Novel human histone deacetylases

• US 20030161830A1
• Filed: 06/14/2002
• Published: 08/28/2003
• Est. Priority Date: 06/14/2001
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Attar, Ricardo M., Jackson, Donald G., Lorenzi, Matthew V., Gottardis, Marco
• Granted Patent: US 7,199,227 B2
• Time in Patent Office: Days
• US Class Current: 424/146.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)

Abstract

The present invention relates to newly discovered human histone deacetylases (HDACs), also referred to as histone deacetylase-like polypeptides. The polynucleotide sequences and encoded polypeptides of the novel HDACs are encompassed by the invention, as well as vectors comprising these polynucleotides and host cells comprising these vectors. The invention also relates to antibodies that bind to the disclosed HDAC polypeptides, and methods employing these antibodies. Also related are methods of screening for modulators, such as inhibitors or antagonists, or agonists. The invention also relates to diagnostic and therapeutic applications which employ the disclosed HDAC polynucleotides, polypeptides, and antibodies, and HDAC modulators. Such applications can be used with diseases and disorders associated with abnormal cell growth or proliferation, cell differentiation, and cell survival, e.g., neoplastic cell growth, and especially breast and prostate cancers or tumors.

Claims

1. An isolated polynucleotide encoding a histone deacetylase polypeptide which consists of an amino acid sequence selected from the group consisting of SEQ ID NO:

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

87, SEQ ID NO;

93, and SEQ ID NO;

95.

5. A cell line comprising the isolated polynucleotide according to claim 1.

6. An expression vector comprising the isolated polynucleotide according to claim 1.

7. A host cell comprising the expression vector according to claim 6, wherein the host cell is selected from the group consisting of bacterial, yeast, insect, mammalian, and human cells.

19. A method of screening test agents to identify a candidate bioactive agent comprising:

a. contacting the isolated polynucleotide according to claim 1 with test agents under conditions to allow a test agent to associate with the polynucleotide to form a complex;

b. detecting the complex of (b), wherein detection of the complex indicates identification of a candidate bioactive agent.

2. An isolated polynucleotide consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

12, SEQ ID NO;

19, SEQ ID NO;

88, SEQ ID NO;

94, and SEQ ID NO;

96.

3. An primer consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO:

24-27, SEQ ID NO;

28-35, SEQ ID NO;

39-46, SEQ ID NO;

47-62, SEQ ID NO;

65-66, SEQ ID NO;

67-74, SEQ ID NO;

75-82, and SEQ ID NO;

104-105.

14. A method of diagnosing a cancer selected from the group consisting of breast and prostate cancer comprising:

a. incubating the primer according to claim 3 with a biological sample under conditions to allow the primer to amplify a polynucleotide in the sample to produce a amplification product; and

b. measuring levels of amplification product formed in (a), wherein an alteration in these levels compared to standard levels indicates diagnosis of the cancer.

4. A probe consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO:

36, SEQ ID NO;

63-64, SEQ ID NO;

83-86, SEQ ID N092, and SEQ ID NO;

101-103.

15. A method of diagnosing a cancer selected from the group consisting of breast and prostate cancer comprising:

a. incubating the probe according to claim 4 with a biological sample under conditions to allow the probe to hybridize with a polynucleotide in the sample to form a complex; and

b. measuring levels of hybridization complex formed in (a), wherein an alteration in these levels compared to standard levels indicates diagnosis of the cancer.

17. A method of detecting a histone deacetylase polynucleotide comprising:

a. incubating the probe according to claim 4 with a biological sample under conditions to allow the probe to hybridize with a polynucleotide in the sample to form a complex; and

b. identifying the complex formed in (a), wherein identification of the complex indicates detection of a histone deacetylase polynucleotide.

8. An isolated polypeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

87, SEQ ID NO;

93, and SEQ ID NO;

95.

9. An antibody which binds specifically to the isolated polypeptide according to claim 8, wherein the antibody is selected from the group consisting of polyclonal and monoclonal antibodies.

16. A method of diagnosing a cancer selected from the group consisting of breast and prostate cancer comprising:

a. contacting the antibody according to claim 9 with a biological sample under conditions to allow the antibody to associate with a polypeptide in the sample to form a complex; and

b. measuring levels of complex formed in (a), wherein an alteration in these levels compared to standard levels indicates diagnosis of the cancer.

18. A method of detecting a histone deacetylase polypeptide comprising:

a. incubating the antibody according to claim 9 with a biological sample under conditions to allow the antibody to associate with a polypeptide in the sample to form a complex; and

b. identifying the complex formed in (a), wherein identification of the complex indicates detection of a histone deacetylase polypeptide.

20. A method of screening test agents to identify a candidate bioactive agent comprising:

a. contacting the isolated polypeptide according to claim 8 with test agents under conditions to allow a test agent to associate with the polypeptide to form a complex;

b. detecting the complex of (b), wherein detection of the complex indicates identification a candidate bioactive agent.

10. An antisense polynucleotide which consists of a nucleotide sequence selected from the group consisting of SEQ ID NO:

36, SEQ ID NO;

63-64, and SEQ ID NO;

83-86.

11. An expression vector comprising the antisense polynucleotide according to claim 10.

12. A pharmaceutical composition selected from the group consisting of:

a. a pharmaceutical composition comprising a monoclonal antibody that specifically binds to an isolated polypeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

87, SEQ ID NO;

93, and SEQ ID NO;

95, and a physiologically acceptable carrier, diluent, or excipient;

b. a pharmaceutical composition comprising an antisense polynucleotide which consists of a nucleotide sequence selected from the group consisting of SEQ ID NO;

36, SEQ ID NO;

63-64, and SEQ ID NO;

83-86, and a physiologically acceptable carrier, diluent, or excipient; and

c. a pharmaceutical composition comprising an expression vector comprising an isolated polynucleotide encoding a histone deacetylase polypeptide which consists of an amino acid sequence selected from the group of SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

87, SEQ ID NO;

93, and SEQ ID NO;

95, and a physiologically acceptable carrier, diluent, or excipient.

13. A method of treating a cancer selected from the group consisting of breast and prostate cancer comprising administering the pharmaceutical composition according to claim 12 in an amount effective for treating the cancer.