Adenovirus formulations
View Patent ↗The invention relates to viral formulations and related pharmaceutical products for use in gene therapy and/or vaccine applications. Especially preferred viral formulations disclosed herein are liquid adenovirus formulations, which show improved stability when stored in about the 2-8° C. range while also being compatible with parenteral administration. These formulations comprise a buffer, a sugar, a salt, a divalent cation, a non-ionic detergent, as well as a free radical scavenger and/or a chelating agent to inhibit free radical oxidation.
1. A virus formulation, comprising:
a) a purified adenovirus;
b) at least one inhibitor of free radical oxidation selected from the group consisting of ethanol, EDTA, histidine, triethanolamine, and sodium citrate; and,
c) at least one sugar at a combined weight to volume percentage from about 7.5% to about 25% w/v and optionally a salt up to a combined concentration of about 100 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L,
wherein the formulation is buffered at a pH from about 6.0 to 7.0.
2. A virus formulation of claim 1 which further comprises a non-ionic detergent selected from the group consisting of Polysorbate-80, Polysorbate-40 and Polysorbate-20 at a concentration range from about 0.001% to about 2%.
3. A virus formulation of claim 1 which further comprises a non-ionic detergent selected from the group consisting of Polysorbate-80, Polysorbate-40 and Polysorbate-20 at a concentration range from about 0.00 1% to about 2% and a divalent cation selected from the group consisting of MgCl 2 and CaCl 2 in an amount from about 0.1 mM to about 5 mM.
4. A virus formulation of claim 1 , wherein said formulation has an adenovirus concentration in the range from about 1×10 7 vp/mL to about 1×10 13 vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L, which is buffered by histidine, and which further comprises sucrose in a weight to volume range from greater than 5% to about 25% (w/v), MgCl 2 at about 1 mM to 2 mM, either Polysorbate-80, Polysorbate-40 or Polysorbate-20 at a concentration from about 0.001% to about 2%, EDTA from about 1 μM to about 1000 μM and ethanol from about 0.1 to 1.0% (w/v).
5. An adenovirus formulation of claim 4 selected from the group consisting of formulation number A423, A424, A428, and A429.
6. A method of preserving a subgroup B adenovirus which comprises generating a formulation of claim 3 .
7. A method of preserving a subgroup B adenovirus which comprises generating a formulation of claim 4 .
8. A method of preserving a subgroup B adenovirus which comprises generating a formulation of claim 5 .
9. A virus formulation of claim 3 , wherein said formulation has an adenovirus concentration in the range from about 1×10 7 vp/mL to about 1×10 13 vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L, which is buffered by histidine, and which further comprises sucrose in a weight to volume range from greater than 5% to about 10% (w/v), NaCl up to about 50 mM, MgCl 2 at about 1 mM to 2 mM, either Polysorbate-80, Polysorbate-40 or Polysorbate-20 at a concentration from about 0.001% to about 2%, EDTA from about 1 μM to about 1000 μM and ethanol from about 0.1 to 1.0% (w/v).
10. An adenovirus formulation of claim 9 having 10 mM histidine, pH 6.6, 35 mM NaCl, 7.5% (w/v) sucrose, 1 mM MgCl 2 , 0.1% (w/v) PS-80, 0.1 mM EDTA, and 0.5% ethanol (v/v).
11. A method of preserving a subgroup B, C or D adenovirus which comprises generating a formulation of claim 9 .
12. A method of preserving a subgroup B, C or D adenovirus which comprises generating a formulation of claim 10 .
13. A virus formulation of claim 3 having an adenovirus concentration in the range from about 1×10 7 vp/ml to about 1×10 13 vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L, which is buffered with Tris, further comprising sucrose in a weight to volume range from greater than 5% to about 25% (w/v), NaCl up to about 50 mM, MgCl 2 at about 1 mM to 2 mM, either Polysorbate-80, Polysorbate-40 or Polysorbate-20 at a concentration from about 0.001% to about 2%, EDTA from about 1 μM to about 1000 μM, ethanol from about 0.1 to 1.0% (w/v) and histidine from about 5 mM to about 10 mM.
14. A virus formulation of claim 13 which further comprises mannitol at a concentration up to about 250 mM.
15. A virus formulation of claim 14 which is formulation A442.
16. A method of preserving a subgroup D adenovirus which comprises generating a formulation of claim 14 .
17. A method of preserving a subgroup D adenovirus which comprises generating a formulation of claim 15 .
18. A virus formulation of claim 13 which further comprises sorbitol at a concentration up to about 250 mM.
19. A virus formulation of claim 18 which is formulation A443.
20. A method of preserving a subgroup D adenovirus which comprises generating a formulation of claim 18 .
21. A method of preserving a subgroup D adenovirus which comprises generating a formulation of claim 19 .
22. A virus formulation of claim 13 which further comprises triethanolamine at a concentration up to about 5 mM.
23. A virus formulation of claim 22 which is formulation A448.
24. A method of preserving a subgroup D adenovirus which comprises generating a formulation of claim 22 .
25. A method of preserving a subgroup D adenovirus which comprises generating a formulation of claim 23 .
26. A method of preserving adenovirus preparations representing serotype B, C and/or D, comprising:
a) purifying adenovirus preparations separately from a subgroup B, C and/or D adenovirus;
b) adding at least one inhibitor of free radical oxidation selected from the group consisting of ethanol, EDTA, an EDTA/ethanol combination, histidine, triethanolamine, and sodium citrate;
c) adding sucrose at a weight to volume percentage up to about 10% and sodium chloride up to about 50 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L; and,
d) adding MgCl 2 at about 1 mM to 2 mM, Polysorbate-80, Polysorbate-40 or Polysorbate-20 at about 0.1%, EDTA at about 100 μM, ethanol at about 0.5% (w/v), wherein the formulation is buffered by histidine at a p1-1 of about 6.0 to 7.0.