Human papillomavirus polypeptides and immunogenic compositions

• US 20060014926A1
• Filed: 10/02/2003
• Published: 01/19/2006
• Est. Priority Date: 10/03/2002
• Status: Active Grant

• Current Assignee: Wyeth Holdings Corporation (Pfizer Inc.)
• Original Assignee: Wyteth Holdings Corporation
• Inventors: Smith, Larry, Pullen, Jeffrey K., McElhiney, Susan P., Cassetti, Maria C.
• Granted Patent: US 7,223,408 B2
• Time in Patent Office: Days
• US Class Current: 530/300
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/585 wherein the target is cancer
• CPC Class Codes: A61K 2039/892 Reproductive system [uterus, ovaries, cervix, testes]
• CPC Class Codes: A61K 38/162 from virus
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/20022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2710/20032 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2710/20034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2710/24143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2770/36143 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The present invention provides immunogenic and pharmaceutical compositions for the treatment and prevention of human papillomavirus (HPV)-associated cancers and in particular, cervical cancer. In particular, this invention relates to fusion proteins, and the nucleic acids encoding these fusion proteins, used to generate immune responses against HPV. Specifically, this invention provides for fusions of HPV E6 and E7 in which the E6 and/or E7 contains one or more mutations. These mutations abrogate the transformation activity of these oncogenic proteins and, thus, confer safety to the E6/E7 fusions. In addition, these fusions maintain or increase the immunogenic efficacy of E6 and E7. Any gene or protein delivery method can be used to deliver or package the immunogenic compositions of the present invention.

Claims

1. A polypeptide comprising human papillomavirus E6 and E7 polypeptides, wherein the E7 polypeptide has mutations at any one or more of the amino acids corresponding to amino acids 24, 26 or 91 of SEQ ID NO:

14 and the E6 polypeptide has no mutations or has mutations at any one or more of the amino acids corresponding to amino acids 63 or 106 of SEQ ID NO;

13.

2. The polypeptide of claim 1 wherein the mutated amino acids are mutated to glycine.

3. The polypeptide of claim 1 wherein the E7 polypeptide precedes the E6 polypeptide.

4. The polypeptide of claim 2 wherein the E7 polypeptide precedes the E6 polypeptide.

5. An isolated nucleic acid encoding the polypeptide of claim 1.

6. The nucleic acid of claim 5 wherein the nucleotide sequence of E7 precedes the nucleotide sequence of E6.

7. An expression vector comprising the nucleic acid sequence of claim 5 under the control of an expression control sequence.

8. A host cell comprising the nucleic acid of claim 5.

9. A host cell which expresses the polypeptide of claim 1.

10. A host cell comprising the expression vector of claim 7.

11. An immunogenic composition comprising:

(a) the polypeptide of claim 1;

and

(b) a pharmaceutically acceptable carrier.

12. The immunogenic composition of claim 11 further comprising adjuvant.

13. An immunogenic composition comprising the nucleic acid of claim 5.

14. A recombinant virus comprising the nucleic acid of claim 5.

15. The recombinant virus of claim 14, wherein the virus is a modified Venezuelan equine encephalitis virus.

16. A method for producing an immune response in an individual, which method comprises administering to the individual the immunogenic composition of claim 11 in an amount sufficient to produce the immune response.

17. A method of treating cervical cancer, which method comprises administering to a patient diagnosed with cervical cancer the immunogenic composition according to claim 11 in an amount sufficient to produce a protective immune response.

18. A method of preventing cervical cancer, which method comprises administering to an individual the immunogenic composition of claim 11 in an amount sufficient to produce a protective immune response.

19. A method of preventing cervical cancer, which method comprises administering to an individual the expression vector of claim 7 in an amount sufficient to produce a protective immune response.

20. A method of treating cervical cancer, which method comprises administering to a patient diagnosed with cervical cancer the expression vector of claim 7 in an amount sufficient to produce a protective immune response.

21. The isolated polypeptide of claim 1 wherein the E7 polypeptide has mutations in at least two of amino acids corresponding to amino acids 24, 26 and 91 of SEQ ID NO:

14 and the E6 polypeptide has one or more mutations at amino acids corresponding to amino acids 63 and 106 of SEQ ID NO;

13.

22. An isolated polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

3, SEQ ID NO;

5, SEQ ID NO;

9, or SEQ ID NO;

11

23. An isolated nucleic acid encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

3, SEQ ID NO;

5, SEQ ID NO;

9 or SEQ ID NO;

11.

24. The isolated nucleic acid of claim 23 having the nucleotide sequence as set forth in SEQ ID NO:

4, SEQ ID NO;

6, SEQ ID NO;

10 or SEQ ID NO;

12.

25. An expression vector comprising the nucleic acid sequence of claim 23 under the control of an expression control sequence.

26. An expression vector comprising the nucleic acid sequence of claim 24 under the control of an expression control sequence.