Compositions and methods for stabilization of active agents

Abstract

Provided herein are methods and compositions for stabilization of active agents. The active agents are distributed, mixed or embedded in a silk fibroin matrix, thereby retaining the bioactivity of the active agents upon storage and/or transportation. In some embodiments, the storage-stable vaccine-silk compositions are also provided herein.

1 .- 238 . (canceled) 
     
     
         239 . A storage-stable composition comprising a silk fibroin matrix and an active agent distributed therein, wherein the active agent is an immunogen selected from the group consisting of a live, attenuated virus; an inactivated virus; a viral subunit; a viral vector; a live, attenuated bacterium; a killed bacterium; an inactivated bacterium; a bacterial subunit; and an inactivated toxin from a pathogen; and the active agent retains at least about 30% of its original bioactivity when the composition is (a) subjected to at least one freeze-thaw cycle, or (b) maintained for at least about 24 hours at a temperature above 0° C., or (c) both (a) and (b). 
     
     
         240 . The composition of  claim 239 , wherein the silk fibroin matrix is a solution. 
     
     
         241 . The composition of  claim 239 , wherein the silk fibroin matrix is a solid-state form. 
     
     
         242 . The composition of  claim 241 , wherein the solid-state form is a film, a fiber, a particle, a gel, a hydrogel, or a composite thereof. 
     
     
         243 . The composition of  claim 239 , wherein the composition is lyophilized. 
     
     
         244 . The composition of  claim 239 , wherein the composition is micronized. 
     
     
         245 . The composition of  claim 244 , wherein the micronized composition comprises nanoparticles or microparticles. 
     
     
         246 . The composition of  claim 245 , wherein the nanoparticles or microparticles have a size of about 10 nm to about 1000 μm. 
     
     
         247 . The composition of  claim 239 , further comprising an additive distributed in the silk fibroin matrix. 
     
     
         248 . The composition of  claim 247 , wherein the additive is selected from a stabilizing agent, a pharmaceutically acceptable carrier, or any combinations thereof. 
     
     
         249 . The composition of  claim 248 , wherein the stabilizing agent is selected from the group consisting of a saccharide, a sugar alcohol, an ion, a surfactant, and any combinations thereof. 
     
     
         250 . The composition of  claim 249 , wherein the saccharide is sucrose. 
     
     
         251 . The composition of  claim 239 , wherein the immunogen is derived from hepatitis B virus,  Haemophilus influenzae  Type B, poliovirus,  Neisseria meningitides  C, influenza,  Varicella , or  Mycobacteria tuberculosis  bacille Calmette-Guerin, tetanus toxoid, diphtheria toxoid, and  Bordetella pertussis.    
     
     
         252 . The composition of  claim 239 , wherein the immunogen is a combination immunogen selected from the group consisting of DTaP, DTwP, DTwP hepB, DTP hep B Hib, DTaP hep B Hib IPV, and any combinations thereof. 
     
     
         253 . The composition of  claim 239 , wherein the immunogen is a live, attenuated virus. 
     
     
         254 . The composition of  claim 253 , wherein the live, attenuated virus is an enveloped virus. 
     
     
         255 . The composition of  claim 254 , wherein the enveloped virus is selected from the group consisting of Paramyxoviridae, Togaviridae, Orthomyxoviridae, Flaviviridae, Herpesviridae, Rhabdovirus, Retroviridae, and any combinations thereof. 
     
     
         256 . The composition of  claim 253 , wherein the virus is  varicella.    
     
     
         257 . The composition of  claim 253 , wherein the virus is influenza. 
     
     
         258 . The composition of  claim 253 , wherein the live, attenuated virus causes measles, mumps, or rubella. 
     
     
         259 . The composition of  claim 239 , wherein the live, attenuated virus or inactivated virus is an enveloped virus selected from the group consisting of  varicella , measles virus, mumps virus, German measles virus, respiratory syncytial virus, yellow fever virus, and influenza virus. 
     
     
         260 . The composition of  claim 239 , wherein the immunogen is a live, attenuated, non-enveloped virus. 
     
     
         261 . The composition of  claim 260 , wherein the non-enveloped virus is rotavirus, reovirus, hepatitis virus, rabies virus or poliovirus. 
     
     
         262 . The composition of  claim 239 , wherein the immunogen is a live, attenuated bacterium; a killed bacterium; or an inactivated bacterium. 
     
     
         263 . The composition of  claim 262 , wherein the bacterium is  Mycobacteria tuberculosis  bacilli Calmette-Guerin or  Bordetella pertussis.    
     
     
         264 . The composition of  claim 239 , wherein the immunogen is a bacterial subunit. 
     
