# US 11,299,715 B2 · Use of iNOS inhibitors to increase viral yield in culture

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- Number: US 11,299,715 B2
- Document ID: US11299715B2
- Status: Granted
- Kind: B2
- Application number: 14/758,785
- Filed: 2014-01-07
- Published: 2015-12-10
- Granted: 2022-04-12
- Earliest claimed: 2013-01-08
- Assignees: Genzyme Corporation
- Applicants: Genzyme Corporation
- Inventors: Peter Pechan; Jeffery Ardinger; Abraham Scaria; Samuel Wadsworth
- Web page: https://patentgrep.com/patent/US11299715B2

## 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)
- C12N 2710/16643: viral genome or elements thereof as genetic vector
- C12N 2710/16651: Methods of production or purification of viral material
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The use of iNOS inhibitors, including aurintricarboxylic acid, dexamethasone and valproic acid, to increase the yield of a variety of viruses in culture, including recombinant herpesviruses is described.

## Claims

### Claim 1

A method for producing herpes simplex-1 virus (HSV-1) comprising culturing cells infected with said herpes simplex-1 virus in a cell culture medium that comprises aurintricarboxylic acid and serum, wherein the aurintricarboxylic acid is unfractionated, wherein aurintricarboxylic acid is at a concentration of about 5 μM to about 75 μM, and wherein the cells are 293 or Vero cells.

### Claim 2 (depends on 1)

The method of claim 1, wherein said HSV-1 is a wild type HSV-1.

### Claim 3 (depends on 1)

The method of claim 1, wherein said HSV-1 is a recombinant HSV-1 .

### Claim 4 (depends on 3)

The method of claim 3, wherein said recombinant HSV-1 is HSV-1 d27.1.

### Claim 5 (depends on 1)

The method of claim 1, wherein said Vero cells are V27 cells.

### Claim 6

A method for producing HSV-1 d27.1 comprising:
(a) infecting V27 cells with said HSV-1 d27.1; and
(b) culturing said infected V27 cells in a cell culture medium comprising serum, and aurintricarboxylic acid, wherein aurintricarboxylic acid is at a concentration of about 5 μM to about 75 μM, and wherein aurintricarboxylic acid is unfractionated.

### Claim 7 (depends on 6)

The method of claim 6, wherein the serum is fetal bovine serum.

### Claim 8

A cell culture comprising aurintricarboxylic acid, serum and 293 or Vero cells infected with HSV-1, wherein aurintricarboxylic acid is unfractionated, wherein aurintricarboxylic acid is at a concentration of about 5 μM to about 75 μM.

### Claim 9 (depends on 8)

The cell culture of claim 8 wherein said Vero cells are V27 cells.