Live attenuated vaccines for influenza viruses

Patent metadata

Field	Value
Publication number	US-9827304-B2
Application number	US-201514940307-A
Country	US
Kind code	B2
Filing date	Nov 13, 2015
Priority date	Nov 13, 2014
Publication date	Nov 28, 2017
Grant date	Nov 28, 2017

Abstract

The subject invention pertains to attenuated influenza viruses, and related vaccines and methods, comprising a genetically modified viral genome. The genetically modified viral genome comprises a disruption in the non-structural (NS1) coding segment and one or more base substitution in the matrix membrane protein coding segment. The genetic modifications result in viruses that lose NS1 functionality, yet remain replication competent.

First claim

We claim:

1. An attenuated influenza virus, comprising a genetically modified viral genome comprising:
a deletion of an exon region in the non-structural (NS1) coding segment, wherein the deletion results in a knockout of the encoded NS1 protein; and a base substitution in a matrix membrane protein coding segment, wherein the base substitution is selected from a G917A substitution, an A14U substitution, and a combination thereof, wherein base numbering is in accordance with the base numbering of SEQ ID NO: 17.

2. The virus of claim 1 , wherein the attenuated influenza virus is attenuated influenza virus strain H7N9.

3. The virus of claim 1 , wherein the attenuated influenza virus is attenuated influenza virus strain H1N1.

4. The virus of claim 1 , wherein the attenuated influenza virus is attenuated influenza virus strain H5N1.

5. The virus of claim 1 , wherein the base substitution is an A14U substitution.

6. The virus of claim 1 , wherein the base substitution is a G917A substitution.

7. A vaccine formulation comprising:
an attenuated influenza virus according to claim 1 , and
a physiologically acceptable carrier.

8. The vaccine formulation of claim 7 , wherein the attenuated influenza virus is attenuated H7N9.

9. The vaccine formulation of claim 7 , wherein the attenuated influenza virus is attenuated H1N1.

10. The vaccine formulation of claim 7 , wherein the attenuated influenza virus is attenuated H5N1.

11. The vaccine formulation of claim 7 , wherein the base substitution is an A14U substitution.

12. The vaccine formulation of claim 7 , wherein the base substitution is a G917A substitution.

13. A method for generating an attenuated influenza virus according to claim 1 , the method comprising:
introducing an influenza virus NS1 gene in which the coding sequence exon has been deleted, into a cell or egg;
introducing a matrix membrane coding sequence of an influenza virus into the cell or egg, wherein the matrix membrane coding sequence comprises one or more base substitutions selected from a G917A substitution, an A14U substitution, and a combination thereof; and wherein the cell or egg comprises the remaining influenza virus gene segments and viral proteins required to produce influenza virus particles; and
culturing the cell or egg, wherein the attenuated influenza virus is replicated.

14. The method of claim 13 , wherein the influenza virus being attenuated is H7N9.

15. The method of claim 13 , wherein the influenza virus being attenuated is H1N1.

16. The method of claim 13 , wherein influenza virus being attenuated is H5N1.

17. The method of claim 13 , wherein the base substitution is an A14U substitution.

18. The method of claim 13 , wherein the base substitution is a G917A substitution.

Assignees

• Univ Hong Kong
• Versitech Ltd

Inventors

• Chen Honglin
• Zheng Min
• Yuen Kwok-Yung

Classifications

• C12N2760/16134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N2760/16121

  Viruses as such, e.g. new isolates, mutants or their genomic sequences · CPC title

• C12N2760/16171

  Demonstrated in vivo effect · CPC title

• A61K2039/5254

  avirulent or attenuated · CPC title

• C12N7/04

  Inactivation or attenuation; Producing viral sub-units · CPC title