Polypeptides and antibodies for treating HBV infection and related diseases
The present invention relates to epitope peptides (or mutants thereof) for treating hepatitis B virus infection, recombinant proteins comprising such epitope peptides (or mutants thereof) and carrier proteins, and uses of such epitope peptides (or mutants thereof) and recombinant proteins. The present invention also relates to antibodies against such epitope peptides, cell lines producing said antibodies, and uses thereof. Furthermore, the present invention relates to vaccines or pharmaceutical compositions for treating or alleviating one or more symptoms associated with hepatitis B virus infection, which comprise the recombinant proteins or antibodies according to the invention, respectively.
1. A method for reducing serum level of HBV DNA and/or HBsAg in a subject, comprising administering to a subject infected with HBV and in need of reducing serum level of HBV DNA and/or HBsAg an effective amount that results in significant reductions of serum level of HBV DNA and/or HBsAg of:
(a) an isolated epitope peptide consisting of 4-38 consecutive amino acid residues of HBsAg protein and comprising amino acid residues from positions 121 to 124 of HBsAg protein, or a mutant thereof, wherein the mutant differs from the epitope peptide merely by conservative substitution of one or several amino acid residues and retains the biological function of the epitope peptide;
(b) a recombinant protein comprising the epitope peptide or mutant thereof of (a) and a carrier protein;
(c) an isolated nucleic acid molecule comprising a nucleotide sequence encoding the epitope peptide or mutant thereof of (a) or the recombinant protein;
(d) a vector comprising the isolated nucleic acid molecule of (c); or
(e) a pharmaceutical composition comprising the epitope peptide or mutant thereof of (a) or the recombinant protein of (b) or the isolated nucleic acid molecule of (c) or the vector of (d), and a pharmaceutically acceptable carrier and/or excipient.
2. The method according to claim 1 , wherein the epitope peptide consists of 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5 or 4 consecutive amino acid residues of HBsAg protein, and comprises amino acid residues from positions 121 to 124 of HBsAg protein.
3. The method according to claim 1 , wherein the amino acid residues from positions 121 to 124 of HBsAg protein are as shown in SEQ ID NO: 10.
4. The method according to claim 1 , wherein the epitope peptide consists of 7-38 consecutive amino acid residues of HBsAg protein and comprising amino acid residues from positions 119 to 125 of HBsAg protein.
5. The method according to claim 1 , wherein the epitope peptide consists of any of the following:
(1) the amino acid residues from positions 119-125 of HBsAg protein;
(2) the amino acid residues from positions 113-127 of HBsAg protein;
(3) the amino acid residues from positions 115-125 of HBsAg protein;
(4) the amino acid residues from positions 113-135 of HBsAg protein; and
(5) the amino acid residues from positions 111-148 of HBsAg protein.
6. The method according to claim 1 , wherein the mutant differs from the epitope peptide merely by conservative substitution of 1, 2, 3 or 4 amino acid residues.
7. The method according to claim 1 , wherein the epitope peptide or mutant thereof has an amino acid sequence selected from the group consisting of SEQ ID NO:1-7 and 10.
8. The method according to claim 1 , wherein the recombinant protein is not a naturally occurring protein.
9. The method according to claim 1 , wherein in the recombinant protein, the epitope peptide or mutant thereof is linked to the carrier protein, optionally via a linker.
10. The method according to claim 9 , wherein the linker is a rigid or flexible linker.
11. The method according to claim 1 , wherein the carrier protein is CRM197 protein or a fragment thereof, and wherein the epitope peptide or mutant thereof is linked to the N-terminus or C-terminus of the CRM197 protein or fragment thereof, optionally via a linker.
12. The method according to claim 11 , wherein the fragment of the CRM197 protein comprises or consists of aa 1-190 of CRM197, or comprises or consists of aa 1-389 of CRM197.
13. The method according to claim 1 , wherein the carrier protein is HBcAg or a fragment thereof, wherein the amino acids from positions 79 to 81 of the HBcAg are replaced with the epitope peptide or mutant thereof, and wherein the epitope peptide or mutant thereof is linked to the HBcAg or fragment thereof optionally via a linker.
14. The method according to claim 13 , wherein the fragment of HBcAg comprises or consists of aa 1-149 of HBcAg.
15. The method according to claim 1 , wherein the carrier protein is WHcAg or a fragment thereof (such as aa 1-149 of WHcAg), wherein the amino acids from positions 79 to 81 of the WHcAg are replaced with the epitope peptide or mutant thereof, and wherein the epitope peptide or mutant thereof is linked to the WHcAg or fragment thereof, optionally via a linker.
16. The method according to claim 15 , wherein the fragment of WHcAg comprises or consists of aa 1-149 of WHcAg.
17. The method according to claim 1 , wherein the recombinant protein has an amino acid sequence selected from a group consisting of SEQ ID NO: 47-53, 56, and 58-97.
18. The method according to claim 1 , wherein the pharmaceutically acceptable carrier and/or excipient is an adjuvant.