Patent US4446126A successfully refreshed from Google Patents.
US4446126A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Antithrombin-heparin complex and method for its production
Assignee: Bayer Corp
•
Filing: 1982-03-12
•
Granted: 1984-05-01
•
Expires: 2002-03-12
Abstract
Abstract A complex of antithrombin and high activity heparin is provided for use as a potent anticoagulant for humans. The complex is prepared by reversibly immobilizing it on a lectin-containing, water-insoluble gel matrix and then removing it from the matrix.
Inventors
- 👤 Robert E. Jordan
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 12:22
CPC Classifications
A; A61; A61K; A61K31/00; A61K31/70; A61K31/715; C; C08; C08B; C08B37/00; C08B37/006; C08B37/0063; C08B37/0075
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
88 words
A method for preparing a composition having as its sole effective anticoagulant agent an antithrombin-heparin complex, comprising the steps of: (a) reversibly binding antithrombin to a lectin-containing water-insoluble matrix; (b) contacting said matrix and said antithrombin bound thereto with heparin, whereby an active portion of said heparin complexes with said antithrombin; (c) washing said matrix to remove portions of heparin not complexed to said antithrombin; (d) displacing said antithrombin-heparin complex from said matrix; and (e) thereby recovering said antithrombin-heparin complex substantially free from uncomplexed antithrombin and uncomplexed heparin.
Claim 2
Independent Claim
88 words
A method for preparing a composition having as its sole effective anticoagulant agent an antithrombin-heparin complex, comprising the steps of: (a) reversibly binding antithrombin to a lectin-containing water-insoluble matrix; (b) contacting said matrix and said antithrombin bound thereto with heparin, whereby an active portion of said heparin complexes with said antithrombin; (c) washing said matrix to remove portions of heparin not complexed to said antithrombin; (d) displacing said antithrombin-heparin complex from said matrix; and (e) thereby recovering said antithrombin-heparin complex substantially free from uncomplexed antithrombin and uncomplexed heparin.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein the lectin is Concanavalin A.
Claim 4
Dependent Claim
13 words
The method of claim 1 wherein the water-insoluble matrix is an agarose gel.
Claim 5
Dependent Claim
42 words
The method of claim 1 wherein the unbound heparin is separated from the matrix by treating the matrix with an aqueous salt solution having an ionic strength sufficient to remove unbound heparin but insufficient to remove high activity heparin from the matrix.
Claim 6
Dependent Claim
32 words
The method of claim 1 wherein the matrix is contacted with an aqueous carbohydrate solution, the carbohydrate being present in an amount sufficient to separate the reversibly bound complex from the matrix.
Claim 7
Dependent Claim
11 words
The method of claim 5 wherein the carbohydrate is a monosaccharide.
Claim 8
Dependent Claim
20 words
The method of claim 6 wherein the carbohydrate is selected from the group consisting of glucose, mannose, galactose, and fructose.
Claim 9
Dependent Claim
11 words
The method of claim 5 wherein the carbohydrate is a dissacharide.
Claim 10
Dependent Claim
19 words
The method of claim 8 wherein the carbohydrate is selected from the group consisting of maltose, sucrose, and lactose.
Claim 11
Independent Claim
124 words
A method of preparing a complex of antithrombin and high activity heparin, which comprises (a) contacting a lectin-containing, water-insoluble matrix with antithrombin and heparin for a period of time and at a temperature sufficient to allow the antithrombin to bind reversibly to the lectin portion of the matrix and the high activity heparin component of the heparin to complex with the antithrombin, (b) washing the matrix with a solution having an ionic strength sufficient to remove unbound heparin from the matrix but insufficient to remove the antithrombin-high activity heparin complex therefrom, (c) contacting the matrix with a solution of a carbohydrate having the ability to displace the complex from the matrix and (d) recovering said complex substantially free of uncomplexed antithrombin and uncomplexed heparin.
Claim 12
Independent Claim
124 words
A method of preparing a complex of antithrombin and high activity heparin, which comprises (a) contacting a lectin-containing, water-insoluble matrix with antithrombin and heparin for a period of time and at a temperature sufficient to allow the antithrombin to bind reversibly to the lectin portion of the matrix and the high activity heparin component of the heparin to complex with the antithrombin, (b) washing the matrix with a solution having an ionic strength sufficient to remove unbound heparin from the matrix but insufficient to remove the antithrombin-high activity heparin complex therefrom, (c) contacting the matrix with a solution of a carbohydrate having the ability to displace the complex from the matrix and (d) recovering said complex substantially free of uncomplexed antithrombin and uncomplexed heparin.
Claim 13
Dependent Claim
24 words
The method of claim 10 wherein 50-100 parts of matrix are contacted in Step (a) with one part of antithrombin on a weight basis.
Claim 14
Dependent Claim
26 words
The method of claim 10 wherein 1-100 parts of heparin per part of antithrombin on a weight basis are contacted with the matrix in Step (a).
Claim 15
Dependent Claim
36 words
The method of claim 10 wherein the matrix prior to contact with antithrombin and heparin is equilibrated with a buffer solution containing no greater than 0.25 M sodium chloride and having a pH of about 6-8.5.
Claim 16
Dependent Claim
25 words
The method of claim 10 wherein the wash solution of Step (b) has an ionic strength of about 0.1-0.4 and a pH of about 6-8.5.
Claim 17
Dependent Claim
22 words
The method of claim 10 wherein the temperature in Step (a) is greater than 5° C. but no greater than 37° C.
Claim 18
Dependent Claim
17 words
The method of claim 10 wherein the time of contact in Step (a) is about 0.1-2 hours.
Claim 19
Dependent Claim
27 words
The method of claim 10 wherein the matrix is contacted with a solution in Step (c) containing 0.02-0.5 M carbohydrate and having a pH of about 6-8.5.
Claim 20
Dependent Claim
23 words
The method of claim 10 wherein the carbohydrate is selected from the group consisting of glucopyranosides, mannopyranosides, fructofuranosides, monosaccharides, dissacharides, and sugar alcohols.
Claim 21
Dependent Claim
25 words
The method of claim 10 which further includes the step of reducing the carbohydrate concentration of the complex displaced from the matrix in Step (c).
Claim 22
Dependent Claim
24 words
The method of claim 10 wherein the lectincontaining, water-insoluble matrix in Step (a) is formed by covalently attaching a lectin to a water-insoluble matrix.
Claim 23
Dependent Claim
59 words
A pharmaceutical preparation as an anticoagulant comprising a complex of antithrombin and high activity heparin prepared by the method of claim 10 wherein the activity of the high activity heparin in the complex is greater than about 300 Units per milligram and wherein the high activity heparin in the complex is derived from heparin not previously fractioned by size.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-05-01 | 1987-10-31 | 1988-05-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-05-01 | 1991-10-31 | 1992-05-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-05-01 | 1995-10-31 | 1996-05-01 | $7700.00 | Window Closed / Lapsed |