# US 2014/0154285 A1 · Fusion proteins for use as immunogenic enhancers for inducing antigen-specific t cell responses

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- Number: US 2014/0154285 A1
- Document ID: US20140154285A1
- Status: Published application
- Kind: A1
- Application number: 14/095,760
- Filed: 2013-12-03
- Published: 2014-06-05
- Earliest claimed: 2012-12-05
- Assignees: TheVax Genetics Vaccine Co., Ltd.
- Applicants: TheVax Genetics Vaccine Co., Ltd.
- Inventors: Chia-Mao WU; Hsiu-Kang Chang
- Web page: https://patentgrep.com/patent/US20140154285A1

## CPC

- C07K 14/005: from viruses
- A61K 39/12: Viral antigens
- A61K 39/135: Foot- and mouth-disease virus
- A61K 39/17: Newcastle disease virus
- A61K 39/21: Retroviridae, e.g. equine infectious anemia virus
- A61K 39/29: Hepatitis virus
- A61K 39/292: {Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen}
- A61K 39/385: Haptens or antigens, bound to carriers
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 15/00: Drugs for genital or sexual disorders (for disorders of sex hormones A61P5/24); Contraceptives
- A61P 17/00: Drugs for dermatological disorders
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/04: Antibacterial agents
- A61P 31/12: Antivirals
- A61P 31/14: for RNA viruses
- A61P 31/16: for influenza or rhinoviruses
- A61P 31/18: for HIV
- A61P 31/20: for DNA viruses
- A61P 31/22: for herpes viruses
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- A61P 37/00: Drugs for immunological or allergic disorders
- A61P 37/04: Immunostimulants
- C07K 14/70596: {Molecules with a "CD"-designation not provided for elsewhere}
- C07K 14/81: Protease inhibitors
- C12N 9/1077: {Pentosyltransferases (2.4.2)}
- C12N 9/14: Hydrolases (3)
- C12Y 204/02036: NAD(+)--diphthamide ADP-ribosyltransferase (2.4.2.36)
- C12Y 306/05002: Small monomeric GTPase (3.6.5.2)
- A61K 2039/57: {characterised by the type of response, e.g. Th1, Th2}
- A61K 2039/585: {wherein the target is cancer}
- A61K 2039/6031: {Proteins}
- C07K 14/21: from Pseudomonadaceae (F)
- C07K 2319/02: containing a signal sequence
- C07K 2319/04: containing an ER retention signal such as a C-terminal HDEL motif
- C07K 2319/095: containing a nuclear export signal
- C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation
- C07K 2319/55: containing a fusion with a toxin, e.g. diphteria toxin
- C12N 2710/20022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2710/20034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2730/10122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2730/10134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2750/10022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2750/10034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2760/18134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/24222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/24234: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/24334: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/32134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

A fusion protein for use as an immunogen enhancer for enhancing antigen-specific T cell responses is disclosed. The fusion protein comprises: (a) an antigen-presenting cell (APC)-binding domain or a CD91 receptor-binding domain; (b) a protein transduction domain; and (c) an antigen of a pathogen, wherein the APC-binding domain or the CD91 receptor-binding domain is located at the N-terminus of the fusion protein, and the antigen of the pathogen is located at the C-terminus of the protein transduction domain. The protein transduction domain is selected from the group consisting of: (i) a fusion polypeptide, comprising a T cell sensitizing signal-transducing peptide, a linker, and a translocation peptide; (ii) a T cell-sensitizing signal-transducing peptide; and (iii) a translocation peptide of 34-112 amino acid residues in length.

## Claims

### Claim 1

A fusion protein comprising:
(a) an antigen-presenting cell (APC)-binding domain or a CD91 receptor-binding domain, located at the N-terminus of the fusion protein;
(b) a protein transduction domain, located at the C-terminus of the APC-binding domain or the CD91 receptor-binding domain, the protein transduction domain being selected from the group consisting of:
(i) a fusion polypeptide, comprising a T cell sensitizing signal-transducing peptide, a linker, and a translocation peptide, wherein:
(1) the T cell sensitizing signal-transducing peptide is located at the N-terminus of the fusion polypeptide;
(2) the linker comprises SEQ ID NO: 15, linking the T cell sensitizing signal-transducing peptide and the translocation peptide; and
(3) the translocation peptide has 34-112 amino acid residues in length and comprises the amino acid sequence that is at least 90% identical to SEQ ID NO: 3, 20 or 4;
(ii) a T cell-sensitizing signal-transducing peptide; and
(iii) a translocation peptide of 34-112 amino acid residues in length, comprising the amino acid sequence that is at least 90% identical to SEQ ID NO: 3, 20 or 4; and
(c) an antigen of a pathogen, located at the C-terminus of the protein transduction domain;
wherein:
the T cell-sensitizing signal-transducing peptide has 28-53 amino acid residues in length and comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 31, in which Xaa8 is I or L; Xaa10 is V, F or A, Xaa11 is M or L, Xaa17 is L or I; and
the APC-binding domain or the CD91 receptor-binding domain is free of the amino acid sequence of Pseudomonas exotoxin A (PE) binding domain II if the protein transduction domain is the translocation peptide (biii).

