COMPOSITIONS AND METHODS OF USE OF AN ONCOLYTIC VESICULAR STOMATITIS VIRUS

• US 20100172877A1
• Filed: 01/07/2010
• Published: 07/08/2010
• Est. Priority Date: 01/08/2009
• Status: Active Grant

• Current Assignee: Yale University
• Original Assignee: Yale University
• Inventors: van den Pol, Anthony N., Wollmann, Guido
• Granted Patent: US 8,481,297 B2
• Time in Patent Office: Days
• US Class Current: 424/93.600
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2760/20222 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/20232 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent

Abstract

Oncolytic VSV viruses have been developed as a strategy for combating cancer. The present invention includes mutant VSV that have one or more mutations in the nucleic acid sequence encoding the viral genome that increase the oncolytic potential of the virus. Pharmaceutical compositions including oncolytic virus disclosed herein are also provided. Pharmaceutical compositions containing virus and one or more excipients may be for systemic or local administration. Methods of administering an effective amount of the compositions for treating cancer are disclosed. Preferred routes of administration include intratumeral and intravenous injection, and intranasal delivery. Administration of the disclosed compositions containing oncolytic viruses may be coupled with surgical, radiologic, other therapeutic approaches to treatment of cancer. Methods of manufacturing mutant VSV viruses exhibiting desired properties include applying selective pressure, and through directed or random mutagenesis.

Claims

1. An isolated oncolytic VSV virus comprising one or more changes in nucleic acid sequence resulting in substitution of the serine at codon 126 SEQ ID NO. 3, with an amino acid selected from the group consisting of alanine (A), arginine (R), asparagine (N), aspartic acid (D), cysteine (C), glutamine (Q), glutamic acid (E), glycine (G), histidine (H), isoleucine (I), leucine (L), lysine (K), methionine (M), phenylalanine (F), proline (P), tryptophan (W), or valine (V) and substitution of aspartic acid (D) at codon 223 of SEQ ID NO. 5 with an amino acid selected from the group consisting of serine (S), threonine (T), or tyrosine (Y).

2. The isolated oncolytic VSV virus of claim 1, wherein the serine (S) at codon 126 SEQ ID NO. 3, is substituted with leucine (L) and the aspartic acid (D) at codon 223 of SEQ ID NO. 5 is substituted with a tyrosine (Y).

3. An isolated oncolytic VSV virus with SEQ ID NO:

6.

4. An isolated oncolytic vesicular stomatitis virus (VSV) virus, comprising one or more mutations in the nucleic acid sequence encoding SEQ ID NO:

2 or SEQ ID NO;

4 that alter the post translational phosphorylation of the protein encoded by SEQ ID NO;

3 or SEQ ID;

5, or a combination thereof, in the presence of a kinase capable of phosphorylating amino acids in SEQ ID NO;

3 or SEQ ID;

5.

5. The isolated oncolytic VSV virus of claim 4, wherein the one or more changes in nucleic acid sequence increase the post translational phosphorylation of the protein encoded by SEQ ID NO:

3 or SEQ ID;

5, or a combination thereof.

6. The isolated oncolytic VSV virus of claim 4, wherein the one or more changes in nucleic acid sequence decrease the post translational phosphorylation of the protein encoded by SEQ ID NO:

3 or SEQ ID;

5, or a combination thereof.

7. The isolated oncolytic VSV virus of claim 4, wherein the one or more changes in nucleic acid sequence increase the post translational phosphorylation of the protein encoded by SEQ ID NO:

3 and decrease the post translational phosphorylation of the protein encoded by SEQ ID;

5.

8. The isolated oncolytic VSV virus of claim 4, wherein the one or more changes in nucleic acid sequence decrease the post translational phosphorylation of the protein encoded by SEQ ID NO:

3 and increase the post translational phosphorylation of the protein encoded by SEQ ID;

5.

9. An isolated oncolytic VSV virus, wherein the virus is engineered to express one or more additional genes encoding proteins selected from the group consisting of targeting proteins and therapeutic proteins.

10. An isolated oncolytic VSV virus, wherein one or more viral genes are rearranged, deleted, or inserted to alter the oncolytic potential of the virus.

11. The isolated oncolytic VSV virus of claim 10, wherein a gene not encoded by SEQ ID NO. 2 or SEQ ID NO. 4, is substituted for a variant gene that alters the oncolytic potential of the virus.

12. A sterile pharmaceutical dosage unit composition comprising an effective amount of an oncolytic virus to reduce tumor burden.

13. The dosage unit of claim 12, in an amount of virus effective to systemically treat an individual having metastastic cancer.

14. The dosage unit of claim 12, comprising an amount of virus effective to locally or regionally treat a tumor.

15. A method for treating cancer comprising administering to a subject in need thereof an effective amount of the viral dosage unit of claims 12.

16. The method of claim 15, wherein the cancer is selected from the group consisting of bone, bladder, brain, breast, cervical, colo-rectal, esophageal, kidney, liver, lung, nasopharangeal, pancreatic, prostate, skin, stomach, and uterine.

17. The method of claim 15, wherein the cancer is a vascular cancer such as multiple myeloma, an adenocarcinomas or a sarcoma.

18. The method of claim 16, comprising administering a viral dosage unit effective to treat a glioma.

19. The method of claim 15, wherein the viral dosage unit is administered intratumorally or parenterally.

20. The method of claims 15, wherein the viral dosage unit is administered in combination with a second therapeutic agent selected from the group consisting of immunosuppressants, anticancer agents, and therapeutic proteins.