ZA200303729B

ZA200303729B - Compounds specific to adenosine A1, A2A, and A3 receptor and uses thereof.

Info

Publication number: ZA200303729B
Application number: ZA200303729A
Authority: ZA
Inventors: Arlindo L Castelhano; Bryan Mckibben; David J Witter
Original assignee: Osi Pharmaceutical Inc
Priority date: 2000-12-01
Filing date: 2003-05-14
Publication date: 2004-05-14
Also published as: YU42703A; NO20032482L; CA2430577A1; BR0115847A; HUP0400692A3; MXPA03004717A; IL155962A0; MEP35308A; OA13295A; AP2003002807A0; NO20032482D0; CN1489590A; PL363245A1; HUP0400692A2; EP1347980A4; NZ525885A; CN1263757C; AU2002248151B2; JP2004517896A; EA007254B1

Description

COMPOUNDS SPECIFIC TO i ADENOSINE A, BA,,, AND A, RECEPTORS AND USES THEREOF

This application is a continuation-in-part and claims priority of U.S. Serial Nos. 09/728,316, filed December 1, 2000, 09/728,607, filed December 1, 2000, and 09/728,616, filed December 1, 2000, each of which is hereby incorporated by reference in its entirety.

Throughout this application, reference is made to compounds that specifically bind to i) adenosine A; receptors (such as inter alia, pages 4-76, 130-175, and 257-287), ii) adenosine

A,, receptors (such as inter alia, pages 176-201, and pages 288-293), and adenosine A; receptors (such as inter alia, pages 202-256 and 294-300).

Background of the Invention

Adenosine is an ubiquitous modulator of numerous physiological activities, particularly within the cardiovascular and nervous systems. The effects of adenosine appear to be mediated by specific cell surface receptor proteins. Adenosine modulates diverse physiological functions including induction of sedation, vasodilation, suppression of cardiac rate and contractility, inhibition of platelet aggregability, stimulation of gluconeogenesis and inhibition of lipolysis. In addition to its effects on adenylate cyclase, adenosine has been shown to open potassium channels, reduce flux through calcium channels, and inhibit ’ or stimulate phosphoinositide turnover through receptor- mediated mechanisms (See for example, C.E. Muller and B. ' Stein "Adenosine Receptor Antagonists: Structures and

Potential Therapeutic Applications," Current Pharmaceutical

Design, 2:501 (1996) and C.E. Muller "Aj-Adenosine Receptor

Antagonists," Exp. Opin. Ther. Patents 7(5):419 (1997)). /

Adenosine receptors belong to the superfamily of purine receptors which are currently subdivided into P, (adenosine) and P, (ATP, ADP, and other nucleotides) receptors. Four receptor subtypes for the nucleoside adenosine have been cloned so far from various species including humans. Two receptor subtypes (A; and A,,) exhibit affinity for adenosine in the nanomolar range while two other known subtypes A, , and

A; are low-affinity receptors, with affinity for adenosine in the low-micromolar range. A; and A; adenosine receptor activation can lead to an inhibition of adenylate cyclase activity, while A,, and A, activation causes a stimulation of adenylate cyclase.

A few A; antagonists have been developed for the treatment of cognitive disease, renal failure, and cardiac arrhythmias.

It has been suggested that A,, antagonists may be beneficial for patients suffering from Morbus Parkinson (Parkinson's disease). Particularly in view of the potential for local delivery, adenosine receptor antagonists may be valuable for treatment of allergic inflammation and asthma. Available information (for example, Nyce & Metzger "DNA antisense

Therapy for Asthma in an Animal Model" Nature (1997) 385: 721-5)indicates that in this pathophysiologic context, A, antagonists may block contraction of smooth muscle underlying respiratory epithelia, while A, or Aj receptor antagonists may block mast cell degranulation, mitigating the release of histamine and other inflammatory mediators. A, receptors have been discovered throughout the gastrointestinal tract, especially in the colon and the intestinal epithelia. It has . been suggested that A, receptors mediate cAMP response (Strohmeier et al., J. Bio. Chem. (1995) 270:2387-94).

