SMALL-MOLECULE COMPOUND HAVING A2A ADENOSINE RECEPTOR ANTAGONISM

Abstract

The present application provides a small-molecule compound, represented by general Formula (I), having A.sub.2A adenosine receptor antagonistic activity and a pharmaceutical composition containing same. The compound and composition can be used as A.sub.2A adenosine receptor antagonistic agents.

##STR00001##

Claims

1. A compound represented by Formula (I), a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof, ##STR00019## wherein: R.sub.1 is selected from the group consisting of aryl, cycloalkyl, heteroaryl, heterocycloalkyl, aryl substituted by one or more substituents R.sup.10, cycloalkyl substituted by one or more substituents R.sup.10, heteroaryl substituted by one or more substituents R.sup.10, heterocycloalkyl substituted by one or more substituents R.sup.10, wherein R.sup.10 is selected from the group consisting of halogen, C.sub.1-6 alkoxy, C.sub.1-6 alkyl, hydroxyl, amino, cyano, nitro, halogenated C.sub.1-6 alkyl, —NHC(O)R.sup.11, benzyloxy, halogenated phenylmethoxy, aryl, heteroaryl, R.sup.11 is C.sub.1-6 alkyl.

2. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sub.1 is selected from the group consisting of phenyl, cyclopentyl, cyclohexyl, pyridyl, thiazolyl, pyrrolyl, imidazolyl, furyl, phenyl substituted by one or more substituents R.sup.10, cyclopentyl substituted by one or more substituents R.sup.10, cyclohexyl substituted by one or more substituents R.sup.10, pyridyl substituted by one or more substituents R.sup.10, thiazolyl substituted by one or more substituents R.sup.10, pyrrolyl substituted by one or more substituents R.sup.10, imidazolyl substituted by one or more substituents R.sup.10, and furanyl substituted by one or more substituents R.sup.10.

3. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sub.1 is selected from the group consisting of phenyl, cyclopentyl, cyclohexyl, pyridin-2-yl, thiazol-5-yl, phenyl substituted by one or more substituents R.sup.10, cyclopentyl substituted by one or more substituents R.sup.10, cyclohexyl substituted by one or more substituents R.sup.10, pyridin-2-yl substituted by one or more substituents R.sup.10, and thiazol-5-yl substituted by one or more substituents R.sup.10.

4. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sup.10 is selected from the group consisting of halogen, C.sub.1-4 alkoxy, C.sub.1-4 alkyl, hydroxyl, amino, cyano, nitro, halogenated C.sub.1-4 alkyl, —NHC(O)R.sup.11, benzyloxy, halogenated phenylmethoxy, aryl, and heteroaryl, R.sup.11 is C.sub.1-4 alkyl.

5. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sub.1 is 4-acetamidophenyl, cyclopentyl, 3,4-dibenzyloxyphenyl, 4-(4-fluorobenzyloxy)phenyl, 3 -benzyloxyphenyl, 2,4-di(trifluoromethyl)phenyl, 4-(pyridin-2-yl)phenyl, 4-phenylphenyl, 4-propoxyphenyl, 4-trifluoromethylphenyl, 5-bromopyridin-2-yl, thiazol-5-yl or cyclohexyl.

6. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sub.1 is phenyl or phenyl substituted by one or more substituents R.sup.10, R.sup.10 is selected from the group consisting of halogen, —NHC(O)R.sup.11, C.sub.1-6 alkoxy, C.sub.1-6 alkyl, hydroxyl, amino, cyano, nitro, halogenated C.sub.1-6 alkyl, benzyloxy, halogenated phenylmethoxy, aryl, and heteroaryl, R.sup.11 is C.sub.1-6 alkyl.

7. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sub.1 is selected from the group consisting of cyclopentyl, thiazolyl and phenyl para- or meta-substituted by substituent R.sup.10, wherein R.sup.10 is selected from the group consisting of —NHC(O)R.sup.11, C.sub.1-6 alkoxy, benzyloxy and halogenated benzyloxy, R.sup.11 is C.sub.1-6 alkyl.

8. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein the compound is selected from the group consisting of: N-{4-{7-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-7H-[1,2,4]triazolo[3,4-i]purin-3-yl}phenyl}acetamide; (2R,3R,4 S, 5R)-2-{3-cyclopentyl-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3R,4S,5R)-2-{3-[3,4-di(benzyloxy)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3R,4S,5R)-2-{3-{4-[(4-fluorobenzyl)oxy]phenyl}-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3R,4S,5R)-2-{3-[3-(benzyloxy)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3R,4S,5R)-2-{3-[2,4-di(trifluoromethyl)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3 S,4R, 5R)-2-{3-[4-(pyridin-2-yl)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3R,4S,5R)-2-{3-{[1,1′-biphenyl]-4-yl}-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3 S,4R, 5R)-2-{3 -(4-propoxyphenyl)-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3S,4R,5R)-2-{3-[4-(trifluoromethyl)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3R,4S,5R)-2-{3-(5-bromopyridin-2-yl)-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; (2R,3 S,4R,5R)-2-{3-(thiazol-5-yl)-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol; and (2R,3R,4S,5R)-2-{3-cyclohexyl-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol.

9. A method for preparing the compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, comprising: ##STR00020## allowing a compound represented by Formula IV to undergo a cyclization reaction to obtain the compound represented by Formula (I), wherein the definition of R.sub.1 is the same as that described in claim 1.

10. A pharmaceutical composition comprising at least one of the compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, and one or more pharmaceutically acceptable carriers or excipients.

11. (canceled)

12. (canceled)

13. A method for preventing and/or treating a human pathological condition or symptom, comprising administering to a patient in need thereof a therapeutically effective amount of the compound represented by Formula (I), a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein the human pathological condition or symptom is improved by antagonizing A.sub.2A adenosine receptor.

14. (canceled)

15. The method according to claim 13, wherein the human pathological condition or symptom is selected from the group consisting of ischemia, supraventricular arrhythmia, atrial fibrillation, acute renal failure, myocardial reperfusion injury, disease caused by fluid retention, allergic reaction, scleroderma, arthritis, inflammatory bowel disease, diabetes, obesity, Parkinson's disease, Huntington's disease, dystonia, dyskinesia, congestive heart failure, hypertension, dialysis hypotension, dementia, anxiety disorder, glaucoma, and alcoholism.

16. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sup.10 is selected from the group consisting of fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, tert-pentyl, neopentyl, hexyl, trifluoromethyl, difluoromethyl, fluoromethyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentyloxy, n-hexyloxy, hydroxyl, amino, cyano, nitro, acetamido, formylamino, propionamido, benzyloxy, fluorophenylmethoxy, phenyl, pyridyl, pyrrolidinyl, cyclopentyl, cyclohexyl, morpholinyl, imidazolyl, and thiazolyl.

17. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sup.10 is selected from the group consisting of fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, hydroxyl, amino, cyano, nitro, acetamido, formylamino, propionamido, benzyloxy, 4-fluorophenylmethoxy, phenyl, pyridin-2-yl, 5-bromopyridin-2-yl, cyclopentyl, and cyclohexyl.

18. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 1, wherein R.sup.11 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, tert-pentyl, neopentyl or hexyl.

19. The compound, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable hydrate or solvate, or a pharmaceutically acceptable ester thereof according to claim 7, wherein R.sub.1 is 4-acetamidophenyl, cyclopentyl, 4-(4-fluorobenzyloxy)phenyl, 3-benzyloxyphenyl, 4-propoxyphenyl, or thiazol-5-yl.

20. The method according to claim 9, wherein the cyclization reaction is carried out in an acetic acid solution and in the presence of NBS, and the cyclization reaction is carried out at room temperature.

21. The method according to claim 9, wherein the compound represented by Formula IV is obtained by reacting a compound represented by Formula II with a substituted formaldehyde represented by Formula III, ##STR00021## wherein the definition of R.sub.1 is the same as that described in claim 9.

22. The method according to claim 21, wherein the compound represented by Formula II reacts with the substituted formaldehyde represented by Formula III in a methanol solution under microwave at 70° C. to 90° C.

23. The method according to claim 22, wherein the compound represented by Formula II is obtained by the hydrazinolysis reaction of 6-chloroadenosine represented by Formula V with hydrazine hydrate, ##STR00022## and the hydrazinolysis reaction is carried out at 60° C. to 80° C.

