# US 7,410,774 B1 · Compounds modulating sister chromatid separation and method for identifying same

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

- Number: US 7,410,774 B1
- Document ID: US7410774B1
- Status: Granted
- Kind: B1
- Application number: 09/500,991
- Filed: 2000-02-15
- Granted: 2008-08-12
- Earliest claimed: 1999-02-15
- Assignees: Boehringer Ingelheim International GmbH
- Inventors: Frank Uhlmann; Kim Nasmyth; Jan-Michael Peters; Irene Waizenegger; Sara Buonomo; Rosemary Clyne
- Web page: https://patentgrep.com/patent/US7410774B1

## CPC

- G01N 33/5011: {for testing antineoplastic activity}
- A61P 5/24: of the sex hormones
- A61P 35/00: Antineoplastic agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- G01N 2800/7023: (Hyper)proliferation
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Method for identifying compounds that interfere with or modulate sister chromatid separation in animal or plant cells by modulating a protease with separin-like cysteine endopeptidase activity. Inhibitors of separin activity are useful in cancer therapy, to prevent birth defects and to increase ploidy in plants.

## Claims

### Claim 1

A method for identifying a compound that has the activity of inhibiting sister chromatid separation in eukaryotic cells, said method comprising:
(a) incubating with a test compound a separin in the presence of a separin substrate, wherein said substrate is a peptide or polypeptide comprising an amino acid sequence EXXR (SEQ ID NO:3), wherein X is any amino acid, and the substrate is capable of being cleaved by the separin; and
(b) determining the inhibiting effect of the test compound on the proteolytic activity of the separin,
wherein a compound determined in (b) to inhibit the proteolytic activity of the separin has the activity of inhibiting sister chromatid separation in eukaryotic cells.

### Claim 2 (depends on 1)

The method of claim 1, wherein said eukaryotic cell is an animal cell.

### Claim 3 (depends on 1)

The method of claim 1, which is high-throughput.

### Claim 4 (depends on 1)

The method of claim 1, wherein said separin is recombinant.

### Claim 5 (depends on 1)

The method of claim 1, wherein said separin is human separin.

### Claim 6 (depends on 1)

The method of claim 1, wherein said substrate is a protein recombinantly produced in baculovirus in the presence of a phosphatase inhibitor.

### Claim 7 (depends on 1)

The method of claim 1, wherein said substrate is human SCC1, or a fragment thereof that can be cleaved by separin or having a separin cleavage site.

### Claim 8 (depends on 7)

The method of claim 7, wherein said substrate is a polypeptide comprising the amino acid sequence of SEQ ID NO:1, or a fragment thereof that can be cleaved by separin or having a separin cleavage site.

### Claim 9 (depends on 1)

The method of claim 1, wherein said substrate comprises a label which generates a detectable signal proportional to the amount of the cleavage product of the proteolytic activity, and wherein the signal is measured in the presence and in the absence of the test compound.

### Claim 10 (depends on 9)

The method of claim 9, wherein said label is fluorescent.

### Claim 11 (depends on 1)

The method of claim 1, wherein said substrate is human SCC1.

### Claim 12 (depends on 7)

The method of claim 7, wherein said substrate is a polypeptide comprising the amino acid sequence of SEQ ID NO:1.