# US 8,541,230 B2 · Polynucleotides encoding antigenic HIV type C polypeptides, polypeptides and uses thereof

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- Number: US 8,541,230 B2
- Document ID: US8541230B2
- Status: Granted
- Kind: B2
- Application number: 13/108,900
- Filed: 2011-05-16
- Published: 2011-09-29
- Granted: 2013-09-24
- Earliest claimed: 1998-12-31
- Assignees: Novartis Vaccines and Diagnostics, Inc.
- Applicants: Susan W. Barnett; Jan Zur Megede
- Inventors: Susan W. Barnett; Jan Zur Megede
- Web page: https://patentgrep.com/patent/US8541230B2

## CPC

- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A61P 31/18: for HIV
- A61P 37/02: Immunomodulators
- C07K 14/005: from viruses
- C12N 15/86: Viral vectors
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K 2039/5156: {expressing foreign proteins}
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/5258: {Virus-like particles}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/54: {characterised by the route of administration}
- C12N 2740/16023: Virus like particles [VLP]
- C12N 2740/16052: relating to complementing cells and packaging systems for producing virus or viral particles
- C12N 2740/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2740/16222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2740/16322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/24222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2800/108: episomal vectors
- C12N 2840/203: having an IRES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- 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 polynucleotides encoding immunogenic HIV type C Gag- and/or Env-containing polypeptides. Uses of the polynucleotides in applications including DNA immunization, generation of packaging cell lines, and production of Gag- and/or Env-containing proteins are also described.

## Claims

### Claim 1

An expression cassette comprising a polynucleotide sequence operably linked to a promoter, wherein the polynucleotide sequence comprises a nucleotide sequence having at least 90% sequence identity to full-length SEQ ID NO: 6 and encodes an immunogenic HIV Env polypeptide that elicits an Env-specific immune response.

### Claim 2

An expression cassette comprising a polynucleotide sequence operably linked to a promoter, wherein the polynucleotide sequence comprises a nucleotide sequence having at least 90% sequence identity to full-length SEQ ID NO:12 and encodes an immunogenic HIV Env polypeptide that elicits an Env-specific immune response.

### Claim 3 (depends on 1)

The expression cassette of claim 1, wherein the nucleotide sequence encoding said HIV Env polypeptide comprises SEQ ID NO:6.

### Claim 4 (depends on 2)

The expression cassette of claim 2, wherein the nucleotide sequence encoding said HIV Env polypeptide consists of SEQ ID NO:12.

### Claim 5 (depends on 1, 2)

The expression cassette of claim 1 or claim 2, wherein said polynucleotide sequence further includes a nucleotide sequence encoding an HIV protease polypeptide.

### Claim 6 (depends on 1, 2)

The expression cassette of claim 1 or claim 2, wherein said polynucleotide sequence further includes a nucleotide sequence encoding an HIV polymerase polypeptide.

### Claim 7 (depends on 1, 2)

The expression cassette of claim 1 or claim 2, wherein said polynucleotide sequence further includes a nucleotide sequence encoding an HIV polymerase polypeptide, wherein the nucleotide sequence encoding the HIV polymerase polypeptide is modified by deletions of coding regions encoding reverse transcriptase and integrase.

### Claim 8 (depends on 7)

The expression cassette of claim 7, wherein said polynucleotide sequence encodes a polypeptide comprising an epitope of a T-helper cell or a cytotoxic T cell (CTL).

### Claim 9 (depends on 1, 2)

A recombinant expression system for use in a selected host cell, comprising, the expression cassette of claim 1 or claim 2, and wherein said polynucleotide sequence is operably linked to control elements compatible with expression in the selected host cell.

### Claim 10 (depends on 9)

The recombinant expression system of claim 9, wherein said control elements are selected from the group consisting of a transcription promoter, a transcription enhancer element, a transcription termination signal, polyadenylation sequences, sequences for optimization of initiation of translation, and translation termination sequences.