Description

EXAMPLE 1

Synthesis of N-{4-{7-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl]-7H-[1,2,4]triazolo[3,4-i]purin-3-yl}phenyl}acetamide (Compound 1)

[0108] ##STR00006##

[0109] 1.1 Synthesis of (2R,3R,4S,5R)-2-(6-hydrazino-9H-purin-9-yl)-5-(hydroxymethyl) tetrahydrofuran-3,4-diol (Compound of Formula II)

[0110] 5 g (0.018 mol) of (2R,3R,4S,5R)-2-(6-chloro-9H-purin-9-yl)-5-(hydroxymethyl) tetrahydrofuran-3,4-diol (Compound of Formula V) was added to 10 ml of hydrazine hydrate (65 wt % aqueous solution), heated to 70° C. under stirring, continuously heated for 2 hours until the reactant (Compound of Formula V) disappeared, the progress of the reaction was monitored by TLC (CH.sub.2Cl.sub.2:MeOH=3:1 (v/v)). Then the reaction mixture was heated to 25° C., and 2-propanol (50 ml) was added for dilution under stirring overnight. The separated precipitate was filtered to obtain 4.8 g of (2R,3R,4S,5R)-2-(6-hydrazino-9H-purin-9-yl)-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound of Formula II) as a white solid, which was directly used in the next reaction.

[0111] 1.2 Synthesis of N-{4-{(E)-{2-{9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl]-9H-purin-6-yl}hydrazono}methyl}phenyl}acetamide (Compound of Formula IV)

[0112] 0.5 g (0.0018 mol) of (2R,3R,4S,5R)-2-(6-hydrazino-9H-purin-9-yl)-5-(hydroxymethyl) tetrahydrofuran-3,4-diol (Compound of Formula II) and 0.33 g (0.002 mol) of 4-acetamidobenzaldehyde (Compound of Formula III, wherein R.sub.1 was 4-acetamidophenyl) (1.1 equivalents) were mixed in methanol (20 ml) and heated by microwave at 70° C. for 30 minutes. The crude product was precipitated from methanol. After filtration, the crude product was further purified to obtain 323 mg of white solid (Compound of Formula IV), which was directly used in the next reaction.

[0113] 1.3 Synthesis of N-{4-{7-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl]-7H-[1,2,4]triazolo[3,4-i]purin-3-yl}phenyl}acetamide (Compound 1) 0.5g (0.0012 mol) of N-{4-{(E)-{2-{9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl]-9H-purin-6-yl}hydrazono}methyl}phenyl}acetamide (Compound of Formula IV) was added to 10 ml of acetic acid and stirred to dissolve, then 0.4 g (0.0024 mol) of NBS (N-bromosuccinimide) (2.0 equivalents) was slowly added in batch, continuously stirred for 1 hour. The resulting solution was distilled under reduced pressure to remove acetic acid, and the residue was purified by flash chromatography, in which a mixed solvent of CH.sub.2Cl.sub.2 and MeOH (CH.sub.2Cl.sub.2:MeOH=10:1 (v/v)) was used as the eluent, and 224 mg of white solid (Compound 1) was obtained. m.p. 132° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 10.26(s, 1H), 9.26(s, 1H), 8.70(s, 1H), 7.93(d, 2H J=8.4Hz), 7.85(d, 2H J=8.4Hz),6.08(d, 1H, J=5.6Hz), 5.58(d, 1H, J=6.0Hz), 5.28(d, 1H, J=4.8Hz), 5.08(t, 1H, J=5.6Hz), 4.20(dd, 1H, J=4.0Hz,4.8Hz), 4.00(dd, 1H, J=4.0Hz,4.0Hz), 3.74-3.59(m, 2H), 2.12(s, 3H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.19H.sub.19N.sub.7O.sub.5: 426.1520; found: 426.1520.

[0114] The following Compounds 2 to 13 could be prepared by referring to the method of Example 1, using different reactants (various substituted formaldehydes represented by Formula III) instead of 4-acetamidobenzaldehyde in step 1.2.

