Compositions and methods for stabilizing alphaviruses with improved formulations

Abstract

Embodiments herein relate to compositions and methods for stabilizing live alphaviruses. Other embodiments relate to compositions and methods for reducing degradation of live, attenuated alphaviruses. Certain embodiments relate to providing a stabilizing composition while reducing immune reaction in a subject to excipients that stabilize the live alphaviruses by providing improved formulations. Yet other embodiments relate to uses of compositions disclosed herein in kits for portable applications and methods wherein the compositions reduce degradation of the live alphaviruses.

1 . A live attenuated alphavirus composition comprising:
 one or more live alphaviruses;   
       one or more amino acids that include histidine; 
       two or more carbohydrate agents; and 
       wherein the composition does not include gelatin and wherein the composition stabilizes live alphaviruses. 
     
     
         2 . The alphavirus composition according to  claim 1 , further comprising HEPES buffer wherein the HEPES buffer concentration is 1.0 mM to 200.0 mM. 
     
     
         3 . The alphavirus composition according to  claim 1 , wherein the one or more live alphaviruses comprise Chikungunya virus (CHIK), o'nyong'nyong virus, Ross River virus, Eastern equine encephalitis and Western equine encephalitis, other Semliki Forest virus, or other Togavirus and combinations thereof. 
     
     
         4 . The alphavirus composition according to  claim 1 , wherein the one or more live alphaviruses are chikungunya (CHIK) viruses. 
     
     
         5 . (canceled) 
     
     
         6 . The alphavirus composition according to  claim 1 , wherein the composition is in aqueous form. 
     
     
         7 . The alphavirus composition according to  claim 1 , wherein the composition is partially or wholly dehydrated. 
     
     
         8 . The alphavirus composition according to  claim 1 , wherein the two or more carbohydrate agents are selected from the group consisting of one or more of trehalose, galactose, fructose, sucrose, chitosan, sorbitol, mannitol and a combination thereof. 
     
     
         9 . The alphavirus composition according to  claim 1 , wherein the two or more carbohydrate agents comprise sucrose and mannitol. 
     
     
         10 . (canceled) 
     
     
         11 . The alphavirus composition according to  claim 2 , wherein the HEPES buffer concentration is from 10.0 mM to 30.0 mM; the two or more carbohydrate agent concentration is from 10.0 to 100.0 mg/ml; and the amino acid concentration is from 1.0 mg/ml to 20.0 mg/ml. 
     
     
         12 . (canceled) 
     
     
         13 . The alphavirus composition according  claim 1 , wherein the alphavirus composition comprises HEPES buffer, histidine, sucrose, and mannitol. 
     
     
         14 . The alphavirus composition according to  claim 13 , wherein the HEPES buffer concentration is 15.0 mM, histidine concentration is 1.0 mg/ml to 20.0 mg/ml; sucrose concentration is 20.0 mg/ml to 90.0 mg/ml; and mannitol concentration is 10.0 mg/ml to 50.0 mg/ml. 
     
     
         15 .- 16 . (canceled) 
     
     
         17 . The alphavirus composition according to  claim 1 , further comprising a salt. 
     
     
         18 .- 25 . (canceled) 
     
     
         26 . The alphavirus composition according to  claim 17 , wherein the salt is selected from the group consisting of: sodium chloride, monosodium glutamate (MSG), sodium phosphate, potassium phosphate, potassium chloride, and a combination thereof. 
     
     
         27 .- 30 . (canceled) 
     
     
         31 . A method for decreasing inactivation of a live alphaviruses comprising, combining one or more live alphaviruses with a composition comprising: two or more carbohydrate agents; and one or more amino acids that include histidine, wherein the composition does not include gelatin and the composition decreases inactivation of the live alphavirus. 
     
     
         32 . The method according to  claim 31 , further comprising HEPES buffer wherein the HEPES buffer concentration is 1.0 mM to 200.0 mM. 
     
     
         33 . The method according to  claim 31 , wherein the live alphaviruses comprise chikungunya virus (CHIK), o'nyong'nyong virus, Ross River virus, other Semliki Forest virus complexes, Eastern equine encephalitis and Western equine encephalitis and combinations thereof. 
     
     
         34 . The method according to  claim 31 , further comprising partially or wholly dehydrating the combination. 
     
