genes 0.000 description 5
- 239000003292 glue Substances 0.000 description 5
- 230000001112 coagulant Effects 0.000 description 4
- 239000000701 coagulant Substances 0.000 description 4
- 239000004365 Protease Substances 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- OZAIFHULBGXAKX-UHFFFAOYSA-N precursor Substances N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000000087 stabilizing Effects 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- NKCXQMYPWXSLIZ-PSRDDEIFSA-N (2S)-2-[[(2S)-1-[(2S)-5-amino-2-[[2-[[(2S)-6-amino-2-[[2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S,3R)-2-amino-3-hydroxybutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-3-hydroxybutanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]-3-m Chemical compound O=C([C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCCCN)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@@H](N)[C@@H](C)O)[C@@H](C)O)C(C)C)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O NKCXQMYPWXSLIZ-PSRDDEIFSA-N 0.000 description 2
- 210000004369 Blood Anatomy 0.000 description 2
- 229940110715 ENZYMES FOR TREATMENT OF WOUNDS AND ULCERS Drugs 0.000 description 2
- 206010018987 Haemorrhage Diseases 0.000 description 2
- 229940024999 Proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 2
- 108090000190 Thrombin Proteins 0.000 description 2
- 229940075469 Tissue adhesives Drugs 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminum Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 238000010876 biochemical test Methods 0.000 description 2
- 230000000740 bleeding Effects 0.000 description 2
- 231100000319 bleeding Toxicity 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 238000004590 computer program Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 235000002639 sodium chloride Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000007711 solidification Methods 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 238000009662 stress testing Methods 0.000 description 2
- 229960004072 thrombin Drugs 0.000 description 2
- KWTQSFXGGICVPE-WCCKRBBISA-N (2S)-2-amino-5-(diaminomethylideneamino)pentanoic acid;hydron;chloride Chemical compound Cl.OC(=O)[C@@H](N)CCCN=C(N)N KWTQSFXGGICVPE-WCCKRBBISA-N 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K Aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- 108010051456 Plasminogen Proteins 0.000 description 1
- 102000013566 Plasminogen Human genes 0.000 description 1
- 102000014961 Protein Precursors Human genes 0.000 description 1
- 108010078762 Protein Precursors Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229940046010 Vitamin K Drugs 0.000 description 1
- 229930003448 Vitamin K Natural products 0.000 description 1
- 229940019697 Vitamin K containing hemostatics Drugs 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000000721 bacterilogical Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000000903 blocking Effects 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000003247 decreasing Effects 0.000 description 1
- 230000004059 degradation Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001419 dependent Effects 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000003480 fibrinolytic Effects 0.000 description 1
- 239000003527 fibrinolytic agent Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 230000018341 negative regulation of fibrinolysis Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000002195 synergetic Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000001225 therapeutic Effects 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
Abstract
A stabilized solution of fibrinogen containing samples, in particular a stabilized solution of the biological active component (BAC) which is a solution of proteins derived from blood plasma comprising fibrinogen, tranexamic acid and arginine or lysine or mixtures of arginine and lysine, their pharmaceutically acceptable salts, as well as, optionally, substances forming a buffered solution in aqueous medium.
Description
A STABILIZED MIXTURE THAT COMPRISES FIBRINOGEN
DESCRIPTION OF THE INVENTION
The invention pertains to a mixture comprising fibrinogen, particularly to a stabilized solution of the Biological Active Component (BAC), a two-component tissue glue comprising separately components A and B as well as a fibrin clot obtainable by mixing component A and component B. The fibrinogen contained in the samples can be used for various applications, for example in two-component tissue adhesives or fibrin glue. The Biologically Active Component (BAC) is a viscous solution of proteins that normally contains fibrinogen in amounts of approximately 50 mg of fibrinogen per ml, which has been derived from the cryoprecipitation of plasma. Such BAC was described in WO 94/22503. Since the procedures employed for virus inactivation and concentration result in an increase in proteolytic activity in cryoprecipitation, stabilization of the final product with antiproteolytic agents is required. Some of these enzymes
Ref .: 30741 proteolytics are in a zymogen form (proenzyme) and their activation is promoted by the minute amount of activated enzymes present in the cryoprecipitation. The temperature interval employed during the separation of the cryoprecipitation has been found to improve the activation of the precursors of the proteolytic enzymes. This activation presumably takes place by means of a "cascade" sequence, by means of which the proteases activate the precursor forms of other proteolytic enzymes, including the generation of the fibrinolytic plasmid of the plasminogenated precursor. Inhibition of fibrinolysis and other proteolytic activity has been found to reduce the degradation of factor VIII and other coagulated and adhesive proteins. It may happen that the BAC coagulants spontaneously already after storage for a day of 2 6 ° C. At lower temperatures this process is slower; Following storage for several months at -18 ° C, BACs without further treatment form a solid clot after melting and can not be reconstituted. The same phenomenon has been observed with cryoprecipitate without purification. During the production process, plasminogen and other proteases dependent on vitamin K are removed by absorption of aluminum hydroxide. However, some proteases are left in the final product. It is desirable to stabilize the fibrinogen comprised in a mixture, in particular a BAC solution in order to ensure a careful use of the product when