CA1128859A - Tissue adhesive - Google Patents

Tissue adhesive

Info

Publication number
CA1128859A
CA1128859A CA344,717A CA344717A CA1128859A CA 1128859 A CA1128859 A CA 1128859A CA 344717 A CA344717 A CA 344717A CA 1128859 A CA1128859 A CA 1128859A
Authority
CA
Canada
Prior art keywords
fibrinogen
tissue adhesive
set forth
inhibitor
cryoprecipitate
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired
Application number
CA344,717A
Other languages
French (fr)
Inventor
Otto Schwarz
Yendra Linnau
Franz Loblich
Thomas Seelich
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Oesterreichisches Institut fuer Haemoderivate
Original Assignee
Immuno AG fuer Chemisch Medizinische Produkte
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=3508534&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=CA1128859(A) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Immuno AG fuer Chemisch Medizinische Produkte filed Critical Immuno AG fuer Chemisch Medizinische Produkte
Application granted granted Critical
Publication of CA1128859A publication Critical patent/CA1128859A/en
Expired legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/16—Blood plasma; Blood serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/005—Enzyme inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A61L24/10—Polypeptides; Proteins
    • A61L24/106—Fibrin; Fibrinogen
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
    • Y10S530/827—Proteins from mammals or birds
    • Y10S530/829—Blood
    • Y10S530/83—Plasma; serum

Abstract

ABSTRACT OF THE DISCLOSURE:

A tissue adhesive on the basis of human or animal pro-tein contains factor XIII and at least 33 % by weight of fibrinogen, has a ratio of factor XIII to fibrinogen, ex-pressed in units of factor XIII per gram of fibrinogen, of at least 80, contains fibrinogen and albumin in the overall protein at a ratio of 33 to 90 : 5 to 40, contains plasmino-gen-activator-inhibitor or plasmin inhibitor in an amount of 250 to 25,000 KIU per g of fibrinogen and has been lyophilized.

Description

1~8~

The invention relates to a tissue adhesive on the basis of human or animal proteins, containing fibrinogen and factor XIII.
It has been known for long to use blood coagulating substances for stopping bleedings and for sealing wounds.
According to the first proposals of this kind, fibrin tampons and fibrin platelets, respectively, were used.
During the Second World War, tissue adherences by means of blood plasma were suggested.
In recent times, a tissue adhesive on the basis of fibrinogen and factor XIII for seamless interfascicular nerve transplantations in animal experiments has been described by H. Matras et al. in "Wiener Medizinischen Wochenschrift", 1972, page 517.
A further study was carried out by Spangler et al.
in "Wiener Klinischen Wochenschrift", 1973, pages 1 to 7.
Also there, the possibility was shown in animal experiments of carrying out a tissue adherence with the aid of fibri-nogen as a cryoprecipitate and thrombin.
The known preparatlons have not yet proved satis-factory, since they do not sufficiently meet the demands set to a tissue adhesive, i.e.
a) high straining capacity of the adherences and wound sealings as well as safe and permanent blood stopping, i.e. good adhering capacity of the adhesive to the wound and tissue surfaces, as well as hi~h internal strength of the same, ; b) controllable durability of the adherences in the body, c) complete absorbability of the adhesive in the course 3~

8~3S~

of the wound healing process, (d) wound healing stimulating properties.
This may partly be due to the fact that, in the known preparat-ions the coagulation factors necessary for blood stopping have not been present in an optimal proportion to one another, and also to the fact that the fibrinolytic activity in the area of adher-ence has not been sufficiently under control. Premature disso-lutions of the tissue adherences frequently occurred due to enzymatic influence.
The invention aims at avoiding these disadvantages and difficulties and has as its object to provide a lyophilized tissue adhesive of human or animal origin, which meets the demandsset out further above and which is present in a lyophili-zed form, which is desir~d for its longer durability and better transporting and storing properties.
Accordingly, the invention provides a tissue adhesive on the basis of human or animal proteins and containing fibrino-gen and factor XIII, which tissue adhesive is characterized in that (a) it contains at least 33% by weight of fibrinogen, (b)- the ratio of factor XIII to fibrinogen, expressed in units of factor XIII per gram of fibrinogen, amounts to at least 80, (c) fibrinogen and albumin are contained in the total protein at a ratio of 33 to 90 : 5 to ~0, (d) it has a content of plasminogen-activator-inhibitor or plasmin-inhibitor, preferably aprotinin, in an amount of 250 to 25,000 Kallikrein-inactivator-units (KIU) per gram of fibrinogen, (e) the preparation has been lyophilized.
According to a preferred embodiment the tissue adhe-~L%88~i~