     
         35 . The method according to  claim 31 , further comprising partially or wholly re-hydrating the composition prior to administration. 
     
     
         36 . (canceled) 
     
     
         37 . The method according to  claim 31 , wherein the HEPES buffer concentration is from 1.0 to 30.0 mM; the two or more carbohydrate agent concentration is from 1.0 to about 100.0 mg/ml each; and the amino acid concentration is from 1.0 to 40.0 mg/ml. 
     
     
         38 . The method according to  claim 32 , wherein the composition comprises HEPES, histidine, sucrose, and mannitol. 
     
     
         39 . (canceled) 
     
     
         40 . The method according to  claim 31 , further comprising a salt agent. 
     
     
         41 . The method according to  claim 31 , wherein the live alphavirus composition is further formulated for use as a medicament for administration to a subject to reduce the onset of or prevent an alphavirus infection. 
     
     
         42 .- 51 . (canceled) 
     
     
         52 . A kit for decreasing inactivation of a live alphaviruses comprising:
 at least one container;   a composition comprising 0.1 mM to 40.0 mM HEPES buffer, two or more carbohydrate agents that include sucrose, and one or more amino acids that include histidine; and   one or more live alphaviruses.   
     
     
         53 . The kit according to  claim 52 , wherein the composition comprises HEPES, histidine, sucrose, and mannitol. 
     
     
         54 . The kit according to  claim 52 , wherein the composition comprises a pharmaceutical composition comprising a pharmaceutically acceptable excipient. 
     
     
         55 . (canceled) 
     
     
         56 . The kit of  claim 52 , further comprising a salt selected from the group consisting of: sodium chloride, monosodium glutamate, sodium phosphate, potassium phosphate, and potassium chloride. 
     
     
         57 . (canceled) 
     
     
         58 . The kit according to  claim 52 , wherein the one or more live alphaviruses comprise chikungunya (CHIK) virus, o'nyong'nyong virus, Ross River virus, Semliki Forest virus complexes, Eastern equine encephalitis and Western equine encephalitis and combinations thereof. 
     
     
         59 . A pharmaceutical composition according to  claim 1 , wherein the alphavirus composition is a live, attenuated alphavirus composition and further comprises a pharmaceutically acceptable excipient. 
     
     
         60 .- 63 . (canceled) 
     
     
         64 . The pharmaceutical composition of  claim 59 , wherein the Alphavirus composition comprises live attenuated Chikungunya virus. 
     
     
         65 . (canceled) 
     
     
         66 . A live attenuated alphavirus composition comprising:
 one or more live alphaviruses;   two or more carbohydrate agents including sucrose and mannitol;   a salt agent; and   
       wherein the composition includes human serum albumin and wherein the composition stabilizes live alphaviruses. 
     
     
         67 . The alphavirus composition of  claim 66 , wherein the salt agent comprises sodium phosphate, sodium chloride, monosodium glutamate, potassium phosphate, potassium chloride, or mixtures thereof. 
     
     
         68 . The alphavirus composition according to  claim 66 , wherein the one or more live alphaviruses comprise Chikungunya virus (CHIK), o'nyong'nyong virus, Ross River virus, Eastern equine encephalitis and Western equine encephalitis, other Semliki Forest virus, or other Togavirus, or combinations thereof. 
     
     
         69 . The alphavirus composition according to  claim 68 , wherein the one or more live alphaviruses are chikungunya (CHIK) viruses. 
     
     
         70 . The alphavirus composition according to  claim 67 , wherein the composition is in aqueous form. 
     
     
         71 . The alphavirus composition according to  claim 67 , wherein the composition is partially or wholly dehydrated. 
     
     
         72 . The alphavirus composition according to  claim 67 , wherein the sucrose concentration is from 1.0% to 15.0% (w/v); mannitol concentration is 15.0 to 40.0 mg/mL; sodium chloride concentration is 0.1% to 1.5% (w/v); monosodium glutamate concentration is from 0.05% to 0.5% (w/v); sodium phosphate concentration is from 0.01 to 0.15% (w/v); potassium phosphate concentration is from 0.01% to 0.1% (w/v); and potassium chloride concentration is from 0.01% to 0.1% (w/v), and the human serum albumin ranges from 0.1% (w/v) to 5%.