US4957903A
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Pharmaceutical and clinical compositions of desAA fibrin monomers and the tetrapeptide gly-pro-arg-pro
Assignee: Biopool AB
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Filing: 1986-03-27
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Granted: 1990-09-18
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Expires: 2006-03-27
Abstract
Abstract The invention relates to a fully solubilizable fibrin based composition, which is characterized by the combination that the fibrin is desAA-fibrin or desAA-fibrin from which the C-terminal portions of the α-chains have been removed by enzymatic digestion, and that the solubilizing agent is a tetrapeptide containing the amino acid sequence -L-prolyl-L-arginyl-, preferably glycyl-L-prolyl-L-arginyl-L-prolin. The full solubility of the fibrin makes possible new uses within the area of determination of important fibrinolytical parameters, and the invention also relates to three such important alternative uses. A first use according to the invention is the use of the composition in connection with detection or quantification of the activity of the enzyme tissue plasminogen activator. A first alternative use means that the composition is usd for the detection of soluble fibrin in bioligical fluids, and a second alternative use means that the composition is used for measuring or studying fibrinolytic activity in vitro or in vivo through the addition of the composition and registration of the degradation of the fibrin.
Inventors
- 👤 Mats G. Ranby
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 05:07
CPC Classifications
C; C07; C07K; C07K5/00; C07K5/04; C07K5/10; C07K5/1002; C07K5/1005; C07K5/1008; C12; C12Q; C12Q1/00; C12Q1/56; G; G01; G01N; G01N2333/00; G01N2333/435; G01N2333/745; G01N2333/75; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/972; G01N2333/9726; Y; Y10; Y10S; Y10S530/00; Y10S530/806; Y10S530/827; Y10S530/829
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
45 words
A solubilizable, fibrin based composition, consisting of: (a) approximately 0.1 to 20 mg/ml desAA-fibrin, and (b) approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, wherein the peptide prevents polymerization of said desAA-fibrin, thus solubilizing said desAA-fibrin to form a stable, non-polymerizing composition.
Claim 2
Dependent Claim
45 words
A solubilizable, fibrin based composition, consisting of: (a) approximately 0.1 to 20 mg/ml desAA-fibrin, and (b) approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, wherein the peptide prevents polymerization of said desAA-fibrin, thus solubilizing said desAA-fibrin to form a stable, non-polymerizing composition.
Claim 3
Dependent Claim
23 words
The composition of claim 1, wherein the desAA-fibrin has been obtained by a selective removal of fibrinopeptide A from fibrinogen with an enzyme.
Claim 4
Dependent Claim
12 words
The composition of claim 1, wherein the peptide concentration is 0.6-4 mg/ml.
Claim 5
Dependent Claim
11 words
The composition of claim 1 which additionally includes a physiological buffer.
Claim 6
Dependent Claim
10 words
The composition of claim 2, wherein said enzyme is batroxobin.
Claim 7
Dependent Claim
13 words
The composition of claim 1, wherein the peptide concentration is about 2 mg/ml.
Claim 8
Independent Claim
81 words
A method for the detection or quantification of tissue plasminogen activator in a sample comprising the steps of: (a) incubating the sample, plasminogen, a plasmin substrate and a soluble fibrin composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin and approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, so that any tissue plasminogen activator in the sample causes the formation of plasmin which causes cleavage of the plasmin substrate; and (b) measuring the amount of plasmin substrate cleaved.
Claim 9
Independent Claim
81 words
A method for the detection or quantification of tissue plasminogen activator in a sample comprising the steps of: (a) incubating the sample, plasminogen, a plasmin substrate and a soluble fibrin composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin and approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, so that any tissue plasminogen activator in the sample causes the formation of plasmin which causes cleavage of the plasmin substrate; and (b) measuring the amount of plasmin substrate cleaved.
