# US 7,745,230 B2 · Estrogen receptors and methods of use

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- Number: US 7,745,230 B2
- Document ID: US7745230B2
- Status: Granted
- Kind: B2
- Application number: 10/591,199
- Filed: 2005-03-10
- Published: 2007-11-08
- Granted: 2010-06-29
- Earliest claimed: 2004-03-10
- Assignees: Creighton University
- Inventors: Zhao Yi Wang
- Web page: https://patentgrep.com/patent/US7745230B2

## CPC

- C07K 14/721: {Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor}
- A61P 15/00: Drugs for genital or sexual disorders (for disorders of sex hormones A61P5/24); Contraceptives
- A61P 15/06: Antiabortive agents; Labour repressants
- A61P 15/12: for climacteric disorders
- A61P 15/18: Feminine contraceptives
- A61P 19/10: for osteoporosis
- A61P 35/00: Antineoplastic agents
- C07K 16/2869: {against hormone receptors (for antibodies against neuromediator receptors C07K16/286)}
- G01N 33/743: {Steroid hormones}
- 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 2039/505: {comprising antibodies}
- G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
- 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)
- 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)
- 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)

## Abstract

The present invention provides isolated polypeptides having an amino acid sequence having at least 70% identity to SEQ ID NO:20, wherein the polypeptide has ER-α36 activity. The invention further provides methods for identifying agents that bind to such polypeptides, methods for detecting such polypeptides, and methods for altering the activity of such polypeptides. Also provided are antibodies that specifically bind to an amino acid sequence depicted at SEQ ID NO:1, or an immunogenic fragment thereof, and methods for making and using such antibodies.

## Claims

### Claim 1

An isolated monoclonal antibody that specifically binds to an amino acid sequence depicted at SEQ ID NO:1.

### Claim 2 (depends on 1)

The antibody of claim 1 wherein the antibody is covalently attached to a compound.

### Claim 3 (depends on 2)

The antibody of claim 2 wherein the compound is a chemotherapeutic agent.

### Claim 4 (depends on 2)

The antibody of claim 2 wherein the compound is a detectable marker.

### Claim 5 (depends on 4)

The antibody of claim 4 wherein the detectable marker is a fluorescent marker.

### Claim 6 (depends on 1)

The antibody of claim 1 wherein the antibody specifically binds to an amino acid sequence depicted at amino acids 13-27 of SEQ ID NO:1.

### Claim 7 (depends on 1)

A composition comprising an antibody of claim 1.

### Claim 8 (depends on 7)

The composition of claim 7 further comprising a pharmaceutically acceptable carrier.

### Claim 9

A method for making an antibody comprising administering to an animal a polypeptide comprising an amino acid sequence depicted at amino acids 13-27 of SEQ ID NO:1 or an immunogenic fragment thereof, and isolating the antibody from the animal, wherein the isolated antibody specifically binds to amino acids 13-27 of SEQ ID NO:1.

### Claim 10 (depends on 9)

The method of claim 9 wherein the polypeptide or immunogenic fragment thereof is covalently attached to a carrier polypeptide.

### Claim 11

A method for making an antibody comprising:
administering to an animal a polypeptide comprising an amino acid sequence depicted at amino acids 13-27 of SEQ ID NO:1 or an immunogenic fragment thereof;
obtaining from the animal a cell that produces the antibody;
making a monoclonal antibody-producing hybridoma using the cell; and
isolating a monoclonal antibody from the monoclonal antibody-producing hybridoma, wherein the isolated monoclonal antibody specifically binds to amino acids 13-27 of SEQ ID NO:1.

### Claim 12 (depends on 9)

A polyclonal antibody produced by a method of claim 9.

### Claim 13 (depends on 11)

A monoclonal antibody produced by a method of claim 11.

### Claim 14

A method for detecting a polypeptide comprising:
providing a cell;
analyzing the cell for a polypeptide, wherein the polypeptide has a molecular weight of 36 kDa as measured following electrophoresis on a sodium dodecyl sulfate (SDS)-polyacrylamide gel, and is recognized by an isolated antibody that specifically binds to a polypeptide comprising the amino acid sequence depicted in SEQ ID NO:1; and
determining whether the cell expresses the polypeptide;
wherein analyzing the cell for a polypeptide comprises contacting the cell with an antibody that specifically binds to the amino acid sequence depicted at SEQ ID NO:1 or an immunogenic fragment thereof, and detecting the bound antibody.

### Claim 15 (depends on 14)

The method of claim 14 wherein the cell is ex vivo.

### Claim 16 (depends on 15)

The method of claim 15 wherein the cell is a tumor cell.

### Claim 17 (depends on 16)

The method of claim 16 wherein the tumor is a breast tumor.

### Claim 18 (depends on 14)

The method of claim 14 wherein the cell is in vivo.

### Claim 19 (depends on 18)

The method of claim 18 wherein the cell is a tumor cell.

### Claim 20 (depends on 19)

The method of claim 19 wherein the tumor is a breast tumor.

### Claim 21 (depends on 14)

The method of claim 14 wherein the antibody is covalently attached to a detectable marker.

### Claim 22 (depends on 21)

The method of claim 21 wherein the detectable marker is a fluorescent marker.

### Claim 23 (depends on 1)

A kit comprising the isolated monoclonal antibody of claim 1, and a packaging material.

### Claim 24 (depends on 1)

The isolated monoclonal antibody of claim 1, wherein the monoclonal antibody comprises a humanized monoclonal antibody.

### Claim 25 (depends on 24)

The antibody of claim 24 wherein the antibody is covalently attached to a compound.

### Claim 26 (depends on 25)

The antibody of claim 25 wherein the compound is a chemotherapeutic agent.

### Claim 27 (depends on 25)

The antibody of claim 25 wherein the compound is a detectable marker.

### Claim 28 (depends on 27)

The antibody of claim 27 wherein the detectable marker is a fluorescent marker.

### Claim 29 (depends on 24)

The antibody of claim 24 wherein the antibody specifically binds to an amino acid sequence depicted at amino acids 13-27 of SEQ ID NO:1.

### Claim 30 (depends on 24)

A composition comprising an antibody of claim 24.

### Claim 31 (depends on 30)

The composition of claim 30 further comprising a pharmaceutically acceptable carrier.

### Claim 32 (depends on 14)

The method of claim 14 wherein the isolated antibody comprises a monoclonal antibody.

### Claim 33 (depends on 32)

The method of claim 32 wherein the monoclonal antibody comprises a humanized monoclonal antibody.

### Claim 34 (depends on 23)

The kit of claim 23 wherein the monoclonal antibody comprises a humanized monoclonal antibody.