Process for the preparation of purified bacterial membranal proteoglycans

4,297,272

06/103,202

December 13, 1979

This invention relates to a process for the preparation of purified bacterial membranal proteoglycans. The process comprises at least one step in which crude proteoglycans are treated in aqueous medium with a base or a hypobromite, followed by elimination of the excess reactant and the insoluble residue, the purified proteoglycans being present in aqueous solution. The thus obtained purified proteoglycans may be used as immunity adjuvants in vaccines without causing side-effects such as pyrogenic reactions.

D'Hinterland, Lucien D. (Castres, FRX); Normier, Gerard (Castres, FRX); Pinel, Anne-Marie (Castres, FRX); Durand, Jacques (Castres, FRX)

Pierre Fabre S.A. (FRX)

What is claimed is

1. A process for the preparation of purified water soluble bacterial membranal proteoglycans which comprises at least one step in which crude proteoglycans are treated in an aqueous medium with a reactant selected from the group consisting of bases and hypobromites, followed by removal of the excess reactant and the insoluble residue, the purified proteoglycans being present in aqueous solution.

2. A process as set forth in claim 1, wherein the base used is an alkali hydroxide having a molarity of from 0.05 to 2 M and the treatment time is a few hours at a temperature of about ambient temperature.

3. A process as set forth in claim 1, wherein the hypobromite used is an alkali hypobromite and the treatment time is a few tens of minutes at ambient temperature.

4. A process as set forth in claim 1 wherein the excess reactant is eliminated by dialysis and the insoluble residue is eliminated by centrifuging.

5. A process as claimed in claim 1, which further comprises at least one step in which the membranal proteoglycans are hydrolysed with acetic acid in an aqueous medium at a temperature of from 70.degree. to 100.degree. C. and the thus formed insoluble fraction is eliminated.

6. A process as claimed in claim 5, wherein the acid hydrolysis step is carried out with 1% by volume glacial acetic acid.

7. A process as claimed in claim 1, wherein the proteoglycans are extracted from a gram-negative bacteria.

8. A process as claimed in claim 1, wherein the proteoglycans are extracted from gram-negative bacteria selected from the group consisting of Klebsiella pneumoniae, Serratia marcesens and Escherichia coli.

260/112R; 420/92; 420/177

C07G 700; A61K 39108; A61K 3902

3408440 October 1968 Voss
3956481 May 1976 Jolles et al.
4001395 January 1977 Jolles et al.
4108849 August 1978 Thomas

edspgr.04297272