applied in clinical operations. Surprisingly, this object is achieved by a mixture comprising fibrinogen, 4- (aminomethyl) cyclohexane-carboxylic acid as well as arginine, lysine or combinations thereof. Preferably, the mixture is present as a solution of fibrinogen, tranexamic acid (4- (aminomethyl) -cyclohexane-carboxylic acid) as well as arginine, lysine or combinations thereof. In a preferred embodiment of the invention comprising a solution of the biological active component (BAC) which has been stabilized with a combination of tranexamic acid its physiologically acceptable salts and arginine or lysine or combinations of lysine and arginine. Optionally, the solution is buffered to a physiologically compatible pH value. Tranexamic acid is a protease inhibitor which the scientific name is 4- (aminomethyl) cyclohexane-carboxylic acid. According to the invention, a buffer containing glycine is preferred. Arginine as well as lysine can be used according to the invention as a common salt, for example, as hydrochloride. The BAC is preferably obtained from the concentrated cryoprecipitate after being worked up as described in EP-A-534 178. Arginine has been described in the art as a stabilizer in therapeutic protein concentrates. Nevertheless, the synergistic effect provided by the combination of tranexamic acid and arginine as well as lysine is surprising. Preferably, the amount of tranexamic acid in the BAC solution is from about 1-20% by weight. The amount of arginine hydrochloride or lysine is preferably from about 0.1-4% by weight. More preferred are amounts of tranexamic acid of about 5-15% by weight, particularly preferred from about 8-12% by weight. Normally about 10% by weight can be used. The amount of arginine hydrochloride or lysine is more preferred in the range of about 1-3% by weight, particularly about 2% by weight is preferred. The sample comprises fibrinogen, in particular a solution of BAC is preferably derived from a cryoprecipitate which was concentrated by ultra-distillation as described in Patent OA-94/22503. The BAC preferably comprises a fibrinogen content of from about 15-150 mg / ml in particular from 20-80 mg / ml. The amount of fibrinogen can be measured according to the Clauss Method (Claues, A., "Gerinnungsphysiologische Schnell ethode zur Bestimmung des Fibrinogens", Acta. Haematol., 17, 237-247, 1957). The use of a BAC derived from the concentrated cryoprecipitate is advantageous from such a fraction which also contains fibrinogen also valuable blood components which play an important role for blood coagulation when a proteolytic enzyme such as a human thrombin is bound with a BAC solution. . The valuable components are factor VIII, factor XIII, fibronectin, vitronectin, von Willebrand factor (vWF), etc. Preferably, the components are derived from the cryoprecipitate in particular concentrated cryoprecipitates. However, it is also possible that the fibrinogen components, factor VIII, factor XIII, fibronectin, - the von factor. Willebrand (vWF), the vitronección have been prepared by recombinant methods. Such preparations, for example, for factor XIII are commercially available. It was found that the combination of tranexamic acid and arginine or lysine stabilizes a mixture containing fibrinogen in particular a solution of the biological active component BAC, (2-d ° C for at least 14 days by keeping at least 50% of activated fibrinogen ). When only one of the components was used, the sample is significantly more unstable. Furthermore, it was surprising that the use of tranexamic acid and arginine hydrochloride or lysine did not adversely affect the properties of a blood clot obtained from stabilized BAC according to the invention. For example, the maximum elongation of the clot was also maintained after 14 days of storage of the respective samples as well as the tensile strength remained almost at the same level. Also the activity of factor VIII in BAC was not negatively affected when tranexamic acid and arginine or lysine were added. In particular, the stabilized solution of the biological active component BAC according to the invention is well suited for the preparation of a two-component tissue adhesive. A tissue glue according to the invention is understood as a system which can be applied with or in a patient in need thereof, for example, to avoid severe bleeding during surgical operations. The tissue glue was also handled as a fibrin glue and was basically analogous to the natural blood coagulation cascade. Tissue glue is derived from two components prior to application in surgical operations. A component that contains fibrinogen which upon exposure to a proteolytic enzyme such as human thrombin forms fibrin which is the polymer that forms the basic material of the natural blood clot. During the surgical operations the two components are applied, for example, by two syringes which are emptied simultaneously to mix the two components as fast as possible and avoiding the blocking of the supply lines. The solution according to the invention is advantageous for preparing a two-component fibrin glue since the fibrinogen solution has not been freshly prepared but can be stored in a refrigerator at -18 ° C, neither decreasing its coagulant capacity nor its remarkable mechanical properties. The degrees of fibrinogen activity can be balanced by providing a higher amount of fibrinogen in the solution so that the proper use of the fibrinogen-containing solution (BAC) is not impeded. t Therefore, the object of the present invention is also a two-component tissue glue comprising separately components A and B wherein the component A comprises a solution according to the embodiments described in claims 1 to 10 and a component B which comprises a solution of a proteolytic enzyme which is capable of reacting with fibrinogen (or BAC respectively) to the fibrin form. Preferably, the proteolytic enzyme is human thrombin in particular has an activity from about 2 to 4,000 IU / ml. The activity of thrombin is measured according to the coagulant assay (European Pharmacopoeia). It is understood by a skilled person that a fibrin glue can be defined by its coagulable protein content instead of the definition based on coagulable fibrinogen. In order to provide a balanced solution of the mixed components A and B it may be advantageous to add to the component B approximately the same concentration of tranexamic acid and arginine.
