# US 10,765,731 B2 · Fusion of heterooligomeric mycobacterial antigens

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- Number: US 10,765,731 B2
- Document ID: US10765731B2
- Status: Granted
- Kind: B2
- Application number: 15/110,514
- Filed: 2015-01-09
- Published: 2016-11-17
- Granted: 2020-09-08
- Earliest claimed: 2014-01-09
- Assignees: Transgene S.A.
- Applicants: Transgene SA
- Inventors: Jean-Baptiste Marchand; Nathalie Silvestre; François Penin
- Web page: https://patentgrep.com/patent/US10765731B2

## CPC

- A61K 39/04: Mycobacterium, e.g. Mycobacterium tuberculosis
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 31/04: Antibacterial agents
- A61P 31/06: for tuberculosis
- A61P 37/04: Immunostimulants
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/35: from Mycobacteriaceae (F)
- C07K 16/1289: Mycobacteriaceae (F)
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/57: {characterised by the type of response, e.g. Th1, Th2}
- C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- C07K 2319/00: Fusion polypeptide
- C07K 2319/035: containing a signal for targeting to the external surface of a cell, e.g. to the outer membrane of Gram negative bacteria, GPI- anchored eukaryote proteins
- C12N 2710/24141: Use of virus, viral particle or viral elements as a vector
- 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
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates generally to novel immunogenic combinations comprising or encoding at least two heterooligomeric mycobacterial antigens and preferably a fusion polypeptide comprising said two heterooligomeric mycobacterial antigens, where the mycobacterial antigens are selected from the group of Esx, PE and PPE antigens of a Mycobacterium species, particularly a Mycobacterium of the tuberculosis complex such as Mycobacterium tuberculosis (Mtb). The present invention also relates to vectors, host cells and compositions comprising or encoding said immunogenic combination as well as to methods for expressing and producing it. The present invention also relates to methods of using said immunogenic combination, fusion polypeptide, vector, host cell, composition particularly for inducing or stimulating an immune response with the goal of providing a protective response against a Mycobacterium infection or any disease caused by or associated with a Mycobacterium infection.

## Claims

### Claim 1

A viral vector comprising:
a nucleic acid molecule encoding a fusion of the heterooligomeric mycobacterial antigens ESAT6 with CFP10, and
nucleic acid molecule(s) encoding one or more additional mycobacterial antigen(s) selected from the group consisting of Ag85B (Rv1886), RpfB, RpfD, Rv1813, Rv2626, Rv3407, and a mycobacterial antigen comprising an amino acid sequence as set forth in SEQ ID NO: 3 or 4.

### Claim 2 (depends on 1)

The viral vector of claim 1, wherein said vector is a poxvirus vector selected from the group consisting of fowlpox, canarypox and vaccinia virus vector.

### Claim 3 (depends on 1)

The viral vector according to claim 1 which is selected from the group consisting of:
(i) A viral vector comprising:
a) a nucleic acid molecule encoding a fusion comprising heterooligomeric mycobacterial antigens ESAT-6 and CFP10,
b) a nucleic acid molecule encoding Ag85B, and
c) a nucleic acid molecule encoding Rv2626; and
(ii) A viral vector comprising:
a) a nucleic acid molecule encoding a fusion comprising heterooligomeric mycobacterial antigens ESAT-6 and CFP10;
b) a nucleic acid molecule encoding a fusion comprising heterooligomeric mycobacterial antigens TB10.4 and TB9.8,
c) a nucleic acid molecule encoding a fusion polypeptide comprising the additional mycobacterial antigens Rv2626 and Ag85B;
d) a nucleic acid molecule encoding a fusion polypeptide comprising the additional mycobacterial antigens RpfB and RpfD; and
e) a nucleic acid molecule encoding a fusion polypeptide comprising the additional mycobacterial antigens Rv3407 and Rv1813.

### Claim 4 (depends on 1)

The viral vector according to claim 1, wherein said nucleic acid molecules are placed under the transcriptional control of a one or more promoter(s) suitable for ensuring expression of the encoded fusion and additional mycobacterial antigen(s) in a mammalian cell.

