US 11,850,305 B2
Method of making lipid formulations with RNA encoding immunogens
Andrew Geall, Littleton, MA (US)
Assigned to GLAXOSMITHKLINE BIOLOGICALS SA, Rixensart (BE)
Filed by GLAXOSMITHKLINE BIOLOGICALS SA, Rixensart (BE)
Filed on Aug. 8, 2023, as Appl. No. 18/231,697.
Application 18/231,697 is a division of application No. 17/560,059, filed on Dec. 22, 2021, granted, now 11,766,401.
Application 17/560,059 is a division of application No. 13/808,080, abandoned, previously published as PCT/US2011/043105, filed on Jul. 6, 2011.
Claims priority of provisional application 61/378,837, filed on Aug. 31, 2010.
Claims priority of provisional application 61/361,830, filed on Jul. 6, 2010.
Prior Publication US 2023/0381107 A1, Nov. 30, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 9/127 (2006.01); A61K 39/12 (2006.01); C12N 15/86 (2006.01); A61K 31/7088 (2006.01); A61K 39/00 (2006.01); A61K 39/39 (2006.01)
CPC A61K 9/127 (2013.01) [A61K 9/1272 (2013.01); A61K 31/7088 (2013.01); A61K 39/00 (2013.01); A61K 39/12 (2013.01); C12N 15/86 (2013.01); A61K 39/39 (2013.01); A61K 2039/53 (2013.01); A61K 2039/55555 (2013.01); C12N 2710/16134 (2013.01); C12N 2760/18534 (2013.01); C12N 2770/36134 (2013.01)] 30 Claims
 
1. A method of obtaining a pharmaceutical composition, the pharmaceutical composition comprising ribonucleic acid (RNA) molecules and lipid particles; the RNA molecules comprising a sequence that encodes an immunogen; the immunogen comprising an Epstein-Barr virus (EBV) immunogen, a cytomegalovirus (CMV) immunogen, a coronavirus spike polypeptide immunogen, an influenza virus A immunogen, a Varicella zoster virus (VZV) immunogen, or a flavivirus immunogen; the lipid particles comprising lipids comprising: (a) from 40 mole % to 60 mole % cationic lipid comprising a tertiary amine, (b) from 1 mole % to 6 mole % polyethylene glycol-conjugated (PEG-conjugated) lipid, and (c) from 35 mole % to 50 mole % cholesterol; at least 80% of the lipid particles having a diameter from 20 nm to 220 nm; the lipid particles encapsulating at least half of the RNA molecules; the pharmaceutical composition being immunogenic in vivo by eliciting an antibody response against the immunogen in vivo; the method comprising the steps of:
(i) mixing the lipids and ethanol, thereby obtaining an ethanolic lipid mixture;
(ii) mixing the RNA molecules and an aqueous buffer, thereby obtaining an aqueous RNA mixture;
(iii) mixing the ethanolic lipid mixture and the aqueous RNA mixture, thereby obtaining an intermediate mixture; and
(iv) purifying the intermediate mixture, thereby obtaining the pharmaceutical composition.