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.
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