# US 12,758,601 B2 · Oncolytic adenovirus with replication selectivity based on status of P53 transcriptional activity

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- Number: US 12,758,601 B2
- Document ID: US12758601B2
- Status: Granted
- Kind: B2
- Application number: 18/151,081
- Filed: 2023-01-06
- Published: 2023-07-13
- Granted: 2026-10-13
- Earliest claimed: 2020-07-06
- Assignees: The Salk Institute for Biological Studies
- Applicants: Salk Institute for Biological Studies
- Inventors: William Partlo; Clodagh O'Shea
- Web page: https://patentgrep.com/patent/US12758601B2

## CPC

- A61K 35/761: Adenovirus
- 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 2710/10321: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2710/10332: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
- C12N 2830/005: repressible enhancer/promoter combination, e.g. KRAB
- C12N 2830/008: cell type or tissue specific enhancer/promoter combination
- C12N 2830/15: chimeric enhancer/promoter combination
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Oncolytic adenoviruses capable of selectively replicating in tumor cells deficient in p53 transcriptional activity are described. Recombinant adenovirus genomes that encode the p53-selective adenoviruses are also described. The recombinant adenoviruses and adenovirus genomes, or compositions thereof, can be used, for example, to reduce or inhibit tumor progression, or reduce tumor volume in a subject with a tumor having dysregulated p53 activity.

## Claims

### Claim 1

A recombinant adenovirus genome, comprising:
an E1B region encoding a modified 55k protein, wherein p53 degradation activity of the modified 55k protein is reduced compared to a wild-type 55k protein;
an E2A region comprising a deletion of the DNA binding protein (DBP) open reading frame (ORF);
an E3 region comprising an adenovirus death protein (ADP) ORF and comprising a deletion of the 12.5k, 6.7k, 19k, RIDα, RIDβ and 14.7k ORFs;
an E4 region;
L1, L2, L3, L4 and L5 regions;
a first exogenous nucleic acid sequence comprising a CMV-Tet-O promoter operably linked to an adenovirus DBP ORF; and
a second exogenous nucleic acid sequence comprising a p53-responsive promoter operably linked to a tetracycline repressor (TetR) protein ORF.

### Claim 2 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the first and second exogenous nucleic acid sequences are located between the L5 and E4 regions of the adenovirus genome.

### Claim 3 (depends on 2)

The recombinant adenovirus genome of claim 2, wherein the first exogenous nucleic acid sequence precedes the second exogenous nucleic acid sequence.

### Claim 4 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the first exogenous nucleic acid sequence further comprises a first heterologous polyA sequence following the DBP ORF.

### Claim 5 (depends on 4)

The recombinant adenovirus genome of claim 4, wherein the second exogenous nucleic acid sequence further comprises a second heterologous polyA sequence following the TetR ORF.

### Claim 6 (depends on 5)

The recombinant adenovirus genome of claim 5, further comprising a third heterologous polyA sequence following the L5 region and preceding the first and second exogenous nucleic acid sequences.

### Claim 7 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the p53-responsive promoter is prMinRGC.

### Claim 8 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the modified 55k protein comprises a mutation selected from H260R, H260A, H260D, R240A, R240E and R240H with respect to SEQ ID NO: 6.

### Claim 9 (depends on 1)

The recombinant adenovirus genome of claim 1, further comprising a reporter gene.

### Claim 10 (depends on 9)

The recombinant adenovirus genome of claim 9, wherein the reporter gene is operably linked to and in the same reading frame as a self-cleaving peptide coding sequence and the ADP ORF.

### Claim 11 (depends on 1)

The recombinant adenovirus genome of claim 1, comprising at least one modification to detarget an adenovirus from the liver.

### Claim 12 (depends on 11)

The recombinant adenovirus genome of claim 11, further comprising one or more binding sites for a liver-specific microRNA.

### Claim 13 (depends on 12)

The recombinant adenovirus genome of claim 12, wherein the liver-specific microRNA is miR-122.

### Claim 14 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the genome encodes a chimeric fiber protein comprising a fiber shaft from a first adenovirus serotype and a fiber knob from a second adenovirus serotype.

### Claim 15 (depends on 14)

The recombinant adenovirus genome of claim 14, wherein the first adenovirus serotype is Ad5 and the second adenovirus serotype is Ad3, Ad9, Ad11, Ad12, Ad34 or Ad37.

### Claim 16 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the genome encodes a fiber protein modified to include an RGD peptide.

### Claim 17 (depends on 1)

The recombinant adenovirus genome of claim 1, wherein the nucleotide sequence of the genome is at least 95% identical to SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5.

### Claim 18 (depends on 1)

An isolated cell comprising the recombinant adenovirus genome of claim 1.

### Claim 19 (depends on 1)

A composition comprising the recombinant adenovirus genome of claim 1 and a pharmaceutically acceptable carrier.

### Claim 20 (depends on 1)

An isolated adenovirus comprising the recombinant adenovirus genome of claim 1.

### Claim 21 (depends on 20)

A composition comprising the adenovirus of claim 20 and a pharmaceutically acceptable carrier.

### Claim 22 (depends on 20)

A method of reducing or inhibiting tumor progression, reducing tumor volume, or both, in a subject having a tumor deficient in p53 transcriptional activity, comprising administering to the subject a therapeutically effective amount of the recombinant adenovirus of claim 20, thereby reducing or inhibiting tumor progression, reducing tumor volume, or both, in the subject.

### Claim 23 (depends on 20)

A method of treating a cancer in a subject having a cancer deficient in p53 transcriptional activity, comprising administering to the subject a therapeutically effective amount of the recombinant adenovirus of claim 20, thereby treating cancer in the subject.