US5929049A
Expired / Lapsed
US
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Polysaccharide conjugates of biomolecules
Assignee: Siemens Healthcare Diagnostics Products GmbH
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Filing: 1997-08-08
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Granted: 1999-07-27
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Expires: 2017-08-08
Abstract
Abstract Compounds that are modified polysaccharides having pendant aldehyde functionalities are disclosed. Each of the aldehyde functionalities is attached through a linker to a position corresponding to a hydrogen atom of a different hydroxyl group of unmodified polysaccharide. Also disclosed is a method for introducing an amine-reactive functionality into a dextran. The method comprises (a) reacting the dextran with an alkylating agent having a functionality that reacts with an hydroxyl group of the dextran thereby forming an alkylated dextran wherein the alkylating agent has an olefin group and (b) treating the alkylated dextran to convert the olefin group to an amine-reactive functionality. A polysaccharide can be conjugated to a biomolecule by carrying out the above method and reacting the amine-reactive functionality with an amine functionality on the biomolecule to produce polysaccharide conjugated to the biomolecule.
Inventors
- 👤 Rajendra Singh
- 👤 Harshvardhan Mehta
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 17:06
CPC Classifications
C; C08; C08B; C08B37/00; C08B37/0006; C08B37/0009; C08B37/0021; A; A01; A01N; A01N43/00; A01N43/02; A01N43/04; A01N43/14; A01N43/16; C08H; C08H1/00; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/564; G01N2333/00; G01N2333/90; G01N2333/988
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
190 words
A compound comprising a polymer having a sequence of repeating monosaccharide units, each of which is independently selected from monosaccharide units of the formula: ##STR4## wherein one of the A's is a bond to the C1 glycosidic carbon of another of said units and the other A's are independently selected from the group consisting of H and QL, wherein L is a linking group linking O and Q and Q is C(Z)═D wherein D is O or CR 1 R 2 wherein R 1 and R 2 are independently H, alkyl or aryl and Z is H or C(Y)═O wherein Y is R 3 , OR 3 or NR 3 R 4 wherein R 3 and R 4 are independently H, alkyl or aryl and wherein any of L, R 1 , R 2 , R 3 and R 4 may be taken together to form a ring with the proviso that at least two of the A's in said polymer are QL wherein said compound is produced by a reaction linking said polymer with a polypeptide selected from the group consisting of avidin, streptavidin, protein A and Protein G.
Claim 2
Dependent Claim
190 words
A compound comprising a polymer having a sequence of repeating monosaccharide units, each of which is independently selected from monosaccharide units of the formula: ##STR4## wherein one of the A's is a bond to the C1 glycosidic carbon of another of said units and the other A's are independently selected from the group consisting of H and QL, wherein L is a linking group linking O and Q and Q is C(Z)═D wherein D is O or CR 1 R 2 wherein R 1 and R 2 are independently H, alkyl or aryl and Z is H or C(Y)═O wherein Y is R 3 , OR 3 or NR 3 R 4 wherein R 3 and R 4 are independently H, alkyl or aryl and wherein any of L, R 1 , R 2 , R 3 and R 4 may be taken together to form a ring with the proviso that at least two of the A's in said polymer are QL wherein said compound is produced by a reaction linking said polymer with a polypeptide selected from the group consisting of avidin, streptavidin, protein A and Protein G.
Claim 3
Dependent Claim
17 words
The compound of claim 1 comprising a polymer having from about 50 to 50,000 repeating monosaccharide units.
Claim 4
Dependent Claim
20 words
The compound of claim 1 comprising a polymer wherein Q is C(Z)═D wherein D is O and Z is H.
Claim 5
Dependent Claim
23 words
The compound of claim 1 comprising a polymer wherein Q is C(Z)═D wherein D is CR 1 R 2 and Z is H.
Claim 6
Dependent Claim
24 words
The compound of claim 1 comprising a polymer wherein Q is C(Z)═D wherein D is O and Z is C(Y)═O wherein Y is OH.
Claim 7
Dependent Claim
33 words
The compound of claim 1 comprising a polymer wherein at least two of the A's are QL in at least one of said monosaccharide units for every 20 monosaccharides units of said polymer.
Claim 8
Dependent Claim
26 words
A compound according to claim 1 wherein said compound is produced by the reductive amination of the polymer with the polypeptide and wherein D is O.
Claim 9
Dependent Claim
158 words
A compound comprising: ##STR5## wherein n is 50 to 50,000, A 1 is independently selected from the group consisting of H and Q 1 L 1 , wherein L 1 is a linking group linking O and Q 1 and Q 1 is C(Z)═D wherein D is O or CR 1 R 2 wherein R 1 and R 2 are independently H, alkyl or aryl and Z is H or C(Y)═O wherein Y is R 3 , OR 3 or NR 3 R 4 wherein R 3 and R 4 are independently H, alkyl or aryl and wherein any of L 1 , R 1 , R 2 , R 3 and R 4 may be taken together to form a ring with the proviso that at least two of the A's in said compound are Q 1 L 1 and conjugated to a polypeptide selected from the group consisting of avidin, streptavidin, protein A and Protein G.
