Compositions, reagents and kits for and methods of diagnosing, monitoring and treating hormonal imbalance

• US 20070099275A1
• Filed: 11/01/2006
• Published: 05/03/2007
• Est. Priority Date: 11/03/2005
• Status: Active Grant

• Current Assignee: Liat Mintz
• Original Assignee: Liat Mintz
• Inventors: Mintz, Liat
• Granted Patent: US 7,858,744 B2
• Time in Patent Office: Days
• US Class Current: 435/69.4
• 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: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin G01N2333/665, corticotropin G01N2333/695
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures

Abstract

The present invention concerns ten novel variants of alternative splicing of the hormonal imbalance related genes.

Claims

1. An isolated nucleic acid sequence selected from the group consisting of SEQ ID NO:

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, and a complement thereof.

2. An isolated nucleic acid sequence having 90% identity to the isolated nucleic acid sequence of claim 1.

3. An isolated nucleic acid fragment comprising at least a 15 contiguous base pair segment of the isolated nucleic acid sequence of claim 1.

4. The isolated nucleic acid fragment of claim 3 comprising a 15-30 contiguous base pair segment of the isolated nucleic acid sequence of claim 1.

5. A primer comprising the isolated nucleic acid fragment of claim 3.

6. A probe comprising the isolated nucleic acid fragment of claim 3.

7. The isolated nucleic acid sequence of claim 1, wherein at least one preferred codon replaces at least one codon of said sequence.

8. An expression vector comprising the isolated nucleic acid sequence of claim 1, said sequence operably-linked to control elements for the expression of said sequence in a host cell.

9. A host cell transfected with the expression vector of claim 8.

10. An antisense oligonucleotide comprising a 10-40 contiguous base pair segment of a reverse complement of a nucleic acid sequence selected from the group consisting of SEQ ID NO:

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, and SEQ ID NO;

19.

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

21, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, and SEQ ID NO;

38.

12. An isolated amino acid sequence having 90% identity to the isolated amino acid sequence of claim 11.

13. An isolated antibody which specifically binds to an epitope on an amino acid sequence selected from the group consisting of SEQ ID NO:

21, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, and SEQ ID NO;

38.

14. An isolated peptide comprising at least a 10 contiguous amino acid segment of an amino acid sequence selected from the group consisting of SEQ ID NO:

21, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

34, SEQ ID NO;

36, and SEQ ID NO;

38.

15. The isolated peptide of claim 14 comprising a 10-20 contiguous amino acid segment of the amino acid sequence selected from the group consisting of SEQ ID NO:

21, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

34, SEQ ID NO;

36, and SEQ ID NO;

38.

16. A method for detecting the presence of at least one variant nucleic acid sequence of hormonal imbalance related genes in a biological sample comprising the steps of:

(a) hybridizing to nucleic acid material of the biological sample an isolated nucleic acid sequence selected from the group consisting of SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, and a complement thereof; and

(b) detecting a hybridization complex produced by step (a);

wherein the presence of the hybridization complex correlates with the presence of at least one variant nucleic acid sequence in the biological sample.

17. A method for determining the level of variant nucleic acid sequences in a biological sample comprising the steps of:

(a) hybridizing to nucleic acid material of the biological sample an isolated nucleic acid sequence selected from the group consisting of SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, and a complement thereof;

(b) determining the amount of hybridization complexes produced by step (a); and

(c) normalizing the amount of hybridization complexes to provide a level of variant nucleic acid sequences in the biological sample.

18. A method for determining the ratio between the level of a nucleic acid sequence of an hormonal imbalance related gene variant in a first biological sample and a variant produced by alternative splicing in a second biological sample comprising the steps of:

(a) determining the level of a first nucleic acid sequence of an hormonal Imbalance related gene variant in a first biological sample;

(b) determining the level of a second nucleic acid sequence of an alternative splicing form of the variant in a second biological sample; and

(c) comparing the levels obtained in step (a) and step (b) to give a ratio.

19. The method of claim 18, wherein the first biological sample and the second biological sample are the same sample.

20. The method of claim 18, wherein the first nucleic acid sequence and the second nucleic acid sequence are mRNA transcripts.

21. The method of claim 20, wherein the first nucleic acid sequence and the second nucleic acid sequence are deposed on a nucleic acid chip.

22. A method for the identification of compounds capable of affecting the binding affinity of hormonal imbalance related proteins to the receptors of said proteins comprising the steps of:

(a) providing an amino acid sequence selected from the group consisting of SEQ ID NO;

21, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, and SEQ ID NO;

38;

(b) contacting a candidate compound with the amino acid sequence in the presence of at least one receptor of an hormonal Imbalance related gene;

(c) determining the effect of the candidate compound on the binding of the amino acid sequence to the at least one receptor; and

(d) selecting a compound capable of affecting the binding affinity of hormonal Imbalance related proteins to the receptors of said proteins.

23. A method for determining the ratio between the level of an hormonal imbalance related protein variant in a first biological sample and a variant produced by alternative splicing in a second biological sample comprising the steps of:

(a) determining the level of a first amino acid sequence of an hormonal imbalance related gene variant in a first biological sample;

(b) determining the level of a second amino acid sequence of an alternative splicing form of the variant in a second biological sample; and

(c) comparing the levels obtained in step (a) and step (b) to give a ratio.

24. A method of detecting a specific hormonal imbalance related nucleic acid sequence by polymerase chain reaction comprising the steps of:

(a) amplifying a specific hormonal imbalance related nucleic acid sequence with a primer pair, wherein at least one of the primers comprises an at least 15 contiguous base pair segment of a nucleic acid sequence selected from the group consisting of SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

15, SEQ ID NO;

17, SEQ ID NO;

19, and a complement thereof; and

(b) detecting the nucleic acid product of step (a).

25. A method of treating hormonal imbalance comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising:

(a) an isolated amino acid sequence selected from the group consisting of SEQ ID NO;

21, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

32, SEQ ID NO;

34, SEQ ID NO;

36, SEQ ID NO;

38, and a peptide comprising at least a 10 contiguous amino acid segment thereof;

and

(b) a carrier.