Polysaccharide conjugates of biomolecules

• US 6,489,309 B1
• Filed: 05/18/1999
• Issued: 12/03/2002
• Est. Priority Date: 08/08/1997
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: DADE Behring Beteiligungs GmbH (Siemens AG)
• Inventors: Mehta, Harshvardhan, Singh, Rajendra
• Primary Examiner(s): Gitomer, Ralph
• Assistant Examiner(s): Khare, Devesh
• Application Number:
  US09/314,055
• Publication Number:
  US 20020193586A1
• Time in Patent Office: 1,295 Days
• Field of Search: 536/51, 514/54, 514/59, 514/367, 524/54
• US Class Current: 514/54
• CPC Class Codes: A01N 43/16 with oxygen as the ring hetero atom
• CPC Class Codes: C08B 37/0021 Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran
• CPC Class Codes: C08H 1/00 Macromolecular products derived from proteins food proteins A23; glue, gelatine C09H
• CPC Class Codes: G01N 2333/988 Lyases (4.), e.g. aldolases, heparinase, enolases, fumarase
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9

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.

Claims

1. A polymer comprising a sequence of repeating monosaccharide units, each of which is independently selected from monosaccharide units of the formula:

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 CR¹R² wherein R¹ and R² are independently H, alkyl or aryl and Z is H or C(Y)═

O wherein Y is R³, OR³ or NR³R⁴ wherein R³ and R⁴ are independently H, alkyl or aryl and wherein any of L, R¹, R², R³ and R⁴ may be taken together to form a ring with the proviso that at least two of the A'"'"'s in said polymer are QL.

2. The polymer of claim 1 having from about 50 to 50,000 repeating monosaccharide units.

3. The polymer of claim 1 wherein Q is C(Z)═

D wherein D is O and Z is H.

4. The polymer of claim 1 wherein Q is C(Z)═

D wherein D is CR¹R² and Z is H.

5. The polymer of claim 1 wherein Q is C(Z)═

D wherein D is O and Z is C(Y)═

O wherein Y is OH.

6. The polymer of claim 1 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.

7. A compound comprising:

wherein n is 50 to 50,000, A'"'"'s is independently selected from the group consisting of H and Q¹L¹, wherein L¹ is a linking group linking O and Q¹ and Q¹ is C(Z)═

D wherein D is O or CR¹R² wherein R¹ and R² are independently H, alkyl or aryl and Z is H or C(Y)═

O wherein Y is R³, OR³ or NR³R⁴ wherein R³ and R⁴ are independently H, alkyl or aryl and wherein any of L, R¹, R², R³ and R⁴ may be taken together to form a ring with the proviso that at least two of the A'"'"'s in said compound are Q¹L¹.

8. The compound of claim 7 wherein Q¹ is C(Z)═

D wherein D is O and Z is H.

9. The compound of claim 7 wherein Q¹ is C(Z)═

D wherein D is CR¹R² and Z is H.

10. The compound of claim 7 wherein Q¹ is C(Z)═

D wherein D is O and Z is C(Y)═

O wherein Y is OH.

11. The compound of claim 7 wherein at least two of the A¹'"'"'s are Q¹L¹ in at least one of said monosaccharide-units for every 20 monosaccharides units of said compound.