Rotavirus vaccine formulations

The present invention provides liquid and lyophilized formulations of vaccines against rotavirus infection and methods of their preparation. The formulations include buffering agents appropriate for oral administration of rotavirus vaccines. The formulations also include compounds to stabilize of the vaccine compositions against loss of potency.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG.

1. Effect of buffer combinations on rotavirus stability at 37° C. for 1 week. Data for the G1 reassortant are shown in panel A and the P1 reassortant in panel B. All values are expressed as pfu/mL normalized to the reference, or 0 day, sample. The buffer combinations are represented as follows: 0.05 M sodium citrate +0.15 M sodium bicarbonate (□), 0.05 M sodium citrate+0.15 M sodium phosphate (∘), 0.05 M lactic acid+0.15 M sodium bicarbonate (), 0.05 M lactic acid+0.15 M sodium phosphate (∇) and 0.20 M sodium succinate+0.05 M sodium phosphate (⋄). All formulations have pH values of 7.

FIG.

2. Acid neutralizing ability of formulation buffers compared to bicarbonate. One mL of each buffer was titrated with 0.01 N HCl. Symbols: 0.4 M sodium bicarbonate (•), 0.1 M sodium citrate+0.3 M sodium phosphate (∘), 0.1 M sodium citrate+0.3 M sodium bicarbonate (+), and 0.2 M sodium succinate+0.1 M sodium phosphate (⋄).

FIG.

3. Stability data for reassortant rotavirus in liquid formulations of 5% sucrose/0.1 M sodium succinate/0.05 M sodium phosphate after storage at various temperatures. Data for G1 rotavirus is shown in panel A and for P1 rotavirus in panel B. Titers are shown per milliliter.

FIG.

4. Stability data for reassortant rotavirus in liquid formulations of 50% sucrose/0.1 M sodium succinate/0.05 M sodium phosphate after storage at various temperatures. Data for G1 rotavirus is shown in panel A and for P1 rotavirus in panel B. Titers are shown per milliliter.

FIG.

5. Stability data for G1 rotavirus liquid formulations with higher buffer, sucrose, and hydrolyzed gelatin concentrations at various temperatures. Panel A shows data for G1 rotavirus in Williams' E media (“WE”), 50% sucrose, 0.2 M sodium succinate, and 0.1 M sodium phosphate. Stability data for vaccine in Williams' E media, 70% sucrose, 0.2 M sodium succinate, and 0.1 M sodium phosphate is shown in panel B. Panel C shows data for G1 rotavirus in 50% sucrose, 0.1 M sodium citrate, and 0.3 M sodium phosphate; panel D shows data for G1 rotavirus in Williams' E media, 50% sucrose, 0.2 M sodium succinate, 0.1 M sodium phosphate, and 5% hydrolyzed gelatin. Titers are shown per milliliter. The 4° C. data points are obscured by the −70° C. and 15° C. data points.

FIG.

6. Stability data for P1 rotavirus liquid formulations with higher buffer, sucrose, and hydrolyzed gelatin concentrations at various temperatures. Panel A shows data for P1 rotavirus in Williams' E media, 50% sucrose, 0.2 M sodium succinate, and 0.1 M sodium phosphate. Stability data for vaccine in Williams' E media, 70% sucrose, 0.2 M sodium succinate, and 0.1 M sodium phosphate is shown in panel B. Panel C shows data for P1 rotavirus in 50% sucrose, 0.1 M sodium citrate, and 0.3 M sodium phosphate; panel D shows data for P1 rotavirus in Williams' E media, 50% sucrose, 0.2 M sodium succinate, 0.1 M sodium phosphate, and 5% hydrolyzed gelatin. Titers are shown per milliliter.

FIG.

7. Stability data for rotavirus liquid formulations in 50% sucrose, 0.1 M sodium succinate, and 0.05 M sodium phosphate after storage at various temperatures. Data for G2 rotavirus is shown in panel A and for G3 in panel B. Titers are shown per milliliter.