EXAMPLE 2

Synthesis of (2R,3R,4S,5R)-2-{3-cyclopentyl-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 2)

[0115] ##STR00007##

[0116] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with cyclopentanecarbaldehyde, the title compound was obtained as 152 mg of white solid (Compound 2). m.p. 84° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.30(s, 1H), 8.66(s, 1H), 6.06(d, 1H J=5.6Hz), 5.60(br, 1H), 5.33(br, 1H), 5.11(br, 1H), 4.57(t, 1H, J=5.2Hz), 4.19(t, 1H, J=4.0Hz), 4.00(dd, 1H, J=4.0Hz,3.6Hz), 3.81-3.72(m, 1H), 3.73-3.58(m, 2H), 2.24-2.18(m, 2H), 2.03-1.93(m, 2H), 1.80-1.68(m, 4H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.16H.sub.20N.sub.6O.sub.4: 361.1619; found: 361.1619.

EXAMPLE 3

Synthesis of (2R,3R,4S,5R)-2-{3-[3,4-di(benzyloxy)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 3)

[0117] ##STR00008##

[0118] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 3,4-benzyloxybenzaldehyde, the title compound was obtained as 190 mg of white solid (Compound 3). m.p. 116° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.13(s, 1H), 8.72(s, 1H), 7.68(d, 1H J=1.6Hz), 7.52-7.30(m, 12H), 6.09(d, 1H, J=5.2Hz), 5.61(d, 1H, J=5.6), 5.33(d, 1H, J=5.2Hz),5.27(s, 4H), 5.11(t, 1H, J=5.2Hz), 4.59(dd, 1H, J=5.6Hz, 5.2Hz), 4.21(dd, 1H J=4.0Hz, 4.4Hz), 4.01(dd, 1H, J=3.6Hz,3.6Hz), 3.75-3.58(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.31H.sub.28N.sub.6O.sub.6: 581.2143; found: 581.2141.

EXAMPLE 4:

Synthesis of (2R,3R,4S,5R)-2-{3-{4-[(4-fluorobenzyl)oxy]phenyl}-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 4)

[0119] ##STR00009##

[0120] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 4-(4-fluorobenzyloxy)benzaldehyde, the title compound was obtained as 235 mg of white solid (Compound 4). m.p. 112° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.21(s, 1H), 8.71(s, 1H), 7.93(d, 2H, J=8.8Hz), 7.56(t, 2H, J=8.8Hz), 7.26(t, 4H, J=8.8Hz), 6.08(d, 1H, J=5.6Hz), 5.61(d, 1H, J=5.6), 5.32(d, 1H, J=4.8Hz), 5.22(s, 2H), 5.10(t, 1H, J=5.6Hz), 4.58(d, 1H, J=4.8Hz), 4.20(d, 1H J=4.0Hz), 4.00(d, 1H, J=4.0Hz), 3.74-3.57(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.24H.sub.21FN.sub.6O.sub.5: 493.1630; found: 493.1630.

EXAMPLE 5

Synthesis of (2R,3R,4S,5R)-2-{3-[3-(benzyloxy)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 5)

[0121] ##STR00010##

[0122] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 3-benzyloxybenzaldehyde, the title compound was obtained as 220 mg of white solid (Compound 5). m.p. 76° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.22(s, 1H), 8.73(s, 1H), 7.64-7.27(m, 9H), 6.08(d, 1H, J=5.2Hz), 5.61(d, 1H, J=6.0Hz), 5.32(d, 1H, J=5.2Hz), 5.25(s, 2H), 5.11(t, 1H, J=5.6Hz), 4.59(dd, 1H, J=5.6Hz, 5.2Hz), 4.20(dd, 1H J=4.4Hz, 4.4Hz), 4.00(dd, 1H, J=4.0Hz, 3.6Hz), 3.74-3.57(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.24H.sub.22N.sub.6O.sub.5: 475.1724; found: 475.1724.

EXAMPLE 6

Synthesis of (2R,3R,4S,5R)-2-{3-[2,4-di(trifluoromethyl)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 6)

[0123] ##STR00011##

[0124] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 2,4-di(trifluoromethyl)benzaldehyde, the title compound was obtained as 205 mg of white solid (Compound 6). .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.10(s, 1H), 8.77(s, 1H), 8.41-8.24(m, 3H), 6.08(d, 1H, J=5.6Hz), 5.60(d, 1H, J=6.0), 5.31(d, 1H, J=4.4Hz), 5.11(t, 1H, J=5.2Hz), 4.58(dd, 1H, J=5.6Hz, 5.2Hz), 4.19(d, 1H J=4.0Hz), 4.00(d, 1H, J=4.0Hz), 3.73-3.57(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.19H.sub.14F.sub.6N.sub.6O.sub.4: 505.1053; found: 505.1052.

