# US 4,397,838 A · Preparations of purified bacterial membranal proteoglycans

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- Number: US 4,397,838 A
- Document ID: US4397838A
- Status: Granted
- Kind: A
- Application number: 06/280,379
- Filed: 1981-07-06
- Granted: 1983-08-09
- Earliest claimed: 1978-12-19
- Assignees: Pierre Fabre S.A.
- Inventors: Lucien D. d'Hinterland; Gerard Normier; Anne-Marie Pinel; Jacques Durand
- Web page: https://patentgrep.com/patent/US4397838A

## CPC

- A61K 35/74: Bacteria (therapeutic use of a bacterial protein A61K38/00)
- A61K 39/0266: {Klebsiella}
- A61P 37/04: Immunostimulants
- C12P 21/005: {Glycopeptides, glycoproteins}
- A61K 2039/522: {avirulent or attenuated}
- A61K 2039/53: {DNA (RNA) vaccination}
- Y10S 435/849: Escherichia coli
- Y10S 435/852: Klebsiella
- Y10S 435/881: Serratia marcescens
- Y10S 530/825: Bacteria
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

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.

## Claims

### Claim 1

An immunity adjuvant for vaccines comprising purified water-soluble proteoglycans obtained by a process which comprises at least one step in which crude membranal proteoglycans of Klebsiella, Serratia or Escherichia 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, said purified water-soluble proteoglycans having the following composition by weight:
______________________________________ hexoses 24-42% proteins 31-53% lipids 12-18% hexosamines 2-6% RNA <0.5% DNA <0.2% LPS <0.001% ______________________________________

### Claim 2 (depends on 1)

An adjuvant as set forth in claim 1 wherein the purified water-soluble proteoglycans have the following composition by weight:
______________________________________ hexoses 37 .+-. 5% proteins 36.5 .+-. 5% lipids 15 .+-. 3% hexosamines 5 .+-. 2% RNA <0.5% DNA <<0.2% LPS <0.001% ______________________________________

### Claim 3 (depends on 1)

An adjuvant as set forth in claim 1 wherein the purified water-soluble proteoglycans have the following composition by weight:
______________________________________ hexoses 29 .+-. 5% proteins 48 .+-. 5% lipids 15 .+-. 3% hexosamines 4 .+-. 2% RNA <0.5% DNA <<0.2% LPS <0.001% ______________________________________

### Claim 4 (depends on 1)

A vaccine based on bacterial ribosome and/or RNA as immunogenic agent containing as immunity adjuvant an effective amount of an adjuvant of the type set forth in claim 1.

### Claim 5 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomes of Klebsiella pneumoniae 3.5 .mu.g ribosomes of Streptococcus pneumoniae 3.0 .mu.g ribosomes of Streptococcus pyogenes A.sub.12 3.0 .mu.g ribosomes of Hemophilus influenzae 0.5 .mu.g proteoglycans of Klebsiella pneumoniae 15.0 .mu.g ______________________________________

### Claim 6 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomal RNA of Klebsiella pneumoniae 2.45 .mu.g ribosomal RNA of Streptococcus pneumoniae 2.10 .mu.g ribosomal RNA of Streptococcus pyogenes A.sub.12 2.10 .mu.g ribosomal RNA of Hemophilus influenzae 0.35 .mu.g proteoglycans of Klebsiella pneumoniae 10.50 .mu.g ______________________________________

### Claim 7 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomes of Rothia dentocariosus 1.2 .mu.g ribosomes of Actinomyces viscosus 1.2 .mu.g ribosomes of Streptococcus mutans 0.8 .mu.g ribosomes of Streptococcus salivarius 2.0 .mu.g ribosomes of Lactobacillus casei 1.2 .mu.g proteoglycans of Klebsiella pneumoniae 9.6 .mu.g ______________________________________

### Claim 8 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomal RNA of Rothia dentocariosus 0.84 .mu.g ribosomal RNA of Actinomyces viscosus 0.84 .mu.g ribosomal RNA of Streptococcus mutans 0.56 .mu.g ribosomal RNA of Streptococcus salivarius 1.40 .mu.g ribosomal RNA of Lactobacillus casei 0.84 .mu.g proteoglycans of Klebsiella pneumoniae 6.72 .mu.g ______________________________________

### Claim 9 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomes of Corynebacterium acnes 2 .mu.g ribosomes of Corynebacterium parvum 2 .mu.g ribosomes of Streptococcus pyogenes 2 .mu.g ribosomes of Staphylococcus epidermidis 2 .mu.g proteoglycans of Serratia marcescens 12 .mu.g ______________________________________

### Claim 10 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomal RNA of Corynebacterium acnes 1.4 .mu.g ribosomal RNA of Corynebacterium parvum 1.4 .mu.g ribosomal RNA of Streptococcus pyogenes 1.4 .mu.g ribosomal RNA of Staphylococcus epidermidis 1.4 .mu.g proteoglycans of Serratia marcescems 8.4 .mu.g ______________________________________

### Claim 11 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomes of Escherichia coli 2 .mu.g ribosomes of Streptococcus faecalis 2 .mu.g ribosomes of Streptococcus H 2 .mu.g ribosomes of Staphylococcus epidermidis 2 .mu.g ribosomes of Candida albicans 2 .mu.g proteoglycans of Escherichia coli 15 .mu.g ______________________________________

### Claim 12 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomal RNA of Escherichia coli 1.4 .mu.g ribosomal RNA of Streptococcus faecalis D 1.4 .mu.g ribosomal RNA of Streptococcus H 1.4 .mu.g ribosomal RNA of Staphylococcus epidermidis 1.4 .mu.g ribosomal RNA of Candida albicans 1.4 .mu.g proteoglycans of Escherichia coli 10.5 .mu.g ______________________________________

### Claim 13 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomes of Bacterium coli 2 .mu.g ribosomes of Salmonella paratyphi A 2 .mu.g ribosomes of Salmonella paratyphi B 2 .mu.g ribosomes of Shigella dysenteria 2 .mu.g ribosomes of Enterococcus 2 .mu.g proteoglycans of Serratia marcescens 15 .mu.g ______________________________________

### Claim 14 (depends on 4)

A vaccine as set forth in claim 4 having the following composition:
______________________________________ ribosomal RNA of Bacterium coli 1.4 .mu.g ribosomal RNA of Salmonella paratyphi A 1.4 .mu.g ribosomal RNA of Salmonella paratyphi B 1.4 .mu.g ribosomal RNA of Shigella dysenteriae 1.4 .mu.g ribosomal RNA of Enterococcus 1.4 .mu.g proteoglycans of Serratia marcescens 10.5 .mu.g ______________________________________