Patent US4745180A successfully refreshed from Google Patents.
US4745180A
Expired / Lapsed
US
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Solubilization of proteins for pharmaceutical compositions using heparin fragments
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1986-06-27
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Granted: 1988-05-17
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Expires: 2006-06-27
Abstract
Abstract A pharmaceutical composition is prepared wherein a biologically active conjugated protein which is β-interferon, interleukin-2, or an immunotoxin is dissolved in an aqueous carrier medium without the presence of a solubilizing agent. The unconjugated protein, which is poorly or not at all water-soluble at pH 6-8 without such solubilizing agent, is selectively conjugated to at least one heparin fragment having a terminal 2,5-anhydro-D-mannose residue which has an aldehyde not involved in intramolecular hemiacetal formation.
Inventors
- 👤 Margaret Moreland
- 👤 Danute E. Nitecki
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 23:16
CPC Classifications
A; A61; A61K; A61K38/00; A61K38/16; A61K38/17; A61K38/19; A61K38/20; A61K38/2013; A61K47/00; A61K47/50; A61K47/51; A61K47/56; A61K47/59; A61K47/60; A61K47/61; Y; Y10; Y10S; Y10S435/00; Y10S435/811
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
94 words
A pharmaceutical composition comprising a non-toxic, inert, pharmaceutically acceptable aqueous carrier medium in which is dissolved a biologically active, selectively conjugated, recombinant protein selected from the group consisting of β-interferon, interleukin-2, and an immunotoxin, wherein the protein is covalently conjugated via at least one of its lysine residues to at least one heparin fragment having a terminal 2,5-anhydro-D-mannose residue through the aldehyde group of such residue, wherein said protein in its unconjugated form is normally hydrophobic and poorly soluble in said aqueous carrier medium at pH 6-8 in the absence of a solubilizing agent.
Claim 2
Dependent Claim
94 words
A pharmaceutical composition comprising a non-toxic, inert, pharmaceutically acceptable aqueous carrier medium in which is dissolved a biologically active, selectively conjugated, recombinant protein selected from the group consisting of β-interferon, interleukin-2, and an immunotoxin, wherein the protein is covalently conjugated via at least one of its lysine residues to at least one heparin fragment having a terminal 2,5-anhydro-D-mannose residue through the aldehyde group of such residue, wherein said protein in its unconjugated form is normally hydrophobic and poorly soluble in said aqueous carrier medium at pH 6-8 in the absence of a solubilizing agent.
Claim 3
Dependent Claim
19 words
The composition of claim 1 wherein the medium is at a pH of about 5-8 and contains a buffer.
Claim 4
Dependent Claim
19 words
The composition of claim 1 wherein the medium is at a pH of about 6.5-7.8 and contains a buffer.
Claim 5
Dependent Claim
16 words
The composition of claim 1 wherein the protein is a recombinant protein from a human source.
Claim 6
Dependent Claim
34 words
The composition of claim 4 wherein said protein has the amino acid which is at position 104 or 125 on those at both position in the native protein substituted with a neutral amino acid.
Claim 7
Dependent Claim
34 words
The composition of claim 5 wherein said IL-2 protein is ser 125 IL-2, des-ala 1 Il-2, des-ala 1 ser 125 IL-2, des-ala 1 ala 104 IL-2, or des-ala 1 ala 104 ser 125 IL-2.
Claim 8
Dependent Claim
26 words
The composition of claim 4 wherein said β-IFN protein has the cysteine residue at position 17 in the native protein substituted with a neutral amino acid.
Claim 9
Dependent Claim
13 words
The composition of claim 7 wherein said β-IFN protein is ser 17 IFN-β.
Claim 10
Dependent Claim
16 words
The composition of claim 4 wherein said protein is an immunotoxin with recombinant ricin A chain.
Claim 11
Dependent Claim
83 words
A process for preparing a pharmaceutical composition comprising: (a) preparing a heparin fragment having a terminal 2,5-anhydro-D-mannose residue with an aldehyde not involved in intramolecular hemiacetal formation; (b) reacting a biologically active, normally hydrophobic, poorly water-soluble recombinant protein selected from the group consisting of β-interferon, interleukin-2, and an immunotoxin with said aldehyde of said heparin fragment so as to provide a more water-soluble, biologically active, selectively conjugated recombinant protein; and (c) formulating said protein in a non-toxic, inert, pharmaceutically acceptable aqueous carrier medium.
Claim 12
Dependent Claim
83 words
A process for preparing a pharmaceutical composition comprising: (a) preparing a heparin fragment having a terminal 2,5-anhydro-D-mannose residue with an aldehyde not involved in intramolecular hemiacetal formation; (b) reacting a biologically active, normally hydrophobic, poorly water-soluble recombinant protein selected from the group consisting of β-interferon, interleukin-2, and an immunotoxin with said aldehyde of said heparin fragment so as to provide a more water-soluble, biologically active, selectively conjugated recombinant protein; and (c) formulating said protein in a non-toxic, inert, pharmaceutically acceptable aqueous carrier medium.
Claim 13
Dependent Claim
21 words
The process of claim 10 wherein step (a) is carried out by treating the sodium salt of heparin with nitrous acid.
Claim 14
Dependent Claim
14 words
The process of claim 10 wherein step (b) is carried out by reductive amination.
Claim 15
Dependent Claim
15 words
The process of claim 12 wherein the reaction is carried out using NaCNBH 3 .
Claim 16
Dependent Claim
14 words
The process of claim 10 wherein step (c) is carried out at pH 6-8.
Claim 17
Dependent Claim
15 words
The process of claim 10 wherein said protein is recombinant and from a human source.
Claim 18
Dependent Claim
10 words
The process of claim 15 wherein said protein is interleukin-2.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-05-17 | 1991-11-16 | 1992-05-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-05-17 | 1995-11-16 | 1996-05-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-05-17 | 1999-11-16 | 2000-05-17 | $7700.00 | Window Closed / Lapsed |