Preferably, components A and B are applied in such a way that the equal volumes of the two components are mixed and applied in the patient to the side of the respective wound. Of course, it should be understood that the two tissue adhesives of the component can be used not only during surgical operations but also in other situations where bleeding must stop. The two components are preferably applied in a ratio of 1: 1. The following examples illustrate the advantages of the stabilizing combination of tranexamic acid and arginine. These examples in no way limit but explain the invention in greater detail.
Example 1
Preparation of the test substance
800 ml of a 0.8 g sample of sodium azide (0.1%) will be added as a powder to the volume to control bacteriological growth, immediately after receiving the sample from the production line. The addition of a bacteriostat is necessary to prevent contamination of the high concentration of fibrinogen in the sample that is difficult to filter. Each test concentration will be prepared extemporaneously by adding a mixture of two solid components to 100 ml of an aliquot of the sample. The addition of tranexamic acid and arginine-HCl will be carried out in a bucket with moderate agitation (multipoint magnetic stirring plate) for 10 minutes. All test formulations will be prepared in parallel and their fibrinogen concentration will be adjusted to 50 mg / ml by the addition of buffer B.
After further stirring for 5 minutes, the 10 ml glass jars with silicone will be filled with 5 ml of aliquots. All flasks will be frozen simultaneously and stored at -80 ° C until used. Before experimentation, they will, at a temperature of 2 6 ° C (day "0", reference time). Two bottles of each group will be stored at -80 ° C as positive controls. All the experiments will be carried out in duplicates and the bottles will be labeled according to the stabilizing concentrations as indicated below:
GROUP A: without stabilizer-bottles Ai - A20 GROUP B: 0% tranexamic acid and 2% of arginine monohydrochloride-fragments Bi-B20 GROUP C: 5% of tranexamic acid and 2% of arginine monohydrochloride - Ci - Cao GROUP D: 10% tranexamic acid and 2% of arginine monohydrochloride-phraseos Di-D20 GROUP E: 10% tranexamic acid and 0% of arginine monohydrochloride-phraseos Ei-E20 GROUP F: 10% tranexamic acid and 1 Arginine monohydrochloride-phraseos Fi-F20% GROUP G: 10% tranexamic acid and 4% arginine monohydrochloride-Gi "G-20 The extra amounts of the sample will take place as the retention samples and used if necessary. Before testing all frozen bottles at -80 ° C, they will melt at 37 ° C for 15 minutes and set at 2 - 6 ° C (day "O") .Two bottles from each group will be turned at room temperature for 15 minutes at day 0 and its mechanical properties and biochemical tests will be carried out and evaluated. The procedure will be repeated on days 2, 4, 7, 9, 11, 14 and 30.
Mechanical properties
The elongation test of the fibrin glue will be done in a CHATILLON MODEL TCD-200 tension machine with a CHATILLON Force Gauge of 1,000 g. The data will be handled by a computer program. This instrument is an engine driven tension and compression test designed to test the elasticity, pints of performance and breakthrough strength of various products and materials. A special molding was designed in order to produce standardized clots of the solidified glue in a form which could easily be attached to the stress testing machine. This consists of two conical aluminum molds, each 2.5 cm in height, which are placed one above the other (see figure 1). The liquid components of the glue are injected into the mold where they solidify in a standard 12.7 mm x 5 mm clot. Figure 1 illustrates the special mold designed to retain the liquid glue until solidification. The molds are then anchored to the stress testing machine, and the tensile strength and the elongation of the glue cylinder within the size of the mold by extracting the two molds separately. A computer program checks the elongation and tension at every 0.2 s. The points are plotted on two axes: the elongation against the force in grams in a given time.
