50365, a novel hexokinase family member and uses therefor

• US 20020009779A1
• Filed: 05/21/2001
• Published: 01/24/2002
• Est. Priority Date: 05/19/2000
• Status: Abandoned Application

• Current Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Inventors: Williamson, Mark, Meyers, Rachel A.
• Application Number:
  US09/861,801
• Publication Number:
  US 20020009779A1
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• 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 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
• CPC Class Codes: C12Y 207/01001 Hexokinase (2.7.1.1)
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides isolated nucleic acids molecules, designated 50365 nucleic acid molecules, which encode novel hexokinase members. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing 50365 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a 50365 gene has been introduced or disrupted. The invention still further provides isolated 50365 proteins, fusion proteins, antigenic peptides and anti-50365 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 a nucleotide sequence which is at least 80% identical to the nucleotide sequence of SEQ ID NO;

1, or SEQ ID NO;

3;

b) a nucleic acid molecule comprising a fragment of at least 300 nucleotides of the nucleotide sequence of SEQ ID NO;

1, or SEQ ID NO;

3;

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

2;

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 15 contiguous amino acids of SEQ ID NO;

2; and

e) a nucleic acid molecule which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO;

2, wherein the nucleic acid molecule hybridizes to a nucleic acid molecule comprising SEQ ID NO;

1, 3, or a complement thereof, under stringent conditions.

2. The isolated nucleic acid molecule of claim 1, which is selected from the group consisting of:

a) a nucleic acid comprising the nucleotide sequence of SEQ ID NO;

1, SEQ ID NO;

3;

and

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

2.

3. The nucleic acid molecule of claim 1 further comprising a vector nucleic acid sequence.

4. The nucleic acid molecule of claim 1 further comprising a nucleic acid sequence encoding a heterologous polypeptide.

5. A host cell which contains the nucleic acid molecule of claim 1.

6. The host cell of claim 5 which is a mammalian host cell.

7. A non-human mammalian host cell containing the nucleic acid molecule of claim 1.

12. A method for producing a polypeptide selected from the group consisting of:

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

2;

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

2, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO;

2; and

c) 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 comprising SEQ ID NO;

1, SEQ ID NO;

3, or a complement thereof under stringent conditions;

the method comprising culturing the host cell of claim 5 under conditions in which the nucleic acid molecule is expressed.

16. A method for detecting the presence of a nucleic acid molecule of claim 1 in a sample, comprising the steps of:

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.

17. The method of claim 16, wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe.

18. A kit comprising a compound which selectively hybridizes to a nucleic acid molecule of claim 1 and instructions for use.

26. A method of inhibiting proliferation, or inducing the killing, of a 50365-expressing hyperproliferative cell, comprising contacting the hyperproliferative cell with a compound that modulates the activity or expression of a nucleic acid molecule of claim 1, in an amount which is effective to reduce or inhibit the proliferation of, or induce the killing of, the hyperproliferative cell.

27. The method of claim 26, wherein the compound is an antisense, a ribozyme, or a triple helix molecule.

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

a) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 80% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO;

1 or SEQ ID NO;

3;

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 comprising SEQ ID NO;

1, SEQ ID NO;

3, or a complement thereof under stringent conditions; and

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

2, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO;

2.

9. The isolated polypeptide of claim 8 comprising the amino acid sequence of SEQ ID NO:

2.

10. The polypeptide of claim 8 further comprising a heterologous amino acid sequence.

11. An antibody which selectively binds to a polypeptide of claim 8.

13. A method for detecting the presence of a polypeptide of claim 8 in a sample, comprising:

a) contacting the sample with a compound which selectively binds to a polypeptide of claim 8; and

b) determining whether the compound binds to the polypeptide in the sample.

14. The method of claim 13, wherein the compound which binds to the polypeptide is an antibody.

15. A kit comprising a compound which selectively binds to a polypeptide of claim 8 and instructions for use.

19. A method for identifying a compound which binds to a polypeptide of claim 8 comprising the steps of:

a) contacting a polypeptide, or a cell expressing a polypeptide of claim 8 with a test compound; and

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

20. The method of claim 19, 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 detecting of test compound/polypeptide binding;

b) detection of binding using a competition binding assay;

c) detection of binding using an assay for 50365-mediated catalytic activity.

21. A method for modulating the activity of a polypeptide of claim 8 comprising contacting a polypeptide or a cell expressing a polypeptide of claim 8 with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.

22. A method for identifying a compound which modulates the activity of a polypeptide of claim 8, comprising:

a) contacting a polypeptide of claim 8 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.

23. A method of inhibiting proliferation, or inducing the killing, of a 50365-expressing hyperproliferative cell, comprising contacting the hyperproliferative cell with a compound that modulates the activity or expression of a polypeptide of claim 8, in an amount which is effective to reduce or inhibit the proliferation of, or induce the killing of, the hyperproliferative cell.

24. The method of claim 23, wherein the compound is selected from the group consisting of a peptide, a phosphopeptide, a small organic molecule, a small inorganic molecule and an antibody.

25. The method of claim 23, wherein the compound is an antibody conjugated to a therapeutic moiety selected from the group consisting of a cytotoxin, a cytotoxic agent and a radioactive metal ion.

28. The method of claim 23, wherein the hyperproliferative cell is found in a solid tumor, a soft tissue tumor, or a metastatic lesion.

29. The method of claim 28, wherein the hyperproliferative cell is found in a cancer of the colon or a metastasis of colonic origin.