Adenosine receptors have also been shown to exist on the retinas of various mammalian species including bovine, porcine, monkey, rat, guinea pig, mouse, rabbit and human (See, Blazynski et al., Discrete Distributions of Adenosine

Receptors in Mammalian Retina, Journal of Neurochemistry, . volume 54, pages 648-655 (1990) ; Woods et al., .

Characterization of Adenosine A,-Receptor Binding Sites in : Bovine Retinal Membranes, Experimental Eye Research, volume 53, pages 325-331 (1991); and Braas et al., Endogenous adenosine and adenosine receptors localized to ganglion cells of the retina, Proceedings of the National Academy of

Science, volume 84, pages 3906-3910 (1987)). Recently,

Williams reported the observation of adenosine transport sites in a cultured human retinal cell line (Williams et al.,

Nucleoside Transport Sites in a Cultured Human Retinal Cell

Line Established By SV-40 T Antigen Gene, Current Eye

Research, volume 13, pages 109-118 (1994)).

Compounds which regulate the uptake of adenosine uptake have previously been suggested as potential therapeutic agents for the treatment of retinal and optic nerve head damage. In

U.S. Patent No. 5,780,450 to Shade, Shade discusses the use of adenosine uptake inhibitors for treating eye disorders.’

Shade does not disclose the use of specific A, receptor inhibitors. The entire contents of U.S. Patent No. 5,780,450 are hereby incorporated herein by reference.

Additional adenosine receptor antagonists are needed as pharmacological tools and are of considerable interest as drugs for the above-referenced disease states and/or conditions.

Summary of the Invention

The present invention is based on compounds which selectively bind to adenosine A; receptor, thereby treating a disease associated with A: adenosine receptor in a subject by administering to the subject a therapeutically effective amount of such compounds. The disease to be treated are associated with cognitive disease, renal failure, cardiac arrhythmias, respiratory epithelia, transmitter release, sedation, vasoconstriction, bradycardia, negative cardiac inotropy and dromotropy, branchoconstriction, neutropil chemotaxis, reflux condition, or ulcerative condition.

The present invention is based, at least in part, on the discovery that certain N-6 substituted 7-deazapurines, : 15 described infra, can be used to treat a N-6 substituted 7- deazapurine responsive state. Examples of such states include those in which the activity of the adenosine . receptors is increased, €.g., bronchitis, gastrointestinal disorders, or asthma. These states can be characterized in that adenosine receptor activation can lead to the inhibition or stimulation of adenylate cyclase activity. Compositions and methods of the invention include enantiomerically or diastereomerically pure N-6 substituted 7-deazapurines.

Preferred N-6 substituted 7-deazapurines include those which have an acetamide, carboxamide, substituted cyclohexyl, e.g., cyclohexanol, or a urea moiety attached to the N-6 nitrogen through an alkylene chain. . The present invention pertains to methods for modulating an adenosine receptor(s) in a mammal by administering to the . mammal a therapeutically effective amount of a N-¢ substituted 7-deazapurine, such that modulation of the adenosine receptor's activity occurs. Suitable adenosine receptors include the families of A;, A;, or A; In a preferred embodiment, the N-6 substituted 7-deazapurine is a adenosine receptor antagonist.

3 o

The invention further pertains to methods for treating N-6 . substituted 7-deazapurine disorders, e.g., asthma, bronchitis, allergic rhinitis, chronic obstructive pulmonary disease, renal . disorders, gastrointestinal disorders, and eye disorders, in a mammal by administering to the mammal a therapeutically effective amount of a N-6 substituted 7-deazapurine, such that treatment of the disorder in the mammal occurs. Suitable N-6 substituted 7 deazapurines include those illustrated by the general formula I:

Ri

Re

XX

Rs Nd \

R4 (I) and pharmaceutically acceptable salts thereof. Ry; and R, are each independently a hydrogen atom or a substituted or unsubstituted alkyl, aryl, or alkylaryl moiety or together form a substituted or unsubstituted heterocyclic ring. R3 is a substituted or unsubstituted alkyl, aryl, or alkylaryl moiety.

