Compositions, methods and uses for improved human papilloma virus constructs

Patent metadata

Field	Value
Publication number	US-10413603-B2
Application number	US-201515320277-A
Country	US
Kind code	B2
Filing date	Jun 19, 2015
Priority date	Jun 19, 2014
Publication date	Sep 17, 2019
Grant date	Sep 17, 2019

Abstract

Embodiments of the present invention provide compositions and methods for recombinantly generating tagless constructs of proteins or peptides. In certain embodiments, recombinant proteins or peptides disclosed herein concern human papilloma virus (HPV). Other embodiments concern using these constructs in compositions to elicit immune responses in a subject to one or more HPV types. Therapeutic and prophylactic vaccines for the prevention and treatment of viral infections are also disclosed. Nucleic acids and expression vectors coding for constructs contemplated herein are provided. In certain embodiments, an HPV capsid protein generated is devoid of any fusion tags. In addition, truncated forms of HPV L1 are contemplated.

First claim

What is claimed:

1. A bacterial plasmid construct comprising:
a target Human Papilloma Virus (HPV) DNA molecule comprising a transcription start codon, wherein the target HPV DNA molecule encodes one or more truncated HPV L1 proteins or peptides having at least 3 amino acids and no more than 24 amino acids deleted at the N-terminus, and up to 29 amino acids deleted at the C-terminus;
a promoter associated with the target HPV DNA molecule;
a restriction site positioned between the promoter and the target HPV DNA molecule;
a resistance gene capable of conferring resistance to a selection agent; and
the bacterial plasmid construct is devoid of any tag or tracking agent, making the bacterial construct tagless.

2. The bacterial plasmid construct of claim 1 , wherein the target HPV DNA molecule encodes a single protein or a chimeric protein.

3. The bacterial plasmid construct of claim 1 , wherein the target HPV DNA molecule encodes one or more proteins or peptides of use in an immunogenic composition against a target HPV.

4. The bacterial plasmid construct of claim 1 , wherein the encoded one or more proteins or peptides are from one or more of HPV6, HPV6a, HPV6b, HPV11, HPV16, HPV18, HPV30, HPV31, HPV33, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV54, HPV55, HPV56, HPV58, and HPV70.

5. The bacterial plasmid construct of claim 1 , wherein the target HPV DNA molecule encodes a truncated HPV L1 protein or peptide of 3, 5, 9, 13, 15, 16, or 20 amino acids deleted at the N-terminus.

6. The bacterial plasmid construct of claim 5 , wherein the encoded truncated HPV L1 protein can form part of a capsomere.

7. The bacterial plasmid construct of claim 1 , wherein the promoter is a Tac promoter.

8. A pharmaceutical composition comprising one or more proteins or peptides expressed from one or more bacterial constructs according to claim 1 and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition of claim 8 , wherein the one or more bacterial constructs each comprises an HPV DNA molecule encoding a protein or fragment thereof from at least one of HPV16, HPV18, HPV31, HPV33, HPV35, HPV45, HPV52 and HPV58.

10. A method for eliciting an immune response to at least one HPV type in a subject, the method comprising administering to the subject a composition comprising one or more proteins or peptides expressed from the one or more bacterial constructs according to claim 1 and eliciting an immune response in the subject to the at least one HPV type.

11. The method of claim 10 , wherein the one or more bacterial constructs each comprises an HPV DNA molecule encoding a protein or fragment thereof from at least one of HPV16, HPV18, HPV31, HPV33, HPV35, HPV45, HPV52 and HPV58.

12. A prophylactic method of treatment for eliciting an immune response to at least one HPV type in a subject comprising, administering to the subject, a prophylactically effective amount of a composition comprising one or more proteins or peptides expressed from the one or more bacterial constructs according to claim 1 and reducing or preventing infection to the at least one HPV type in the subject.

13. The method of claim 12 , wherein the one or more bacterial constructs each comprises an HPV DNA molecule encoding an L1 protein or peptide from at least one of HPV16, HPV18, HPV31, HPV33, HPV35, HPV45, HPV52 and HPV58.

14. A therapeutic treatment method for eliciting an immune response to at least one HPV type infection in a subject comprising, administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising one or more proteins or peptides expressed from one or more bacterial constructs according to claim 1 and treating the infection in the subject.

15. The method of claim 14 , wherein the one or more bacterial constructs each comprises an HPV DNA molecule encoding a protein or peptide from at least one of HPV16, HPV18, HPV31, HPV33, HPV35, HPV45, HPV52 and HPV58.

16. An isolated cell comprising the bacterial plasmid construct of claim 1 .

17. An isolated preparation of expressed inclusion bodies comprising a polypeptide encoded by the bacterial plasmid construct of claim 1 .

18. The bacterial plasmid construct of claim 1 , wherein the target HPV DNA molecule encodes a truncated HPV L1 protein.

19. The bacterial plasmid construct of claim 1 , wherein the target HPV DNA molecule encodes a truncated HPV L1 protein or peptide with 3 amino acids deleted at the N-terminus and up to 29 amino acids deleted at the C-terminus.

20. The bacterial plasmid construct of claim 1 , wherein the target HPV DNA molecule encodes a truncated HPV L1 protein or peptide with 29 amino acids deleted at the C-terminus.

21. The bacterial plasmid construct of claim 1 , wherein the bacterial construct is void of a GST tag.

22. The bacterial plasmid construct of claim 1 , wherein the bacterial constructs associate to form a pentamer.

23. The bacterial plasmid construct of claim 1 , wherein the bacterial construct is not a fusion protein or peptide.

Assignees

• Univ Colorado Regents

Inventors

• Garcea Robert L
• Macejak Dennis G

Classifications

• A61P31/20

  for DNA viruses · CPC title

• C12N2800/22

  Vectors comprising a coding region that has been codon optimised for expression in a respective host · CPC title

• C12N2800/101

  for bacteria · CPC title

• A61K2039/53

  DNA (RNA) vaccination · CPC title

• C12N2710/20022

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title