Recombinant protein carrying human papillomavirus epitopes inserted in an adenylate cyclase protein or fragment thereof therapeutic uses thereof

Patent metadata

Field	Value
Publication number	US-9387243-B2
Application number	US-201314106921-A
Country	US
Kind code	B2
Filing date	Dec 16, 2013
Priority date	Mar 18, 2004
Publication date	Jul 12, 2016
Grant date	Jul 12, 2016

Abstract

The invention relates to a recombinant protein comprising one or several polypeptides bearing one or several epitopes of one or several HPV antigens, said polypeptides being inserted in the same or different permissive sites of an adenylate cyclase (CyaA) protein or of a fragment thereof, wherein said CyaA fragment retains the property of said adenylate cyclase protein to target Antigen Presenting Cells. It also concerns polynucleotides encoding the same. The recombinant protein or the polynucleotide can be used for the design of therapeutic means against HPV infection or against its malignant effects.

First claim

The invention claimed is:

1. A therapeutic vaccine composition, comprising a recombinant Bordetella pertussis adenylate cyclase (CyaA) protein comprising:
the HPV16-E7 fragment GQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKP (SEQ ID NO: 25) inserted at a first insertion site within the CyaA protein; and
the HPV16-E7 fragment MHGDTPTLHEYMLDLQPETTDLYCYEQLN (SEQ ID NO: 26) inserted at a second insertion site within the CyaA protein;
wherein the enzymatic activity of the recombinant CyaA protein has been genetically inactivated; and
wherein administering the therapeutic vaccine composition to a mammalian subject infected with HPV prevents HPV-induced tumor growth in the subject.

2. The therapeutic vaccine composition of claim 1 , wherein administering the therapeutic vaccine composition to the mammalian subject causes regression of an HPV induced tumor in the subject.

3. The therapeutic vaccine composition of claim 1 , wherein administering the therapeutic vaccine composition to the mammalian subject protects against the onset of malignant transformation by HPV in the subject.

4. The therapeutic vaccine composition of claim 1 , wherein the subject is a human.

5. The therapeutic vaccine composition of claim 1 , wherein the enzymatic activity of the recombinant CyaA protein has been genetically inactivated by insertion of a dipeptide between residues 188 and 189 of the native CyaA protein.

6. The therapeutic vaccine composition of claim 5 , wherein the dipeptide is the LQ dipeptide.

7. The therapeutic vaccine composition of claim 1 , wherein the first insertion site is between residues 224 and 235 of the native CyaA protein.

8. The therapeutic vaccine composition of claim 1 , wherein the second insertion site is between residues 319 and 320 of the native CyaA protein.

9. The therapeutic vaccine composition of claim 1 , wherein the first insertion site is between residues 224 and 235 of the native CyaA protein, and wherein the second insertion site is between residues 319 and 320 of the native CyaA protein.

10. The therapeutic vaccine composition of claim 6 , wherein the first insertion site is between residues 224 and 235 of the native CyaA protein, and wherein the second insertion site is between residues 319 and 320 of the native CyaA protein.

11. The therapeutic vaccine composition of claim 1 , wherein the recombinant CyaA protein is encoded by the insert contained in plasmid pTRACE5-HPV16E7 Δ30 _ 42 (C.N.C.M. 1-3190).

12. The therapeutic vaccine composition of claim 1 , further comprising at least one of an adjuvant, a surfactant, and an immunomodulating substance.

13. A therapeutic vaccine composition, comprising:
A) a first recombinant Bordetella pertussis adenylate cyclase (CyaA) protein comprising:
the HPV16-E7 fragment GQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKP (SEQ ID NO: 25) inserted at a first insertion site within the CyaA protein; and
the HPV16-E7 fragment MHGDTPTLHEYMLDLQPETTDLYCYEQLN (SEQ ID NO: 26) inserted at a second insertion site within the CyaA protein;
wherein the enzymatic activity of the first recombinant CyaA protein has been genetically inactivated; and
B) a second recombinant Bordetella pertussis adenylate cyclase (CyaA) protein comprising:
amino acid residues 43 to 105 of HPV 18-E7 inserted at a first insertion site within the CyaA protein; and
amino acid residues 1 to 31 of HPV 18-E7 inserted at a second insertion site within the CyaA protein;
wherein the enzymatic activity of the second recombinant CyaA protein has been genetically inactivated; and
wherein administering the therapeutic vaccine composition to a mammalian subject infected with HPV prevents HPV-induced tumor growth in the subject.

