Immunogenic peptides from the HPV E7 protein

• US 6,013,258 A
• Filed: 10/09/1997
• Issued: 01/11/2000
• Est. Priority Date: 10/09/1997
• Status: Expired due to Term

• Current Assignee: Eisai Incorporated (Eisai Medical Research Incorporated)
• Original Assignee: Zycos, Inc. (Eisai Medical Research Incorporated)
• Inventors: Urban, Robert G., Hedley, Mary Lynne, Chicz, Roman M., Collins, Edward J.
• Primary Examiner(s): Eisenschenk, Chris
• Assistant Examiner(s): SALIMI, ALI REZA
• Application Number:
  US08/948,378
• Time in Patent Office: 824 Days
• Field of Search: 424/199.1, 424/204.1, 424/185.1, 424/186.1, 424/489, 424/450, 435/235.1, 435/326, 435/320.1, 436/829, 514/44, 536/23.72
• US Class Current: 424/186.1
• CPC Class Codes: A61K 2039/51 comprising whole cells, viruses or DNA/RNA
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• 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: Y10S 436/829 Liposomes, e.g. encapsulation

Abstract

The invention provides immunogenic peptides from the HPV type 16 E7 protein that comprise overlapping class I restricted T cell epitopes. Also disclosed are methods of administering DNA molecules encoding these peptides to a host mammal.

Claims

1. A nucleic acid comprising a coding sequence coding for expression of a peptide less than 19 amino acids in length, wherein the peptide comprises the amino acid sequence Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys (SEQ ID NO:

16).

2. The nucleic acid of claim 1, wherein the amino acid sequence of the peptide comprises 13 amino acid residues.

3. The nucleic acid of claim 1, wherein the amino acid sequence of the peptide consists of 13 amino acid residues.

4. The nucleic acid of claim 2, wherein the amino acid sequence of the Peptide comprises Xaa Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys, wherein Xaa is Ala, Ser, Arg, Lys, Gly, Gln, Asp, or Glu (SEQ ID NO:

19).

5. The nucleic acid of claim 4, wherein the amino acid sequence of the peptide comprises Ala Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys (SEQ ID NO:

4).

6. The nucleic acid of claim 1, wherein the amino acid sequence of the peptide comprises Leu Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys (SEQ ID NO:

3).

7. A plasmid comprising the nucleic acid of claim 4.

8. A microparticle comprising a polymeric matrix and the plasmid of claim 7.

9. A nucleic acid comprising a coding sequence coding for expression of a polypeptide comprising a first peptide and a second peptide linked by a peptide bond, the first peptide comprising a trafficking sequence, and the second peptide consisting of a sequence 12-18 amino acids in length comprising the sequence Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys (SEQ ID NO:

16).

10. The nucleic acid of claim 9, wherein the first peptide is a signal sequence.

11. The nucleic acid of claim 9, wherein the amino acid sequence of the first peptide comprises Met Ala Ile Ser Gly Val Pro Val Leu Gly Phe Phe Ile Ile Ala Val Leu Met Ser Ala Gln Glu Ser Trp Ala (SEQ ID NO:

18).

12. The nucleic acid of claim 9, wherein the amino acid sequence of the second peptide is Xaa Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys, Xaa being Leu, Ala, Ser, Arg, Lys, Gly, Gln, Asp, or Glu (SEQ ID NO:

19).

13. The nucleic acid of claim 9, wherein the amino acid sequence of the second peptide is Leu Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys (SEQ ID NO:

3).

14. The nucleic acid of claim 9, wherein the amino acid sequence of the second peptide is Ala Leu Met Gly Thr Leu Gly Ile Val Cys Pro Ile Cys (SEQ ID NO:

4).

15. The nucleic acid of claim 9, wherein the first peptide trafficks the polypeptide to the intracellular compartment to which the HLA-DRα

leader sequence (SEQ ID NO;

18) trafficks a linked peptide.

16. A plasmid comprising the nucleic acid of claim 15.

17. A plasmid comprising the nucleic acid of claim 11.

18. A plasmid comprising the nucleic acid of claim 13.

19. A microparticle comprising a polymeric matrix and the plasmid of claim 16.

20. A microparticle comprising a polymeric matrix and the plasmid of claim 17.

21. A microparticle comprising a polymeric matrix and the plasmid of claim 18.

22. A plasmid comprising the nucleic acid of claim 14.

23. A microparticle comprising a polymeric matrix and the plasmid of claim 22.

24. A plasmid comprising the nucleic acid of claim 9.

25. A microparticle comprising a polymeric matrix and the plasmid of claim 24.

26. The microparticle of claim 25, wherein the polymeric matrix of said microparticle consists essentially of a copolymer of PLGA.

27. The microparticle of claim 25, wherein said microparticle has a diameter of 0.02 to 20 microns.

28. The microparticle of claim 25, wherein said microparticle has a mean diameter of less than about 11 microns.

29. A cell comprising the plasmid of claim 24.

30. The cell of claim 29, wherein said cell is a mammalian B cell or other antigen presenting cell (APC).

31. A DNA comprising the sequence of SEQ ID NO:

5.

32. A DNA comprising the sequence of nucleotides 3219-3624 of SEQ ID NO:

7.

33. A DNA comprising the sequence of nucleotides 3290-3413 of SEQ ID NO:

7.

34. A plasmid DNA comprising the sequence of SEQ ID NO:

7.

35. A microparticle comprising a polymeric matrix and a nucleic acid, wherein the polymeric matrix consists essentially of poly-lactic-co-glycolic acid (PLGA) and the nucleic acid comprises the sequence of SEQ ID NO:

7.