Alphavirus and Alphavirus Replicon Particle Formulations and Methods

Abstract

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 said 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 dried composition with rehydration and storage stability comprising (1) alphaviruses or alphavirus replicon particles, (2) a salt, (3) a surfactant, and (4) a hydrogen-bonding sugar, sugar alcohol or other polyol. 
     
     
         2 . The composition of  claim 1  further comprising a plasticizer. 
     
     
         3 . A dried composition with rehydration and storage stability comprising (1) a alphaviruses or alphavirus replicon particles, (2) a salt, (3) a surfactant, (4) a hydrogen-bonding sugar or sugar alcohol or other polyol, and (5) a bulking agent. 
     
     
         4 . The composition of  claim 2  further comprising a plasticizer. 
     
     
         5 . The composition of  claim 1 , wherein the pH of the composition prior to drying is maintained at a pH from about 7 to about 9. 
     
     
         6 . The composition of  claim 5 , wherein boric acid and a pharmaceutically acceptable salt of tetraborate are used to buffer the composition prior to drying. 
     
     
         7 . The composition of  claim 1 , wherein the alphaviruses or alphavirus replicon particles are produced in Vero cell culture, CHO cell culture, BHK cell culture, or 293 cell culture. 
     
     
         8 . The composition of  claim 1 , wherein the alphavirus is Venezuelan Equine Encephalitis Virus (VEE). 
     
     
         9 . The composition of  claim 1 , wherein the salt is provided at an ionic strength sufficient to reduce, minimize or eliminate aggregation in the population of alphaviruses or alphavirus replicon particles. 
     
     
         10 . The composition of  claim 9 , wherein the salt is sodium sulfate, sodium citrate, sodium glutamate, magnesium sulfate or sodium sucrose octasulfate. 
     
     
         11 . The composition of  claim 1 , wherein the surfactant is a protein, Tween 20 ™ (polyoxyethylene (20) sorbitan monolaurate), Tween 80™ (Polyoxyethylenesorbitan monooleate), Brij 35™ (poly(ethoxethylene-23) lauryl ether), poloxamer 188 (triblock copolymer of the form poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)) or gelatin. 
     
     
         12 . The composition of  claim 11 , wherein the protein is human serum albumin. 
     
     
         13 . The composition of  claim 12 , wherein the human serum albumin is produced recombinantly. 
     
     
         14 . The composition of  claim 2  wherein the bulking agent is hydroxyethyl starch, dextran, mannitol, glycine, Ficoll or polyvinylpyrrolidone. 
     
     
         15 . The composition of any of  claim 1 , wherein there is residual moisture content from about 0.5% to about 10%. 
     
     
         16 . The composition of  claim 15  wherein the residual moisture content is from about 2 to about 7%. 
     
     
         17 . The composition of  claim 16 , wherein the residual moisture content is from about 3 to about 5%. 
     
     
         18 . The composition of  claim 2 , wherein the plasticizer is from about 0.05% to about 2% glycerol. 
     
     
         19 . A method for preparing the composition of  claim 1  which comprises the steps of: (a) preparing an aqueous solution or dispersion comprising an alphavirus or alphavirus replicon particle, a salt, a surfactant, and a hydrogen-bonding sugar or sugar alcohol; and (b) drying the aqueous dispersion to obtain a composition wherein the alphavirus or alphavirus replicon particle is dispersed in an amorphous glassy matrix comprising the surfactant and sugar or sugar alcohol. 
     
     
         20 . The method of  claim 19  wherein the infectivity titer of the dried composition is no more than one log less than the infectivity titer of the aqueous solution. 
     
     
         21 . The method of  claim 19 , wherein the aqueous solution or dispersion further comprises a plasticizer. 
     
     
         22 . The method of  claim 20 , wherein the aqueous solution or dispersion further comprises a bulking agent. 
     
     
         23 . The method of  claim 20 , wherein the aqueous solution or dispersion further comprises a plasticizer and a bulking agent.