FIG.

8. Stability data for G1 rotavirus lyophilized formulations after storage at various temperatures. Panel A shows data for G1 rotavirus dialyzed prior to lyophilization into 1% sucrose, 4% mannitol, and 10 mM sodium phosphate. Stability data for vaccine dialyzed prior to lyophilization into 1% lactose, 4% mannitol, and 10 mM sodium phosphate is shown in panel B. Panel C shows data for G1 rotavirus diluted phosphate prior to lyophilization into 1% sucrose, 4% mannitol, and 75 mM sodium phosphate. Titers are shown per milliliter.

FIG.

9. Stability data for P1 rotavirus lyophilized formulations after storage at various temperatures. Panel A shows data for P1 rotavirus dialyzed prior to lyophilization into 1% sucrose, 4% mannitol, and 10 mM sodium phosphate. Stability data for vaccine dialyzed prior to lyophilization into 1% lactose, 4% mannitol, and 10 mM sodium phosphate is shown in panel B. Panel C shows data for P1 rotavirus diluted prior to lyophilization into 1% sucrose, 4% mannitol, and 75 mM sodium phosphate. Titers are shown per milliliter.

FIG.

10. Stabilizing effect of recombinant human albumin (rHA) on the stability of G1 rotavirus in liquid formulation at 30° C. Potency change values are expressed as log pfu compared to −70° C. samples. The stabilizer is 50% sucrose/0.1 M phosphate/0.2 M citrate/tissue culture medium/pH 6.2 in the presence or absence of rHA.

1. A liquid rotavirus vaccine formulation comprising:

at least one component selected from the group consisting of

2. The formulation of claim 1 wherein said at least one carboxylate is selected from the group consisting of succinate, citrate, fumarate, tartarate, maleate and lactate.

3. The formulation according to claim 1 wherein said sugar is selected from the group consisting of sucrose, mannitol, lactose, sorbitol, dextrose, fucose, trehalose, polyaspartic acid, inositol hexaphosphate (phytic acid), sialic acid or N-acetyineuraminic acid-lactose.

4. The liquid vaccine formulation of claim 1 further comprising: g)at least one diluent selected from the group consisting of Tissueculture medium, saline and water to volume.

5. The formulation of claim 1 wherein the concentration of sugar is between about to about 70%; the concentration of phosphate is between about 0.05 to about 0.3 M; and said at least one carboxylic acid is citrate or succinate at a concentration between about 0.05 to about 0.7 M.

6. The formulation according to claim 1 wherein the pH is between about pH 5.0 to about pH 8.0.

7. The formulation according to claim 1 wherein said phosphate is selected from the group consisting of monophosphates, polyphosphates and phosphorylated compounds.

8. The formulation according to claim 7 wherein said phosphorylated compounds are phosphorylated sugars.

9. The formulation according to claim 1 wherein the non ionic surfactant is selected from the group consisting of polysorbates, polyoxyethylene alkyl ethers, nonaethylene glycol octylphenol ethers, hepatethylene glycol octylphenyl ethers, sorbitan trioleates, and polyoxyethylene-polyoxypropylene block copolymers.

10. The formulation according to claim 1 wherein the concentration of surfactant is from about 0.005% to about 0.1%.

11. The rotavirus vaccine formulation of claim 1 wherein the non ionic surfactant is a polysorbate.

12. The rotavirus vaccine formulation of claim 11 wherein the non ionic surfactant is polysorbate 80.

13. A method of preparing rotavirus vaccine formulations, comprising:

14. The method of claim 13 further comprising the step of lyophilizing the vaccine solution.

15. A method of administering an oral rotavirus vaccine formulation to an individual comprising treatment of the individual with a compound having sufficient buffering capacity to neutralize stomach acid, wherein the treatment is before, coincident with or subsequent to the administration of the vaccine;

16. The method of claim 15 wherein the non ionic surfactant is a polysorbate.

17. The method of claim 16 wherein the non ionic surfactant is polysorbate 80.