25869, a novel human carboxylesterase and uses thereof

• US 20020076786A1
• Filed: 06/29/2001
• Published: 06/20/2002
• Est. Priority Date: 06/29/2000
• Status: Abandoned Application

• Current Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Inventors: Curtis, Rory A.J., Logan, Thomas Joseph
• Application Number:
  US09/895,860
• Publication Number:
  US 20020076786A1
• Time in Patent Office: Days
• US Class Current: 435/196
• 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: C12N 9/18 Carboxylic ester hydrolases (3.1.1)

Abstract

The invention provides isolated nucleic acid molecules, designated COE-1 nucleic acid molecules, which encode novel carboxylesterase family members. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing COE-1 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a COE-1 gene has been introduced or disrupted. The invention still further provides isolated COE-1 proteins, fusion proteins, antigenic peptides and anti-COE-1 antibodies. Diagnostic methods utilizing compositions of the invention are also provided.

Claims

1. An isolated nucleic acid molecule selected from the group consisting of:

(a) a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO;

1; and

(b) a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO;

3.

2. An isolated nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

2.

3. An isolated nucleic acid molecule comprising the nucleotide sequence contained in the plasmid deposited with ATCC®

as Accession Number ______.

4. An isolated nucleic acid molecule which encodes a naturally-occurring allelic variant of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

2.

5. An isolated nucleic acid molecule selected from the group consisting of:

(a) a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to the nucleotide sequence of SEQ ID NO;

1 or 3, or a complement thereof;

(b) a nucleic acid molecule comprising a fragment of at least 30 nucleotides of a nucleic acid comprising the nucleotide sequence of SEQ ID NO;

1 or 3, or a complement thereof;

(c) a nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence at least about 60% identical to the amino acid sequence of SEQ ID NO;

2; and

(d) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO;

2, wherein the fragment comprises at least 10 contiguous amino acid residues of the amino acid sequence of SEQ ID NO;

2.

6. An isolated nucleic acid molecule which hybridizes to the nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5 under stringent conditions.

7. An isolated nucleic acid molecule comprising a nucleotide sequence which is complementary to the nucleotide sequence of the nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5.

8. An isolated nucleic acid molecule comprising the nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5, and a nucleotide sequence encoding a heterologous polypeptide.

9. A vector comprising the nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5.

10. The vector of claim 9, which is an expression vector.

11. A host cell transfected with the expression vector of claim 10.

12. A method of producing a polypeptide comprising culturing the host cell of claim 11 in an appropriate culture medium to, thereby, produce the polypeptide.

13. An isolated polypeptide selected from the group consisting of:

a) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO;

2, wherein the fragment comprises at least 10 contiguous amino acids of SEQ ID NO;

2;

b) a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO;

2, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule consisting of SEQ ID NO;

1 or 3 under stringent conditions;

c) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO;

1 or 3; and

d) a polypeptide comprising an amino acid sequence which is at least 60% identical to the amino acid sequence of SEQ ID NO;

2.

14. The isolated polypeptide of claim 13 comprising the amino acid sequence of SEQ ID NO:

2.

15. The polypeptide of claim 13, further comprising heterologous amino acid sequences.

16. An antibody which selectively binds to a polypeptide of claim 13.

17. A method for detecting the presence of a polypeptide of claim 13 in a sample comprising:

a) contacting the sample with a compound which selectively binds to the polypeptide; and

b) determining whether the compound binds to the polypeptide in the sample to thereby detect the presence of a polypeptide of claim 13 in the sample.

18. The method of claim 17, wherein the compound which binds to the polypeptide is an antibody.

19. A kit comprising a compound which selectively binds to a polypeptide of claim 13 and instructions for use.

20. A method for detecting the presence of a nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5 in a sample comprising:

a) contacting the sample with a nucleic acid probe or primer which selectively hybridizes to the nucleic acid molecule; and

b) determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample to thereby detect the presence of a nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5 in the sample.

21. The method of claim 20, wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe.

22. A kit comprising a compound which selectively hybridizes to a nucleic acid molecule of any one of claims 1, 2, 3, 4, or 5 and instructions for use.

23. A method for identifying a compound which binds to a polypeptide of claim 13 comprising:

a) contacting the polypeptide, or a cell expressing the polypeptide with a test compound; and

b) determining whether the polypeptide binds to the test compound.

24. The method of claim 23, wherein the binding of the test compound to the polypeptide is detected by a method selected from the group consisting of:

a) detection of binding by direct detection of test compound/polypeptide binding;

b) detection of binding using a competition binding assay; and

c) detection of binding using an assay for COE-1 activity.

25. A method for modulating the activity of a polypeptide of claim 13 comprising contacting the polypeptide or a cell expressing the polypeptide with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.

26. A method for identifying a compound which modulates the activity of a polypeptide of claim 13 comprising:

a) contacting a polypeptide of claim 13 with a test compound; and

b) determining the effect of the test compound on the activity of the polypeptide to thereby identify a compound which modulates the activity of the polypeptide.

27. A method of identifying a subject having a carboxylesterase-associated disorder, or at risk for developing a carboxylesterase-associated disorder comprising:

a) contacting a sample obtained from said subject comprising nucleic acid molecules with a hybridization probe comprising at least 25 contiguous nucleotides of SEQ ID NO;

1; and

b) detecting the presence of a nucleic acid molecule in said sample that hybridizes to said probe, thereby identifying a subject having a carboxylesterase-associated disorder.

28. A method of identifying a subject having a carboxylesterase-associated disorder, or at risk for developing a carboxylesterase-associated disorder comprising:

a) contacting a sample obtained from said subject comprising nucleic acid molecules with a first and a second amplification primer, said first primer comprising at least 25 contiguous nucleotides of SEQ ID NO;

1 and said second primer comprising at least 25 contiguous nucleotides from the complement of SEQ ID NO;

1;

b) incubating said sample under conditions that allow nucleic acid amplification; and

c) detecting the presence of a nucleic acid molecule in said sample that is amplified, thereby identifying a subject having a carboxylesterase-associated disorder, or at risk for developing a carboxylesterase-associated disorder.

29. A method of identifying a subject having a carboxylesterase-associated disorder, or at risk for developing a carboxylesterase-associated comprising:

a) contacting a sample obtained from said subject comprising polypeptides with a COE-1 binding substance; and

b) detecting the presence of a polypeptide in said sample that binds to said COE-1 binding substance, thereby identifying a subject having a carboxylesterase-associated disorder, or at risk for developing a carboxylesterase-associated disorder.

30. A method for identifying a compound capable of treating a carboxylesterase-associated disorder characterized by aberrant COE-1 nucleic acid expression or COE-1 polypeptide activity comprising assaying the ability of the compound to modulate COE-1 nucleic acid expression or COE-1 polypeptide activity, thereby identifying a compound capable of treating a carboxylesterase-associated disorder characterized by aberrant COE-1 nucleic acid expression or COE-1 polypeptide activity.

31. A method for treating a subject having a carboxylesterase-associated disorder characterized by aberrant COE-1 polypeptide activity or aberrant COE-1 nucleic acid expression comprising administering to the subject a COE-1 modulator, thereby treating said subject having a carboxylesterase-associated disorder.

32. The method of claim 31, wherein the carboxylesterase-associated disorder is a lipid homeostasis disorder.