Claim 10
Dependent Claim
158 words
A compound comprising: ##STR5## wherein n is 50 to 50,000, A 1 is independently selected from the group consisting of H and Q 1 L 1 , wherein L 1 is a linking group linking O and Q 1 and Q 1 is C(Z)═D wherein D is O or CR 1 R 2 wherein R 1 and R 2 are independently H, alkyl or aryl and Z is H or C(Y)═O wherein Y is R 3 , OR 3 or NR 3 R 4 wherein R 3 and R 4 are independently H, alkyl or aryl and wherein any of L 1 , R 1 , R 2 , R 3 and R 4 may be taken together to form a ring with the proviso that at least two of the A's in said compound are Q 1 L 1 and conjugated to a polypeptide selected from the group consisting of avidin, streptavidin, protein A and Protein G.
Claim 11
Dependent Claim
18 words
The compound of claim 8 wherein Q 1 is C(Z)═D wherein D is O and Z is H.
Claim 12
Dependent Claim
21 words
The compound of claim 8 wherein Q 1 is C(Z)═D wherein D is CR 1 R 2 and Z is H.
Claim 13
Dependent Claim
22 words
The compound of claim 8 wherein Q l is C(Z)═D wherein D is O and Z is C(Y)═O wherein Y is OH.
Claim 14
Dependent Claim
35 words
The compound of claim 8 wherein at least two of the A 1 's are Q 1 L 1 in at least one of said monosaccharide units for every 20 monosaccharides units of said compound.
Claim 15
Dependent Claim
30 words
A compound according to claim 8 wherein said compound is produced by the reductive amination of the polymer with the polypeptide and wherein D is O and Z is H.
Claim 16
Independent Claim
103 words
A method for conjugating a polysaccharide as in claims 1 or 8 to a polypeptide, said method comprising: (a) reacting said polysaccharide with an alkylating agent having a functionality that reacts with a hydroxy group of said polysaccharide thereby forming an alkylated polysaccharide wherein said alkylating agent has an olefin group, (b) treating said alkylated polysaccharide to convert said olefin group to an amine-reactive functionality, and (c) reacting said amine-reactive functionality with an amine functionality on said polypeptide to produce a polysaccharide conjugated to said polypeptide, wherein said polypeptide is selected from the group consisting of Protein A, Protein G, avidin and streptavidin.
Claim 17
Dependent Claim
14 words
The method of claim 14 wherein step (b) comprises ozonolysis of said alkylated polysaccharide.
Claim 18
Dependent Claim
10 words
The method of claim 14 wherein said polysaccharide is dextran.
Claim 19
Dependent Claim
88 words
The method of claim 14 wherein said alkylating agent having the structure: L.sup.3 (Z.sup.1)C═CR.sup.5 R.sup.6 wherein L 3 is a group reactive with a hydroxyl group of said dextran, R 5 and R 6 are independently H, alkyl, aryl or may be taken together with each other or with L 3 to form a ring and Z 1 is H or C(W)═O wherein W is R 7 , OR 7 , or NR 7 R 8 wherein R 7 and R 8 are independently H, alkyl or aryl.
Claim 20
Dependent Claim
19 words
The method of claim 17 wherein L 3 having an epoxide or an alkyl radical comprising a leaving group.
Claim 21
Dependent Claim
39 words
The method of claim 17 wherein L 3 is an alkyl radical having a leaving group and said leaving group is selected from the group consisting of bromide, chloride, iodide, aryl sulfonates, alkyl sulfonates, aryl sulfonates and alkyl sulfates.
Claim 22
Dependent Claim
29 words
The method of claim 17 wherein L 3 is CH 2 O CH 2 (C 2 H 3 O) wherein (C 2 H 3 O) is an oxiranyl radical.
Claim 23
Dependent Claim
12 words
The method of claim 17 wherein L 3 is CH 2 Br.
Claim 24
Dependent Claim
12 words
The method of claim 14 wherein said amine-reactive functionality is an aldehyde.
Claim 25
Dependent Claim
17 words
The method of claim 22 wherein said olefin is subjected to ozonolysis to yield said aldehyde functionality.
Claim 26
Dependent Claim
15 words
The method of claim 14 wherein said amine-reactive functionality is an alpha-keto carboxylic acid functionality.
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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) | 2002-07-26 | 2003-01-25 | 2003-07-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2006-07-26 | 2007-01-25 | 2007-07-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2010-07-26 | 2011-01-25 | 2011-07-27 | $7700.00 | Window Closed / Lapsed |