Method for identifying and using A2B adenosine receptor antagonists to mediate mammalian cell proliferation

• US 20020002142A1
• Filed: 02/16/2001
• Published: 01/03/2002
• Est. Priority Date: 02/17/2000
• Status: Active Grant

• Current Assignee: Gilead Palo Alto Incorporated (Gilead Sciences Inc.), University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Belardinelli, Luiz, Grant, Maria B.
• Granted Patent: US 6,894,021 B2
• Time in Patent Office: Days
• US Class Current: 514/44
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/522 having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 2500/10 involving cells

Abstract

This invention concerns methods for identifying A_2B adenosine receptor agonists and antagonists as well as methods for using A_2B. adenosine receptor antagonists to treat cell proliferation orders mediated by the A_2B adenosine receptor.

Claims

1. A method for inhibiting the proliferation of mammalian cells that express the A_2B adenosine receptor comprising administering a therapeutically effective amount of an A_2B adenosine receptor antagonist to the mammal.

2. The method of claim 1 wherein the cells that express the A_2B adenosine receptor are vascular endothelial cells.

3. The method of claim 2 wherein the vascular endothelial cells that express the A_2B adenosine receptor are selected from the group consisting of coronary endothelial cells, endothelial cells from the vascular bed.

4. The method of claim 3 wherein the vascular bed endothelial cells are selected from the group consisting of tumor endothelial cells, retinal endothelial cells, dermal endothelial cells, and brain endothelial cells.

5. The method of claim 1 wherein the endothelial cells are retinal endothelial cells.

6. The method of claim 1 wherein the A_2B adenosine receptor antagonist inhibits the expression of vascular endothelial cell growth factor (VEGF).

7. The method of claim 1 wherein the A_2B adenosine receptor antagonist is an A_2B adenosine receptor antisense oligonucleotide.

8. The method of claim 1 wherein the A_2B adenosine receptor antagonist is an A_2B-specific ribozyme.

9. The method of claim 1 wherein the A_2B adenosine receptor antagonist is a non-selective adenosine receptor antagonist.

10. The method of claim 1 wherein the A_2B adenosine receptor antagonist is a selective A_2B adenosine receptor antagonist.

11. The method of claim 1 wherein the A_2B adenosine receptor antagonist is administered in an amount raging from about 1 microgram/kg to about 50 miligrams/kg.

12. The method of claim 1 wherein the adenosine A_2B adenosine receptor antagonist is administered in an amount ranging from about 1 microgram/kg to about 10 miligrams/kg.

13. The method of claim 1 wherein the A_2B adenosine receptor antagonist is administered by a method selected from the group consisting of orally, nasally, transdermally, by bolus, intravenously, in eye drops, by inhalation, and by using micropumps.

14. The method of claim 1 wherein the A_2B adenosine receptor agonist is administered in eye drops.

15. The method of claim 1 wherein the mammal is a human.

16. A method for assaying compounds to determine if they are A_2B adenosine receptor antagonists or A_2B adenosine receptor agonists comprising the steps of:

a. preparing a first and second sample of human retinal endothelial cells;

b. adding a compound to be tested to the first sample of human retinal endothelial cells and allowing the compound to remain in contact with the first sample of human retinal endothelial cells for a defmed period of time; and

c. comparing the number of new cells grown in the first sample with the number of new cells grown in the second sample.

17. An A_2B adenosine receptor antagonist compound identified by the method of claim 16 wherein the compound caused fewer new cells to grow in the first sample in comparison to the second sample.

18. An A_2B adenosine receptor agonist compound identified by the method of claim 16 wherein the compound caused more new cells to grow in the first sample in comparison to the second sample.