US 12,419,943 B2
Methods for producing Streptococcus pneumoniae capsular polysaccharide carrier protein conjugates
Patrick Leonard Ahl, West Point, PA (US); Akhilesh Bhambhani, Doylestown, PA (US); Christopher Jon Farrell, Wayne, PA (US); Patrick McHugh, Pipersville, PA (US); Morrisa C. Jones, Upper Darby, PA (US); Daniel D. Roth, Coopersburg, PA (US); Jessica R. Sinacola, Collegeville, PA (US); Justin Stanbro, Sellersville, PA (US); Matthew P. Watson, Ottsville, PA (US); Emily Wen, Harleysville, PA (US); and Michael A. Winters, Doylestown, PA (US)
Assigned to Merck Sharp & Dohme LLC, Rahway, NJ (US)
Filed by Merck Sharp & Dohme LLC, Rahway, NJ (US)
Filed on Apr. 17, 2024, as Appl. No. 18/637,589.
Application 18/637,589 is a division of application No. 18/049,507, filed on Oct. 25, 2022, granted, now 11,992,521.
Application 18/049,507 is a continuation of application No. 17/049,245, abandoned, previously published as PCT/US2019/029048, filed on Apr. 25, 2019.
Claims priority of provisional application 62/664,590, filed on Apr. 30, 2018.
Prior Publication US 2024/0261384 A1, Aug. 8, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 39/02 (2006.01); A61K 39/09 (2006.01); A61K 47/61 (2017.01); A61K 47/62 (2017.01); A61K 39/00 (2006.01)
CPC A61K 39/092 (2013.01) [A61K 47/61 (2017.08); A61K 47/62 (2017.08); A61K 2039/6037 (2013.01); A61K 2039/64 (2013.01); A61K 2039/70 (2013.01)] 15 Claims
 
1. 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, wherein reconstituting requires eight minutes or less;
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, wherein mixing comprises 120 minutes or less;
(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.