# US 7,303,898 B2 · Defective sindbis viral vectors

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- Number: US 7,303,898 B2
- Document ID: US7303898B2
- Status: Granted
- Kind: B2
- Application number: 11/392,926
- Filed: 2006-03-28
- Published: 2007-01-25
- Granted: 2007-12-04
- Earliest claimed: 2005-03-29
- Assignees: New York University
- Inventors: Alicia Hurtado; Daniel Meruelo
- Web page: https://patentgrep.com/patent/US7303898B2

## 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)
- A61K 35/76: Viruses; Subviral particles; Bacteriophages
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/005: from viruses
- C12N 15/86: Viral vectors
- A01K 2267/0331: Animal model for proliferative diseases
- C12N 2770/36122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/36132: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
- C12N 2770/36143: viral genome or elements thereof as genetic vector
- C12N 2770/36161: Methods of inactivation or attenuation
- 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)
- 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)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS

## Abstract

Disclosed herein are new defective Sindbis viral vectors made from wild type Ar-339 Sindbis virus, with differences in replicase and envelope proteins between JT vectors and consensus Sindbis virus sequences, and also between JT and Ar-339 vectors. Also disclosed are plasmids used for the production of the vectors, methods for producing the vectors, methods for treating mammals suffering from tumors and pharmaceutical formulations for use in the treatment methods.

## Claims

### Claim 1

A purified, isolated nucleic acid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO. 39 (SP6-H-I3-K70-E181-Val 251).

### Claim 2

A purified, isolated nucleic acid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO. 36 (SP6-R).

### Claim 3

A method for producing defective Sindbis viral vectors comprising the steps of
(a) providing a linearized replicon plasmid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO: 36 and a linearized Helper plasmid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO: 39;
(b) transcribing said replicon plasmid and said Helper plasmid to produce RNA;
(c) collecting the RNA transcribed in step (b) from said replicon plasmid and said Helper plasmid and transfecting cells with said RNA;
(d) incubating said transfected cells for a time and at a temperature effective for producing defective Sindbis viral vectors; and
(e) collecting said defective Sindbis viral vectors from the medium of said transfected.