Methods for determining isolated p27 protein levels and uses thereof

• US 6,316,208 B1
• Filed: 02/03/1997
• Issued: 11/13/2001
• Est. Priority Date: 01/07/1994
• Status: Expired due to Term

• Current Assignee: Fred Hutchinson Cancer Research Center (Fred Hutchinson Cancer Research Center|Argus Genetics|Mars), GPC Biotech Inc. (GPC AG - Genome Pharmaceuticals Corp.), Memorial Sloan-Kettering Cancer Center
• Original Assignee: Fred Hutchinson Cancer Research Center (Fred Hutchinson Cancer Research Center|Argus Genetics|Mars), Memorial Sloan-Kettering Cancer Center
• Inventors: Polyak, Kornelia, Koff, Andrew, Porter, Peggy L., Massague, Joan, Roberts, James M.
• Primary Examiner(s): Chan, Christina Y.
• Assistant Examiner(s): HAYES, ROBERT CLINTON
• Application Number:
  US08/794,002
• Time in Patent Office: 1,744 Days
• Field of Search: 435/7.8, 435/7.1, 435/7.21, 435/7.4
• US Class Current: 435/7.21
• CPC Class Codes: 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
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4738 Cell cycle regulated proteins, e.g. cyclin, CDC, INK-CCR cell cycle dependent kinases C12N9/12
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 2333/4704 Inhibitors; Supressors
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/574 for cancer

Abstract

The subject invention is directed to the discovery of a protein involved in regulation of cell-cycle progression, and includes reagents and methods related thereto.

Claims

1. A method for determining the relative amount of p27 protein in a cell or sample of cells (cell sample) comprising (i) determining the level of a p27 protein having a sequence of one of SEQ ID Nos. 2, 4, and 6 in the cell sample, and (ii) comparing the level of the p27 protein in the cell sample with the level of a wild-type p27 protein determined for a control cell, wherein the wild-type p27 protein inhibits activation of a cyclin E-Cdk2 (cyclin-dependent kinase-2) complex and has an amino acid sequence of one of SEQ ID Nos. 2, 4, and 6.

5. The method of any one of claims 1, 2 or 3, wherein the level of p27 protein is determined by chromatography or electrophoresis.

2. A method for diagnosing a hyperproliferative disorder characterized by abnormal cell proliferation in a patient, comprising:

(i) ascertaining the level of a p27 protein having an amino acid sequence of one of SEQ ID Nos. 2, 4, and 6 in a sample of cells from the patient; and

(ii) diagnosing the presence or absence of a hyperproliferative disorder including utilizing the ascertained level of the p27 protein, wherein a reduced level of the p27 protein in the sample, relative to a level of a wild-type p27 protein in a control sample of normal cells, correlates with the presence of a hyperproliferative disorder, and the wild-type p27 protein inhibits activation of a cyclin E-Cdk2 (cyclin-dependent kinase) complex and has an amino acid sequence of one of SEQ ID Nos. 2, 4, and 6.

6. The method of any one of claims 2 or 3, wherein the patient is a human.

7. The method of claim 2, wherein the hyperproliferative disorder is cancer.

3. A method for evaluating a cancer patient'"'"'s prognosis, comprising (i) ascertaining the level of a p27 protein having an amino acid sequence of one of SEO ID Nos. 2, 4, and 6 in a sample of cancer cells from the patient;

and (ii) comparing the ascertained level of the p27 protein to a level of wild-type p27 protein in a normal control sample of cells, wherein a reduced level of the p27 protein in the sample correlates with an increased risk of recurrence of a cancer, and wherein the wild-type p27 protein inhibits activation of a cyclin E-Cdk2 (cyclin-dependent kinase-2) complex and has an amino acid sequence of one of SEQ ID Nos. 2, 4, and 6.

8. The method of claim 3 or 7, wherein the cancer is a cancer of an epithelial tissue.

9. The method of claim 3 or 7, wherein the cancer is a cancer selected from the group consisting of carcinomas, melanomas and sarcomas.

10. The method of claim 9, wherein the cancer is a carcinoma of a tissue selected from the group consisting of breast, ovaries, lung, intestinal, pancreas, prostate, testis, liver, skin, stomach, renal, cervical and colorectal.

11. The method of claim 9, wherein the cancer is a breast carcinoma.

12. The method of claim 11, wherein the breast carcinoma is a ductal carcinoma, a mucinous carcinoma or a lobular carcinoma.

13. The method of claim 3, wherein the cancer is a node-negative breast carcinoma.

4. A method for determining the relative amount of p27 protein in a cell or sample of transformed cells (cell sample) from a patient comprising (i) determining the level of p27 protein having an amino acid sequence of one of SEQ ID Nos. 2, 4, and 6 in the cell sample, and (ii) comparing the level of p27 protein in the cell sample with the level of p27 protein determined for a control cell, wherein the p27 protein inhibits activation of a cyclin E-Cdk2 (cyclin-dependent kinase) complex and has an amino acid sequence of one of SEQ ID Nos. 2, 4, and 6.