# US 5,705,342 A · Interaction of BCL-2 and R-RAS

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

- Number: US 5,705,342 A
- Document ID: US5705342A
- Status: Granted
- Kind: A
- Application number: 08/484,945
- Filed: 1995-06-07
- Granted: 1998-01-06
- Earliest claimed: 1993-08-20
- Assignees: Onyx Pharmaceuticals, Inc.
- Inventors: James R. Bischoff; Maria Jose Fernandez-Sarabia
- Web page: https://patentgrep.com/patent/US5705342A

## CPC

- C07K 14/82: Translation products from oncogenes
- C07K 14/4702: {Regulators; Modulating activity}
- 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)
- 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)

## Abstract

The invention provides compositions and methods for screening for agents which are modulators of bcl-2 function and can modulate bcl-2-mediated apoptosis and/or modulate neoplastic and immune conditions dependent upon bcl-2 function. The invention also provides a composition comprising a substantially pure protein complex comprising a R-ras polypeptide and a bcl-2 polypeptide.

## Claims

### Claim 1

A method of screening for candidate bcl-2 modulating agents, comprising:
performing a heterodimerization assay which includes combining substantially purified full length R-ras polypeptide with a substantially purified full length bcl-2 polypeptide and an agent; and
determining whether the agent inhibits the heterodimerization.

### Claim 2

A method for screening for candidate bcl-2-modulating agents, comprising:
performing a heterodimerization assay which includes combining a substantially purified R-ras polypeptide, said R-ras polypeptide having at least 60 C-terminal domain amino acids of a naturally occurring R-ras polypeptide, with a substantially purified full length bcl-2 polypeptide, and an agent; and
determining whether the agent inhibits the heterodimerization.

### Claim 3 (depends on 1, 2)

A method of claim 1, or claim 2 wherein the heterodimerization assay comprises an in vitro binding reaction.

### Claim 4 (depends on 1, 2)

A method of claim 1, or claim 2 wherein the heterodimerization assay comprises a two-hybrid assay in yeast.

### Claim 5 (depends on 1, 2)

A method of claim 1 or claim 2 wherein the two-hybrid assay comprises a human R-ras polypeptide sequence fused to a GAL4 protein and a human bcl-2 polypeptide sequence fused to a GAL4 protein.

### Claim 6 (depends on 3)

A method of claim 3, wherein a R-ras or bcl-2 polypeptide is immobilized.

### Claim 7 (depends on 1, 2)

A method for altering apoptosis of an isolated cell, comprising administering to said cell an agent that inhibits R-ras:bcl-2 heterodimerization formation as described in claim 1 or claim 2.

### Claim 8 (depends on 7)

A method of claim 7, wherein said cell is derived from the hematopoietic lineage.

### Claim 9 (depends on 7)

A method of claim 7, wherein the cell is a neoplastic cell.

### Claim 10 (depends on 8)

A method of claim 8, wherein the cell is a leukocyte.