     
         265 . The composition of  claim 264 , wherein the bacterial subunit is derived from  Neisseria meningitides  type C,  Haemophilus influenzae  type B,  Streptococcus pneumoniae , or Group B  streptococcus.    
     
     
         266 . The composition of  claim 264 , wherein the bacterial subunit is a polysaccharide. 
     
     
         267 . The composition of  claim 239 , wherein the immunogen is a viral subunit. 
     
     
         268 . The composition of  claim 267 , wherein the viral subunit is derived from Hepatitis B virus or Human Papillomavirus. 
     
     
         269 . The composition of  claim 239 , wherein the immunogen is recombinant. 
     
     
         270 . The composition of  claim 239 , wherein the immunogen is a vaccine product selected from the group consisting of Anthrax vaccine (BioThrax); BCG (Bacillus Calmette-Guerin) (Tice, Mycobax); DTaP (Daptacel); DTaP (Infanrix); DTaP (Tripedia); DTaP/Hib (TriHIBit); DTaP-IPV (Kinrix); DTaP-HepB-IPV (Pediarix); DtaP-IPV/Hib (Pentacel); DT (diphtheria vaccine plus tetanus vaccine) (Sanofi); Hib vaccine (ACTHib); DT (Massachusetts); Hib (PedvaxHib); Hib/Hep B (Comvax); Hep A (Havrix), Hepatitis A vaccine; Hep A (Vaqta), Hepatitis A vaccine; Hep B (Engerix-B), Hepatitis B vaccine; Hep B (Recombivax), Hepatitis B vaccine; HepA/HepB vaccine (Twinrix); Human Papillomavirus (HPV) (Gardasil); Influenza vaccine (Afluria); Influenza vaccine (Fluarix); Influenza vaccine (Flulaval); Influenza vaccine (Fluvirin); Influenza vaccine (Fluzone); Influenza vaccine (FluMist); IPV (Ipol), Polio vaccine; Japanese encephalitis vaccine (JE-Vax); Japanese encephalitis vaccine (Ixiaro); Meningococcal vaccine (Menactra); MMR vaccine (MMR-11); MMRV vaccine (ProQuad); Pneumococcal vaccine (Pneumovax); Pneumococcal vaccine (Prevnar); Poliovirus inactivated (Poliovax), Polio vaccine; Rabies vaccine (Imovax); Rabies vaccine (RabAvert); Rotavirus vaccine (RotaTeq); Rotavirus vaccine (Rotarix); Td vaccine (Decavac); Td vaccine (Massachusetts); Tdap vaccine (Adacel); Tdap vaccine (Boostrix); Typhoid (inactivated—Typhim Vi), Typhus vaccine; Typhoid (oralTy21a), Typhus vaccine; Vaccinia (ACAM2000);  Varicella  vaccine (Varivax); Yellow fever vaccine (YF-Vax); Zoster vaccine (Zostavax); and any combinations thereof. 
     
     
         271 . A method for preparing a storage-stable composition of  claim 241 , the method comprising the steps of:
 a. providing a silk fibroin solution comprising at least one active agent; and   b. drying the silk fibroin solution of step (a) to form a solid-state silk fibroin,   
       thereby obtaining a composition in which the at least one active agent retains at least about 30% of its original bioactivity upon storage. 
     
     
         272 . The method of  claim 271 , further comprising lyophilizing the solid-state silk fibroin from step (b). 
     
     
         273 . The method of  claim 271 , further comprising post-treatment of the composition. 
     
     
         274 . The method of  claim 273 , wherein the post-treatment alters the crystallinity of the composition. 
     
     
         275 . The method of  claim 273 , wherein the post-treatment comprises contacting the composition with methanol or ethanol, subjecting the composition to shear stress, subjecting the composition to an electric field, subjecting the composition to pressure, or contacting the composition with salt. 
     
     
         276 . The method of  claim 271 , further comprising reducing the solid-state silk fibroin of step (b) by a mechanical means to obtain micronized particles. 
     
     
         277 . A method of stabilizing an immunogen comprising mixing an immunogen and a silk fibroin solution, wherein the immunogen is selected from the group consisting of a live, attenuated virus; an inactivated virus; a viral subunit; a viral vector; a live, attenuated bacterium; a killed bacterium; an inactivated bacterium; a bacterial subunit; and an inactivated toxin from a pathogen. 
     
     
         278 . The method of  claim 277 , further comprising forming a solid-state form from the silk fibroin solution. 
     
     
         279 . The method of  claim 278 , wherein the solid-state form is selected from the group consisting of a film, a fiber, a hydrogel, a scaffold, a mat, a particle, a lyophilized matrix, a needle, and a composite thereof.