Attenuated mutant dengue viruses comprising a mutation in the NS5 non-structural protein

Patent metadata

Field	Value
Publication number	US-11318196-B2
Application number	US-201916684103-A
Country	US
Kind code	B2
Filing date	Nov 14, 2019
Priority date	May 22, 2001
Publication date	May 3, 2022
Grant date	May 3, 2022

Abstract

A menu of mutations was developed that is useful in fine-tuning the attenuation and growth characteristics of dengue virus vaccines.

First claim

What is claimed is:

1. A dengue virus comprising an A to U mutation at nucleic acid position 7849, wherein the numbering is based on the prototypic isolate DEN4 Dominica 1981 and wherein the dengue virus has a phenotype selected from temperature sensitivity in Vero cells or human liver cell line HuH-7 and attenuation in mice.

2. The dengue virus of claim 1 , further comprising the A30 mutation.

3. The dengue virus of claim 1 , wherein the dengue virus is a dengue virus type 1, a dengue virus type 2, a dengue virus type 3, or a dengue virus type 4.

4. The dengue virus of claim 1 , wherein the dengue virus is a chimeric virus.

5. The chimeric virus of claim 4 , having a dengue 1 backbone, a dengue 2 backbone, a dengue 3 backbone, or a dengue 4 backbone.

6. A pharmaceutical composition comprising a pharmacologically acceptable vehicle and a dengue virus of claim 1 .

7. A kit comprising the pharmaceutical composition of claim 6 in a pack or dispenser device and instructions for administration.

8. A method of producing neutralizing antibodies against dengue virus in a subject, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a pharmacologically acceptable vehicle and the dengue virus of claim 1 to the subject.

9. The method of claim 8 , wherein administration is by subcutaneous, intradermal, or intramuscular injection.

10. An immunogenic composition comprising a pharmacologically acceptable vehicle and a dengue virus of claim 1 .

11. A live attenuated vaccine comprising a pharmacologically acceptable vehicle and a dengue virus of claim 1 .

12. The live attenuated vaccine of claim 11 in unit dosage form having from about 10 2 -10 9 plaque forming units (PFU)/ml.

13. An inactivated vaccine comprising a pharmacologically acceptable vehicle and a dengue virus of claim 1 .

14. The inactivated vaccine of claim 13 in unit dosage form having from about 0.1 to 50 μg of E protein/ml.

15. A method of preparing a dengue virus comprising:
(a) synthesizing full-length viral genomic RNA in vitro using a cDNA molecule that encodes a dengue virus of claim 1 ;
(b) transfecting cultured cells with the viral genomic RNA to produce virus; and
(c) isolating the virus from the cultured cells.

16. A method of making a pharmaceutical composition comprising:
isolated the dengue virus of claim 1 ; and
combining the isolated dengue virus with a pharmacologically acceptable vehicle.

Assignees

• Us Health

Inventors

• Whitehead Stephen S
• Murphy Brian R
• Hanley Kathryn A
• Blaney Joseph E
• Lai Ching-Juh

Classifications

• C12N7/00 Primary

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title

• A61K2039/5252

  inactivated (killed) · CPC title

• C12N2770/24161

  Methods of inactivation or attenuation · CPC title

• C12N2770/24151

  Methods of production or purification of viral material · CPC title