Polypeptide carrier for presenting target polypeptide and uses thereof

Patent metadata

Field	Value
Publication number	US-10987418-B2
Application number	US-201816484366-A
Country	US
Kind code	B2
Filing date	Feb 5, 2018
Priority date	Feb 17, 2017
Publication date	Apr 27, 2021
Grant date	Apr 27, 2021

Abstract

The invention relates to a polypeptide carrier for presenting a target polypeptide, and use thereof. In particular, the invention relates to a nucleic acid molecule, comprising a nucleotide sequence encoding a polypeptide carrier, and being used for insertion of a nucleotide sequence encoding a target polypeptide. In addition, the invention further relates to a recombinant protein comprising the polypeptide carrier and a target polypeptide. Furthermore, the invention further relates to use of the nucleic acid molecule and the recombinant protein. In addition, the invention further relates to a vaccine or a pharmaceutical composition useful for preventing, alleviating or treating HBV infection or a disease associated with HBV infection (e.g., hepatitis B), comprising a recombinant protein comprising the polypeptide carrier of the invention and an epitope from HBV.

First claim

The invention claimed is:

1. A nucleic acid molecule, comprising a nucleotide sequence encoding a polypeptide carrier, wherein the polypeptide carrier is selected from the following polypeptide carriers:
(1) RBHBcAg-T carrier, which differs from roundleaf bat HBV core antigen protein (RBHBcAg protein) by difference comprising the following:
(1a) one or more amino acid residues of the amino acid residues from positions 78-83 at N-terminus of RBHBcAg protein are deleted or substituted with a linker;
(1b) one or more amino acid residues of the amino acid residues from positions 18-27, one or more amino acid residues of the amino acid residues from positions 50-69 and/or one or more amino acid residues of the amino acid residues from positions 120-140 at N-terminus of RBHBcAg protein are each independently substituted with a human T cell epitope; and
(1c) optionally, 1-5, 5-10, 10-15, 15-20, 20-25, 25-30, 30-35, or 35-40 amino acid residues are deleted at C-terminus of RBHBcAg protein;

(2) TBHBcAg-T carrier, which differs from tent-making bat HBV core antigen protein (TBHBcAg protein) by difference comprising the following:
(2a) one or more amino acid residues of the amino acid residues from positions 80-84 at N-terminus of TBHBcAg protein are deleted or substituted with a linker;
(2b) one or more amino acid residues of the amino acid residues from positions 18-27, one or more amino acid residues of the amino acid residues from positions 54-73 and/or one or more amino acid residues of the amino acid residues from positions 124-144 at N-terminus of TBHBcAg protein are each independently substituted with a human T cell epitope; and
(2c) optionally, 1-5, 5-10, 10-15, 15-20, 20-25, 25-30, or 30-35 amino acid residues are deleted at C-terminus of TBHBcAg protein; and

(3) HBHBcAg-T carrier, which differs from horseshoe bat HBV core antigen protein (HBHBcAg protein) by difference comprising the following:
(3a) one or more amino acid residues of the amino acid residues from positions 78-83 at N-terminus of HBHBcAg protein are deleted or substituted with a linker;
(3b) one or more amino acid residues of the amino acid residues from positions 18-27, one or more amino acid residues of the amino acid residues from positions 50-69 and/or one or more amino acid residues of the amino acid residues from positions 120-140 at N-terminus of HBHBcAg protein are each independently substituted with a human T cell epitope; and
(3c) optionally, 1-5, 5-10, 10-15, 15-20, 20-25, 25-30, 30-35, or 35-40 amino acid residues are deleted at C-terminus of HBHBcAg protein.

