Alphavirus and alphavirus replicon particle formulations and methods
Disclosed are methods for preparing dried (preferably lyophilized) preparations comprising a population of alphaviruses or alphavirus replicon particles, a sugar or polyol, a surfactant and a salt and preparations made by these methods, both in the dried form but also as liquids prior to drying or after reconstituting dried preparations. These preparations may further comprise a plasticizer and/or a bulking agent. These preparations are readily reconstituted, with little or no loss in infectivity of the viruses or replicon particles.
1. A method for preparing a dried composition, said method comprising the steps of:
(a) preparing an aqueous solution or dispersion comprising Venezuelan equine encephalitis virus or Venezuelan equine encephalitis virus replicon particles, 10 mM sodium phosphate, 4% (w/v) sucrose, 0.1% (w/v) human serum albumin, 100 mM sodium sulfate, 0.25% (w/v) glycerol, a buffer comprising boric acid and a pharmaceutically acceptable tetraborate salt for maintaining the pH of said aqueous solution or dispersion at 7.4; and
(b) drying the aqueous solution or dispersion to obtain a dried composition containing Venezuelan equine encephalitis virus or Venezuelan equine encephalitis virus replicon particles that are dispersed in an amorphous glassy matrix comprising the sucrose, human serum albumin and sodium sulfate, wherein infectivity titer of the dried composition is no more than one log less than the infectivity titer of the aqueous solution.
2. The method of claim 1 , wherein the human serum albumin is recombinant human serum albumin.
3. The method of claim 1 , wherein the aqueous solution or dispersion further comprises a bulking agent.
4. The method of claim 3 , wherein the bulking agent is hydroxyethyl starch, dextran, mannitol, glycine, Ficoll or polyvinylpyrrolidone.
5. The method of claim 1 , where the aqueous solution or dispersion further comprises polypropylene glycol.
6. The method of claim 1 , wherein the dried composition has residual moisture content from about 0.5% (w/v) to about 10% (w/v).
7. The method of claim 6 , wherein the residual moisture content is from about 2% (w/w) to about 7% (w/w) in the dried composition.
8. The method of claim 7 , wherein the residual moisture content is from about 3 (w/w) to about 5% (w/w) in the dried composition.
9. The method of claim 1 , wherein the Venezuelan equine encephalitis virus or Venezuelan equine encephalitis virus particle is produced in Vero cell culture, CHO cell culture, BHK cell culture, or 293 cell culture.