Glycoconjugation process

Patent metadata

Field	Value
Publication number	US-11117928-B2
Application number	US-202016923154-A
Country	US
Kind code	B2
Filing date	Jul 8, 2020
Priority date	Dec 20, 2012
Publication date	Sep 14, 2021
Grant date	Sep 14, 2021

Abstract

The present disclosure relates generally to methods of preparing glycoconjugates containing a saccharide conjugated to a carrier protein by use of stable nitroxyl radical related agent/oxidant as an oxidizing agent, to immunogenic compositions comprising such glycoconjugates, and to methods for the use of such glycoconjugates and immunogenic compositions.

First claim

The invention claimed is:

1. An immunogenic composition comprising a glycoconjugate comprising a Pn-serotype 12F capsular polysaccharide conjugated to a carrier protein, wherein the amount of free Pn-serotype 12F polysaccharide in the composition is less than 35% after 120 days from when it was prepared, and wherein said glycoconjugate is prepared by a process comprising:
a) reacting a Pn-serotype 12F capsular polysaccharide with a stable nitroxyl radical compound and an oxidant, to produce an activated capsular polysaccharide, wherein said oxidant is a molecule bearing a N-halo moiety which selectively oxidizes primary alcohols in the presence of a nitroxyl radical compound to generate aldehyde groups, wherein said stable nitroxyl radical compound is a molecule bearing a TEMPO or a PROXYL (2,2,5,5-tetramethyl-1-pyrrolidinyloxy) moiety, having the ability to selectively oxidize primary alcohols in the presence of an oxidant, to generate aldehyde groups without affecting secondary hydroxyl groups; and
b) reacting the activated capsular polysaccharide with the carrier protein comprising one or more amine groups.

2. The immunogenic composition of claim 1 , wherein the amount of free Pn-serotype 12F polysaccharide is less than 30% after 120 days from when it was prepared.

3. The immunogenic composition of claim 1 , wherein said nitroxyl radical compound is selected from the group consisting of TEMPO, 2,2,6,6-Tetramethyl-4-(methylsulfonyloxy)-1-piperidinooxy, 4-Phosphonooxy-TEMPO, 4-Oxo-TEMPO, 4-Methoxy-TEMPO, 4-Isothiocyanato-TEMPO, 4-(2-lodoacetamido)-TEMPO free radical, 4-Hydroxy-TEMPO, 4-Cyano-TEMPO, 4-Carboxy-TEMPO, 4-(2-Bromoacetamido)-TEMPO, 4-Amino-TEMPO, and 4-Acetamido-2,2,6,6-tetramethylpiperidine 1-oxyl.

4. The immunogenic composition of claim 1 , wherein said oxidant is selected from the group consisting of N-Chlorosuccinimide, N-Bromosuccinimide, N-lodosuccinimide, Dichloroisocyanuric acid, 1,3,5-trichloro-1,3,5-triazinane-2,4,6-trione, Dibromoisocyanuric acid, 1,3,5-tribromo-1,3,5-triazinane-2,4,6-trione, Diiodoisocyanuric acid and 1,3,5-triiodo-1,3,5-triazinane-2,4,6-trione.

5. The immunogenic composition of claim 1 , wherein the carrier protein is a toxin from tetanus, diphtheria, pertussis, Pseudomonas, E. coli, Staphylococcus or Streptococcus.

6. The immunogenic composition of claim 1 , wherein the carrier protein is CRM 197 .

7. The immunogenic composition of claim 1 further comprising a pharmaceutically acceptable excipient, carrier, or diluent.

8. The immunogenic composition of claim 1 , further comprising an additional antigen.

9. The immunogenic composition of claim 8 , wherein the additional antigen comprises a protein antigen or a glycoconjugate of a capsular polysaccharide derived from S. pneumonia.

10. The immunogenic composition of claim 9 , wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide selected from Pn-serotypes 1, 4, 5, 6A, 6B, 7F, 8, 9V, 11A, 14, 15B, 18C, 19A, 19F, 22F, and 23F capsular polysaccharides.

11. The immunogenic composition of claim 8 , wherein the additional antigen comprises a protein antigen or a glycoconjugate of a capsular polysaccharide derived from N. meningitidis.

12. The immunogenic composition of claim 11 , wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide selected from serotypes A, C, W135 and Y capsular polysaccharides.

13. The immunogenic composition of claim 11 , wherein the additional antigen comprises a glycoconjugate of serotypes X capsular polysaccharide.

14. The immunogenic composition of claim 8 , wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide derived from Group B Streptococcus (GBS).

15. The immunogenic composition of claim 14 , wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide selected from GBS serotypes Ia, Ib, II, III, IV, V, VI, VII and VIII.

16. The immunogenic composition of claim 1 , further comprising an adjuvant.

17. The immunogenic composition of claim 16 , wherein the adjuvant is an aluminum-based adjuvant.

18. The immunogenic composition of claim 17 , wherein the aluminum-based adjuvant is selected from the group consisting of aluminum phosphate, aluminum sulfate, and aluminum hydroxide.

19. A method of ameliorating a bacterial infection, disease or condition in a subject, comprising administering to the subject an immunologically effective amount of an immunogenic composition of claim 1 .

20. The method of claim 19 , wherein the infection, disease or condition is associated with S. pneumoniae bacteria.

21. A method of inducing a protective immune response in a subject, comprising administering to the subject an immunologically effective amount of an immunogenic composition of claim 1 .

Assignees

• Pfizer

Inventors

• Han Mingming
• Kainthan Rajesh Kumar
• Kim Jin-Hwan
• Prasad Avvari Krishna

Classifications

• A61K39/09 Primary

  {Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus}, streptococcus · CPC title

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title

• C07K1/107

  by chemical modification of precursor peptides · CPC title

• C07H3/06

  Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages · CPC title

• A61K47/50

  the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates · CPC title