TEMPERATURE-RESPONSIVE VIRUS STORAGE SYSTEM
A temperature-responsive virus storage system that allows virus to be stored, such as a non-frozen liquid, and maintain infectivity is described.
1 . A method of treating a subject suffering from bladder cancer, the method comprising administering to the subject a composition comprising an infective replication-deficient adenovirus encoding interferon α-2b;
wherein, prior to administration to the subject, the composition is: (a) stored in a frozen state at a temperature lower or equal to −60° C. for at least one year to preserve the infectivity of the virus; (b) stored in a frozen state at a temperature of about −35° C. to about −10° C. for up to about 3 months; or (c) after thawing to room temperature, stored as a liquid: (i) at a temperature of about 2° C. to about 8° C. for up to 7 days; (ii) at room temperature for up to 24 hours; or (iii) at about 2° C. to about 8° C. for up to 1 day, up to 2 days, up to 3 days, up to 4 days, up to 5 days, up to 6 days, or up to 7 days followed by up to 24 hours at room temperature; or (iv) in a syringe for up to about 6 hours;
wherein the composition comprises the infective virus and a final buffer made by combining one part (w/w) of a buffer formulation with nine parts (w/w) purified water;
further wherein, the infective virus is suspended in the composition at a concentration of about 3×10 11 adenoviral particles per mL of buffer; and
still further wherein, the buffer formulation comprises the following components, in an amount (w/w) of each component expressed as a percent of the weight of tromethamine used to make the buffer:
about 6000% glycerol;
about 1214% sucrose;
about 100% tromethamine;
about 100% sodium dihydrogen phosphate dihydrate;
about 560% hydroxypropyl beta-cyclodextrin;
about 29% magnesium chloride hexahydrate;
about 68% (3α,53,7α,12α)-N-[3-[(4-O-D-galactopyranosy]-D-gluconoyl)aminolpropyl]-3,7,12-trihydroxy-N[3-[[(3α,53,7α,12α)-3,7,12-trihydroxy-24-oxocholan-24-yl]amino]propyl]-cholan-24-amide (NODA)
about 34% polysorbate 80;
about 2.8% sodium citrate dihydrate; and
about 0.71% citric acid monohydrate.
2 . The method of claim 1 , wherein after storage in a frozen state, at a temperature lower or equal to −60° C. for about one year, the particles retain at least about 95% of the initial total particle concentration and at least about 80% of their initial infectious titer as measured as Normalized and Adjusted Standard-Infectious Units (NAS IU).
3 . The method of claim 1 , wherein the temperature is about −60° C. to about −100° C., about −60° C. to about −90° C. or about −60° C. to about −80° C.
4 . The method of claim 1 , wherein the composition is stored in a frozen state for at least 18 months, at least 24 months, or at least 36 months.
5 . The method of claim 1 , wherein prior to administration to the subject, the composition is stored in a frozen state at a temperature of about −25° C. to about −15° C.
6 . The method of claim 1 , wherein the bladder cancer is high-grade non-muscle invasive bladder cancer.
7 . The method of claim 1 , wherein the bladder cancer is carcinoma in situ (CIS) with or without high-grade papillary disease.
8 . The method of claim 1 , wherein the bladder cancer is Bacillus Calmette-Guérin (BCG)-resistant bladder cancer.