# US 11,524,065 B2 · Large and small T antigens of merkel cell polyomavirus, nucleic acid constructs and vaccines made therefrom, and methods of using same

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,524,065 B2
- Document ID: US11524065B2
- Status: Granted
- Kind: B2
- Application number: 16/962,886
- Filed: 2019-01-18
- Published: 2020-11-05
- Granted: 2022-12-13
- Earliest claimed: 2018-01-19
- Assignees: The Wistar Institute of Anatomy and Biology
- Applicants: The Wistar Institute of Anatomy and Biology
- Inventors: David B. Weiner; Elizabeth Duperret
- Web page: https://patentgrep.com/patent/US11524065B2

## CPC

- A61K 39/12: Viral antigens
- A61P 31/20: for DNA viruses
- A61P 35/00: Antineoplastic agents
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/572: {cytotoxic response}
- A61K 2039/575: {humoral response}
- A61K 2039/585: {wherein the target is cancer}
- A61K 2039/70: {Multivalent vaccine}
- C12N 2710/22034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Nucleic acid molecules and compositions comprising one or more nucleotide sequences that encode a consensus Merkel Cell Polyomavirus (MCV) T antigen. Immunomodulatory methods and methods of inducing an immune response against MCV are disclosed. Method of treating infection by MCV and methods of treating or preventing Merkel Cell Carcinoma associated with MCV are disclosed. Modified consensus MCV T antigens are disclosed.

## Claims

### Claim 1

A nucleic acid molecule encoding at least one modified Merkel Cell Polyomavirus (MCV) T antigen, wherein the T antigen comprises at least one mutation that disrupts at least one oncogenic feature of a native MCV T antigen selected from the group consisting of CR1 binding, DnaJ binding, phophatase pp2A binding, Rb binding, ATPase activity, helicase activity, chaperone protein binding, hVam6p binding, Fbxw7 binding, origin binding, and transformation; wherein the nucleic acid molecule comprises a sequence selected from the group consisting of:
a) a nucleotide sequence encoding an amino acid sequence comprising at least 98% identity to SEQ ID NO:4;
b) a nucleotide sequence encoding an amino acid sequence comprising at least 90% identity to SEQ ID NO:6;
c) a nucleotide sequence encoding an amino acid sequence selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:6;
d) a nucleotide sequence having at least about 90% identity over an entire length of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5; and
e) a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5.

### Claim 2 (depends on 1)

The nucleic acid molecule of claim 1, wherein at least one mutation is a mutation at an amino acid selected from the group consisting of D44, W209, E216, L142, L91, K92, D93, Y94 and M95.

### Claim 3 (depends on 1)

The nucleic acid molecule of claim 1, wherein at least one mutation is selected from the group consisting of a D44N mutation, a W209A, an E216K mutation, an L142A mutation, an L91A mutation, a K92A mutation, a D93A mutation, a Y94A mutation and a M95A mutation.

### Claim 4 (depends on 1)

The nucleic acid molecule of claim 1, wherein the MCV T antigen is selected from the group consisting of a large T antigen (LTAg), a small t antigen (STAg), and a combination thereof.

### Claim 5 (depends on 1)

The nucleic acid molecule of claim 1, wherein the nucleic acid molecule is selected from the group consisting of a DNA molecule and an RNA molecule.

### Claim 6 (depends on 1)

The nucleic acid molecule of claim 1, wherein the nucleic acid molecule comprises
a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5.

### Claim 7 (depends on 1)

The nucleic acid molecule of claim 1, wherein the encoded peptide is operably linked to at least one regulatory sequence selected from the group consisting of a start codon, an IgE leader sequence and a stop codon.

### Claim 8 (depends on 7)

The nucleic acid molecule of claim 7, wherein the nucleic acid molecule
comprises a sequence selected from the group consisting of:
a) a nucleotide sequence encoding an amino acid sequence comprising at least 98% identity to SEQ ID NO:4,
b) a nucleotide sequence encoding an amino acid sequence comprising at least 90% identity to SEQ ID NO:6,
c) a nucleotide sequence encoding an amino acid sequence selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:6,
d) a nucleotide sequence having at least about 90% identity over an entire length of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5; and
e) a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5,
operably linked to an amino acid sequence as set forth in SEQ ID NO:7.

### Claim 9 (depends on 7)

The nucleic acid molecule of claim 7, wherein the nucleic acid molecule comprises
a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5,
operably linked to a nucleotide sequence encoding SEQ ID NO:7.

### Claim 10 (depends on 1)

The nucleic acid molecule of claim 1, wherein the nucleic acid molecule comprises an expression vector.

### Claim 11 (depends on 1)

The nucleic acid molecule of claim 1, wherein the nucleic acid molecule comprises a viral particle.

### Claim 12 (depends on 1)

An immunogenic composition comprising at least one nucleic acid molecule of claim 1.

### Claim 13 (depends on 12)

The immunogenic composition of claim 12, further comprising at least one selected from the group consisting of a pharmaceutically acceptable excipient and an adjuvant.

### Claim 14 (depends on 12)

A method of inducing an immune response against a MCV T antigen in a subject in need thereof, the method comprising administering an immunogenic composition of claim 12 to the subject.

### Claim 15 (depends on 1)

A method of treating or preventing a MCV associated pathology in subject in need thereof, the method comprising administering an immunogenic composition of claim 1 to the subject.

### Claim 16 (depends on 15)

The method of claim 15, wherein the MCV associated pathology is at least one of MCV infection and Merkel Cell Carcinoma.

### Claim 17

A peptide comprising an amino acid sequence selected from the group consisting of
a) an amino acid sequence comprising at least 98% identity to SEQ ID NO:4;
b) an amino acid sequence comprising at least 90% identity to SEQ ID NO:6; and
c) an amino acid sequence selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:6.

### Claim 18 (depends on 17)

An immunogenic composition comprising a peptide of claim 17.