Heat stable liquid rotavirus vaccine
The invention is directed to an oral vaccine composed of a micronized freeze-dried rotavirus particle emulsion with buffering excipients in a non-aqueous liquid. This IVT-06 formulation has imparted heat stability by protecting the virus at temperatures of 30° C. and 40° C. for at least twelve months. Extrapolations from the 12-month stability data indicate a shelf life of more than two years at 30° C., and six months at 50° C. In addition, for ease of administration, the formulated dose has a volume of 0.5 mL.
1. A method for the manufacture of a vaccine comprising:
combining a buffering agent, sucrose, and arginine with a virus-containing composition forming a mixture, wherein:
the buffering agent comprises a HEPES buffer;
the virus-containing composition comprises a rotavirus
and the ratio of arginine to sucrose is about 1:1.1 to 1:1.5;
the virus titer of the composition comprises 10 5.9 FFU/mL or higher;
lyophilizing the mixture to form a lyophilized composition containing less than or equal to about 0.8% moisture; and
milling the mixture to an approximate uniform particle size of about 5 μm or less;
wherein the particles comprise the vaccine.
2. The method of claim 1 , wherein the rotavirus-containing composition comprises multiple serotypes of rotavirus.
3. The method of claim 1 , wherein the pH does not significantly change upon lyophilization.
4. The method of claim 1 , wherein the buffering agent maintains a pH of the mixture at about 7.0-8.0.
5. The method of claim 1 , wherein the vaccine further comprises a bulking agent.
6. The method of claim 1 , wherein the mixture comprises approximately 2% arginine.
7. The method of claim 1 , wherein the mixture comprises approximately 4% arginine.
8. The method of claim 1 , wherein the mixture contains medium-chain triglyceride (MCT).
9. The method of claim 1 , wherein the vaccine can be stored at about room temperature or higher for at least about three months without significant loss of efficacy.
10. The method of claim 1 , wherein the vaccine can be stored at about 30° C. or higher for at least six months without significant loss of efficacy.
11. The method of claim 1 , wherein the vaccine can be stored at about 30° C. or higher for at least twelve months without significant loss of efficacy.
12. The method of claim 1 , wherein the vaccine is apportioned to individual doses containing virus at about 10 8 FFU/mL and a volume of about 0.5 mL.
13. A vaccine manufactured according to claim 1 .
14. A formulation comprised of a composition lyophilized with a HEPES buffer containing an attenuated rotavirus strain, a bulking agent containing glucose, sucrose or both glucose and sucrose, and an excipient containing at least arginine, wherein the ratio of arginine to bulking agent is about 1:1.1 to 1:1.5, which has a thermo-stability at about 30° C. for at least about 2 years and a thermo-stability at about 50° C. for at least about 3 months.
15. The formulation of claim 14 , which has a uniform particle size of less than or equal to about 5 μm.
16. The formulation of claim 14 , which is homogenized with medium-chain triglyceride (MCT).
17. The formulation of claim 14 , which contains multiple doses of virus at about 10 S9 FFU/mL or higher in a volume of about 0.5 mL.
18. The formulation of claim 14 , which contains citrate and calcium carbonate, Tris, HEPES, phosphates and/or histidine as a formulation buffering agent.
19. A method for the manufacture of a vaccine comprising:
combining a buffering agent, sucrose, and an excipient containing at least arginine with a virus-containing composition forming a mixture, wherein:
the virus-containing composition comprises a rotavirus
the ratio of arginine to sucrose is about 1:1.1 to 1:1.5,
the buffering agent comprises a HEPES buffer; and
the virus titer of the composition comprises 10 8 FFU/mL or higher;
lyophilizing the mixture to form a lyophilized composition containing less than or equal to about 0.8% moisture;
milling the mixture to an approximate uniform particle size of about 5 μm or less; and
homogenizing the particles with medium-chain triglyceride (MCT) to form the vaccine.
20. The method of claim 19 , wherein the vaccine is thermo-stability at about 30° C. for at least about 2 years without significant loss of efficacy.
21. The method of claim 19 , wherein the mixture comprises from approximately 2% to 6% arginine.
22. The composition of claim 19 , wherein the mixture comprises approximately 7.5% sucrose.
23. The composition of claim 19 , wherein the mixture comprises approximately 6% arginine and approximately 7.5% sucrose.
24. A composition of particles lyophilized with a HEPES buffering agent containing:
one or more attenuated strains of a rotavirus;
sucrose;
an excipient containing at least arginine;
medium-chain triglycerides; and
a buffering agent, wherein
the particles are about 5 μm or less in size,
the composition contains less than about 0.8% moisture, and
the ratio of arginine to sucrose is from about 1:1.1 to about 1:1.5.
25. The composition of claim 23 , wherein the mixture comprises from approximately 2% to 6% arginine.
26. The composition of claim 23 , wherein the mixture comprises approximately 7.5% sucrose.
27. The composition of claim 23 , wherein the mixture comprises approximately 6% arginine and approximately 7.5% sucrose.
28. The composition of claim 23 , which has a thermo-stability at about 30° C. for at least about 2 years without significant loss of efficacy.
29. The composition of claim 23 , which has a thermo-stability at about 50° C. for at least about 3 months without significant loss of efficacy.