Ry is a hydrogen atom or a substituted or unsubstituted alkyl, aryl, or alkylaryl moiety. Rg and Rg are each independently a halogen atom, e.g., chlorine, fluorine, or bromine, a hydrogen atom or a substituted or unsubstituted alkyl, aryl, or alkylaryl moiety, or Rg is carboxyl, esters of carboxyl, or carboxamides, or Ry; and Rg or Rg and Rg together form a substituted or unsubstituted heterocyclic or carbocyclic ring.

In certain embodiments, R, and R, can each independently be a : substituted or unsubstituted cycloalkyl or heteroarylalkyl moieties. In other embodiments, R3 is a hydrogen atom or a substituted or unsubstituted heterocaryl moiety. In still other embodiments, R,, R: and R¢ can each be independently 2 . heteroaryl moieties. In a preferred emdbodimen:, KR; 1s a hydrogen atom, R, is a cyclohexanol, e.c., trans- . cyclohexanol, Ry is phenyl, R; is a hydrogen a:zom, R: is a methyl group and Rg is a methyl group. In still ancther embodiment, R; is a hydrogen atom, R, is

O

~ NHMe

Ry is phenyl, R; is a hydrogen atom and Rg and R; are methyl groups.

I5

The invention further pertains to pharmaceutical compositions for treating a N-6 substituted 7-deazapurine responsive state in a mammal, e.g., asthma, bronchitis, allergic rhinitis, chronic obstructive pulmonary disease, renal disorders, gastrointestinal disorders, and eye disorders. The pharmaceutical «composition includes a therapeutically effective amount of a N-6 substituted 7-deazapurine and a pharmaceutically acceptable carrier.

The present invention also pertains to packaged pharmaceutical compositions for treating a N-6 substituted 7- deazapurine responsive state in a mammal. The packaged pharmaceutical composition includes a container holding a therapeutically effective amount of at least one N-6 substituted 7-deazapurine and instructions for using the N-¢ ) substituted 7-deazapurine for treating a N-6 substituted 7- deazapurine responsive state in a mammal.

The invention further pertains to compounds of formula 1 wherein . Ry is hydrogen;

R, is substituted or unsubstituted cycloalkyl, Subssizuzed or . 5 unsubstituted alkyl, or Ry; and R, together form a substizu:ted or unsubstituted heterocyclic ring;

R3 is unsubstituted or substituted aryl;

Ry is hydrogen; and

Ry and Rg are each independently hydrogen or alkyl, and pharmaceutically acceptable salts thereof. The deazapurines of this embodiment may advantageously be selective A. receptor antagonists. These compounds may be useful for numerous therapeutic uses such as, for example, the treatment of asthma, kidney failure associated with heart failure, and glaucoma. In a particularly preferred embodiment, the deazapurine is a water soluble prodrug that is capable of

EE being metabolized in vivo to an active drug by, for example, esterase catalyzed hydrolysis.

In yet another embodiment, the invention features a method for inhibiting the activity of an adenosine receptor (e.g.,

A;) in a cell, by contacting the cell with N-6 substituted 7- deazapurine (e.g., preferably, an adenosine receptor antagonist).

In another aspect, the invention features a method for treating damage to the eye of an animal(e.g., a human) by administering to the animal an effective amount of an N-6§ } substituted 7-deazapurine of formula I. Preferably, the N-6 substituted 7-deazapurine is an antagonist of A, adenosine i} receptors in cells of the animal. The damage is to the retina or the optic nerve head and may be acute or chronic. } The damage may be the result of, for example, glaucoma, edema, ischemia, hypoxia or trauma.