EXAMPLE 7

Synthesis of (2R,3S,4R,5R)-2-{3-[4-(pyridin-2-yl)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 7)

[0125] ##STR00012##

[0126] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetaminobenzaldehyde in step 1.2 with 4-(2-pyridyl)benzaldehyde, the title compound was obtained as 195 mg of white solid (Compound 7). m.p. 172° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.37(s, 1H), 8.75(s, 2H), 8.37(d, 2H, J=8.4Hz), 8.15-8.12(m, 3H), 7.99-7.95(m, 1H), 7.46-7.43(m, 1H), 6.10(d, 1H, J=5.6Hz), 5.66(br,1H), 5.38(br, 1H), 5.14(br, 1H), 4.61(t, 1H, J=5.2Hz), 4.22(t, 1H, J=4.0Hz), 4.20(d, 1H, J=4.0Hz), 3.74-3.59(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.22H.sub.19N.sub.7O.sub.4: 446.1571; found: 446.1571.

EXAMPLE 8

Synthesis of (2R,3R,4S,5R)-2-{3-{[1,1′-biphenyl]-4-yl}-7H-[1,2,4]triazolo[3, 4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 8)

[0127] ##STR00013##

[0128] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 4-biphenylaldehyde, the title compound was obtained as 175 mg of white solid (Compound 8). m.p. 280° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.34(s, 1H), 8.74(s, 1H), 8.10(d, 2H, J=8.4Hz), 7.96(d, 2H, J=8.4Hz), 7.81(d, 2H, J=7.2Hz), 7.55(t, 2H, J=7.2Hz), 7.46(t, 1H, J=7.2Hz), 6.10(d, 1H, J=5.6Hz), 5.62(d, 1H, J=5.6Hz), 5.33(d, 1H, J=4.8Hz), 5.13(t, 1H, J=5.6Hz), 4.60(d, 1H, J=5.2Hz), 4.21(d, 1H J=4.0Hz), 4.01(dd, 1H, J=4.0Hz, 3.6Hz), 3.74-3.58(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.23H.sub.20N.sub.6O.sub.4: 445.1619; found: 445.1619.

EXAMPLE 9

Synthesis of (2R,3S,4R,5R)-2-{3-(4-propoxyphenyl)-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 9)

[0129] ##STR00014##

[0130] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetaminobenzaldehyde in step 1.2 with 4-propoxybenzaldehyde, the title compound was obtained as 185 mg of white solid (Compound 9). m.p. 118° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.21(s, 1H), 8.71(s, 1H), 7.90(d, 2H, J=8.8Hz), 7.18(d, 2H, J=8.8Hz), 6.08(d, 1H, J=5.6Hz), 5.62(d, 1H, J=6.0Hz), 5.32(d, 1H, J=5.2Hz), 5.12(t, 1H, J=5.2Hz), 4.58(dd, 1H, J=5.2Hz, 5.2Hz), 4.20(d, 1H J=3.6Hz),4.06(t, 2H, J=6.8Hz), 4.00(d, 1H, J=3.6Hz), 3.74-3.58(m, 2H), 1.83-1.75(m, 2H), 1.03(t, 3H, J=7.6Hz); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.20H.sub.22N.sub.6O.sub.5: 427.1724; found: 427.1725.

EXAMPLE 10

Synthesis of (2R,3S,4R,5R)-2-{3-[4-(trifluoromethyl)phenyl]-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 10)

[0131] ##STR00015##

[0132] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 4-(trifluoromethyl)benzaldehyde, the title compound was obtained as 165 mg of white solid (Compound 10). m.p. 124° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.37(s, 1H), 8.75(s, 1H), 8.25(d, 2H, J=8.0Hz), 8.02(d, 2H, J=8.0Hz), 6.10(d, 1H, J=5.6Hz), 5.62(d, 1H, J=6.0Hz), 5.33(d, 1H, J=4.8Hz), 5.12(t, 1H, J=5.2Hz), 4.60(dd, 1H, J=5.2Hz, 5.6Hz), 4.21(dd, 1H J=3.6Hz, 4.8Hz), 4.01(d, 1H, J=4.0Hz), 3.75-3.58(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.18H.sub.16F.sub.3N.sub.6O.sub.4: 437.1180; found: 437.1179.