Claim 10
Independent Claim
73 words
A method for the detection or quantification of plasminogen in a sample comprising the steps of: (a) incubating the sample, a tissue plasminogen substrate and a soluble fibrin composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin and approximately 0.4 to 10 mg/ml of a peptide consisting of glycl-L-prolyl-L-arginyl-L-proline, so that any tissue plasminogen activator in the sample causes cleavage of the tissue plasminogen substrate; and (b) measuring the amount of substrate cleaved.
Claim 11
Independent Claim
73 words
A method for the detection or quantification of plasminogen in a sample comprising the steps of: (a) incubating the sample, a tissue plasminogen substrate and a soluble fibrin composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin and approximately 0.4 to 10 mg/ml of a peptide consisting of glycl-L-prolyl-L-arginyl-L-proline, so that any tissue plasminogen activator in the sample causes cleavage of the tissue plasminogen substrate; and (b) measuring the amount of substrate cleaved.
Claim 12
Dependent Claim
13 words
The method of claim 8, wherein the plasmin substrate is a paranitro anilide.
Claim 13
Dependent Claim
11 words
The method of claim 9, wherein the paranitro anilide is H-D-norleucyl-L-hexahydrotyrosyl-L-lysinyl-paranitro-anilide.
Claim 14
Dependent Claim
12 words
The method of claim 8, wherein the product is cleaved plasmin substrate.
Claim 15
Independent Claim
78 words
A method for the detection or quantification of tissue plasminogen activator in a sample comprising the steps of: (a) incubating the sample with a synthetic tissue plasminogen activator substrate, and a soluble fibrin composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin and approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, so that any tissue plasminogen activator in the sample causes cleavage of the tissue plasminogen substrate; and (b) measuring the amount of substrate cleaved.
Claim 16
Independent Claim
78 words
A method for the detection or quantification of tissue plasminogen activator in a sample comprising the steps of: (a) incubating the sample with a synthetic tissue plasminogen activator substrate, and a soluble fibrin composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin and approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, so that any tissue plasminogen activator in the sample causes cleavage of the tissue plasminogen substrate; and (b) measuring the amount of substrate cleaved.
Claim 17
Dependent Claim
14 words
The method of claim 12, wherein the product is cleaved tissue plasminogen activator substrate.
Claim 18
Dependent Claim
66 words
A solubilizable, fibrin based composition, consisting of: (a) approximately 0.1 to 20 mg/ml desAA-fibrin from which the C-terminal portions of the α-chains have been removed from some of the desAA-fibrin by enzymatic digestion with plasmin, and (b) approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, wherein, the peptide prevents polymerization of said desAA-fibrin, thus solubilizing said desAA-fibrin to form a stable, non-polymerizing composition.
Claim 19
Dependent Claim
66 words
A solubilizable, fibrin based composition, consisting of: (a) approximately 0.1 to 20 mg/ml desAA-fibrin from which the C-terminal portions of the α-chains have been removed from some of the desAA-fibrin by enzymatic digestion with plasmin, and (b) approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline, wherein, the peptide prevents polymerization of said desAA-fibrin, thus solubilizing said desAA-fibrin to form a stable, non-polymerizing composition.
Claim 20
Dependent Claim
23 words
The composition of claim 14, wherein the desAA-fibrin has been obtained by a selective removal of fibrinopeptide A from fibrinogen with an enzyme.
Claim 21
Dependent Claim
10 words
The composition of claim 15, wherein the enzyme is batroxobin.
Claim 22
Dependent Claim
12 words
The composition of claim 14, wherein the peptide concentration is 0.6-4 mg/ml.
Claim 23
Dependent Claim
13 words
The composition of claim 17, wherein the peptide concentration is approximately 2 mg/ml.
Claim 24
Dependent Claim
11 words
The composition of claim 14, which additionally includes a physiological buffer.
Claim 25
Independent Claim
98 words
A method for the detection or quantification of tissue plasminogen activator in a sample comprising the steps of: (a) incubating the sample with plasminogen, a plasmin substrate, and a stable, soluble composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin from which the C-terminal portions of the α chain have been removed from some of the desAA-fibrin by enzymatic digestion with plasmin, and approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline so that any tissue plasminogen activator in the sample causes cleavage of the plasmin substrate; and (b) measuring the amount of plasmin substrate cleaved.