Claims

What is claimed is:

1. A compound having the structure: R R ~N N pa NTO N\ Rs FG N N H wherein R;NR, together form a ring having the structure: HO NH, or N NH, \ N lo] OH or Ry is H and R, is: - wherein when R;NR; together are Amended Sheet - 2004-07-27

HO NH, or \ NH, 0 N 0) Rg is H, —-CH, (NCsHg) (OH) (CeHs) , -CH,0CHj3, ~ (CH) 2C(0O)OH, -CH,0CH,C(O)NH,, -C(O)OH, -CHj, ~CH,0 (CH;) 2OH, -CH,OCH,C (0) OCH3, or -CH,OCH,C(O)OH; or OH when R1 is H and R2 is Rs is -CH,(N;C3H3), -C(O)OH, -CH,0(CH;),0H, - C (0) OCH3;, or -C (O) NH, or a pharmaceutically acceptable salt thereof.

Amended Sheet - 2004-07-27

2. The compound of claim 1, having the structure: ony NH2 OH YY Sa) SZ N~ N Cy

3. The compound of claim 1, having the structure: Lo~" N Oo DR ~ N (oy N

4. The compound of claim 1, having the structure: Lo~" N 0 PY ~ N (yr N Amended Sheet —- 2004-07-27

5. The compound of claim 1, having the structure: HO, Law N ©) LD ~~ N (yr N

6. The compound of claim 1, having the structure: LA~"" N 0 Oo N° N OH

N

7. The compound of claim 1, having the structure: LAN N 5 _CONH, 82 ~ N Cy N Amended Sheet —- 2004-07-27

8. The compound of claim 1, having the structure: Lo~" N 0) Pay _ N oO oh N

9. The compound of claim 1, having the structure: LA~™ N 0 DR ~ N Cy N

10. The compound of claim 1, having the structure: S) “Ann, N=\_O Ooh J \ OOH NIN H Amended Sheet - 2004-07-27

11. The compound of claim 1, having the structure: OH N HN ¢ J SH > N N or :

12. The compound of claim 1, having the structure: OH § HN OH 8 AN N 0] or Amended Sheet - 2004-07-27

13. The compound of claim 1, having the structure: OH § HN Na 0) I > N oh H

14. The compound of claim 1, having the structure: OH ) HN 0O— en SN 0 N oh H

15. The compound of claim 1, having the structure: S, “IE, N= 0] Q J « JN 00 NT Sn H Amended Sheet - 2004-07-27

16. The compound of claim 1, having the structure: S NZNO A « JN oon NTN H

17. The compound of claim 1, having the structure: OH ) HN NH x 0] N oh H

18. A pharmaceutically acceptable salt of the compound of claim 6, 8, 12, 15, or 16 wherein the salt contains a cation selected from the group consisting of sodium, calcium and ammonium.

19. A pharmaceutical composition comprising a therapeutically effective amount of the compound of any one of claims 1-17, and a pharmaceutically acceptable carrier. Amended Sheet - 2004-07-27

20. A pharmaceutical composition comprising the compound of any of claims 1-17, a pharmaceutically acceptable carrier and at least one of a steroid, {2 agonist, glucocorticoid, leukotriene antagonist, or an anticholinergic agonist.

21. A water-soluble prodrug of the compound of any of claims 1-17, wherein the water-soluble prodrug ‘is metabolized in vivo to produce an active drug which selectively inhibits Ai adenosine receptor.

22. Use of the compound of any of «claims 1-17 for manufacturing a medicament useful for treating a disease associated with an A1 adenosine receptor in a subject.

23. The use of «claim 22, wherein said A1 adenosine receptor 1s associated with cognitive disease, renal failure, cardiac arrhythmias, respiratory epithelia, transmitter release, sedation, vasoconstriction, bradycardia, negative cardiac inotropy and dromotropy, bronchoconstriction, neutrophil chemotaxis, reflux condition, or ulcerative condition.

