Methods of identifying inverse agonists of the serotonin 2A receptor

• US 20050148018A1
• Filed: 03/04/2005
• Published: 07/07/2005
• Est. Priority Date: 10/07/1999
• Status: Abandoned Application

• Current Assignee: David Weiner, Mark R. Brann
• Original Assignee: David Weiner, Mark R. Brann
• Inventors: Brann, Mark R., Weiner, David
• Application Number:
  US11/073,313
• Publication Number:
  US 20050148018A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: G01N 2500/10 involving cells
• 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 33/942 Serotonin, i.e. 5-hydroxy-tryptamine

Abstract

The present invention relates to a method of identifying compounds which act as inverse agonists of the 5-HT2A receptor, the method comprising contacting a constitutively active 5-HT2A receptor with at least one test compound and determining any decrease in the level of basal activity of the receptor so as to identify a test compound which is an inverse agonist of the 5-HT2A receptor. Such inverse agonists may be used in the treatment of schizophrenia and related psychoses.

Claims

1. A method of identifying a compound which is an inverse agonist of the 5-HT2A receptor, the method comprising:

(a) contacting a constitutively active 5-HT2A receptor with at least one test compound; and

(b) determining any decrease in basal activity level of the 5-HT2A receptor so as to identify a test compound which is an inverse agonist of the 5-HT2A receptor.

2. A method of identifying a compound which is an inverse agonist of the 5-HT2A receptor, the method comprising:

(a) culturing cells which express a constitutively active 5-HT2A receptor;

(b) incubating the cells with at least one test compound; and

(c) determining any decrease in basal activity level of the 5-HT2A receptor so as to identify a test compound which is an inverse agonist of the 5-HT2A receptor.

3. The method of claim 2, wherein the cells of step (a) overexpress said 5-HT2A receptor.

4. The method of claim 2, wherein the identified inverse agonist is selective for the 5-HT2A receptor.

5. A method of identifying mutation(s) in a gene encoding a constitutively active 5-HT2A receptor, the method comprising:

(a) extracting nucleic acid from a biological sample obtained from an individual having a disorder or condition putatively associated with constitutive activity of the 5-HT2A receptor;

(b) preparing cDNA from the extracted nucleic acid;

(c) selecting from the cDNA in step (b) cDNA encoding the 5-HT2A receptor;

(d) transfecting a cell with an expression vector comprising said selected cDNA;

(e) selecting a cell expressing constitutively active 5-HT2A receptor; and

(f) sequencing the cDNA in said selected cell to detect the mutation(s).

6. The method of claim 5, wherein said biological sample comprises blood, platelets or brain tissue.

7. The method of claim 5, wherein the extracted nucleic acid is RNA.

8. The method of claim 5, further comprising prior to said transfecting step amplifying the cDNA.

9. The method of claim 5, wherein said cDNA selection is performed using one or more oligodeoxynucleotide probes specific to the gene encoding the 5-HT2A receptor.

10. A method of diagnosing a disorder or condition, or a susceptibility thereto, associated with constitutive activity of the 5-HT2A receptor, the method comprising:

(a) obtaining a biological sample from an individual putatively affected by or susceptible to a disorder or condition associated with constitutive activity of the 5-HT2A receptor;

(b) isolating from said biological sample a nucleic acid sequence encoding said receptor, or a portion of said nucleic acid sequence corresponding to the portion of the gene identified to include mutation(s) by the method of claim 5;

and

(c) detecting the presence or absence of mutation(s) in said nucleic acid sequence or said portion thereof.

11. The method of claim 10, wherein said biological sample comprises blood, platelets or brain tissue.

12. A test kit for detecting mutation(s) in a gene encoding a constitutively active 5-HT2A receptor, the test kit comprising a nucleic acid sequence corresponding to a portion of the gene identified by the method of claim 5 to include at least one mutation.

13. A method of decreasing the basal activity level of the 5-HT2A receptor in a subject in need thereof, the method comprising contacting a 5-HT2A receptor in said subject with an inverse agonist of the 5-HT2A receptor in an amount effective to substantially decrease the basal activity level of said receptor.

14. The method of claim 13, wherein the inverse agonist is selective for the 5-HT2A receptor.

15. The method of claim 13, wherein the inverse agonist has little or substantially no anti-dopaminergic activity.

16. A method of ameliorating symptoms of schizophrenia or psychosis in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an inverse agonist of the 5-HT2A receptor.

17. Use of an inverse agonist of the 5-HT2A receptor for the preparation of a medicament for substantially decreasing the basal activity level of a constitutively active 5-HT2A receptor.

18. Use according to claim 17, wherein the inverse agonist is selective for the 5-HT2A receptor.

19. Use according to claim 17, wherein the inverse agonist has little or substantially no anti-dopaminergic activity.

20. Use of a 5-HT2A receptor to identify a compound acting as an inverse agonist at said receptor.

21. Use of a 5-HT2A receptor to identify a compound acting as an inverse agonist at said receptor and useful in the treatment of schizophrenia or psychosis.