EP0376189A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Kit or reagent for assaying the cellular integrity of blood platelets and pharmaceutical compositions containing a PKA inhibitor
Assignee: Yeda Research and Development Co Ltd
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Filing: 1989-12-22
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Granted: —
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Expires: 2009-12-22
Abstract
Abstract There is provided a kit or reagent for assaying the cellular integrity (intactness) of blood platelets comprising means for determining the quantity of PKA released by a number of platelets into a certain volume of plasma. The assay is based on determining cAMP-dependent phosphorylation of vitronectin (protein S) in plasma using radioactively labelled) [γ³²P]ATP as phosphate donor. The assay can be based on the specific inhibition of the phosphorylation in blood fluid by protein kinase inhibitors. The invention further relates to pharmaceutical compositions for the treatment of pathological conditions associated with impaired platelet function in humans, which comprises administering to a person in need thereof a physiologically active quantity of a PKA inhibitor adapted to decrease or prevent vitronectin phosphorylation. The inhibitor can be a synthetic or genetically engineered peptide.
Inventors
- 👤 Shmuel Shaltiel
- 👤 Beatriz Grodzicki
- 👤 Daniel Chain
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 06:42
CPC Classifications
C; C07; C07K; C07K7/00; C07K7/04; C07K7/06; C07K14/00; C07K14/435; C07K14/78; C07K16/00; C07K16/18; C07K16/38; C12; C12Q; C12Q1/00; C12Q1/48; C12Q1/56; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/86; A; A61; A61K; A61K38/00; G01N2333/00; G01N2333/435; G01N2333/745; G01N2333/78; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/975
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
34 words
A kit or reagent for assaying the cellular integrity (intactness) of blood platelets which comprises means for determining the quantity of PKA released by a number of platelets into a certain volume of plasma.
Claim 2
Dependent Claim
34 words
A kit or reagent for assaying the cellular integrity (intactness) of blood platelets which comprises means for determining the quantity of PKA released by a number of platelets into a certain volume of plasma.
Claim 3
Dependent Claim
43 words
A kit or reagent according to claim 1, wherein the cAMP-dependent phosphorylation of vitronectin (protein S) is determined in plasma using radioactively labelled [γ³²P]ATP as phosphate donor, optionally in the presence of heparin, comprising means for determining the radioactivity of the PKA-phosphorylated vitronectin.
Claim 4
Dependent Claim
26 words
A kit or reagent according to claim 1 or 2, wherein the specific inhibition of the phosphorylation in blood fluid by protein kinase inhibitors is used.
Claim 5
Dependent Claim
32 words
A kit or reagent according to claim 1 for carrying out a radioimmunoassay, which comprises means for monitoring the release of PKA in plasma by means of anti-PKA, anti-phosphovitronectin or anti-vitronectin antibodies.
Claim 6
Dependent Claim
40 words
A kit or reagent according to any one of claims 1 to 4, wherein the substrate of PKA is purified vitronectin, an animal protein with at least one of the activities of vitronectin, or a biologically active fragment of vitronectin.
Claim 7
Dependent Claim
25 words
A kit or reagent according to claim 5, wherein the substrate of PKA is a genetically engineered protein which exhibits at least one vitronectin-like activity.
Claim 8
Dependent Claim
20 words
A kit or reagent according to claim 6, wherein the substrate of PKA is the heparin binding domain of vitronectin.
Claim 9
Dependent Claim
31 words
A kit or reagent according to claim 6, wherein the substrate of PKA is a synthetic or genetically engineered peptide which contains the sequence Arg-Arg-Pro-X-Arg where X is Ser or Thr.
Claim 10
Dependent Claim
22 words
A kit or reagent according to claim 8, wherein one, two or all three of the Arg residues is replaced by Lys.
Claim 11
Dependent Claim
19 words
A kit or reagent according to claim 8, wherein the PKA substrate is a peptide containing the sequence Arg-Arg-Pro-Ser-Arg.
Claim 12
Dependent Claim
46 words
A kit or reagent according to any one of the claims 1 to 10 for assaying the biological functionality of isolated blood platelets which comprises means for determining their ability to release PKA when stimulated with a physiological platelet agonist, preferably with thrombin, collagen or ADP.
Claim 13
Dependent Claim
9 words
PKA inhibitor adapted to decrease or prevent vitronectin phosphorylation.
Claim 14
Dependent Claim
9 words
PKA inhibitor adapted to decrease or prevent vitronectin phosphorylation.
Claim 15
Dependent Claim
29 words
PKA inhibitor according to claim 12, which is a synthetic or genetically engineered peptide containing any of the sequences wherein R is a protecting group or a hydrogen atom.
Claim 16
Dependent Claim
20 words
PKA inhibitor according to claim 13, wherein one, two or all three of the Arg residues is replaced by Lys.
Claim 17
Dependent Claim
26 words
A pharmaceutical composition containing a PKA inhibitor according to any one of claims 12 to 14 and optionally in addition a pharmaceutically acceptable carrier or diluent.
Claim 18
Dependent Claim
18 words
A pharmaceutical composition according to claim 15 for the treatment of pathological conditions associated with impaired platelet function.
Claim 19
Dependent Claim
21 words
A composition according to claim 15 or 16 additionally containing an effective quantity of heparin, heparan sulfate or an equivalent thereof.
Claim 20
Independent Claim
30 words
A method for determining the integrity and/or biological functionality of blood platelets which comprises determining the quantity of PKA released by such platelets into plasma and evaluating the quantity released.
Claim 21
Independent Claim
30 words
A method for determining the integrity and/or biological functionality of blood platelets which comprises determining the quantity of PKA released by such platelets into plasma and evaluating the quantity released.
Claim 22
Independent Claim
64 words
A method for the determination of the cellular integrity and/or biological functionality of blood platelets based on the determination of cAMP dependent phosphorylation of vitronectin in plasma and/or the inhibition of such phosphorlyation with radioactively labelled ATP as phosphate donor, and determining the degree of phosphorylation, and/or its prevention which is indicative of the quantity of PKA released by the platelets into the plasma.
Claim 23
Independent Claim
64 words
A method for the determination of the cellular integrity and/or biological functionality of blood platelets based on the determination of cAMP dependent phosphorylation of vitronectin in plasma and/or the inhibition of such phosphorlyation with radioactively labelled ATP as phosphate donor, and determining the degree of phosphorylation, and/or its prevention which is indicative of the quantity of PKA released by the platelets into the plasma.
References Cited (Backward Citations)
No citation records indexed.
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