# US 2018/0022781 A1 · Polypeptides for engineering integrase chimeric proteins and their use in gene therapy

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- Number: US 2018/0022781 A1
- Document ID: US20180022781A1
- Status: Published application
- Kind: A1
- Application number: 15/549,531
- Filed: 2016-02-12
- Published: 2018-01-25
- Earliest claimed: 2015-02-13
- Assignees: Inserm (Institut National De La Sante Et De La Recherche Medicale); Centre National de la Recherche Scientifique (CNRS); Universite Paris Diderot—Paris 7; Commissariat A L'energie Atomique Et Aux Energies Alternatives; CNAM - Conservatoire National Des Arts Et Metiers
- Applicants: Inserm (Institut National De La Sante Et De La Recherche Medicale); Centre National De La Recherche Scientifique (Cnrs); Universite Paris Diderot - Paris 7; Commissariat A L'energie Atomique Et Aux Energies Alternatives; Cnam - Conservatoire National Des Arts Et Metiers
- Inventors: Antoine BRIDIER-NAHMIAS; Michel WERNER; Pascale LESAGE
- Web page: https://patentgrep.com/patent/US20180022781A1

## CPC

- C07K 14/005: from viruses
- C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]}
- C12N 15/62: DNA sequences coding for fusion proteins
- C12N 15/867: Retroviral vectors
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- C07K 19/00: Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes}
- C07K 2319/00: Fusion polypeptide
- C12N 2740/13022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2740/15022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- 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)
- 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)

## Abstract

The present invention relates to polypeptide for engineering integrase chimeric proteins and their use in gene therapy. In particular, the present invention relates to a polypeptide which comprises the amino acid sequence ranging from the amino acid residue at position 617 to the amino acid residue at position 622 in SEQ ID NO: 1 or a function conservative thereof.

## Claims

### Claim 1

A polypeptide which comprises an amino acid sequence ranging from the amino acid residue at position 617 to the amino acid residue at position 622 in SEQ ID NO:1 or a function conservative thereof.

### Claim 2 (depends on 1)

The polypeptide of claim 1 wherein the function-conservative variant comprises an amino acid sequence having 50; 51; 52; 53; 54; 55; 56; 57; 58; 59; 60; 61; 62; 63; 64; 65; 66; 67; 68; 69; 70; 71; 72; 73; 74; 75; 76; 77; 78; 79; 80; 81; 82; 83; 84; 85; 86; 87; 88; 89; 90; 91; 92; 93; 94; 95; 96; 97; 98; or 99% identity with the amino acid sequence ranging from the amino acid residue at position 617 to the amino acid residue at position 622 in SEQ ID NO:1.

### Claim 3 (depends on 1)

The polypeptide of claim 1 wherein an amino acid at position 618 or 620 is substituted by any amino acid.

### Claim 4 (depends on 1)

The polypeptide of claim 1 wherein an amino acid residue at position 618 is substituted by an alanine (A).

### Claim 5 (depends on 1)

The polypeptide of claim 1 wherein an amino acid residue at position 620 is substituted by an aspartic acid residue (D).

### Claim 6 (depends on 1)

The polypeptide of claim 1 which comprises an amino acid sequence as set forth in SEQ ID NO:3 (KEMDSL).

### Claim 7 (depends on 1)

The polypeptide of claim 1 which comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:12 and amino acid sequences having at least 50% of identity with SEQ ID NO:2 to SEQ ID NO:12.

### Claim 8 (depends on 1)

The polypeptide of claim 1 which is fused to a heterologous polypeptide.

### Claim 9 (depends on 1)

A chimeric integrase which comprises the polypeptide of claim 1.

### Claim 10 (depends on 9)

The chimeric integrase of claim 9 which comprises a retroviral integrase selected from the group consisting of feline immunodeficiency virus (FIV) integrases, Foamy virus (FV) integrases, murine leukemia virus (MLV) integrases, and lentivirus integrases.

### Claim 11 (depends on 10)

The chimeric integrase of claim 10 wherein the retroviral integrase has a sequence having at least 80% of identity with SEQ ID NO: 13 or SEQ ID NO:14.

### Claim 12 (depends on 9)

The chimeric integrase of claim 9 wherein the polypeptide is fused at the C-terminal end of the integrase protein or at the N-terminal end of the integrase protein.

### Claim 13 (depends on 9)

The chimeric integrase of claim 9 wherein the polypeptide is made part of a retroviral integrase.

### Claim 14 (depends on 1)

A nucleic acid molecule which encodes a polypeptide of claim 1 or a chimeric integrase comprising the polypeptide.

### Claim 15 (depends on 9)

A retroviral vector which comprises a nucleic acid molecule encoding the chimeric integrase of claim 9.

### Claim 16 (depends on 15)

The retroviral vector of claim 15 which is derived from a retrovirus selected from the group consisting of alpharetroviruses, betaretroviruses, gammaretroviruses, deltaretroviruses, epsilonretroviruses, lentiviruses and spumaviruses.

### Claim 17 (depends on 15)

The retroviral vector of claim 15 which comprises a transgene.

### Claim 18 (depends on 17)

A method for expressing a transgene from the genome of a plurality of cells comprising infecting the plurality of cells with the retroviral vector of claim 17 under conditions which i) result in integration of the retroviral vector into the genome of the plurality of cells and ii) permit expression of the transgene from the integrated retroviral vector.

### Claim 19 (depends on 17)

A method of performing gene therapy in a subject in need thereof, comprising
providing the subject with a transgene by administering to the subject the retroviral vector of claim 17, under conditions that allow expression of the transgene within the subject.