DNA vaccines against foot-and-mouth disease virus

Patent metadata

Field	Value
Publication number	US-10865389-B2
Application number	US-201916369177-A
Country	US
Kind code	B2
Filing date	Mar 29, 2019
Priority date	Sep 8, 2016
Publication date	Dec 15, 2020
Grant date	Dec 15, 2020

Abstract

This application is directed generally to foot-and-mouth disease virus (FMDV) 3C proteases that have been modified by mutating a polynucleotide sequence coding for the FMDV 3C protease. The modified FMDV proteases exhibit proteolytic activity on FMDV P1 precursor protein and exhibit a reduction in one or more toxic or inhibitory properties associated with an unmodified FMDV 3C protease on a host cell used to recombinantly produce it. Vectors carrying polynucleotides encoding modified FMDV 3C protease sequences can induce production of FMDV virus-like particles in a host cell when expressed in the host cell. The modified FMDV 3C proteases can generally be used to produce immunogenic FMDV preparations capable of inducing an immune response against FMDV.

First claim

What is claimed:

1. A DNA vaccine against foot-and-mouth disease virus (FMDV), comprising:
a polynucleotide encoding a FMDV P1 precursor polypeptide; and
a polynucleotide encoding a modified FMDV 3C protease, wherein the modified FMDV 3C protease comprises a L127 amino acid substitution of a wild-type FMDV 3C protease.

2. The DNA vaccine of claim 1 , wherein the polynucleotide encoding a FMDV P1 precursor polypeptide and the polynucleotide encoding a modified FMDV 3C protease are contained in a single vector or polynucleotide construct.

3. The DNA vaccine of claim 1 , comprising a first vector or polynucleotide construct comprising the polynucleotide encoding a FMDV P1 precursor polypeptide, and a second vector or polynucleotide construct comprising the polynucleotide encoding a modified FMDV 3C protease.

4. The DNA vaccine of claim 1 , comprising a polynucleotide encoding a fusion protein of the modified FMDV 3C protease and the FMDV P1 precursor polypeptide.

5. The DNA vaccine of claim 1 , wherein the polynucleotide encoding a FMDV P1 precursor polypeptide and the polynucleotide encoding a modified FMDV 3C protease together comprise a single open reading frame encoding both the FMDV P1 precursor polypeptide and the modified FMDV 3C protease.

6. The DNA vaccine of claim 1 , formulated against at least one of FMDV 0 serotype, A serotype, C serotype, Asia 1 serotype, SAT1 serotype, SAT2 serotype or SAT3 serotype.

7. The DNA vaccine of claim 1 , formulated against two or more different strains of FMDV.

8. The DNA vaccine of claim 1 , wherein the modified FMDV 3C protease further comprises a C142 substitution.

9. The DNA vaccine of claim 1 , formulated for injection, oral administration, intranasal administration, topical administration, electroporation, gene gun administration, transfection, or liposome-mediated delivery.

10. The DNA vaccine of claim 1 , wherein the vaccine is a marker vaccine or a DIVA vaccine (Differentiating Infected from Vaccinated Animals).

11. The DNA vaccine of claim 1 , wherein the polynucleotide encoding a FMDV P1 precursor polypeptide and the polynucleotide encoding a modified FMDV 3C protease are contained in at least one of a minicircle vector, a replication deficient adenovirus vector or a vaccinia virus vector.

12. The DNA vaccine of claim 1 , wherein the modified FMDV 3C protease comprises a L127P substitution.

13. The DNA vaccine of claim 12 , wherein the modified FMDV 3C protease further comprises a C142T substitution.

14. A pharmaceutical composition, comprising:
a polynucleotide encoding a FMDV P1 precursor polypeptide,
a polynucleotide encoding a modified FMDV 3C protease, wherein the modified FMDV 3C protease comprises a L127 amino acid substitution of a wild-type FMDV 3C protease; and
a pharmaceutically acceptable carrier.

15. A method for inducing an immune response against foot-and-mouth disease virus (FMDV) in a subject, vaccinating a subject against FMDV, or reducing severity of an FMDV infection in a subject, comprising administering to said subject an effective amount of a composition comprising:
a polynucleotide encoding a FMDV P1 precursor polypeptide; and
a polynucleotide encoding a modified FMDV 3C protease, wherein the modified FMDV 3C protease comprises a L127 amino acid substitution of a wild-type FMDV 3C protease, wherein the FMDV P1 precursor polypeptide is expressed and proteolytically processed by the co-expressed modified FMDV 3C protease.

16. The method of claim 15 , wherein the modified FMDV 3C protease further comprises a C142T substitution.

17. The method of claim 15 , wherein the composition is administered by subcutaneous injection, intradermal injection, intramuscular injection, jet injection, orally, intranasally, topically, by electroporation, gene gun, transfection, or liposome-mediated delivery.

18. The method of claim 15 , wherein the composition is administered prophylactically to prevent or ameliorate the effects of a future infection, therapeutically to treat or empower the immune system of an infected subject, or both.

19. The method of claim 15 , wherein the composition is administered to the subject in a single dose, a two dose schedule, or a multiple dose schedule.

20. The method of claim 15 , wherein the FMDV is one or more of FMDV 0 serotype, A serotype, C serotype, Asia 1 serotype, SAT1 serotype, SAT2 serotype or SAT3 serotype.

21. The method of claim 15 , wherein the subject is a cow, pig, sheep, goat, water buffalo, yak, reindeer, deer, elk, llama, alpaca, bison, moose, camel, chamois, giraffe, hog, warthog, kudu, antelope, gazelle or wildebeest.

22. The method of claim 15 , wherein the modified FMDV 3C protease comprises a L127P substitution.

23. The method of claim 22 , wherein the modified FMDV 3C protease further comprises a C142T substitution.

Assignees

• The Government Of The Us Secretary Of Homeland Security

Inventors

• Puckette Michael
• Rasmussen Max V

Classifications

• C07K16/106

  Picornaviridae (F), e.g. hepatitis A virus · CPC title

• A61P31/14

  for RNA viruses · CPC title

• G01N33/56972

  White blood cells · CPC title

• C12N2770/32134

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

• A61K39/135

  Foot- and mouth-disease virus · CPC title