Test procedure
Bottles containing BAC were incubated
37 ° C for 15 minutes and then turned at room temperature for 15 minutes. The particular design of the aluminum molds (as described above) were filled with a combined solution of 0.5 ml of BAC and 0.5 ml of thrombin (8 IU / ml).
The molds were left at an ambient temperature for 45 minutes to allow the solidification of the glue, and then they were mounted on the indicator and the resulting clots were tested for the maximum elongation capacity and tensile strength (the tensile force required for break the clot).
Performing biochemical tests
The following proteins were tested for stability by the methods described:
The activity of the coagulant Fibrinogen: The fibrinogen was measured quantitatively by the coagulation method according to Clauss.
Example 2 - The Effect of Various Concentrations of Tranexamic Acid and Arginine-HCl on Force / mm (Declination) of the Clot
or.
Conclusion: In freezing either the Arg. Or TEA improves the decline of the coagulum (the clot is stronger than without the stabilizers but TEA has a stabilizing effect on the coagulum strength over time when incubated at 4 - 8 ° C
Example 3 The Effect of Various Concentrations of Tranexamic Acid and Arginine-HCl on the Content of Coagulable Fibrinogen
s.
Conclusion: Stabilized coagulable fibrinogen of TEA
Example 4, The Effect of Various Concentrations of Tranexamic Acid and Arginine-HCl on Factor VIII Activity
Conclusion: The stabilized Factor VIII of Arg. In freezing and the stabilized Factor VIII of TEA during the incubation of 4 - 8 ° C.
It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers. Having described the invention as above, it is -claimed as property contained in the following:
Claims (2)
1. A mixture characterized in that it comprises fibrinogen, (4-aminomethyl) -cyclohexane-carboxylic acid (tranexamic acid) as well as arginine, lysine or combinations thereof, wherein the amount of tranexamic acid is about 1% -20% by weight and the amount of arginine or lysine is about 0.1% - 4% by weight of arginine hydrochloride or lysine 2. The mixture according to claim 1, characterized in that it comprises a solution of fibrinogen, tranexamic acid as well as arginine, lysine and / or combinations thereof. 3. The mixture according to claim 2, characterized in that the solution comprises a biological active component (BAC) which is a protein solution derived from the blood plasma comprising fibrinogen, tranexamic acid and arginine or lysine or mixtures or arginine and lysine, its pharmaceutically acceptable salts, as well as, optionally, substances that form a buffer solution in an aqueous medium. 4. The mixture according to any of claims 2 - 3, characterized in that the amount of tranexamic acid is approximately 5% - 15% by weight and the amount of lysine or arginine hydrochloride is about 1% - 3% by weight. 5. The mixture according to claim 3, characterized in that the BAC was derived from a concentrated cryoprecipitate. 6. The compliance solution csn any of claims 2 to 5, characterized in that the BAC comprises a fibrinogen content from about 15 to 150 mg / ml 7. The mixture according to any of claims 2 to 6, characterized in that the BAC further comprises factor VIII, factor XIII, fibronectin, von Willebrand factor (vWF), vitronectin. 8. The mixture according to claim 7, characterized in that factors VIII, XIII, fibronectin, von Willebrand factor (vWF), vitronectin have been prepared by recombinant methods. 9. The solution according to any of claims 2 to 8, characterized in that the buffer solution is a glycine buffer. 10. A two-component tissue glue comprising separately components A and B, characterized in that component A comprises a mixture of one of claims 2 to 9, and component B comprises a solution of a proteolytic enzyme which is capable of forming fibrin when reacted with fibrinogen. 11. The two-component tissue adhesive according to claim 10, characterized in that component B comprises human thrombin in an activity of approximately 2 4000 international units per ml measured according to the Clauss method. 12. The tissue glue of two components according to any of claim 11 or 12, characterized in that the component B is stabilized with tranexamic acid and arginine.
2 . The fibrin clot characterized in that it is obtained by mixing component A and component B in an appropriate ratio, preferably 1: 1.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP97101569 | 1997-01-31 |
Publications (1)
Publication Number | Publication Date |
---|---|
MXPA99006754A true MXPA99006754A (en) | 2000-06-01 |
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