Activated matrix and method of activation

• US 4,224,439 A
• Filed: 02/02/1978
• Issued: 09/23/1980
• Est. Priority Date: 02/08/1977
• Status: Expired due to Term

• Current Assignee: Development Finance Corporation Of New Zealand
• Original Assignee: Development Finance Corporation Of New Zealand
• Inventors: Hancock, William S., Hearn, Milton T. W., Ayers, John S., Bethell, Geoffrey S.
• Primary Examiner(s): Marquis, Melvyn I.
• Application Number:
  US05/874,628
• Time in Patent Office: 964 Days
• Field of Search: 536/32, 536/43, 536/44, 536/48, 536/49, 536/50, 536/107, 536/108, 536/110, 536/115, 536/120, 536/112 R, 260/112.5 R, 260/6
• US Class Current: 536/32
• CPC Class Codes: C08B 15/06 containing nitrogen , e.g. carbamates
• CPC Class Codes: C08B 37/0039 Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose
• CPC Class Codes: C12N 11/10 the carrier being a carbohydrate
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/548 Carbohydrates, e.g. dextran
• CPC Class Codes: Y10S 530/813 Carrier is a saccharide

Abstract

Carbonylation of a polysaccharide and the product of that carbonylation. The carbonylated product can be used to prepare an affinity chromatography matrix which is an uncharged species of varying pH'"'"'s. The carbonylated product is also useful for the preparation of other compounds. Typical polysaccharides are agarose, starch, dextran, cellulose and regenerated cellulose and typical carbonylating agents are N,N'"'"'-carbonyl diimidazole; N,N'"'"' carbonyl di-1,2,3-benzotriazole; and N,N'"'"'-carbonyl di-1,2,4-triazole.

Claims

1. A compound of the general formula III ##STR8## wherein MATRIX is a water and organic solvent insoluble radical selected from the group consisting of:

agarose, starch, dextran, cellulose, regenerated cellulose, hydroxy C₂ -C₄ alkylated cellulose and hydroxy C₂ -C₄ lower alkylated regenerated cellulose; and

Y is a member selected from the group consisting of imidazoyl, 1,2,4-triazoyl and 1,2,3-benzotriazoyl.

2. The compound according to claim 1 wherein MATRIX in formula III is cross-linked.

3. The compound according to claims 1 or 2, wherein Y is imidazoyl.

4. The compound according to claim 1 or 2 wherein MATRIX is agarose.

5. The compound according to claim 1 or 2 wherein MATRIX is cross-linked hydrozypropylated regenerated cellulose.

6. A process for the preparation of a compound of the general formula III ##STR9## wherein MATRIX is a water and organic solvent insoluble radical selected from the group consisting of:

agarose, starch, dextran, cellulose, regenerated cellulose, hydroxy C₂ -C₄ alkylated cellulose and hydroxy C₂ -C₄ lower alkylated regenerated cellulose; and

Y is a member selected from the group consisting of imidazoyl, 1,2,4-triazoyl and 1,2,3-benzotriazoyl which process comprises;

carbonylating MATRIX as defined above, in an organic solvent selected from the group consisting of dimethyl formamide, dioxan and acetone at a temperature of 0°

C. to 80°

C., the upper temperature limit being below the degradation temperature of MATRIX;

with a carbonylating agent ##STR10## wherein Y is selected from the group consisting of a diimidazole, a 1,2,4-triazole or a 1,2,3-benzotriazole.

7. A process according to claim 6 in which said carbonylating agent is selected from the group consisting of N,N'"'"'-carbonyl diimidazole;

N,N'"'"'-carbonyl di-1,2,3-benzotriazole; and

N,N'"'"'-carbonyl di-1,2,4-triazole.

8. The process according to claim 6 wherein MATRIX is cross-linked.

9. The process according to one of claims 6 or 8 wherein said carbonylating agent is N,N'"'"'-carbonyl di-1,2,4-triazole and the reaction is conducted for at least 5 minutes.

10. The process according to one of claims 6 or 8 wherein the carbonylating agent is N,N'"'"'-carbonyl di-1,2,3-benzotriazole, and is conducted in dioxan for a period of 7 days.

11. The process according to one of claims 6 or 8 wherein the carbonylating agent is N,N'"'"'-carbonyl diimidazole and the reaction is conducted in dioxan solution for at least 5 minutes.

12. A process for the preparation of a compound of the general formula V:

##STR11## wherein MATRIX is a cross linked water and organic solvent insoluble radical selected from the group consisting of agarose, starch, dextran, cellulose, regenerated cellulose, hydroxy C₂ -C₄ alkylated cellulose and hydroxy C₂ -C₄ lower alkylated regerated cellulose, R⁵ is selected from the group consisting of hydrogen, and an alkyl, and R⁶ being selected from the group consisting of --(CH₂)_n NH₂, and --(CH₂)_n CO₂ H wherein n is an integer of from 2 to 12,

which process comprises reacting a compound of the formula ##STR12## wherein MATRIX is defined as above and Y is a member selected from the group consisting of imidazoyl, 1,2,4-triazoyl and 1,2,3-benzotriazoyl

with an amine of the general formula NHR⁵ R⁶ wherein R⁵ and R⁶ are as defined above.

13. The process as claimed in claim 12 wherein when in the compound of formula V, R⁵ or R⁶ is --(CH₂)_n COOH, n being from 2 to 12 the product is further reacted with a primary or secondary amine.

14. The process as claimed in claim 12 wherein when, in the compound of Formula V, R R⁵ or R⁶ is --(CH₂)_n NH₂, n being from 2 to 12, the product is further reacted with a carboxylic acid.

15. A compound of the general formula V:

##STR13## wherein MATRIX is a cross linked water and organic solvent insoluble radical selected from the group consisting of agarose, starch, dextran, cellulose, regenerated cellulose, hydroxy C₂ -C₄ alkylated cellulose and hydroxy C₂ -C₄ lower alkylated regenerated cellulose, R¹ is selected from the group consisting of hydrogen, and an alkyl, and R² being selected from the group consisting of --(CH₂)_n NH₂, and --(CH₂)_n CO₂ H, wherein n is an integer of from 2 to 12.