24. The use of claim 22, wherein said disease is asthma, chronic obstructive pulmonary disease, allergic rhinitis, or an upper respiratory disorder. Amended Sheet - 2004-07-27

25. The use of claim 22, wherein the disease is asthma, allergic rhinitis, or chronic obstructive pulmonary disease.

26. The use of claim 22, wherein the A1 adenosine receptor is associated with congestive heart failure.

27. Use of the compound of any of claims 1-17 for manufacturing a medicament useful for treating a disease associated with an A: adenosine receptor in a subject 1n need of such treatment, wherein the disease is antidiuresis, bradycardia, bronchitis, bronchoconstriction, Alzheimer’s disease, cardiac arrythmias, cardiac hypoxia, hypertension, inflammation, negative cardiac inotropy and dromotropy, renal failure, sedation or is associated with transmitter release, respiratory epithelia, contraction of smooth muscle underlying respiratory epithelia, vasoconstriction or mast cell degranulation.

28. Use of the compound of any of claims 1-17 for enhancing the memory of a subject.

29. Use of the compound of any of claims 1-17 for manufacturing a medicament useful for inhibiting the activity of an Al adenosine receptor in a cell. Amended Sheet - 2004-07-27

30. A packaged pharmaceutical composition for treating a disease associated with an Al adenosine receptor in a subject, comprising: (a) a container holding a therapeutically effective amount of the compound of any of claims 1-17; and (b) instructions for using said compound for treating said disease in a subject.

31. A compound having the structure: (R) NH, ig N O ==N I \ 2 on NTN / H or a pharmaceutically acceptable salt.

32. A compound having the structure: CP N 0] ~=N QO — - I \ 4 CO N N Vi H or a pharmaceutically acceptable salt. Amended Sheet - 2004-07-27

33. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 31 or 32 and a pharmaceutically acceptable carrier.

34. The pharmaceutical composition of claim 33, wherein said therapeutically effective amount is effective to treat Parkinson’s disease and diseases associated with locomotor activity, vasodilation, platelet inhibition, neutrophil superoxide generation, cognitive disorder, or senile dementia.

35. The pharmaceutical composition of claim 33, wherein said pharmaceutical composition 1s an ophthalmic formulation.

36. The pharmaceutical composition of claim 19 or 33, wherein said pharmaceutical composition is an periocular, retrobulbar or intraocular injection formulation.

37. The pharmaceutical composition of claim 19 or 33, wherein said pharmaceutical composition is a systemic formulation.

38. The pharmaceutical composition of claim 19 or 33, wherein said pharmaceutical composition is a surgical irrigating solution. Amended Sheet - 2004-07-27

39. A pharmaceutical composition comprising the compound of claim 31 or 32, a pharmaceutically acceptable carrier and at least one dopamine enhancer.

40. A pharmaceutical composition comprising the compound of claim 31 or 32, a pharmaceutically acceptable carrier and at least one cytotoxic agent.

41. A pharmaceutical composition comprising the compound of claim 31 or 32, a pharmaceutically acceptable carrier and at least one of a prostaglandin agonist, a muscrinic agonist, or a B-2 antagonist.

42. A water-soluble prodrug of the compound of claim 31 or 32, wherein said water-soluble prodrug is metabolized in vive to an active drug which selectively inhibits the A;, adenosine receptor.

43. The prodrug of claim 42, wherein said prodrug is metabolized in vivo by esterase catalyzed hydrolysis.

44. A pharmaceutical composition comprising the prodrug of claim 21 or 42 and a pharmaceutically acceptable carrier.

45. Use of the compound of «claim 31 or 32 for manufacturing a medicament useful for treating a disease associated with an A2a adenosine receptor in a

. subject. Amended Sheet -~ 2004-07-27

46. The use of claim 22 or 45, wherein the subject 1is human.

47. The use of claim 45, wherein said A,, adenosine receptor 1s associated with locomotor activity, vasodilation, platelet inhibition, neutrophil superoxide generation, cognitive disorder, senile dementia, or Parkinson’s disease.

48. The use of claim 45, wherein the compound treats said diseases by stimulating adenylate cyclase.

49. Use of the compound of «claim 31 or 32 for manufacturing a medicament useful for treating a disease associated with an A2a adenosine receptor in a subject in need of such treatment, wherein the disease associated with the A2a adenosine receptor is Parkinson’s disease or glaucoma.