### Claim 2 (depends on 1)

The fusion protein of claim 1, wherein the APC-binding domain or the CD91 receptor-binding domain is a polypeptide comprising an amino acid sequence that is at least 90% identical to the sequence selected from the group consisting of SEQ ID NOs: 5, 9, 6, 7, and 8.

### Claim 3 (depends on 1)

The fusion protein of claim 1, further comprising an endoplasmic reticulum retention sequence located at the C-terminus of the fusion protein.

### Claim 4 (depends on 3)

The fusion protein of claim 3, wherein the endoplasmic reticulum retention sequence comprises the amino acid sequence of Lys-Asp-Glu-Leu (SEQ ID NO: 14).

### Claim 5 (depends on 1)

The fusion protein of claim 1, wherein the fusion protein is free of an endoplasmic reticulum retention sequence at C-terminus thereof if the antigen contains 10 or more epitopes.

### Claim 6 (depends on 1)

The fusion protein of claim 1, wherein the protein transduction domain is the fusion polypeptide (bi).

### Claim 7 (depends on 1)

The fusion protein of claim 1, wherein the protein transduction domain is the T cell-sensitizing signal-transducing peptide (bii).

### Claim 8 (depends on 7)

The fusion protein of claim 7, further comprising an additional linker between the protein transduction domain and the antigen, the additional linker comprising SEQ ID NO: 15.

### Claim 9 (depends on 1)

The fusion protein of claim 1, wherein the protein transduction domain is the translocation peptide (biii).

### Claim 10 (depends on 9)

The fusion protein of claim 9, further comprising an additional linker between the APC-binding domain or the CD91 receptor-binding domain and the translocation peptide, the additional linker comprising SEQ ID NO: 15.

### Claim 11 (depends on 1)

The fusion protein of claim 1, wherein the protein transduction domain comprises the sequence of SEQ ID NO: 30.

### Claim 12 (depends on 1)

A vaccine composition comprising:
(a) a therapeutically effective amount of the fusion protein of claim 1; and
(b) an adjuvant.

### Claim 13 (depends on 1)

The fusion protein of claim 1, wherein the APC-binding domain or the CD91 receptor-binding domain is a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 9, 6, 7, and 8.

### Claim 14 (depends on 1)

The fusion protein of claim 1, wherein the T cell sensitizing signal-transducing peptide comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 1 or 2.

### Claim 15 (depends on 14)

The fusion protein of claim 14, wherein the T cell sensitizing signal-transducing peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1 and 2.

### Claim 16 (depends on 1)

The fusion protein of claim 1, wherein the translocation peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 3, 20, and 4.

### Claim 17 (depends on 1)

A method for inducing enhanced pathogen antigen-specific T cell responses, comprising:
administering a vaccine composition comprising a therapeutically effective amount of the fusion protein of claim 1 to a subject in need thereof, and thereby inducing enhanced pathogen antigen-specific T cell responses.

### Claim 18 (depends on 1)

A method for killing a disease cell that presents an antigen via class I MHC molecules on the cell membrane of the disease cell, comprising:
administering a vaccine composition comprising a therapeutically effective amount of the fusion protein of claim 1 to a subject in need thereof, and thereby killing the disease cell that that presents the antigen via class I MHC molecules on the cell membrane of the disease cell.

### Claim 19 (depends on 18)

The method of claim 18, wherein the disease cell is a cancer cell.

### Claim 20 (depends on 1)

A method for preventing, treating infection caused by a pathogen, and/or minimizing symptoms caused by the infection, comprising:
administering a vaccine composition comprising a therapeutically effective amount of the fusion protein of claim 1 to a subject in need thereof, and thereby preventing, treating infection caused by the pathogen, and/or minimizing symptoms caused by the infection.