# US 7,745,212 B2 · Melanoma antigens and their use in diagnostic and therapeutic methods

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- Number: US 7,745,212 B2
- Document ID: US7745212B2
- Status: Granted
- Kind: B2
- Application number: 11/932,338
- Filed: 2007-10-31
- Published: 2008-11-20
- Granted: 2010-06-29
- Earliest claimed: 1994-04-22
- Assignees: The United States of America as represented by the Department of Health and Human Services
- Inventors: Yutaka Kawakami; Steven A. Rosenberg
- Web page: https://patentgrep.com/patent/US7745212B2

## CPC

- C07K 14/47: from mammals
- A61K 39/00119: {Melanoma antigens}
- A61K 39/001191: {Melan-A/MART}
- A61K 39/001192: {Glycoprotein 100 [Gp100]}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/4271: {Melanoma antigens}
- A61K 40/4272: {Melan-A/MART}
- A61K 40/4273: {Glycoprotein 100 [Gp100]}
- A61P 35/00: Antineoplastic agents
- A61P 37/04: Immunostimulants
- C07K 14/4748: {Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE}
- G01N 33/5751: of the skin, e.g. melanoma
- 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)
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- 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)
- 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
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present invention provides a nucleic acid sequence encoding a melanoma antigen recognized by T lymphocytes, designated MART-1. This invention further relates to bioassays using the nucleic acid sequence, protein or antibodies of this invention to diagnose, assess or prognoses a mammal afflicted with melanoma or metastata melanoma. This invention also provides immunogenic peptides derived from the MART-1 melanoma antigen and a second melanoma antigen designated gp100. This invention further provides immunogenic peptides derived from the MART-1 melanoma antigen or gp100 antigen which have been modified to enhance their immunogenicity. The proteins and peptides provided can serve as an immunogen or vaccine to prevent or treat melanoma.

## Claims

### Claim 1

An isolated or purified recombinant expression vector comprising a nucleic acid encoding a MART-1 protein (SEQ ID NO: 2) or an immunogenic MART-1 peptide, wherein the peptide consists of 5-20 contiguous amino acids of SEQ ID NO: 2 and reacts with T-lymphocytes.

### Claim 2 (depends on 1)

A host organism transformed or transfected with the recombinant expression vector according to claim 1 in a manner to allow expression of said protein encoded by said recombinant expression vector.

### Claim 3

A method of producing a recombinant MART-1 protein (SEQ ID NO: 2), comprising:
(a) inserting a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1 into an expression vector;
(b) transferring the expression vector into a host cell;
(c) culturing the host cell under conditions appropriate for amplification of the vector and expression of the protein; and
(d) harvesting the protein.

### Claim 4 (depends on 3)

The method of claim 3, wherein the expression vector is a eukaryotic expression vector or a prokaryotic expression vector.

### Claim 5 (depends on 4)

The method of claim 4, wherein the expression vector is a baculovirus vector.

### Claim 6 (depends on 3)

The method of claim 3, wherein the host cell is a eukaryotic cell or prokaryotic cell.

### Claim 7 (depends on 6)

The method of claim 6, wherein the eukaryotic cell is an insect cell.