EXAMPLE 11

Synthesis of (2R,3R,4S,5R)-2-{3-(5-bromopyridin-2-yl)-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 11)

[0133] ##STR00016##

[0134] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with 5-bromo-2-pyridinecarbaldehyde, the title compound was obtained as 155 mg of yellow solid (Compound 11). m.p. 222° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 10.29(s, 1H), 8.97(s, 1H), 8.77(s, 1H), 8.38-8.32(m, 2H), 6.11(d, 1H, J=5.6Hz), 5.64(d, 1H, J=6.0Hz), 5.33(d, 1H, J=5.2Hz), 5.12(t, 1H, J=5.6Hz), 4.63(dd, 1H, J=5.2Hz, 5.6Hz), 4.23(dd, 1H J=4.0Hz, 4.8Hz), 4.02(dd, 1H, J=4.0Hz, 3.6Hz), 3.76-3.59(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.16H.sub.14BrN.sub.7O.sub.4: 448.0363; found: 448.0361.

EXAMPLE 12

Synthesis of (2R,3S,4R,5R)-2-{3-(thiazol-5-yl)-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 12)

[0135] ##STR00017##

[0136] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with thiazole-5-carbaldehyde, the title compound was obtained as 190 mg of white solid (Compound 12). m.p. 216° C.; .sup.1H NMR (DMSO-d.sub.6): Δ (ppm) 9.52(s, 1H), 9.39(s, 1H), 8.86(s, 1H), 8.76(s, 1H), 6.10(d, 1H, J=5.6Hz), 5.64(d, 1H, J=5.6Hz), 5.34(d, 1H, J=4.8Hz), 5.12(s, 1H), 4.59(dd, 1H, J=5.2Hz, 5.6Hz), 4.21(d, 1H J=4.0Hz), 4.01(dd, 1H, J=4.0Hz, 4.0Hz), 3.75-3.59(m, 2H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.14H.sub.13N.sub.7O.sub.4S: 376.0822; found: 376.0820.

EXAMPLE 13

Synthesis of (2R,3R,4S,5R)-2-{3-cyclohexyl-7H-[1,2,4]triazolo[3,4-i]purin-7-yl}-5-(hydroxymethyl)tetrahydrofuran-3,4-diol (Compound 13)

[0137] ##STR00018##

[0138] Referring to the method of steps 1.2 and 1.3 in Example 1, replacing the 4-acetamidobenzaldehyde in step 1.2 with cyclohexanecarbaldehyde, the title compound was obtained as 215 mg of white solid (Compound 13). m.p. 130° C.; .sup.1H NMR (DMSO-d.sub.6): δ (ppm) 9.37(s, 1H), 8.65(s, 1H), 6.06(d, 1H, J=5.6Hz), 5.60(br, 1H), 5.32(br, 1H), 5.10(s, 1H), 4.57(t, 1H, J=5.2Hz), 4.19(d, 1H J=4.4Hz), 4.00(dd, 1H, J=3.6Hz, 4.0Hz), 3.72-3.58(m, 2H), 2.09-1.31(m, 10H); HRMS (ESI+) m/z [M+H].sup.+ calculated for C.sub.17H.sub.22N.sub.6O.sub.4: 375.1775; found: 375.1776.

EXAMPLE 14

Radioligand Binding Test

[0139] 1) Experimental Materials

[0140] [3H] CGS21680(2-[p-(2-carboxyethyl)phenylethylamino]-5′-N-ethylcarboxamidoadenosine, [carboxy-1-ethyl-3H(N)]-; 250 μCi) was purchased from PerkinElmer Research Products (Boston, Mass.).

[0141] Cell membranes stably transfected with (human) A.sub.2A adenosine receptor were prepared in HEK-293 cells. The cell membranes were obtained from PerkinElmer Research Products (Boston, Mass.).

[0142] CGS 21680 (2-[p-(2-carboxyethyl)phenylethylamino]-5′-ethylcarboxamidoadenosine) was purchased from Selleck (Shanghai, CN).

