Oncolytic viruses and methods for treating neoplastic disorders

• US 8,679,509 B2
• Filed: 07/09/2010
• Issued: 03/25/2014
• Est. Priority Date: 07/10/2009
• Status: Active Grant

• Current Assignee: The Governors of the University of Alberta (University of Alberta)
• Original Assignee: The Governors of the University of Alberta (University of Alberta)
• Inventors: Evans, David H., Gammon, Don B.
• Primary Examiner(s): CHEN, SHIN LIN
• Application Number:
  US13/383,396
• Publication Number:
  US 20120114612A1
• Time in Patent Office: 1,355 Days
• Field of Search: 424/93.2, 424/205.1, 424/232.1, 435/320.1, 435/455
• US Class Current: 424/232.1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/17 having the group >N—C(O)—N< or >N—C(S)—N<, e.g. urea, thiourea, carmustine isoureas, isothioureas A61K31/155; sulfonylureas A61K31/64
• CPC Class Codes: A61K 31/7068 having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
• CPC Class Codes: A61K 31/7076 containing purines, e.g. adenosine, adenylic acid
• CPC Class Codes: A61K 35/768 Oncolytic viruses not provided for in groups A61K35/761 - A61K35/766
• CPC Class Codes: A61K 38/162 from virus
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 2710/24121 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2710/24132 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2710/24162 by genetic engineering
• 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

The disclosure provides mutant ribonucleotide reductase strains of poxviruses including for example vaccinia viruses. The disclosure also provides methods and for the use of these mutant ribonucleotide reductase strains of vaccinia viruses in oncolytic virotherapy.

Claims

1. An Orthopoxvirus comprising a gene encoding a modified R2 protein comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:

1,

wherein the modified R2 protein expressed by the Orthopoxvirus comprises a substitution of a tyrosine residue, wherein the substituted tyrosine residue corresponds to position 300 of SEQ ID NO;

1, or

wherein the modified R2 protein expressed by the Orthopoxvirus comprises a deletion of all or part of R1 binding domain (R1 BD); and

wherein the Orthopoxvirus does not express a wild type R2 protein.

2. The Orthopoxvirus of claim 1, wherein the modified R2 protein further comprises a substitution of an amino acid, wherein the substituted amino acid corresponds to one of the following positions in SEQ ID NO:

1;

W34, E38, D70, E101, H104, Y108, F167, F171, G181, I193, D196, E197, H200, Y254, and E294.

3. The Orthopoxvirus of claim 1, wherein the modified R2 protein comprises the substitution of the tyrosine residue, wherein the substituted tyrosine residue corresponds to position 300 of SEQ ID NO:

1.

4. The Orthopoxvirus of claim 1, wherein the modified R2 protein comprises the deletion of all or part of the R1BD.

5. The Orthopoxvirus of claim 4, wherein the modified R2 protein comprises a deletion of 2 amino acids of the R1BD.

6. The Orthopoxvirus of claim 4, wherein the modified R2 protein comprises a deletion of 7 amino acids of the R1BD.

7. The Orthopoxvirus of claim 1, wherein the poxvirus is vaccinia virus.

8. The Orthopoxvirus of claim 1, wherein the poxvirus further comprises a functionally inactivated R1 gene, thymidine kinase gene, vaccinia virus growth factor gene, or a nucleotide metabolism-related gene.

9. A composition comprising the Orthopoxvirus of claim 1, and a pharmaceutically acceptable diluent or carrier, optionally wherein the diluent is phosphate buffered saline.

10. A vector comprising the gene encoding the modified R2 protein of claim 1.

11. An in vitro method of making a recombinant Orthopoxvirus comprising:

introducing the vector of claim 10 into cells infected with an Orthopoxvirus, under conditions suitable for recombination; and

isolating a recombinant Orthopoxvirus comprising the gene.