# US 7,744,899 B2 · Recombinant herpes simplex virus useful for treating neoplastic disease

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- Number: US 7,744,899 B2
- Document ID: US7744899B2
- Status: Granted
- Kind: B2
- Application number: 11/546,233
- Filed: 2006-10-11
- Published: 2007-02-08
- Granted: 2010-06-29
- Earliest claimed: 2006-10-11
- Assignees: The University of Chicago; The UAB Research Foundation
- Inventors: Richard J. Whitley; Bernard Roizman
- Web page: https://patentgrep.com/patent/US7744899B2

## 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 35/763: Herpes virus
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61P 35/00: Antineoplastic agents
- C07K 14/5406: {IL-4}
- C07K 14/5428: {IL-10}
- C12N 15/86: Viral vectors
- 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)
- C12N 2710/16632: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
- C12N 2710/16643: viral genome or elements thereof as genetic vector
- 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)

## Abstract

Recombinant herpes simplex viruses comprising DNA encoding cytokines and methods for treating neoplastic diseases using the inventive recombinant viruses are disclosed.

## Claims

### Claim 1

A recombinant herpes simplex virus that preferentially kills tumor cells and not healthy cells, that expresses only one γ134.5 gene copy and that comprises an expressible tumor-growth-inhibiting cytokine-encoding DNA.

### Claim 2 (depends on 1)

The recombinant herpes simplex virus of claim 1 wherein said virus lacks all or part of a γ134.5 gene copy.

### Claim 3 (depends on 2)

The recombinant herpes simplex virus of claim 2 wherein said virus comprises a γ134.5 gene having a deletion of a portion of a coding sequence of said γ134.5 gene, said deletion comprising a Bst EII-Stu I fragment of said γ134.5 gene.

### Claim 4 (depends on 1)

The recombinant herpes simplex virus of claim 1 wherein said virus comprises a γ134.5 gene having a stop codon at a Bst EII site in said γ134.5 gene.

### Claim 5 (depends on 1)

The recombinant herpes simplex virus of claim 1 wherein said expressible tumor-growth-inhibiting cytokine-encoding DNA is under the promoter-regulatory control of a herpes simplex virus gene promoter.

### Claim 6 (depends on 5)

The recombinant herpes simplex virus of claim 5 wherein said herpes simplex virus gene promoter is an EGR-1 promoter.

### Claim 7 (depends on 1)

The recombinant herpes simplex virus of claim 1 wherein said tumor-growth-inhibiting cytokine-encoding DNA is under the promoter-regulatory control of a synthetic herpes simplex virus-derived promoter.

### Claim 8 (depends on 7)

The recombinant herpes simplex virus of claim 7 wherein said synthetic herpes simplex virus-derived promoter comprises a herpes simplex virus α gene promoter fragment operatively linked 5′ to a herpes simplex virus γ gene promoter fragment.

### Claim 9 (depends on 8)

The recombinant herpes simplex virus of claim 8 wherein said α gene promoter fragment comprises promoter sequences upstream of the transcription initiation site of the α4 gene and said γ gene promoter fragment comprises a transcription initiation site and the 5′ transcribed non-coding sequence of the γ1UL9 gene.

### Claim 10 (depends on 1)

The recombinant herpes simplex virus of claim 1 wherein an unexpressed γ134.5 gene copy is replaced by said expressible tumor-growth-inhibiting cytokine-encoding DNA.

### Claim 11 (depends on 1)

The recombinant herpes simplex virus of claim 1 wherein said virus comprises two or more copies of said tumor-growth-inhibiting cytokine-encoding DNA.

### Claim 12 (depends on 10)

The recombinant herpes simplex virus of claim 10 where said herpes simplex virus is a HSV-1.

### Claim 13 (depends on 1)

The recombinant virus of claim 1 wherein said tumor-growth-inhibiting cytokine-encoding DNA further comprises a polyadenylation signal.

### Claim 14 (depends on 13)

The recombinant virus of claim 13 wherein said polyadenylation signal is a hepatitis B virus-derived polyadenylation signal.

### Claim 15

A method for treating neoplastic disease, the method comprising administering to a target tumor, a recombinant herpes simplex virus capable of expressing only one γ134.5 gene copy and comprising an expressible tumor-growth-inhibiting cytokine-encoding DNA, wherein the expressed cytokine augments tumor cell killing.

### Claim 16 (depends on 15)

The method of claim 15 wherein said recombinant herpes simplex virus lacks all or part of an unexpressed γ134.5 gene copy.

### Claim 17 (depends on 15)

The method of claim 15 wherein said recombinant herpes simplex virus comprises a γ134.5 gene having a stop codon at a Bst EII site in said γ134.5 gene.

### Claim 18 (depends on 15)

The method of claim 15 wherein said recombinant herpes simplex virus comprises a γ134.5 gene lacking a portion of the coding sequence corresponding to a Bst EII/Stu I restriction fragment of said γ134.5 gene.

### Claim 19 (depends on 15)

The method of claim 15 wherein said expressible tumor-growth-inhibiting cytokine-encoding DNA is under the promoter-regulatory control of a herpes simplex virus gene promoter.

### Claim 20 (depends on 19)

The method of claim 19 wherein said herpes simplex virus promoter is an EGR-1 promoter.

### Claim 21 (depends on 15)

The method of claim 15 wherein said tumor-growth-inhibiting cytokine-encoding DNA is under the promoter regulatory control of a synthetic herpes simplex virus-derived promoter.

### Claim 22 (depends on 21)

The method of claim 21 wherein said synthetic herpes simplex virus-derived promoter comprises a herpes simplex virus α gene fragment operatively linked 5′ to a herpes simplex virus γ gene promoter fragment.

### Claim 23 (depends on 22)

The method of claim 22 wherein said α gene promoter fragment comprises a promoter sequence upstream of the transcription initiation site of said α gene promoter fragment comprising the transcription initiation site and the 5′ transcribed non-coding sequence of the UL19 gene.

### Claim 24 (depends on 15)

The method of claim 15 wherein said γ134.5 gene is replaced by said expressible tumor-growth-inhibiting cytokine-encoding DNA.

### Claim 25

A pharmaceutical composition comprising in a pharmaceutically acceptable carrier, diluent, or adjuvant, a recombinant herpes simplex virus expressing only one γ134.5 gene copy, said virus comprising an expressible tumor-growth-inhibiting cytokine-encoding DNA, wherein the expressed cytokine augments tumor cell killing.

### Claim 26 (depends on 15)

The method of claim 15, wherein the target tumor is a tumor of the central nervous system.

### Claim 27 (depends on 3)

The recombinant herpes simplex virus of claim 3, wherein the expressible tumor growth inhibiting cytokine-encoding DNA is selected from DNA encoding IL-1, IL-2, IL-A, IL-6, IL-7, IFN-γ, GM-CSF or TNF-α.

### Claim 28 (depends on 15)

The method of claim 15, wherein the expressible tumor growth inhibiting cytokine-encoding DNA is selected from DNA encoding IL-1, IL-2, IL-4, IL-6, IL-7, IFN-γ, GM-CSF or TNF-α.