[0143] All other reagents were of analytical grade and obtained from commercial sources.

[0144] 2) Experimental Method

[0145] The A.sub.2A adenosine receptors used were all expressed in the cell membranes. The compound was diluted 3 times serially with DMSO (Solarbio, D8371-250 ml) to generate compound source plates with 10 different concentrations (10 μM, 3.3 μM, 1.1 μM, 0.37 μM, 0.12 μM, 0.0412 μM, 0.0137 μM, 0.0046 μM, 0.0015 μM, 0.0005 μM), 250 nL of compound was added to 384-well Opti-plate, sealed with parafilm. 20 U hA.sub.2A HEK-293 cell membrane was diluted with 1 mL of assay buffer (50 mM Tris-HCl, pH 7.4, 10 mM MgCl.sub.2, 1 mM EDTA, 1 μg/mL adenosine deaminase), 0.75 uCi [3H]-CGS 21680 (final 25 nM) was added to the diluted cell membrane and mixed well. 50 μL of the prepared dilution solution of cell membrane was transferred to the 384-well Opti-plate containing compound and incubated at 25° C. for 90 minutes. 100 μL of 0.5% Polyethyleneimine solution (PEI) was added to UNIFILTER-96 GF/B filter plate, and soaked for 90 minutes at 4° C., then 500 μL of washing buffer/well (50 mM Tris-HCl, pH 7.4, 154 mM NaCl) was transferred with Cell Harvester to wash the UNIFILTER-96 GF/B filter plate twice. The mixed system in the Opti-plate was transferred to the washed UNIFILTER-96 GF/B filter plate, 500 μL of washing buffer/well (50 mM Tris-HCl, pH 7.4, 154 mM NaCl) was used to wash the UNIFILTER-96 GF/B filter plate 9 times. Incubation was performed at 37° C. for 3 minutes. 40 μL of ULTIMA GOLD scintillation solution (Perkin Elmer, Cat #77-16061) was added to each well, CPM (count per minute) value was read by a MicroBeta liquid scintillation counter (PerkinElmer). The specific binding percentage of [3H]CGS21680 was calculated according to the CPM value, % specific binding of [3H]CGS21680=(CPM.sub.sample−CPM.sub.Low Control)/(CPM.sub.High Control−CPM.sub.Low Control)*100, in which High Control was 0.5% DMSO, Low Control was 100 μM CGS21680. The IC.sub.50 value was calculated by curve fitting according to the compound concentration and the specific binding percentage of [3H]CGS21680.

[0146] 3) Experimental Results

[0147] The inhibition constant (K.sub.i) value was calculated from the IC.sub.50 value according to the Cheng and Prusoff equation, K.sub.i=IC.sub.50/(1+[S]/K.sub.m), wherein [S] was the concentration of radioligand (25 nM) and K.sub.m was dissociation constant (22 nM) of [3H] CGS21680 binding to human A.sub.2AAR. Table 1 showed the inhibition constant K.sub.i for Compounds 1 to 13 of the present application binding to A.sub.2A adenosine receptor.

TABLE-US-00001 TABLE 1 Test results of compounds binding to A.sub.2A adenosine receptor Compound IC.sub.50 (nM) K.sub.i (nM) Compound 1 6.4 3.0 Compound 2 3.9 1.8 Compound 3 >10,000 >10,000 Compound 4 3.9 1.8 Compound 5 6.2 2.9 Compound 6 >10,000 >10,000 Compound 7 >10,000 >10,000 Compound 8 7509 3515 Compound 9 4.3 2.0 Compound 10 >10,000 >10,000 Compound 11 >10,000 >10,000 Compound 12 3.9 1.8 Compound 13 >10,000 >10,000

EXAMPLE 15

A.SUB.2A .Adenosine Receptor cAMP Test

[0148] 1) Experimental Materials

[0149] Experimental reagents and consumables: DMEM/F12, G418, Penicillin-Streptomycin, Versene Solution, HEPES, Hank's Buffered Saline Solution, PBS (pH 7.4, 1×, sterile), FBS, BSA Stabilizer 7.5%, Rolipram, NECA were separately purchased from Gibico, Hyclone and Sigma. LANCE® Ultra cAMP kit (Eu-cAMP tracer, Ulight-anti-cAMP reagent, cAMP detection buffer) and hADORA2A-HEK293 cells were purchased from PerkinElmer Research Products (Boston, Mass.). All other reagents were of analytical grade and obtained from commercial sources. 384-well polypropylene microplate and 384-well solid white plate were purchased from Labcyte and Corning, respectively.