2. The nucleic acid molecule according to claim 1 , wherein
(1) the polypeptide carrier is the RBHBcAg-T carrier; and optionally, the nucleic acid molecule is characterized by one or more of the following:
(i) the RBHBcAg protein has an amino acid sequence as set forth in SEQ ID NO: 1;
(ii) the amino acid residues from positions 78-83, positions 78-82, positions 78-81, or positions 78-80 at N-terminus of RBHBcAg protein are deleted or substituted with a linker;
(iii) the linker is a flexible linker;
(iv) the human T cell epitope is an MHC I or MHC II restricted human T cell epitope;
(v) the human T cell epitope is selected from human T cell epitopes as set forth in SEQ ID NOs: 87-89;
(vi) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid residues of the amino acid residues from positions 18-27 at N-terminus of RBHBcAg protein are substituted with an MHC I or MHC II restricted human T cell epitope; and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues of the amino acid residues from positions 50-69 at N-terminus of RBHBcAg protein are substituted with an MHC I or MHC II restricted human T cell epitope; and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 amino acid residues of the amino acid residues from positions 120-140 at N-terminus of RBHBcAg protein are substituted with an MHC I or MHC II restricted human T cell epitope;
(vii) the amino acid residues at positions 18-27, 50-69 and 120-140 at N-terminus of the RBHBcAg protein are substituted with human T cell epitopes set forth in SEQ ID NO: 87, 88 and 89, respectively;
(viii) a restriction enzyme cleavage site is introduced at a position of nucleotides encoding the one or more amino acid residues within positions 78-83 at N-terminus of RBHBcAg protein that are deleted;
(ix) a restriction enzyme cleavage site is introduced in the nucleotide sequence encoding the linker, and/or at either or both of the termini thereof;
(x) the polypeptide carrier has an amino acid sequence as set forth in SEQ ID NO: 75 or 78; and
(xi) the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 81 or 84;
or

(2) the polypeptide carrier is the TBHBcAg-T carrier; and optionally, the nucleic acid molecule is characterized by one or more of the following:
(i) the TBHBcAg protein has an amino acid sequence as set forth in SEQ ID NO: 2;
(ii) the amino acid residues from positions 80-84, positions 80-83, or positions 80-82, at N-terminus of TBHBcAg protein are deleted or substituted with a linker;
(iii) the linker is a flexible linker;
(iv) the human cell epitope is an MHC I or MHC II restricted human cell epitope;
(v) the human T cell epitope is selected from human cell epitopes as set forth in SEQ ID NOs: 87-89;
(vi) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid residues of the amino acid residues from positions 18-27 at N-terminus of TBHBcAg protein are substituted with an MHC 1 or MHC II restricted human cell epitope; and/or 1, 2; 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues of the amino acid residues from positions 54-73 at N-terminus of TBHBcAg protein are substituted with an MHC I or MHC II restricted human T cell epitope; and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 amino acid residues of the amino acid residues from positions 124-144 at N-terminus of TBHBcAg protein are substituted with an MHC 1 or MHC II restricted human T cell epitope;
(vii) the amino acid residues at positions 18-27, 54-73 and 124-144 at N-terminus of the TBHBcAg protein are substituted with human T cell epitopes set forth in SEQ ID NO: 87, 88 and 89, respectively;
(viii) a restriction enzyme cleavage site is introduced at a position of nucleotides encoding the one or more amino acid residues within positions 80-84 at N-terminus of TBHBcAg protein that are deleted;
(ix) a restriction enzyme cleavage site is introduced in the nucleotide sequence encoding the linker, and/or at either or both of the termini thereof;
(x) the polypeptide carrier has an amino acid sequence as set forth in SEQ ID NO: 76 or 79; and
(xi) the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 82 or 85;
or

(3) the polypeptide carrier is the HBHBcAg-T carrier; and optionally, the nucleic acid molecule is characterized by one or more of the following:
(i) the HBHBcAg protein has an amino acid sequence as set forth in SEQ NO: 3;
(ii) the amino acid residues from positions 78-83, positions 78-82, positions 78-81, or positions 78-80 at N-terminus of HBHBcAg protein are deleted or substituted with a linker;
(iii) the linker is a flexible linker;
(iv) the human cell epitope is an MHC I or MHC II restricted human cell epitope;
(v) the human T cell epitope is selected from human T cell epitopes as set forth in SEQ ID NOs: 87-89;
(vi) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid residues of the amino acid residues from positions 18-27 at N-terminus of HBHBcAg protein are substituted with an MHC I or WIC II restricted human T cell epitope; and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues of the amino acid residues from positions 50-69 at N-terminus of HBHBcAg protein are substituted wi

Assignees

• Univ Xiamen
• Yang Sheng Tang Co Ltd

Inventors

• Zhang Tianying
• Yuan Quan
• Guo Xueran
• Wei Minxi
• Kang Xiaozhen
• Zhang Jun
• Xia Ningshao

Classifications

• C12N2740/16034

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N2730/10134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K14/70532

  B7 molecules, e.g. CD80, CD86 · CPC title

• C07K14/705

  Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} · CPC title