TEMPERATURE-RESPONSIVE VIRUS STORAGE SYSTEM
A temperature-responsive vims storage system that allows vims to be stored, such as a non-frozen liquid, and maintain infectivity is described.
1 . A composition comprising infectious viral particles, tromethamine and cyclodextrin, wherein the composition comprises about 1×10 9 to about 1×10 12 cyclodextrin molecules per viral particle.
2 . A composition comprising infectious viral particles, cyclodextrin, tromethamine, and sodium phosphate, wherein the composition comprises about 1 to about 1.5 moles of tromethamine per mole of sodium phosphate.
3 . The composition of claim 2 , wherein the composition comprises about 1×10 9 to about 1×10 12 cyclodextrin molecules per viral particle.
4 . The composition of any one of claims 1 - 3 , further comprising a cryoprotective-effective amount of glycerol, sucrose, or both.
5 . The composition of claim 4 , wherein the composition comprises glycerol in a relative amount of about 600 times the amount of tromethamine (w/w), and the composition comprises sucrose in a relative amount of about 120 times the amount of tromethamine (w/w).
6 . The composition of any one of claims 1 - 5 , wherein the cyclodextrin is hydroxypropyl beta-cyclodextrin.
7 . The composition of any one of claims 1 - 6 , wherein the composition comprises hydroxypropyl beta-cyclodextrin in a relative amount of about 6 times the amount of tromethamine (w/w).
8 . The composition of any one of claims 1 - 6 , further comprising (3α,5β, 7α,12α)-N-[3-[(4-O-D-galactopyranosyl-D-gluconoyl)amino]propyl]-3,7,12-trihydroxy-N-[3-[[(3α,5β, 7α, 12α)-3,7,12-trihydroxy-24-oxocholan-24-yl]amino] propyl]-cholan-24-amide (NODA) in a relative amount of about 0.7 times the amount of tromethamine (w/w).
9 . The composition of any one of claims 2 - 8 , wherein the sodium phosphate is sodium dihydrogen phosphate dihydrate.
10 . The composition of any one of claims 1 - 9 , further comprising magnesium chloride, polysorbate 80, sodium citrate, and citric acid.
11 . The composition of any one of claims 1 - 10 , wherein the virus is present in an amount of about 1×10 11 viral particles per milliliter of composition.
12 . The composition of any one of claims 1 - 11 , wherein the composition has a first pH at a first temperature, and a second pH at a second temperature, wherein the first temperature is lower than the second temperature, and the first pH is higher than the second pH.
13 . The composition of claim 12 , wherein the first temperature is about −20° C., and the first pH is a basic pH.
14 . The composition of claim 12 or 13 , wherein the second temperature is about 20° C. to about 25° C., and the second pH is an acidic pH.
15 . The composition of any one of claims 1 - 14 , wherein after storage as a non-frozen liquid, or in a frozen state, at −20° C. for about one year, the viral particles retain at least about 95% of the initial total viral particle concentration and at least about 80% of their initial infectious titer measured as Normalized and Adjusted Standard—Infectious Units (NAS IU).
16 . The composition of any one of claims 1 - 15 , wherein the infectious virus is a lentivirus, adenovirus or adeno-associated virus.
17 . The composition of any one of claims 1 - 16 , wherein the infectious virus is a replication-deficient adenovirus.
18 . A composition comprising sodium dihydrogen phosphate dehydrate, tromethamine, glycerol, sucrose, hydroxypropyl beta-cyclodextrin, NODA, and infectious replication-deficient adenovirus, wherein the composition comprises:
tromethamine in a relative amount of from about 1 to about 1.5 moles of tromethamine per mole of sodium dihydrogen phosphate dehydrate;
glycerol in a relative amount of about 600 times the amount of tromethamine (w/w);
sucrose in a relative amount of about 120 times the amount of tromethamine (w/w);
hydroxypropyl beta-cyclodextrin in a relative amount of about 6 times the amount of tromethamine (w/w);
NODA in a relative amount of about 0.7 times the amount of tromethamine (w/w); and
about 1×10 11 replication-deficient adenovirus particles per milliliter of composition.
19 . A composition comprising infectious viral particles, tromethamine and cyclodextrin, the cyclodextrin present in a relative amount of from about 1×10 9 to about 1×10 12 cyclodextrin molecules per viral particle, the tromethamine able to change pH in response to change in temperature, the tromethamine present in an amount whereby if the composition is stored in a liquid, non-frozen state, or at a frozen state, at −20° C. for one year, the viral particles retain at least about 95% of the initial total viral particle concentration and at least about 80% of their initial infectious titer measured as NAS IU.
20 . The composition of claim 19 , further comprising sodium phosphate present in a relative amount of from about 1 to about 1.5 moles of tromethamine per mole of sodium phosphate.
21 . The composition of claim 20 , the sodium phosphate is sodium dihydrogen phosphate dehydrate.
22 . The composition of any one of claims 19 - 21 , further comprising a cryoprotective-effective amount of glycerol, sucrose, or both.
23 . The composition of claim 22 , wherein the composition comprises glycerol in a relative amount of about 600 times the amount of tromethamine (w/w), and the composition comprises sucrose in a relative amount of about 120 times the amount of tromethamine (w/w).
24 . The composition of any one of claims 19 - 25 , wherein the cyclodextrin is hydroxypropyl beta-cyclodextrin.
25 . The composition of claim 24 , wherein the composition comprises hydroxypropyl beta-cyclodextrin in a relative amount of about 6 times the amount of tromethamine (w/w).
26 . The composition of any one of claims 19 - 25 , wherein the infectious virus is a lentivirus, adenovirus or adeno-associated virus.
27 . The composition of any one of claims 19 - 26 , wherein the infectious virus is a replication-deficient adenovirus.
28 . The composition of any one of claims 19 - 27 , further comprising NODA in a relative amount of about 0.7 times the amount of tromethamine (w/w), and wherein the virus is present in an amount of about 1×10 11 viral particles per milliliter of composition.
29 . The composition of claim 19 , further comprising sodium dihydrogen phosphate dehydrate present in a relative amount of from about 1 to about 1.5 moles of tromethamine per mole of sodium dihydrogen phosphate dehydrate, and further comprising glycerol and sucrose, the glycerol present in a relative amount of about 600 times the amount of tromethamine (w/w) and the sucrose present in a relative amount of about 120 times the amount of tromethamine (w/w), wherein the cyclodextrin comprises hydroxypropyl beta-cyclodextrin in a relative amount of about 6 times the amount of tromethamine (w/w), wherein the infectious virus comprises replication-deficient adenovirus, and further comprising NODA in a relative amount of about one times the amount of tromethamine (w/w), where the virus is present in an amount of about 1×10 11 viral particles per milliliter of composition.
30 . A method of preserving level of infectivity of an infective virus, the method comprising storing the composition of any one of claims 1 - 29 in a liquid, non-frozen state, or in a frozen state, at −20° C. for at least one year.
31 . The method of claim 30 , wherein the viral particles retain at least about 95% of the initial total viral particle concentration and at least about 80% of their initial infectious titer measured as NAS IU.
32 . A method of treating a subject suffering from cancer, the method comprising administering to the subject the composition of any one of claims 1 - 29 , wherein the viral particles are recombinant adenoviral particles encoding human interferon α-2b.