Inactivated dengue virus vaccine

• US 6,254,873 B1
• Filed: 04/17/1995
• Issued: 07/03/2001
• Est. Priority Date: 04/17/1995
• Status: Expired due to Term

• Current Assignee: United States Department of the Army
• Original Assignee: The United States Of America As Represented By The Secretary Of The Army
• Inventors: Dubois, Doris R., Eckels, Kenneth, Putnak, J. Robert
• Primary Examiner(s): Housel, James
• Assistant Examiner(s): Bui, Phuong T.
• Application Number:
  US08/423,338
• Time in Patent Office: 2,269 Days
• Field of Search: 424/218.1, 424/202.1, 536/23.72, 530/300, 530/350, 435/236, 435/238
• US Class Current: 424/218.1
• CPC Class Codes: A61K 2039/5252 inactivated (killed)
• CPC Class Codes: A61K 2039/55505 Inorganic adjuvants
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: C12N 2770/24134 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2770/24161 Methods of inactivation or attenuation
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

An inactivated dengue virus vaccine to immunize and protect humans against dengue fever is described. The vaccine is based on dengue viruses which have been propagated to high titers in suitable cells, purified and inactivated under conditions which destroy infectivity but preserve immunogenicity, a high level of which is demonstrated in animal models.

Claims

1. A vaccine comprising a purified, inactivated dengue virus effective for inducing a protective immune response in primates against dengue virus.

2. The purified, inactivated dengue virus vaccine of claim 1 comprising a first inactivated dengue virus, wherein said inactivated dengue virus is selected from the group consisting of dengue virus type 1, dengue virus type 2, dengue virus type 3, and dengue virus type 4.

3. The purified, inactivated dengue virus vaccine of claim 2, wherein said dengue virus vaccine further comprises of one or more purified inactivated dengue virus serotype(s) different from the first inactivated dengue virus, wherein said purified, inactivated dengue virus is selected from the group consisting of dengue virus type 1, dengue virus type 2, dengue virus type 3 and dengue virus type 4.

4. A method for producing antibodies against dengue virus comprising administering to an individual a purified, inactivated dengue virus vaccine according to claim 1 in an amount sufficient to produce said antibodies wherein said vaccine is chosen from the group consisting of dengue virus type 1 vaccine, dengue virus type 2 vaccine, dengue virus type 3 vaccine and dengue virus type 4 vaccine.

5. A method for producing the purified, inactivated dengue virus vaccine of claim 1 comprising the steps of:

(i) propagating dengue virus in a cell culture;

(ii) harvesting said virus from said cell culture;

(iii) concentrating said virus;

(iv) purifying said virus such that it is essentially free of cell culture proteins and DNA;

(v) inactivating said virus; and

(vi) adding a suitable adjuvant in a pharmaceutically acceptable amount.

6. A method for producing a purified, inactivated dengue virus vaccine according to claim 5, further comprising the step of removing serum protein supplements prior to harvesting said virus according to step (ii).

7. The purified inactivated dengue virus vaccine of claim 1 wherein said inactivated dengue virus is chemically inactivated.

8. The purified, inactivated dengue virus vaccine of claim 7 comprising a first inactivated dengue virus, wherein said first inactivated dengue virus is selected from the group consisting of dengue virus type 1, dengue virus type 2, dengue virus type 3, and dengue virus type 4.

9. The purified, inactivated dengue virus vaccine of claim 8, wherein said dengue virus vaccine further comprises an additional one or more purified inactivated dengue virus serotype(s) different from said first inactivated dengue virus, wherein said additional inactivated dengue virus is selected from the group consisting of dengue virus type 1, dengue virus type 2, dengue virus type 3, and dengue virus type 4.

10. The purified, inactivated dengue virus vaccine according to claim 1 wherein the specific activity of said vaccine is at least about 1×

10⁸ plaque forming units per milligram of total protein.

11. The purified inactivated dengue virus vaccine of claim 1, wherein said dengue virus vaccine is dengue 1 virus vaccine produced from deposited dengue 1 virus having ATCC accession no. VR-2649.

12. The purified inactivated dengue virus vaccine of claim 1, wherein said dengue virus vaccine is dengue 2 virus vaccine produced from deposited dengue 2 virus having ATCC accession no. VR-2650.

13. The purified inactivated dengue virus vaccine of claim 1, wherein said dengue virus vaccine is dengue 3 virus vaccine produced from deposited dengue 3 virus having ATCC accession no. VR-2654.

14. The purified inactivated dengue virus vaccine of claim 1, wherein said dengue virus vaccine is dengue 4 virus vaccine produced from deposited dengue 4 virus having ATCC accession no. VR-2651.

15. A method for inducing an immune response in an individual against dengue virus comprising administering to an individual a purified, inactivated dengue virus vaccine according to claim 1 in an amount sufficient to produce said immune response wherein said vaccine is chosen from the group consisting essentially of dengue virus type 1 vaccine, dengue virus type 2 vaccine, dengue virus type 3 vaccine and dengue virus type 4 vaccine or any combination thereof.

16. A dengue 2 virus vaccine produced by inactivating a dengue 2 virus deposited at ATCC under accession no. VR2650.

17. A multivalent vaccine comprising purified inactivated dengue virus propagated in vertebrate tissue culture cells and effective for inducing a protective immune response in primates against dengue virus wherein said virus is chosen from the group consisting of purified inactivated dengue virus type 1, purified inactivated dengue virus type 2, purified inactivated dengue virus type 3, and purified inactivated dengue virus type 4.

18. An isolated dengue type 1 virus having ATCC accession number VR-2649.

19. An isolated dengue type 1 virus vaccine produced by inactivating a dengue 1 virus of claim 18.

20. An isolated cell infected with the dengue type 1 virus of claim 18.

21. An isolated dengue type 2 virus having ATCC accession number VR-2650.

22. An isolated cell infected with the dengue type 2 virus of claim 21.

23. An isolated dengue type 3 virus having ATCC accession number VR-2654.

24. An isolated dengue type 3 virus vaccine produced by inactivating a dengue 3 virus of claim 23.

25. An isolated cell infected with the dengue type 3 virus of claim 23.

26. An isolated dengue type 4 virus having ATCC accession number VR-2651.

27. An isolated dengue type 4 virus vaccine produced by inactivating a dengue 4 virus of claim 26.

28. An isolated cell infected with the dengue type 4 virus of claim 26.