Stabilizer and Vaccine Composition Comprising One or More Live Attenuated Flaviviruses

Abstract

The present invention relates to stabilizers for compositions, including immunogenic compositions, such as vaccine compositions, comprising one or more live attenuated flaviviruses, to bulk vaccine compositions stabilized with these stabilizers, particularly dry vaccine compositions prepared from these bulk vaccine compositions, and to methods for stabilizing one or more live attenuated flaviviruses.

1 . A stabilizer for compositions comprising one or more live attenuated flaviviruses, which comprises, in an aqueous solution without proteins of animal origin and without added salts having divalent cations,
 a buffer,   2.5% to 6.5% of sorbitol,   2.5% to 13% of sucrose,   0 to 7.5% of trehalose and/or 0 to 7.5% of any other disaccharide or trisaccharide,   0.2% to 0.5% of urea,   0.8% to 2.5% of an amino acid mixture comprising arginine (Arg), cystine (Cys-Cys), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Val), alanine (Ala), asparagine (Asn), aspartic acid (Asp), glutamic acid (Glu), glycine (Gly), proline (Pro) and serine (Ser).   
     
     
         2 . The stabilizer as claimed in  claim 1 , which comprises one or more buffers chosen from the group comprising TRIS (tris(hydroxymethyl)-aminomethane), HEPES (2-(4-(2-hydroxyethyl)-1-piperazinyl)ethane-sulfonic acid) and potassium phosphate and/or sodium phosphate. 
     
     
         3 . The stabilizer as claimed in  claim 2 , wherein the TRIS is present at a concentration of from 5 to 10 mM. 
     
     
         4 . The stabilizer as claimed in  claim 2 , wherein the HEPES is present at a concentration of from 7.5 to 20 mM. 
     
     
         5 . The stabilizer as claimed in  claim 1 , comprising
 3.8% (w/v) of sorbitol,   7.5% (w/v) of sucrose,   5.5% (w/v) of trehalose,   0.25% (w/v) of urea and   1.5% (w/v) of the amino acid mixture.   
     
     
         6 . A stabilized bulk aqueous vaccine composition comprising one or more live attenuated flaviviruses and the stabilizer as claimed in  claim 1 . 
     
     
         7 . The vaccine composition as claimed in  claim 6 , which comprises one or more live attenuated dengue (DEN) virus serotypes. 
     
     
         8 . The vaccine composition as claimed in  claim 1 , which comprises live attenuated yellow fever (YF) viruses. 
     
     
         9 . The vaccine composition as claimed in  claim 1 , which comprises live attenuated West Nile (WN) virus disease viruses. 
     
     
         10 . The vaccine composition as claimed in  claim 1 , which comprises live attenuated Japanese encephalitis (JE) viruses. 
     
     
         11 . The vaccine composition as claimed in  claim 1 , which comprises one or more chimeric live attenuated flaviviruses. 
     
     
         12 . The vaccine composition as claimed in  claim 11 , which comprises one or more serotypes of a chimeric YF-DEN (yellow fever-dengue) virus. 
     
     
         13 . The vaccine composition as claimed in  claim 1 , which comprises a chimeric YF-WN (yellow fever-West Nile virus) virus. 
     
     
         14 . The vaccine composition as claimed in  claim 1 , which comprises a chimeric YF-JE (yellow fever-Japanese encephalitis) virus. 
     
     
         15 . A method for stabilizing one or more live attenuated flaviviruses, comprising the dilution of a purified and concentrated viral harvest comprising one or more live attenuated flaviviruses by adding stabilizer so as to obtain the final concentrations of the stabilizer defined according to  claim 1  in order to obtain a stabilized bulk aqueous vaccine composition comprising one or more live attenuated flaviviruses and the stabilizer. 
     
     
         16 . The stabilization method as claimed in  claim 15 , comprising drying the aqueous composition by means of a method selected from the group of foam-drying, spray-drying or freeze-foam-drying. 
     
     
         17 . The stabilization method as claimed in  claim 15 , comprising drying the aqueous composition by means of the freeze-drying method. 
     
     
         18 . The stabilization method as claimed in  claim 15 , comprising drying the aqueous composition by means of the spray-freeze-drying method. 
     
     
         19 . The stabilization method as claimed in  claim 17 , wherein, in a first step, the aqueous solution is frozen in the form of uniform particles or of beads, and wherein, in a second step, the frozen uniform particles or beads are subjected to drying in order to obtain a stabilized dry product in the form of uniform particles or of beads. 
     
     
         20 . The stabilization method as claimed in  claim 19 , wherein the uniform particles or beads of the stabilized dry product have a diameter of approximately 100 μm to 1500 μm. 
     
     
         21 - 28 . (canceled)