14. The therapeutic vaccine composition of claim 13 , wherein administering the therapeutic vaccine composition to the mammalian subject causes regression of an HPV induced tumor in the subject.

15. The therapeutic vaccine composition of claim 13 , wherein administering the therapeutic vaccine composition to the mammalian subject protects against the onset of malignant transformation by HPV in the subject.

16. The therapeutic vaccine composition of claim 13 , wherein the subject is a human.

17. The therapeutic vaccine composition of claim 13 , wherein the enzymatic activity of the first and second recombinant CyaA proteins has been genetically inactivated by insertion of a dipeptide between residues 188 and 189 of the native CyaA protein.

18. The therapeutic vaccine composition of claim 17 , wherein the dipeptide is the LQ dipeptide.

19. The therapeutic vaccine composition of claim 13 , wherein the first insertion site in each of the first and second recombinant CyaA proteins is between residues 224 and 235 of the native CyaA protein.

20. The therapeutic vaccine composition of claim 13 , wherein the second insertion site in each of the first and second recombinant CyaA proteins is between residues 319 and 320 of the native CyaA protein.

21. The therapeutic vaccine composition of claim 13 , wherein the first insertion site in each of the first and second recombinant CyaA proteins is between residues 224 and 235 of the native CyaA proteins; and wherein the second insertion site in each of the first and second recombinant CyaA proteins is between residues 319 and 320 of the native CyaA proteins.

22. The therapeutic vaccine composition of claim 18 , wherein the first insertion site in each of the first and second recombinant CyaA proteins is between residues 224 and 235 of the native CyaA proteins; and wherein the second insertion site in each of the first and second recombinant CyaA proteins is between residues 319 and 320 of the native CyaA proteins.

23. The therapeutic vaccine composition of claim 13 , wherein the recombinant CyaA protein comprising inserted HPV16-E7 fragments is encoded by the insert contained in plasmid pTRACE5-HPV16E7 Δ30 _ 42 (C.N.C.M. 1-3190).

24. The therapeutic vaccine composition of claim 13 , further comprising at least one of an adjuvant, a surfactant, and an immunomodulating substance.

25. A method for treating a human papilloma virus (HPV) infection in a mammalian subject infected with HPV, comprising:
administering to the mammalian subject infected with HPV a therapeutic vaccine composition according to claim 1 ; and
inducing a cell-mediated immune response against HPV16-E7;
wherein the cell-mediated immune response prevents HPV-induced tumor growth in the subject.

26. The method of claim 25 , wherein the cell-mediated immune response causes regression of an HPV induced tumor in the subject.

27. The method of claim 25 , wherein the cell-mediated immune response protects against the onset of malignant transformation by HPV in the subject.

28. The method of claim 25 , wherein the subject is a human.

29. A method for treating a human papilloma virus (HPV) infection in a mammalian subject infected with HPV, comprising:
administering to the mammalian subject infected with HPV a therapeutic vaccine composition according to claim 13 ; and
inducing a cell-mediated immune response against HPV16-E7 and HPV18-E7;
wherein the cell-mediated immune response prevents HPV-induced tumor growth in the subject.

30. The method of claim 29 , wherein the cell-mediated immune response causes regression of an HPV induced tumor in the subje

Assignees

• Pasteur Institut
• Inst Nat Sante Rech Med
• Centre Nat Rech Scient
• Genticel

Inventors

• Preville Xavier-Edmond-Edouard
• Leclerc Claude
• Ladant Daniel
• Timmerman Benedikt

Classifications

• A61P31/20

  for DNA viruses · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title