US 12,144,855 B2
Methods for producing streptococcus pneumoniae capsular polysaccharide carrier protein conjugates from lyospheres
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)
Appl. No. 17/049,208
Filed by Merck Sharp & Dohme LLC, Rahway, NJ (US)
PCT Filed Apr. 25, 2019, PCT No. PCT/US2019/029034
§ 371(c)(1), (2) Date Oct. 20, 2020,
PCT Pub. No. WO2019/212842, PCT Pub. Date Nov. 7, 2019.
Claims priority of provisional application 62/664,621, filed on Apr. 30, 2018.
Prior Publication US 2021/0236646 A1, Aug. 5, 2021
Int. Cl. A61K 39/02 (2006.01); A61K 9/19 (2006.01); A61K 39/09 (2006.01); A61K 39/116 (2006.01); A61K 47/61 (2017.01); A61K 47/64 (2017.01); A61K 47/68 (2017.01); A61K 47/69 (2017.01); A61K 39/00 (2006.01)
CPC A61K 39/116 (2013.01) [A61K 9/19 (2013.01); A61K 39/092 (2013.01); A61K 47/61 (2017.08); A61K 47/6415 (2017.08); A61K 47/6829 (2017.08); A61K 47/6921 (2017.08); A61K 2039/6031 (2013.01); A61K 2039/6037 (2013.01); A61K 2039/70 (2013.01); A61K 47/646 (2017.08)] 13 Claims
 
1. A method for making a composition comprising one or more Streptococcus pneumoniae polysaccharides covalently linked to a carrier protein, the method comprising:
(a) providing a first lyosphere composition comprising activated Streptococcus pneumoniae polysaccharides from one or more Streptococcus pneumoniae serotypes and a second lyosphere composition comprising carrier protein,
wherein the first and second lyosphere compositions are prepared by a sublimative drying process selected from lyophilization and radiant energy vacuum (REV) dehydration comprising: (i) providing a first aqueous solution comprising activated Streptococcus pneumoniae polysaccharides from one or more Streptococcus pneumoniae serotypes and a second aqueous solution comprising a carrier protein and a buffer, wherein the first aqueous solution comprises about 4% to 6% sucrose and the second aqueous solution comprises about 4% to 8% sucrose, and (ii) subjecting the first and second aqueous solutions to the sublimative drying process to produce the first and second lyosphere compositions:
(b) combining an amount of the first lyosphere composition and an amount of the second lyosphere composition together to provide a lyosphere mixture having a predetermined ratio of the one or more activated polysaccharides to the carrier protein;
(c) reconstituting the lyosphere mixture in an organic solvent and mixing to provide a reconstituted mixture; and
(d) adding a reducing agent to the reconstituted mixture to produce a conjugate solution comprising carrier protein conjugated to the polysaccharides of the one or more Streptococcus pneumoniae serotypes, wherein the conjugated carrier protein has a lysine loss value of greater than five (mole/mole).