Virus preparations and methods

• US 20060141566A1
• Filed: 12/23/2004
• Published: 06/29/2006
• Est. Priority Date: 12/23/2004
• Status: Active Grant

• Current Assignee: AuRx, Inc.
• Original Assignee: AuRx, Inc.
• Inventors: Calton, Gary J., Fishelevich, Rita
• Granted Patent: US 7,344,839 B2
• Time in Patent Office: Days
• US Class Current: 435/69.100
• CPC Class Codes: A61K 2039/525 Virus
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/245 Herpetoviridae, e.g. herpes simplex virus
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: C12N 2710/16634 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

Methods are disclosed for the preparation of herpesvirus, such as herpes simplex virus type 2 for vaccine use. Such viruses can be grown on serum free or serum containing media and can be prepared from the virus containing culture supernatant or virus containing cells. The virus is prepared for subsequent pharmaceutical formulation by methods which may include treatment with solid phase affinity reagents containing sulfate- or sulfonate-comprising binding groups. Such sulfated polysaccharide groups as heparin or dextran sulfate may be used, and eluted with salt solutions. The process can be combined with other culture, harvesting and formulation steps.

Claims

1. A process for preparing herpesvirus for a vaccine or therapy, which comprises:

(a) culturing host cells infected with the virus,

(b) separating of the medium from the infected cells and

(c) using the supernatant to provide a virus preparation suitable for immunotherapy or vaccination of an animal.

2. The process of claim 1 wherein said herpesvirus is an HSV-2 which has the PK domain deleted.

3. The process of claim 1 wherein said herpesvirus is ICP10deltaPK.

4. The process of claim 1 wherein said culture is carried out in a serum free medium.

5. The process of claim 1 wherein said culture is carried out in a medium containing low levels of protein.

6. The process of claim 1 where said culture is carried out in a medium with added epidermal growth factor.

7. The process of claim 1 where said culture is carried out in a medium with added insulin.

8. The process of claim 1 further comprising:

(d) contacting the herpesvirus-containing preparation to be purified, with an affinity binding reagent that comprises a solid phase carrying a sulfate or sulfonate-comprising binding group that can bind materials with affinity for heparin, thereby to bind said herpesvirus to the affinity binding reagent, and

(e) eluting said herpesvirus from said binding reagent.

9. A process according to claim 1, further comprising the step:

(d) formulating said herpesvirus with a pharmaceutically acceptable carrier, excipient or stabilizing agents.

10. A process according to claim 9 further comprising the step:

(e) sterilizing and freezing or freeze-drying the preparation.

11. A process according to claim 1 wherein the herpesvirus comprises infectious herpesvirus.

12. A process according to claim 1 wherein the resulting pharmaceutical formulation is for use as an immunotherapy.

13. A process according to claim 8, wherein said affinity binding reagent carries binding groups containing sulfate and nonionic polar groups.

14. A process according to claim 8, wherein said affinity binding reagent carries heparin or dextran sulfate.

15. A process according to claim 8, wherein said affinity binding reagent carries sulfated polysaccharide groups.

16. A process according to claim 8, wherein said affinity binding reagent carries pendent polyacrylamide chains substituted by sulfoisobutyl groups.

17. A process for growing herpesvirus comprising the step:

(a) culturing host cells infected with the virus in a serum free medium.

18. The process of claim 17 wherein the herpesvirus is an HSV-2 which has the PK domain deleted.