### Claim 11 (depends on 10)

The recombinant expression system of claim 10, wherein said transcription promoter is selected from the group consisting of CMV, CMV+intron A, SV40, RSV, HW-Ltr, MMLV-ltr, and metallothionein.

### Claim 12 (depends on 1, 2)

An isolated cell comprising the expression cassette of claim 1 or claim 2, and wherein said polynucleotide sequence is operably linked to control elements compatible with expression in the selected cell.

### Claim 13 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is a mammalian cell.

### Claim 14 (depends on 13)

The isolated cell of claim 13, wherein the isolated cell is selected from the group consisting of BRK, VERO, HT1080, 293, RD, COS-7, and CHO cells.

### Claim 15 (depends on 14)

The isolated cell of claim 14, wherein said isolated cell is a CHO cell.

### Claim 16 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is an insect cell.

### Claim 17 (depends on 16)

The isolated cell of claim 16, wherein the isolated cell is either Trichoplusia ni (Tn5) or Sf9 insect cells.

### Claim 18 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is a bacterial cell.

### Claim 19 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is a yeast cell.

### Claim 20 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is a plant cell.

### Claim 21 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is an antigen presenting cell.

### Claim 22 (depends on 21)

The isolated cell of claim 21, wherein the isolated antigen presenting cell is selected from the group consisting of macrophage, monocytes, dendritic cells, B-cells, T-cells, stem cells, and progenitor cells thereof.

### Claim 23 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is a primary cell.

### Claim 24 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is an immortalized cell.

### Claim 25 (depends on 12)

The isolated cell of claim 12, wherein the isolated cell is a tumor-derived cell.

### Claim 26 (depends on 1, 2)

A composition for generating an immunological response, comprising: the expression cassette of claim 1 or claim 2 with a pharmaceutically acceptable excipient or vehicle.

### Claim 27 (depends on 26)

The composition of claim 26, further comprising an Env polypeptide.

### Claim 28 (depends on 26)

The composition of claim 26, further comprising an adjuvant.

### Claim 29 (depends on 26)

A method of generating an immune response in a subject, comprising, introducing the composition of claim 26 into said subject such that the expression cassette is expressed in said subject.

### Claim 30 (depends on 29)

The method of claim 29, wherein said expression cassette is introduced using a gene delivery vector.

### Claim 31 (depends on 30)

The method of claim 30, wherein the gene delivery vector is a non-viral vector.

### Claim 32 (depends on 30)

The method of claim 30, wherein said gene delivery vector is a viral vector.

### Claim 33 (depends on 32)

The method of claim 32, wherein said gene delivery vector is a Sindbis-virus derived vector.

### Claim 34 (depends on 32)

The method of claim 32, wherein said gene delivery vector is a retroviral vector.

### Claim 35 (depends on 32)

The method of claim 32, wherein said gene delivery vector is a lentiviral vector.

### Claim 36 (depends on 29)

The method of claim 29, wherein said composition is delivered using a particulate carrier.

### Claim 37 (depends on 29)

The method of claim 29, wherein said composition is coated on a gold or tungsten particle and said coated particle is delivered to said subject using a gene gun.

### Claim 38 (depends on 29)

The method of claim 29, wherein said composition is encapsulated in a liposome preparation.

### Claim 39 (depends on 29)

The method of claim 29, wherein said subject is a mammal.

### Claim 40 (depends on 29)

The method of claim 29, wherein said mammal is a human.

### Claim 41 (depends on 29)

The method of claim 29, where the method further comprises administration of an HIV polypeptide.

### Claim 42 (depends on 41)

The method of claim 41, wherein administration of the polypeptide to the subject is carried out before introducing said expression cassette.

### Claim 43 (depends on 41)

The method of claim 41, wherein administration of the polypeptide to the subject is carried out concurrently with introducing said expression cassette.

### Claim 44 (depends on 41)

The method of claim 41, wherein administration of the polypeptide to the subject is carried out after introducing said expression cassette.