Vaccine candidates for human respiratory syncytial virus (RSV) having attenuated phenotypes

Patent metadata

Field	Value
Publication number	US-10808012-B2
Application number	US-201716335099-A
Country	US
Kind code	B2
Filing date	Sep 22, 2017
Priority date	Sep 23, 2016
Publication date	Oct 20, 2020
Grant date	Oct 20, 2020

Abstract

Reported herein are presumptively de-attenuating mutations that are useful, either individually or in combinations that may include other known mutations, in producing recombinant strains of human respiratory syncytial virus (RSV) exhibiting attenuation phenotypes. Also described herein is a novel RSV construct, Min_L-NPM2-1(N88K)L, which exhibits an attenuated phenotype, is stable and is as immunogenic as wild type RSV. The recombinant RSV strains described here are suitable for use as live-attenuated RSV vaccines. Exemplary vaccine candidates are described. Also provided are polynucleotide sequences capable of encoding the described viruses, as well as methods for producing and using the viruses.

First claim

The invention claimed is:

1. An isolated polynucleotide molecule encoding a recombinant respiratory syncytial virus (RSV) variant having an attenuated phenotype comprising a RSV genome or antigenome sequence, wherein the RSV genome or antigenome is modified by a mutation in the L ORF at a position corresponding to T1166 of the L protein in SEQ ID NO:11.

2. The isolated polynucleotide molecule of claim 1 , wherein the RSV genome or antigenome is further modified by a mutation selected from the group consisting of:
i. a mutation in the M2-1 ORF at a position corresponding to N88 or A73 of the M2-1 protein in SEQ ID NO:9
ii. a mutation in the N ORF at a position corresponding to K136 of the N protein in SEQ ID NO:3;
iii. a mutation in the P ORF at a position corresponding to E114 of the P protein in SEQ ID NO:4; and
iv. combinations thereof.

3. The isolated polynucleotide molecule of claim 1 , wherein the mutation in the L ORF at a position corresponding to T1166 of the L protein in SEQ ID NO:11 is T1166I.

4. The isolated polynucleotide molecule of claim 2 , wherein
a. the mutation in the M2-1 ORF at a position corresponding to N88 of the M2-1 protein in SEQ ID NO:9 is N88K and the mutation in the M2-1 ORF at a position corresponding to A73 of the M2-1 protein in SEQ ID NO:9 is A73S;
b. the mutation in the N ORF at a position corresponding to K136 of the N protein in SEQ ID NO:3 is K136R; and
c. the mutation in the P ORF at a position corresponding to E114 of the P protein in SEQ ID NO:4 is E114V.

5. The isolated polynucleotide molecule of claim 1 , wherein the RSV genome or antigenome comprises a deletion in at least one of the proteins selected from M2-2, NS1 and NS2.

6. The isolated polynucleotide molecule of claim 1 , wherein the RSV genome or antigenome is codon-pair deoptimized.

7. The isolated polynucleotide molecule of claim 1 , wherein the L ORF of the RSV genome or antigenome is codon-pair deoptimized.

8. A vector comprising the isolated polynucleotide molecule of claim 1 .

9. A cell comprising the isolated polynucleotide of claim 1 .

10. A pharmaceutical composition comprising an immunologically effective amount of the recombinant RSV variant encoded by the isolated polynucleotide molecule of claim 1 .

11. A method of vaccinating a subject against RSV comprising administering the pharmaceutical composition of claim 10 .

12. A method of inducing an immune response comprising administering the pharmaceutical composition of claim 10 .

13. The method of claim 11 , wherein the pharmaceutical composition is administered intranasally.

14. The method of claim 13 , wherein the pharmaceutical composition is administered via injection, aerosol delivery, nasal spray or nasal droplets.

15. A live attenuated RSV vaccine comprising the recombinant RSV variant encoded by the isolated polynucleotide of claim 1 .

16. A pharmaceutical composition comprising the RSV vaccine of claim 15 .

17. An isolated polynucleotide molecule encoding a recombinant respiratory syncytial virus (RSV) variant having an attenuated phenotype comprising a RSV genome or antigenome sequence, wherein the RSV genome or antigenome is modified by one or more mutations selected from the positions recited in Tables S1, or S2 or S3.

18. The isolated polynucleotide molecule of claim 17 , wherein the RSV genome or antigenome comprises a deletion in at least one of the proteins selected from M2-2, NS1 and NS2.

19. The isolated polynucleotide molecule of claim 17 , wherein the RSV genome or antigenome is codon-pair deoptimized.

20. An isolated polynucleotide molecule that is at least about 80% identical to the nucleotide sequence of SEQ ID NO:14.

Assignees

• The Usa As Represented By The Secretary Dept Of Health And Human Services
• Codagenix Inc

Inventors

• Lenouen Cyril
• Buchholz Ursula J
• Collins Peter L
• Mueller Steffen

Classifications

• C12N2760/00062

  by genetic engineering · CPC title

• C12N2760/00021

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

• C12N2760/00034

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

• C12N7/04

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

• A61K39/155

  Paramyxoviridae, e.g. parainfluenza virus · CPC title