50. Use of the compound of «claim 31 or 32 for manufacturing a medicament useful for inhibiting the activity of an A, adenosine receptor in a cell.

51. A packaged pharmaceutical composition for treating a disease associated with an A;; adenosine receptor in a subject, comprising: (a) a container holding a therapeutically effective amount of the compound of claim 31 or 32; and Amended Sheet - 2004-07-27

- 317 = (b) instructions for using said compound for treating said disease in a subject.

52. A compound having the structure: N ¢ N H NH OD ~ N or N N or a pharmaceutically acceptable salt thereof.

53. A pharmaceutically acceptable salt of the compound of claim 31, 32 or 52, wherein the pharmaceutically acceptable salt contains an anion selected from the group consisting of maleic, fumaric, tartaric, acetate, phosphate and mesylate.

54. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 52 and a pharmaceutically acceptable carrier.

55. The pharmaceutical composition of claim 54, further comprising one or more compounds selected from the group consisting of beta adrenoceptor antagonists, alpha-2 adrenoceptor agonists, carbonic anhydrase inhibitors, cholinergic agonists, prostaglandins and prostaglandin receptor agonists, angiotensin converting enzyme (ACE) inhibitors, AMPA receptor antagonists, 5-HT agonists, angiogenesis inhibitors, Amended Sheet - 2004-07-27

NMDA antagonists, renin inhibitors, cannabinoid receptor agonists, angiotensin receptor antagonists, hydrochlorothiazide (HCTZ), somatostatin agonists, glucocorticoid antagonists, mast cell degranulation inhibitors, alpha-adrenergic receptor blockers, alpha-2 adrenoceptor antagonists, thromboxane A2 mimetics, protein kinase inhibitors, prostaglandin F ’ derivatives, prostaglandin-2 alpha antagonists, dopamine D1 and 5-HT2 agonists, nitric-oxide- releasing agents, 5-HT 2 antagonists, cyclooxygenase inhibitors, inosine, dopamine D2 receptor and alpha 2 adrenoceptor agonists, dopamine D1 receptor antagonist and D2 receptor agonists, vasopressin receptor antagonists, endothelin antagonists, 1-(3- hydroxy-2-phosphonylmethoxypropyl) cytosine (HPMPC) and related analogs and prodrugs, thyroid hormone receptor ligands, muscarinic M1 agonists, sodium channel blockers, mixed-action ion channel blockers, beta adrenoceptor antagonist and PGF2 alpha agonist combinations, guanylate cyclase activators, nitrovasodilators, endothelin receptor modulators, ethacrynic acid, other phenoxyacetic acid analogs, actin disrupters, calcium channel blockers and neuroprotective agents.

56. The pharmaceutical composition of claim 54, further comprising one or more compounds selected from the group consisting of beta adrenoceptor antagonists, alpha-2 adrenoceptor agonists, carbonic anhydrase inhibitors, cholinergic agonists and prostaglandin receptor agonists. Amended Sheet - 2004-07-27

