Glycoconjugation process

Patent metadata

Field	Value
Publication number	US-11603384-B2
Application number	US-202117400938-A
Country	US
Kind code	B2
Filing date	Aug 12, 2021
Priority date	Dec 20, 2012
Publication date	Mar 14, 2023
Grant date	Mar 14, 2023

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 capsular polysaccharide from Streptococcus pneumoniae conjugated to a carrier protein, wherein the glycoconjugate is prepared by a process comprising:
a) reacting the capsular polysaccharide with a stable nitroxyl radical compound and an oxidant, to produce an activated capsular polysaccharide, wherein the stable nitroxyl radical compound is 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) and the oxidant is N-chlorosuccinimide (NCS); 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 capsular polysaccharide is reacted with 0.1 to 10 molar equivalent of N-chlorosuccinimide.

3. The immunogenic composition of claim 1 , wherein the capsular polysaccharide is reacted with less than about 0.3 molar equivalent of 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO).

4. The immunogenic composition of claim 1 , wherein the capsular polysaccharide is selected from Pn-serotype 3, Pn-serotype 10A, Pn-serotype 12F, and Pn-serotype 33F capsular polysaccharides.

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

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

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

8. The immunogenic composition of claim 7 , wherein the additional antigen comprises a protein antigen or a glycoconjugate of a capsular polysaccharide derived from S. pneumonia.

9. The immunogenic composition of claim 8 , 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.

10. The immunogenic composition of claim 7 , wherein the additional antigen comprises a protein antigen or a glycoconjugate of a capsular polysaccharide derived from N. meningitidis.

11. The immunogenic composition of claim 10 , wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide selected from serotypes A, C, W135 and Y capsular polysaccharides.

12. The immunogenic composition of claim 10 , wherein the additional antigen comprises a glycoconjugate of serotypes X capsular polysaccharide.

13. The immunogenic composition of claim 7 , wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide selected from Group B Streptococcus (GBS) serotypes Ia, Ib, II, III, IV, V, VI, VII and VIII.

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

15. The immunogenic composition of claim 14 , wherein the adjuvant is an aluminum-based adjuvant selected from aluminum phosphate, aluminum sulfate, and aluminum hydroxide.

16. 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 .

17. The method of claim 16 , wherein the infection, disease or condition is associated with S. pneumoniae bacteria.

18. 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 .

19. An immunogenic composition comprising a glycoconjugate comprising a Pn-serotype 12F capsular polysaccharide conjugated to a carrier protein, wherein said glycoconjugate is prepared by a process comprising:
a) reacting said capsular polysaccharide with a stable nitroxyl radical compound and an oxidant, to produce an activated capsular polysaccharide, wherein said stable nitroxyl radical compound is 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) and said oxidant is N-chlorosuccinimide; and
b) reacting the activated capsular polysaccharide with the carrier protein comprising one or more amine groups.

20. The immunogenic composition of claim 19 , further comprising an additional antigen, wherein the additional antigen comprises a glycoconjugate of a capsular polysaccharide selected from Pn-serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 14, 15B, 18C, 19A, 19F, 22F, 23F, and 33F capsular polysaccharides.

Assignees

• Pfizer

Inventors

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

Classifications

• 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

• C07K14/315

  from Streptococcus (G), e.g. Enterococci · CPC title

• A61K2039/6037

  Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT] · CPC title