Claim 26
Independent Claim
98 words
A method for the detection or quantification of tissue plasminogen activator in a sample comprising the steps of: (a) incubating the sample with plasminogen, a plasmin substrate, and a stable, soluble composition consisting of approximately 0.1 to 20 mg/ml desAA-fibrin from which the C-terminal portions of the α chain have been removed from some of the desAA-fibrin by enzymatic digestion with plasmin, and approximately 0.4 to 10 mg/ml of a peptide consisting of glycyl-L-prolyl-L-arginyl-L-proline so that any tissue plasminogen activator in the sample causes cleavage of the plasmin substrate; and (b) measuring the amount of plasmin substrate cleaved.
Claim 27
Dependent Claim
12 words
The method of claim 20, wherein the product is cleaved plasmin substrate.
Claim 28
Dependent Claim
12 words
The composition of claim 14, wherein the plasmin comprises matrix bound plasmin.
Claim 29
Dependent Claim
23 words
The method of claim 20, wherein the desAA-fibrin has been obtained by a selective removal of fibrinopeptide A from fibrinogen with an enzyme.
Claim 30
Dependent Claim
10 words
The method of claim 23, wherein the enzyme is batroxobin.
Claim 31
Dependent Claim
14 words
The method of claim 20, wherein the peptide concentration is 0.6 to 4 mg/ml.
Claim 32
Dependent Claim
12 words
The method of claim 20, wherein the peptide concentration is 2 mg/ml.
Claim 33
Dependent Claim
14 words
The method of claim 20, wherein the soluble composition additionally includes a physiological buffer.
Claim 34
Dependent Claim
12 words
The method of claim 20, wherein the plasmin comprises matrix bound plasmin.
Claim 35
Dependent Claim
23 words
The method of claim 7, wherein the desAA-fibrin has been obtained by a selective removal of fibrinopeptide A from fibrinogen with an enzyme.
Claim 36
Dependent Claim
10 words
The method of claim 29, wherein the enzyme is batroxobin.
Claim 37
Dependent Claim
14 words
The method of claim 7, wherein the peptide concentration is 0.6 to 4 mg/ml.
Claim 38
Dependent Claim
12 words
The method of claim 7, wherein the peptide concentration is 2 mg/ml.
Claim 39
Dependent Claim
15 words
The method of claim 7, wherein the soluble fibrin composition additionally includes a physiological buffer.
Claim 40
Dependent Claim
23 words
The method of claim 8, wherein the desAA-fibrin has been obtained by a selective removal of fibrinopeptide A from fibrinogen with an enzyme.
Claim 41
Dependent Claim
10 words
The method of claim 34, wherein the enzyme is batroxobin.
Claim 42
Dependent Claim
14 words
The method of claim 8, wherein the peptide concentration is 0.6 to 4 mg/ml.
Claim 43
Dependent Claim
12 words
The method of claim 8, wherein the peptide concentration is 2 mg/ml.
Claim 44
Dependent Claim
15 words
The method of claim 8, wherein the soluble fibrin composition additionally includes a physiological buffer.
Claim 45
Dependent Claim
23 words
The method of claim 12, wherein the desAA-fibrin has been obtained by a selective removal of fibrinopeptide A from fibrinogen with an enzyme.
Claim 46
Dependent Claim
10 words
The method of claim 39, wherein the enzyme is batroxobin.
Claim 47
Dependent Claim
14 words
The method of claim 12, wherein the peptide concentration is 0.6 to 4 mg/ml.
Claim 48
Dependent Claim
12 words
The method of claim 12, wherein the peptide concentration is 2 mg/ml.
Claim 49
Dependent Claim
15 words
The method of claim 12, wherein the soluble fibrin composition additionally includes a physiological buffer.
References Cited (Backward Citations)
US3966701A
Cited
US4455290A
Cited
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