sive additionally contains glycine, whereby the resolubility of the lyophilized product is improved.
Furthermore, the tissue adhesive additionally may contain glucose or sucrose, which components also improve the solubility.
The tissue adhesive furthermore may contain 0.2 to 200 International Units (IU) of heparin per gram of fibrinogen, whereby a stabilizing effect is obtained.
The tissue adhesive according to the invention posses-ses characteristic cross-linking prop~ertiés after the dis-solution, which are determinable by the sodium-dodecyl-sul-phate-(SDS)-polyacrylamide-gel-electrophoresis method. The test is carried out in that, after mixing of the tissue ad-hesive with an equal volume of a solution containing 40 ~Moles of CaCl2 and 15 NIH-units (U.S. National Institute of Health units) of thrombin per ml, the mixture is incu-bated at 37C. The cross-linking degree is determined by a gel electrophoresis after stopping of the reaction and reductive splitting of the disulphide bridges contained in the proteins by the addition of a mixture of urea, sodium dodecyl sulphate and ~-mercaptoethanol. Typical of the tissue adhesive according to the invention is a complete cross-linking of the fibrin-~-chains after 3 to 5 minutes, and a cross-linking of at least 35 ~ of the fibrin-~ chains a ter two hours.
Fibrinogen, albumin and cold-insoluble globulin, in the total protein, are to be present in the tissue adhesive ac-cording to the invention at a certain ratio; this ratio amounts to 33 to 90 : 5 to 40 : 0.2 to 15.
The invention moreover comprises a method of producing 81~S~3 the tissue adhesive described from plasma cryoprecipitate, which method comprises:
removing from the cryoprecipitate plasma-protein that is soluble in the cold, by treating the cryoprecipitate with a buffer solution containing sodium citrate, sodium chloride, glycine, glucose, a plasminogen-activator-inhibitor or plasmin-inhibitor, and h~parin, dissolving the purified precipitate, adding human albumin, and lyophilizing the resulting solution.
Advantageously, the cryoprecipitate has been produced of human or animal fresh plasma frozen at -20C. When in-creasing the temperature to 0 to 2C the cryoprecipitate is gained and separated by centrifugation. The precipitate is eluted by a single or repeated elution with the buffer solution having a pH of 6 to 8.0, and centrifuged at 0 to 4C in order to remove the plasma-protein that is soluble in the cold. The treat-ment with the buffer solution is carried out until the desired factor-XIII-fibrinogen ratio is reached.
The purified precipitate is dissolved with a further buffer solution containing human albumin, glycine and, if desired, glucose or sucrose, a plasminogen-activator-inhibitor or plasmin inhibitor as well as heparin, and having a pH of 6.5 to 9.0, and is diluted to a protein concentration of 4.~0 to 9.0 ~. The solution is filtered through a membrane filter having a pore size of down to 0.2 )um, filled into final containers and lyophilized.
The lyophilized tissue adhesive thus obtained can be stored at room temperature or preferably at ~4C; it is ready for use after reconstitution with aqua ad iniectabilia, to which~
if desired, a plasminogen-activator-inhibitor , . ~
,,, l ~1~8~5~

or a plasmin inhibitor, preferably aprotinin, can be added.
When dissolving the lyophilized preparation, attention has to be paid that the solution ready for use contains at least 70 mg of fibrinogen per ml.
The tissue adhesive according to the invention can be applied universally. It can be used for seamless connection of human or animal tissue or organ parts, for sealing wounds and stopping bleedings as well as for stimulating wound healings.
Preferred fields of application in which the tissue adhesive can be successfully used are: indications in the field of ear, nose and throat surgery, oral surgery, dentistry, neurosurgery, plastic surger~7, general sur~ery, abdominal surgery, thorax and vascular surgery, orthopaedics, accident surgery, urology, ophthalmology and gynaecology.
Advantageously,a mixture of thrombin and calcium chloride is added to the adhesive prior to the application of the tissue adhesive according to the invention or is applied onto the tissues to be connected.
The method of the lnvention is explained in more detail by way of the following example:
21 l of human plasma, which had been frozen at -20C, were heated to +2C. The resulting cryoprecipitate (435 ~) was separated by centrifugation at +2C and treated at +2C
with 4.3 l of a buffer solution adjusted at a pH of 6.5 and containing 6.6 g of Na3-citrate.2H20, 3.4 g of NaCl, 10.0 g of glycine, 13.0 of glucose.1H20~ 50,000 KIU of aprotinin and 200 IU of heparin per l, and again centrifuged at +2C.
The separated precipitate was dissolved in a further buffer 3G solution having a pH of 7.9 and containing 35.0 g of human ~z~

