US4870166A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for preparing high-purity dematan sulphate, and pharmaceutical compositions which contain it
Assignee: Mediolanum Farmaceutici SpA
•
Filing: 1987-03-19
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Granted: 1989-09-26
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Expires: 2007-03-19
Abstract
Abstract A process for preparing dermatan sulphate (DS) of pharmaceutical purity from animal organs rich in mucopolysaccharides (MPS), by: (a) stabilizing the fresh organs by freezing them either as such or in the form of powder, (b) micronizing the material containing the MPS with an aqueous CaCl2 solution, (c) digesting the homogenate comprising the raw material and the CaCl2 with proteolytic enzymes at alkaline pH and at low temperature, (d) acidifying, heating and filtering the lysate, (e) treating the filtrate with quaternary ammonium salts able to undergo complexing with and thus precipitate either the DS along or all the MPS selectively, (f) recovering and purifying the DA either directly from the ammonium salt complex containing it or from the complex obtained from the mixture of ammonium salt complexes of all the MPS by fractional solubilization with acetone.
Inventors
- 👤 Rinaldo Del Bono
- 👤 Luigi De Ambrosi
- 👤 Gianni Ferrari
- 👤 Pier L. Rugarli
- 👤 Pier G. Pagella
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 17:44
CPC Classifications
C; C08; C08B; C08B37/00; C08B37/0006; C08B37/0024; C08B37/0027; C08B37/003; C08B37/006; C08B37/0063; C08B37/0069; A; A61; A61P; A61P7/00; A61P7/02
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
120 words
A process for preparing dermatan sulphate (DS) of pharmaceutical purity from fresh animal organs rich in mucopolysaccharides (MPS), comprising: (a) stabilizing the fresh organs by freezing them either as such or in the form of powder, (b) micronizing the stable material containing MPS with an aqueous CaCl 2 solution to form a homgeneate, (c) digesting the homogenate comprising the stable material and the CaCl 2 with proteolytic enzymes at alkaline pH and at low temperature to form a lysate, (d) acidifying, heating and filtering the lysate, (e) treating the filtrate with quaternary ammonium salts to complex with and thus precipitate either the DS alone or all the MPS selectively, and (f) recovering and purifying the DS from the ammonium complex.
Claim 2
Dependent Claim
120 words
A process for preparing dermatan sulphate (DS) of pharmaceutical purity from fresh animal organs rich in mucopolysaccharides (MPS), comprising: (a) stabilizing the fresh organs by freezing them either as such or in the form of powder, (b) micronizing the stable material containing MPS with an aqueous CaCl 2 solution to form a homgeneate, (c) digesting the homogenate comprising the stable material and the CaCl 2 with proteolytic enzymes at alkaline pH and at low temperature to form a lysate, (d) acidifying, heating and filtering the lysate, (e) treating the filtrate with quaternary ammonium salts to complex with and thus precipitate either the DS alone or all the MPS selectively, and (f) recovering and purifying the DS from the ammonium complex.
Claim 3
Dependent Claim
32 words
A process as claimed in claim 1, wherein the micronizing is carried out on frozen stable material mixed with an aqueous 0.01 M CaCl 2 solution in a weight ratio of 1:1.
Claim 4
Dependent Claim
47 words
A process as claimed in claim 1, wherein the micronization is carried out on frozen stable material mixed with an aqueous 1 M CaCl 2 solution in a weight ratio of 1:1, and the micronized organs are repeatedly treated with cold acetone and filtered, and finally dried.
Claim 5
Dependent Claim
48 words
A process as claimed in claim 1, wherein the micronization is carried out on frozen stable material mixed with an aqueous 0.1 M CaCl 2 solution in a weight ratio of 1:1, and the mixture obtained is spray-dried with air at a temperature of less than 150° C.
Claim 6
Dependent Claim
42 words
A process as claimed in claim 1, wherein the micronization is carried out on frozen stable material mixed with an aqueous 1 M CaCl 2 solution containing 10% of sorbitol, in a weight ratio of 1:1, and the mixture obtained is lyophilized.
Claim 7
Dependent Claim
33 words
A process as claimed in claim 1, wherein the digesting with proteolytic enzyme comprising adding the enzyme to the micronized mixture diluted with water, adjusting to pH 7-9 and heating to 40°-55° C.
Claim 8
Dependent Claim
26 words
A process as claimed in claim 6, wherein the enzyme is added in a weight ratio of stable material to enzyme of between 1:0.0001 and 1:0.001.
Claim 9
Dependent Claim
21 words
A process as claimed in claim 6, wherein the pH is adjusted to between 7 and 9 with Ca(OH) 2 .
Claim 10
Dependent Claim
25 words
A process as claimed in claim 1, wherein the lysate is acidified to a pH of between 3 and 6 and heated to 70°-90° C.
Claim 11
Dependent Claim
14 words
A process as claimed in claim 9, wherein the lysate is acidified with HCl.
Claim 12
Dependent Claim
23 words
A process as claimed in claim 1, wherein the filtered lysate is treated with dimethyl-ethyl-cetyl ammonium ethylsulphate to selectively precipitate a DS complex.
Claim 13
Dependent Claim
49 words
A process as claimed in claim 1, wherein the filtered lysate is treated with a quaternary ammonium salt selected from the group consisting of Hyamine, cetyl-trimethyl ammonium bromide and cetyl-dimethyl-ethyl ammonium bromide, at a temperature of 60°-70° C., to precipitate in the form of complexes all the MPS present.
Claim 14
Dependent Claim
16 words
A process as claimed in claim 12, wherein the DS complex is selectively solubilized with acetone.
Claim 15
Dependent Claim
45 words
A process as claim in claim 11, wherein the DS is recovered from the ammonium complex by treating the ammonium complex with an aqueous 2 M CaCl 2 solution containing 10% ethanol, adjusting the pH to between 7 and 9 and heating to 60°-80° C.
Claim 16
Dependent Claim
39 words
A process as claimed in claim 13, wherein the DS is recovered from the ammonium complex by treating with a 3 M saline solution, using a volumetric ratio of acetone extract to saline solution of between 0.25 and 1.
Claim 17
Dependent Claim
25 words
A process as claimed in claim 13, wherein the residue from the acetone treatment is treated with ethanol to selectively solubilize a heparin sulphate complex.
Claim 18
Dependent Claim
25 words
A process as claimed in claim 16, wherein the residue from the treatment with ethanol is treated with methanol to selectively solubilize the heparin complex.
References Cited (Backward Citations)
US3862003A
Cited
US4783447A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-09-25 | 1993-03-27 | 1993-09-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-09-25 | 1997-03-27 | 1997-09-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-09-25 | 2001-03-27 | 2001-09-26 | $7700.00 | Window Closed / Lapsed |