Fusion protein comprising diphtheria toxin non-toxic mutant CRM197 or fragment thereof

Patent metadata

Field	Value
Publication number	US-9764028-B2
Application number	US-201615248850-A
Country	US
Kind code	B2
Filing date	Aug 26, 2016
Priority date	Jun 1, 2011
Publication date	Sep 19, 2017
Grant date	Sep 19, 2017

Abstract

Provided in the present invention are a diphtheria toxin non-toxic mutant CRM197 or a fragment thereof as an adjuvant in a fusion protein and the use thereof to enhance the immunogenicity of a target protein fused therewith, for example, an HEV capsid protein, or an influenza virus M2 protein or an immunogenic fragment thereof. Also provided is a method for enhancing the immunogenicity of a target protein, comprising the fusion expression of the CRM197 or the fragment thereof with the target protein to form a fusion protein. Further provided is a fusion protein comprising the CRM197 or the fragment thereof and a target protein, the CRM197 or the fragment thereof enhancing the immunogenicity of the target protein. The present invention also provides an isolated nucleic acid encoding the fusion protein, a construct and a vector comprising said nucleic acid, and a host cell comprising the nucleic acid.

First claim

The invention claimed is:

1. A fusion protein comprising a fragment of CRM197 and a target protein, wherein said fragment of CRM197 enhances immunogenicity of the target protein, wherein said fragment of CRM197 consists of amino acids 1-389 of SEQ ID NO:2, and wherein said fragment of CRM197 is linked to the N-terminus and/or C-terminus of the target protein, optionally via a linker.

2. The fusion protein of claim 1 , wherein the target protein is HEV capsid protein or an immunogenic fragment thereof.

3. The fusion protein of claim 2 , wherein the immunogenic fragment of HEV capsid protein comprises or is HEV-239 (aa 368-606 of HEV capsid protein), E2 (aa 394-606 of HEV capsid protein) or E2s (aa 455-606 of HEV capsid protein).

4. The fusion protein of claim 2 , wherein the fusion protein comprises (a) the fragment of CRM197, and (b) HEV capsid protein or the immunogenic fragment of HEV capsid protein; wherein (a) and (b) are linked together, optionally via a linker.

5. The fusion protein of claim 2 , wherein the fusion protein has an amino acid sequence as set forth in SEQ ID NO: 8, 10, or 16.

6. The fusion protein of claim 1 , wherein the target protein is influenza virus M2 protein or an immunogenic fragment thereof.

7. The fusion protein of claim 6 , wherein the immunogenic fragment of M2 protein comprises or is M2e (aa 1-24 of M2 protein).

8. The fusion protein of claim 6 , wherein the fusion protein comprises (a) the fragment of CRM197, and (b) influenza virus M2 protein or the immunogenic fragment of M2 protein; wherein (a) and (b) are linked together, optionally via a linker.

9. The fusion protein of claim 6 , wherein the fusion protein has an amino acid sequence as set forth in SEQ ID NO: 36 or 42.

10. A polynucleotide encoding the fusion protein of claim 1 .

11. An expression vector comprising the polynucleotide of claim 10 .

12. A host cell comprising the polynucleotide of claim 10 .

13. A pharmaceutical composition or vaccine comprising the fusion protein of claim 1 and a pharmaceutically acceptable carrier and/or excipient.

14. A method for preventing and/or treating HEV infection or a disease associated with HEV infection, comprising administering an effective amount of the fusion protein of claim 4 or a pharmaceutical composition comprising the fusion protein.

15. The method of claim 14 , wherein the disease associated with HEV infection is Hepatitis E.

16. A method for preventing and/or treating influenza virus infection or a disease associated with influenza virus infection, comprising administering an effective amount of the fusion protein of claim 8 or a pharmaceutical composition comprising the fusion protein.

17. The method of claim 16 , wherein the disease associated with influenza virus infection is influenza.

Assignees

• Univ Xiamen
• Xiamen Innovax Biotech Co Ltd

Inventors

• Li Shaowei
• Song Cuiling
• Yang Chunyan
• Gu Ying
• Luo Wenxin
• Xia Ningshao

Classifications

• A61P31/16

  for influenza or rhinoviruses · CPC title

• A61P31/14

  for RNA viruses · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P37/04

  Immunostimulants · CPC title

• C12N2770/28134

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