Glycoconjugation process

Patent metadata

Field	Value
Publication number	US-10745438-B2
Application number	US-201916505757-A
Country	US
Kind code	B2
Filing date	Jul 9, 2019
Priority date	Dec 20, 2012
Publication date	Aug 18, 2020
Grant date	Aug 18, 2020

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

What is claimed is:

1. A method of making a glycoconjugate comprising a capsular polysaccharide from Streptococcus pneumoniae conjugated to a carrier protein, comprising the steps of:
a) reacting said 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 NI-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-I-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 a carrier protein comprising one or more amine groups.

2. The method 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-Iodoacetamido)-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.

3. The method of claim 1 , wherein said oxidant is selected from the group consisting of N-ChloroSuccinimide, N-Bromosuccinimide, N-Iodosuccinimide, 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, and Diiodoisocyanuric acid and 1,3,5-triiodo-1,3,5-triazinane-2,4,6-trione.

4. The method 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 method of claim 4 , wherein the capsular polysaccharide is a Pn-serotype 12F capsular polysaccharide.

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

7. The method of claim 1 , wherein the carrier protein is CRM 197 .

8. The method of claim 1 , wherein prior to step a), the capsular polysaccharide is hydrolyzed to a molecular weight ranging from 50 to 500 kDa.

9. The method of claim 8 , wherein the capsular polysaccharide is hydrolyzed to a molecular weight ranging from 100 to 350 kDa.

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