Patent US4748156A successfully refreshed from Google Patents.
US4748156A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Thrombin-binding substance and process for preparing the same
Assignee: Kowa Co Ltd
•
Filing: 1987-01-15
•
Granted: 1988-05-31
•
Expires: 2007-01-15
Abstract
Abstract A novel thrombin-binding substance having the characteristics of (a) molecular weight: 47,000±6,000 in reduced condition and 38,000±6,000 in unreduced condition, (b) isoelectric point: pH 4.9-5.7, (c) affinity: strong affinity to thrombin, (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C, (2) prolongs clotting time; and (e) stability: stable to denaturing agents (urea sodium and dodecylsulfate), can be prepared from a human placenta by means of extraction, fractionation using ion exchange chromatography and affinity chromatography, and/or gel filtration, followed by electrophoresis. The substance is useful as a medicine for the treatment of thrombosis.
Inventors
- 👤 Nobuo Aoki
- 👤 Masami Shiratsuchi
- 👤 Shigeru Kimura
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 11:24
CPC Classifications
A; A61; A61K; A61K35/00; A61K35/12; A61K35/48; A61K35/50
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
103 words
A thrombin-binding substance derived from human tissue obtained by (1) extracting human tissue with a buffer containing a non-ionic surfactant, (2) isolating said thrombin-binding substance in pure form from the resulting extract by diisopropylphosphorothrombin affinity chromatography, (3) subjecting isolate to SDS-polyacrylamide electrophoresis and (4) elution of said thrombin-binding substance having the following characteristics: (a) molecular weight: 47,000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate.
Claim 2
Dependent Claim
103 words
A thrombin-binding substance derived from human tissue obtained by (1) extracting human tissue with a buffer containing a non-ionic surfactant, (2) isolating said thrombin-binding substance in pure form from the resulting extract by diisopropylphosphorothrombin affinity chromatography, (3) subjecting isolate to SDS-polyacrylamide electrophoresis and (4) elution of said thrombin-binding substance having the following characteristics: (a) molecular weight: 47,000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate.
Claim 3
Dependent Claim
100 words
A process for preparing a thrombin-binding substance having the following characteristics: (a) molecular weight: 47,000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate, which comprises: (1) extracting human placental tissue with a buffer solution containing a non-ionic surfactant, (2) fractionating said resultant extract by means of affinity chromatography using a thrombin-bound carrier, and (3) further subjecting the thus obtained fractions to electrophoresis.
Claim 4
Dependent Claim
100 words
A process for preparing a thrombin-binding substance having the following characteristics: (a) molecular weight: 47,000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate, which comprises: (1) extracting human placental tissue with a buffer solution containing a non-ionic surfactant, (2) fractionating said resultant extract by means of affinity chromatography using a thrombin-bound carrier, and (3) further subjecting the thus obtained fractions to electrophoresis.
Claim 5
Dependent Claim
17 words
The process of claim 2, wherein said affinity chromatography is carried out upon a resin of diisopropylphosphorothrombin.
Claim 6
Dependent Claim
15 words
The thrombin-binding substance according to claim 1, wherein said human tissue is human placental tissue.
Claim 7
Independent Claim
115 words
A method of promoting the thrombin catalyzed activation of protein C which comprises the administration of a thrombin-binding substance obtained by (1) extracting human tissue with a buffer containing a non-ionic surfactant, (2) isolating said thrombin-binding substance in pure form from the resulting extract by diisopropylphosphorothrombin affinity chromatography, (3) subjecting isolate to SDS-polyacrylamide electrophoresis and (4) elution of said thrombin-binding substance having the following characteristics: (a) molecular weight: 47,000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate.
Claim 8
Independent Claim
115 words
A method of promoting the thrombin catalyzed activation of protein C which comprises the administration of a thrombin-binding substance obtained by (1) extracting human tissue with a buffer containing a non-ionic surfactant, (2) isolating said thrombin-binding substance in pure form from the resulting extract by diisopropylphosphorothrombin affinity chromatography, (3) subjecting isolate to SDS-polyacrylamide electrophoresis and (4) elution of said thrombin-binding substance having the following characteristics: (a) molecular weight: 47,000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate.
Claim 9
Independent Claim
67 words
A method of prolonging the clothing time of blood which comprises the administration of a thrombin-binding substance obtained by (1) extracting human tissue with a buffer containing a non-ionic surfactant, and (2) isolating said thrombin-binding substance in pure form from the resulting extract by diisopropylphosphorothrombin affinity chromatography, (3) subjecting isolate to SDS-polyacrylamide electrophoresis and (4) elution of said thrombin-binding substance having the following characteristics: (a) molecular weight:
Claim 10
Independent Claim
67 words
A method of prolonging the clothing time of blood which comprises the administration of a thrombin-binding substance obtained by (1) extracting human tissue with a buffer containing a non-ionic surfactant, and (2) isolating said thrombin-binding substance in pure form from the resulting extract by diisopropylphosphorothrombin affinity chromatography, (3) subjecting isolate to SDS-polyacrylamide electrophoresis and (4) elution of said thrombin-binding substance having the following characteristics: (a) molecular weight:
Claim 11
Dependent Claim
47 words
000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate.
Claim 12
Dependent Claim
47 words
000±6,000 in reduced condition 38,000±6,000 in unreduced condition (b) isoelectric point: pH 4.9-5.7 (c) affinity: strong affinity to thrombin (d) activity: (1) capable of promoting the thrombin catalyzed activation of protein C (2) prolongs clotting time; and (e) stability: stable to denaturing agents, urea and sodium dodecylsulfate.
References Cited (Backward Citations)
US4254021A
Cited
US4301064A
Cited
US4302385A
Cited
US4368148A
Cited
US4402872A
Cited
US4468345A
Cited
US4500451A
Cited
US4507229A
Cited
US4599318A
Cited
US4594328A
Cited
US4638050A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-05-31 | 1991-11-30 | 1992-05-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-05-31 | 1995-11-30 | 1996-05-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-05-31 | 1999-11-30 | 2000-05-31 | $7700.00 | Window Closed / Lapsed |