Vaccines against HPV
The present invention relates to therapeutic compounds, such as vaccines against human papillomavirus (HPV) and in particular to DNA vaccines against HPV16 or HPV18. The invention further relates to protein construct encoding homodimeric peptides, which peptides may be released from a DNA vaccine or used separately. Further described are pharmaceutical formulations, host cells and methods for producing the vaccines, as well as methods for the treatment of various HPV induced diseases, such as cancers and infectious diseases by application.
1. A nucleic acid molecule, encoding an amino acid chain comprising (1) a signal peptide, (2) a targeting unit, (3) a dimerization motif, and (4) an antigenic unit, said targeting unit comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence 24-93 of SEQ ID NO:1, and an antigenic unit comprising an amino acid sequence having at least 80% sequence identity to early protein E6 (SEQ ID NO: 22) of HPV16 and an amino acid sequence having at least 80% sequence identity to early protein E7 (SEQ ID NO: 23) from HPV16, wherein the antigenic unit comprising an amino acid sequence having at least 80% sequence identity to early protein E6 further comprises the amino acid substitutions: F47R, C63R, and C106R.
2. The nucleic acid molecule according to claim 1 , which nucleic acid molecule is human codon optimized.
3. A nucleic acid molecule comprising nucleotide sequence SEQ ID NO:31.
4. The nucleic acid molecule according to claim 1 formulated for administration to a patient to induce production of the homodimeric protein in said patients.
5. The nucleic acid molecule according to claim 1 being a DNA molecule.
6. The nucleic acid molecule according to claim 1 being a RNA molecule.
7. The nucleic acid molecule according to claim 1 , wherein said targeting unit, dimerization motif and antigenic unit in said amino acid chain are in the N-terminal to C-terminal order of targeting unit, dimerization motif and antigenic unit.
8. The nucleic acid molecule according to claim 1 , wherein said signal peptide consists of an amino acid sequence having at least 80% sequence identity to the amino acid sequence 1-23 of SEQ ID NO:1.
9. The nucleic acid molecule according to claim 1 , wherein the dimerization motif comprises a hinge region and optionally another domain that facilitate dimerization, optionally connected through a linker.
10. The nucleic acid molecule according to claim 9 , wherein the hinge region is an immunoglobulin domain.
11. The nucleic acid molecule according to claim 10 , wherein the hinge region is derived from IgG3.
12. The nucleic acid molecule according to claim 1 , wherein the dimerization motif consist of an amino acid sequence having at least 80% sequence identity to the amino acid sequence 94-237 of SEQ ID NO:3.
13. The nucleic acid molecule according to claim 1 , wherein said antigenic unit and the dimerization motif is connected through a linker being a GLGGL linker or a GLSGL linker.
14. The nucleic acid molecule according to claim 1 , wherein said antigenic unit comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence 243-293 of SEQ ID NO:3.
15. The nucleic acid molecule according to claim 1 , wherein said antigenic unit comprises the amino acid sequence 243-293 of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, or SEQ ID NO:9, or an antigenic fragment thereof.
16. The nucleic acid molecule according to claim 1 , wherein said antigenic unit comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence 243-340 of SEQ ID NO:11.
17. The nucleic acid molecule according to claim 1 , wherein said antigenic unit comprises the amino acid sequence 243-340 of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, or SEQ ID NO:17, or an antigenic fragment thereof.
18. The nucleic acid molecule according to claim 1 , wherein said antigenic unit further comprises one or more amino acid substitutions at a position selected from the list consisting of L50 and I128 of SEQ ID NO:22, or a deletion involving one or more amino acid selected from the list consisting of Y43-L50 of SEQ ID NO:22.
19. The nucleic acid molecule according to claim 1 , wherein said antigenic unit further comprises one or more amino acid substitutions at a position selected from the list consisting of C24, E26, C58, C61, C91, and C94 of SEQ ID NO:23, or a deletion involving one or more amino acid selected from the list consisting of L22-E26 and/or C58-C61 and/or C91-S95 of SEQ ID NO:23.
20. The nucleic acid molecule according to claim 1 , wherein said antigenic unit further comprises one or more amino acid substitutions at a position selected from the list consisting of L50 and I128 of SEQ ID NO:22 and C24, E26, C58, C61, C91, C94 of SEQ ID NO:23.
21. The nucleic acid molecule according to claim 1 , wherein said antigenic unit comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence 243-501 of SEQ ID NO:19, SEQ ID NO:32, or SEQ ID NO:34.
22. The nucleic acid molecule according to claim 1 , wherein said amino acid chain consists of amino acid sequence SEQ ID NO:32.