- Document Number:
20240261384
- Appl. No:
18/637589
- Application Filed:
April 17, 2024
- نبذة مختصرة :
A method is described for producing a pneumococcal capsular polysaccharide protein conjugate in which one or more activated pneumococcal polysaccharides of particular pneumococcal serotypes and carrier protein are separately lyophilized, the separately lyophilized polysaccharides and carrier protein are separately reconstituted in an organic solvent, and the reconstituted polysaccharide and carrier protein are then combined together by Tee-mixing and conjugated together to produce polysaccharide carrier protein conjugates. A plurality of conjugates, each comprising polysaccharides of a particular serotype, may be used to produce multivalent pneumococcal immunogenic compositions having a combination of conjugates for use in vaccines.
- Assignees:
Merck Sharp & Dohme LLC (Rahway, NJ, US)
- Claim:
1-17. (canceled)
- Claim:
18. A method for making a composition comprising two or more conjugates, each conjugate having Streptococcus pneumoniae polysaccharides from one or more serotypes covalently linked to a carrier protein, the method comprising: (a) providing (i) two or more first aqueous solutions, each first aqueous solution comprising activated polysaccharides of a specific Streptococcus pneumoniae serotype, wherein the polysaccharides have been reacted with an oxidizing agent to provide the activated polysaccharides and wherein the two or more first aqueous solutions do not contain the same Streptococcus pneumoniae serotype polysaccharides; or, (ii) two or more first aqueous solutions, each first aqueous solution comprising activated polysaccharides of two or more Streptococcus pneumoniae serotypes wherein the polysaccharides have been reacted with an oxidizing agent to provide the activated polysaccharide and wherein the two or more first aqueous solutions do not contain the same Streptococcus pneumoniae serotype polysaccharides; (b) providing two or more second aqueous solutions, each comprising a carrier protein and a buffer, wherein the quantity of two or more second aqueous solutions corresponds to at least the quantity of the two or more first aqueous solutions; (c) separately drying the two or more first aqueous solutions and the two or more second aqueous solutions in a sublimative drying process to produce two or more first dried compositions, each comprising dried polysaccharides, and two or more second dried compositions, each comprising dried carrier protein; (d) reconstituting separately each of the two or more first dried compositions and each of the two or more second dried compositions in an organic solvent and mixing to provide two or more first homogenous solutions, each independently comprising either (i) polysaccharides of a specific Streptococcus pneumoniae serotype or (ii) polysaccharides of two or more specific Streptococcus pneumoniae serotypes, and two or more second homogenous solutions, each comprising the carrier protein; (e) separately combining each first homogenous solution with a second homogenous solution by Tee-mixing to produce a plurality of mixtures, each mixture comprising a first homogenous solution and a second homogenous solution; (f) adding a reducing agent to each mixture comprising the plurality of mixtures to produce a plurality conjugate solutions, each conjugate solution independently comprising either (i) polysaccharides of a specific Streptococcus pneumoniae serotype or (ii) polysaccharides of two or more specific Streptococcus pneumoniae serotypes, conjugated to the carrier protein; and (g) combining two or more of the plurality of conjugate solutions to produce a composition comprising two or more conjugates, each conjugate having Streptococcus pneumoniae polysaccharides from one or more serotypes covalently linked to a carrier protein.
- Claim:
19. The method of claim 18, wherein at least one conjugate solution is prepared by separately Tee-mixing a first homogenous solution with a second homogenous solution and adding the reducing agent to produce the plurality of conjugate solutions; or, wherein at least one conjugate solution is prepared by separately Tee-mixing a first aqueous solution with a second homogenous solution and adding the reducing agent to produce the plurality of conjugate solutions; or, wherein each conjugate solution is prepared by separately Tee-mixing a first homogenous solution with a second homogenous solution and adding the reducing agent to produce the plurality of conjugate solutions; or, wherein each conjugate solution is prepared by separately Tee-mixing a first aqueous solution with a second homogenous solution and adding the reducing agent to produce the plurality of conjugate solutions.
- Claim:
20. The method of claim 18, wherein the sublimative drying process comprises lyophilization or radiant energy vacuum (REV) dehydration.
- Claim:
21. The method of claim 18, wherein the sublimative drying process comprises freezing the first and second aqueous solutions in the form of cakes or lyosphere beads prior to the sublimative drying process.
- Claim:
22. The method of claim 18, wherein the sublimative drying comprises bulk drying performed in a container selected from the group consisting of metal tray, plastic tray, plastic bag, and class I tubing vials.
- Claim:
23. The method of claim 18, wherein the first and second aqueous solutions comprise about 0.5% (w/v) or more sucrose.
- Claim:
24. The method of claim 18, wherein the organic solvent is dimethyl sulfoxide (DMSO).
- Claim:
25. The method of claim 18, wherein the reconstituting comprises eight minutes or less and the mixing comprises 120 minutes or less.
- Claim:
26. The method of claim 18, wherein each conjugate solution comprises polysaccharide conjugated to the carrier protein at a ratio from about 0.6 to about 1.3 polysaccharide to carrier protein on a weight to weight basis.
- Claim:
27. The method of claim 18, wherein each conjugate solution comprises a free polysaccharide amount that is less than about 15% of the total polysaccharides in the solution.
- Claim:
28. The method of claim 18, wherein the buffer is a Histidine, Succinate, MES, MOPS, HEPES, or Acetate buffer in a pH range of 5.0-7.0.
- Claim:
29. The method of claim 18, wherein the buffer is a Phosphate or Citrate buffer in a pH range of 5.0-7.0.
- Claim:
30. The method of claim 18, wherein the one or more and the two or more Streptococcus pneumoniae serotypes are selected from the group consisting of Streptococcus pneumoniae serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 6E, 6G, 6H, 7F, 7A, 7B, 7C, 8, 9A, 9L, 9N, 9V, 10F, 10A, 10B, 10C, 11F, 11A, 11B, 11C, 11D, 11E, 12F, 12A, 12B, 13, 14, 15F, 15A, 15B, 15C, 16F, 16A, 17F, 17A, 18F, 18A, 18B, 18C, 19F, 19A, 19B, 19C, 20A, 20B, 21, 22F, 22A, 23F, 23A, 23B, 24F, 24A, 24B, 25F, 25A, 27, 28F, 28A, 29, 31, 32F, 32A, 33F, 33A, 33B, 33C, 33D, 33E,34, 35F, 35A, 35B, 35C, 36, 37, 38, 39, 40, 41F, 41A, 42, 43, 44, 45, 46, 47F, 47A, 48, CWPS1, CWPS2, and CWPS3.
- Claim:
31. The method of claim 18, wherein the carrier protein is an inactivated bacterial toxoid selected from the group consisting of tetanus toxoid, diphtheria toxoid, pertussis toxoid, bacterial cytolysins or pneumolysin.
- Claim:
32. The method of claim 31, wherein the diphtheria toxoid is CRM197.
- Claim:
33. The method of claim 18, wherein the conjugate solutions are sterile filtered.
- Claim:
34-37. (canceled)
- Current International Class:
61; 61; 61; 61
- الرقم المعرف:
edspap.20240261384
Processing Request
No Comments.