[0150] Experimental instruments: TECAN automated liquid handling workstation, Echo Acoustic Liquid Handler and EnVison multimode plate reader were purchased from TECAN, Labcyte and Envision, respectively.

[0151] 2) Experimental Method

[0152] The cells stably expressing human adenosine receptor A.sub.2A (hADORA.sub.2A-HEK293 cells) were cultured in DMEM/F12 medium containing 10% FBS, 1× Penicillin-Streptomycin and 400 μg/ml G418 in a 37° C. and 5% CO.sub.2 environment. Before the experiment, the cells were digested with Versene solution, and the cells were collected by centrifugation at 200 g and room temperature for 5 minutes, and finally resuspended with assay buffer (Hank's buffered saline solution, containing 5 mM HEPES, 0.1% BSA stabilizer and 10 μM Rolipram, pH 7.4). The TECAN automated liquid handling workstation was used to dilute the compound in a 384-well polypropylene microplate with DMSO to 11 concentration points in a 3-fold gradient to prepare the compound source plate, in which the 11 concentration points of the compound were 10 mM, 3.33 mM, 1.11 mM., 0.37 mM, 0.12 mM, 0.041 mM, 0.013 mM, 4.57×10.sup.−3 mM, 1.52×10.sup.−3 mM, 5×10−4 mM and 1.7×10.sup.−4 mM. The Echo Acoustic Liquid Handler (Labcyte) was used to transfer the test compound from the compound source plate to an assay plate, in which the transfer volume of the compound was 10 nl/well. The hADORA.sub.2A-HEK293 cell suspension was diluted with assay buffer to 30,000 cells/ml, and the cell suspension was transferred to the assay plate at a volume of 10 μl/well (300 cells/well). The assay plate was centrifuged at 150 g for 1 minute and pre-incubated at room temperature for 30 minutes. The Eu-cAMP tracer working solution (Eu-cAMP tracer 40 μl, cAMP detection buffer 1.96 ml) (5 μl/well) was added to the assay plate, and then the Ulight-anti-cAMP working solution (13 μl of Ulight-anti-cAMP reagent and 1.95 ml of cAMP detection buffer) (5 μl/well) was added to the assay plate. The assay plate was rotated at 150 g for 30 seconds and incubated at room temperature for 30 minutes. The EnVison multimode plate reader (PerkinElmer) was used determine the level of cyclic adenosine monophosphate in the final solution (λ.sub.ex=320 nm, λ.sub.em=665 nm & 615 nm). The EC.sub.50 (nM) value of the compound interacting with the A.sub.2A adenosine receptor to stimulate the production of cyclic adenosine monophosphate was calculated and the titer of the compound function was expressed as the EC.sub.50 (nM) value.

[0153] 3) Experimental Results

[0154] When the test compounds interacted with A.sub.2AAR, the EC.sub.50 (nM) values for stimulating the production of cyclic AMP were shown in Table 2. The results showed that Compounds 1, 2, 4, 5, 9, 12 prepared in the present application were all shown as hA.sub.2AAR antagonists. The cAMP EC.sub.50 values of Compounds 1, 2, 4, 5, 9, 12 were basically all greater than 10 μM, which could not stimulate the production of cyclic adenosine monophosphate, indicating these compounds had no agonistic activity.

TABLE-US-00002 TABLE 2 EC.sub.50 value test results of compound A.sub.2A antagonist function determination Compound cAMP EC.sub.50 (nM) Compound 1 >10000 Compound 2 >10000 Compound 4 >10000 Compound 5 >10000 Compound 9 >10000 Compound 12 >10000

[0155] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that according to all the teachings that have been disclosed, various modifications and substitutions can be made to those details, and these changes are all within the protection scope of the present application. The full scope of the present application is given by the appended claims and any equivalents thereof. The publications, patents and patent documents cited in the present application are incorporated herein by reference.