### Claim 5 (depends on 1)

A host cell comprising the viral vector according to claim 1.

### Claim 6 (depends on 1, 5)

A composition comprising at least one of the viral vector of claim 1 or at least one of the host cell of claim 5.

### Claim 7 (depends on 6)

The composition of claim 6, wherein the composition further comprises a pharmaceutically acceptable vehicle.

### Claim 8 (depends on 6)

A method for treating a Mycobacterium infection, or any disease and pathologic condition caused by or associated with such a Mycobacterium infection, in a subject in need thereof, comprising the step of administering a therapeutically effective amount of the composition of claim 6 to the subject.

### Claim 9 (depends on 6)

A method for delaying the risk of infection with a Mycobacterium in a subject in need thereof, comprising the step of administering a therapeutically effective amount of the composition of claim 6 to the subject.

### Claim 10 (depends on 8)

The method of claim 8, for treating an active disease in a subject infected with a Mycobacterium species.

### Claim 11 (depends on 8)

The method of claim 8, for treating reactivation in a subject latently infected with a Mycobacterium.

### Claim 12 (depends on 8)

The method of claim 8, wherein the subject has previously been immunized with Bacille-Calmette-Guerin (BCG).

### Claim 13 (depends on 8)

The method of claim 8, further comprising the administration of one or more chemotherapeutic drug(s) effective against a Mycobacterium infection.

### Claim 14 (depends on 8)

The method of claim 8, for inducing or enhancing an immune response in the administered subject.

### Claim 15 (depends on 14)

The method of claim 14, wherein said induced or stimulated immune response is a CD4+ and/or CD8+-mediated T cell response directed to a mycobacterial antigen/epitope.

### Claim 16 (depends on 1)

The viral vector according to claim 1, further comprising nucleic acid molecule(s) encoding a fusion of the heterooligomeric mycobacterial antigens TB10.4 (Rv0288) with TB9.8 (Rv0287).

### Claim 17 (depends on 1)

The viral vector according to claim 1, further comprising nucleic acid molecule(s) encoding one or more other additional mycobacterial antigen(s).

### Claim 18 (depends on 1)

The viral vector according to claim 1, wherein said vector comprises a nucleic acid molecule encoding a fusion of the heterooligomeric mycobacterial antigens ESAT6 with CFP10, and
a) nucleic acid molecule(s) encoding mycobacterial antigens Rv2626 and Ag85B; or
b) a nucleic acid molecule(s) encoding a fusion of the heterooligomeric mycobacterial antigens TB10.4 and TB9.8 and nucleic acid molecule(s) encoding mycobacterial antigens Rv2626, Ag85B, RpfB, RpfD, Rv3407 and Rv1813.

### Claim 19 (depends on 1)

The viral vector according to claim 1, wherein said additional mycobacterial antigen(s) are encoded in the form of separate polypeptides and/or in the form of fusion of two.

### Claim 20 (depends on 19)

The viral vector according to claim 19, wherein said fusion polypeptide of additional mycobacterial antigens comprises (i) Rv2626 and Ag85B, (ii) RpfB and RpfD, or (iii) Rv3407 and Rv1813.

### Claim 21 (depends on 20)

The viral vector according to claim 20, wherein said fusion polypeptide of additional mycobacterial antigens comprises:
a) one or more peptide(s) to ensure appropriate folding of said mycobacterial antigens such as a protease cleavage site inserted between the upstream and downstream polypeptides of said fusion polypeptide; or
b) an amino acid sequence exhibiting at least 70% identity with any of the amino acid sequences shown in SEQ ID NO: 8 to SEQ ID NO: 12.

### Claim 22 (depends on 21)

A composition comprising at least one viral vector according to claim 21.

### Claim 23 (depends on 21)

A method of treating a Mycobacterium infection in a subject in need thereof, comprising administering a therapeutically effective amount of a composition comprising the vector of claim 21 to the subject.