57. Use of the compound of claim 52 for manufacturing a medicament useful for treating damage to the eye of a subject.

58. The use of claim 57, wherein the damage comprises retinal or optic nerve head damage.

59. Use of the compound of claim 52 for manufacturing a medicament useful for treating glaucoma in a subject.

60. The use of claim 59, wherein the medicament further comprises one or more compounds selected from the group consisting of beta adrenoceptor antagonists, alpha-2 adrenoceptor agonists, carbonic anhydrase inhibitors, cholinergic agonists, prostaglandins and prostaglandin receptor agonists, angiotensin converting enzyme (ACE) inhibitors, AMPA receptor antagonists, 5-HT agonists, angiogenesis inhibitors, NMDA antagonists, renin inhibitors, cannabinoid receptor agonists, angiotensin receptor antagonists, hydrochlorothiazide (HCTZ), somatostatin agonists , glucocorticoid antagonists, mast cell degranulation inhibitors, alpha-adrenergic receptor blockers, alpha-2 adrenoceptor antagonists, thromboxane A2 mimetics, protein kinase inhibitors, prostaglandin F derivatives, prostaglandin-2 alpha antagonists, dopamine D1 and 5-HT2 agonists, nitric-oxide- releasing agents, 5-~HT 2 antagonists, cyclooxygenase inhibitors, inosine, dopamine D2 receptor and alpha 2 adrenoceptor agonists, dopamine D1 receptor antagonist and D2 receptor agonists, vasopressin Amended Sheet - 2004-07-27 ’

receptor antagonists, endothelin antagonists, 1-{(3- hydroxy-2-phosphonylmethoxypropyl) cytosine (HPMPC) and related analogs and prodrugs, thyroid hormone receptor ligands, muscarinic M1 agonists, sodium channel blockers, mixed-action ion channel blockers, beta adrenoceptor antagonist and PGF2 alpha agonist combinations, guanylate cyclase activators, nitrovasodilators, endothelin receptor modulators, ethacrynic acid, other phenoxyacetic acid analogs, actin disrupters, calcium channel blockers and neuroprotective agents.

61. The use of claim 59, wherein the medicament further comprises one or more compounds selected from the group consisting of beta adrenoceptor antagonists, alpha-2 adrenoceptor agonists, carbonic anhydrase inhibitors, cholinergic agonists and prostaglandin receptor agonists.

62. Use of the compound of claim 52 for manufacturing a medicament useful for treating a disease associated with an A3 adenosine receptor in a subject in need of such treatment, wherein the disease associated with the A3 adenosine receptor is asthma, bronchitis, chronic obstructive pulmonary disorder, or bronchoconstriction.

63. Use of the compound of claim 52 for manufacturing a medicament useful for treating a disease associated with an A3 adenosine receptor in a subject in need of such treatment, wherein the disease associated with Amended Sheet - 2004-07-27 the A3 adenosine receptor 1s myocardial ischemia, bronchitis, or bronchoconstriction.

64. Use of the compound of claim 52 for manufacturing a medicament useful for treating inflammation of the eye associated with an A3 adenosine receptor in a subject.

65. Use of the compound of claim 52 for manufacturing a medicament useful for treating a disease associated with an A3 adenosine receptor in a subject in need of such treatment, wherein the disease associated with the A3 adenosine receptor is associated with mast cell degranulation.

066. Use of the compound of claim 52 for manufacturing a medicament useful for treating a disease associated with an A3 adenosine receptor in a subject in need of such treatment, wherein the disease associated with the A3 adenosine receptor is asthma, glaucoma, retinopathy, ocular ischemia, or macular degeneration.

67. Use of the compound of claim 52 for manufacturing a medicament useful for inhibiting the activity of an A3 adenosine receptor in a cell.

68. The use of claim 29, 50 or 67, wherein the cell is a human cell. Amended Sheet - 2004-07-27

69. A packaged pharmaceutical composition for treating a disease associated with an A3 adenosine receptor in a subject, comprising: (a) a container holding a therapeutically effective amount of the compound of claim 52; and (b) instructions for using said compound for treating said disease in a subject.

70. A method of making a composition which comprises the compound of claim 52, the method comprising admixing the compound with a suitable carrier.

71. A process of manufacturing the compound of claim 52, comprising the steps of: (a) reacting the compound Cl NF | AN NN N ] N H Amended Sheet - 2004-07-27

- 323 -~ with histamine in the presence of dimethyl sulfoxide; (b) heating the reaction mixture under an inert atmosphere; (c) cooling the reaction mixture and separating the organic layer from the aqueous layer; (d) washing the organic layer with brine, drying, and filtering, to yield the compound.

72. The process of claim 71, wherein in step (b) the reaction mixture is heated under a nitrogen atmosphere.

73. The process of claim 71, wherein in step (d) the organic layer is dried with MgSO;. Amended Sheet - 2004-07-27