albumin, 20.0 g of glycine, 50,000 ~IU of aprotinin and 200 IU of heparin per l, and diluted to a concentration of 70 mg of protein per ml.
Then the solution was sterilized by filtration through membrane filters having a pore size of down to 0.2 ~m, fill-ed into final containers at 2.2 ml each, deepfrozen and lyo-philized. After reconstitution of the lyophilized product to a fibrinogen concentration of ~0 mg/ml, the tissue adhesive preparation ready for use showed, in the cross-linking test, complete fibrin-~-cross-linking after S minutes and 66 %
fibrin-~-cross-linking after 2 hours at 37C.
The ratio of the proteins fibrinogen to albumin to cold-insoluble globulin, contained in the tissue adhesive, was determined to be 64.0 : 22.3 : 7.7. The heparin content was 4.5 IU per g of fibrinogen. Aprotinin was contained at a concentration of 1,133 KIU per g of fibrinogen. The con-tent of factor XIII amounted to 161 units per g of fibrino-gen. The total protein content in the lyophilized prepara-t~on was 72.2 %, the content of fibrinogen in the lyophilized preparation was 46.2 %.
The determinations were carried out in the following manner: The determination of the factor-XIII-units was per-formed by means of a cross-linking test, in which factor-XIII-free fibrinogen was used as a substrate and the fibrin cross-linking caused by the addition of the unknown diluted sample served as a measure for the amount of factor XIII
contained therein~ A corresponding calibration curve was obtained with pooled human citrate plasma, 1 ml plasma con-taining 1 unit of factor XIII per definitionem. The quanti-tative protein determinations were carried out by the method ~1~8~5~

according to Kjeldahl.
The determination of the proteins relative to oneanother was also performed by the SDS-polyacrylamide-gel-electrophoresis method, i.e. a) with a non-reduced sample of the tissue adhesive and b) with a sample reduced with ~-mercaptoethanol.

Claims (11)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A tissue adhesive on the basis of human or animal pro-teins and containing fibrinogen and factor XIII, which tissue adhesive is characterized in that a) it contains at least 33 % by weight of fibrinogen, b) the ratio of factor XIII to fibrinogen, expressed in units of factor XIII per gram of fibrinogen, amounts to at least 80, c) fibrinogen and albumin are contained in the total protein at a ratio of 33 to 90 : 5 to 40, d) it contains plasminogen-activator-inhibitor or plasmin inhibitor in an amount of 250 to 25,000 KIU per g of fibrinogen, and e) it is lyophilized.
2. A tissue adhesive as set forth in claim 1, wherein the plasminogen-activator-inhibitor or plasmin-inhibitor is aprotinin.
3. A tissue adhesive as set forth in claim 1, further con-taining glycine.
4. A tissue adhesive as set forth in claim 1, further con-taining glucose.
5. A tissue adhesive as set forth in claim 1, further con-taining sucrose.
6. A tissue adhesive as set forth in claim 1, which con-tains 0.2 to 200 IU of heparin per g of fibrinogen.
7. A tissue adhesive as set forth in claim 1 being capable, after dissolution of the lyophilized preparation of com-plete cross-linking of the fibrin-?-chains after 3 to 5 minutes of incubation, and of at least 35 % cross-linking of the fibrin-?-chains after 2 hours of incubation, deter-mined according to the SDS-polyacrylamide-gel-electrophor-esis method.
8. A tissue adhesive as set forth in claim 1, wherein the ratio of fibrinogen to albumin to cold-insoluble globulin in the total protein is 33 to 90 : 5 to 40 : 0.2 to 15.
9. A method of producing a tissue adhesive as set forth in claim 1 from plasma cryoprecipitate, which method com-prises removing from the cryoprecipitate plasma-protein that is soluble in the cold, by treating the cryoprecipitate with a buffer solution containing sodium citrate, sodium chloride, glycine, glucose, a plasminogen-activator-in-hibitor or plasmin-inhibitor, and heparin, dissolving the purified precipitate, adding human albumin, and lyophilizing the resulting solution.
10. A method as set forth in claim 9, wherein the cryopreci-pitate is treated once with said buffer solution.
11. A method as set forth in claim 9, wherein the cryoprecipitate is treated several times with said buffer solution.
CA344,717A 1979-02-15 1980-01-30 Tissue adhesive Expired CA1128859A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AT118979A AT359652B (en) 1979-02-15 1979-02-15 METHOD FOR PRODUCING A TISSUE ADHESIVE
ATA1189/79 1979-02-15

Publications (1)

Publication Number Publication Date
CA1128859A true CA1128859A (en) 1982-08-03

Family

ID=3508534

Family Applications (1)

Application Number Title Priority Date Filing Date
CA344,717A Expired CA1128859A (en) 1979-02-15 1980-01-30 Tissue adhesive

Country Status (14)

Country Link
US (2) US4362567A (en)
JP (1) JPS55110556A (en)
AT (1) AT359652B (en)
BE (1) BE881466A (en)
CA (1) CA1128859A (en)
CH (1) CH649711A5 (en)
DE (1) DE3002934C2 (en)
DK (1) DK150815B (en)
ES (1) ES8101381A1 (en)
FR (1) FR2448900A1 (en)
GB (1) GB2041942B (en)
IT (1) IT1141211B (en)
NL (1) NL190714B (en)
SE (2) SE453799C (en)

Cited By (2)

* Cited by examiner, † Cited by third party
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US5395923A (en) * 1993-02-23 1995-03-07 Haemacure-Biotech, Inc. Process for the obtention of a biological adhesive made of concentrated coagulation factors by "salting-out"

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