PYRROLOPYRIMIDINE COMPOUNDS USED AS TLR7 AGONIST

Abstract

The present invention relates to a pyrrolopyrimidine compound as TLR7 agonist, and particularly relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, a preparation process thereof, a pharmaceutical composition containing such compounds and use thereof for manufacturing a medicament against viral infection.

##STR00001##

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof ##STR00075## wherein L.sub.1 and L.sub.2 are each independently selected from the group consisting of —O—, —CH.sub.2—, —S—, —NH—, —NHC(═O)—, —C(═O)—, —C(═O)NH—, —S(═O)—, —S(═O).sub.2—, —NHS(═O).sub.2— and —S(═O).sub.2NH—, wherein the above groups are optionally substituted by one or more R.sub.4; R.sub.1 is selected from the group consisting of hydrogen, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl, wherein the above C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl are optionally substituted by one or more R.sup.4; R.sub.2 is selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, thiol, amino, COOH, —CONH.sub.2, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl, wherein the above hydroxyl, thiol, amino, COOH, —CONH.sub.2, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl are optionally substituted by one or more R.sup.4; B is selected from the group consisting of C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl; L.sub.3 is selected from the group consisting of C.sub.0-6 alkylene, imino, —O—, —S—, —S(═O)— and —S(═O).sub.2—, wherein the above C.sub.0-6 alkylene and imino are optionally substituted by one or more R.sub.4; R.sub.3 is selected from the group consisting of hydrogen, amino, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 to alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl, wherein the above amino, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, aryl and heteroaryl are optionally substituted by one or more R.sub.4; or R.sub.3 and L.sub.3 together with the adjacent atom at the ring B form a saturated or unsaturated 5-8 membered ring, the 5-8 membered ring is optionally substituted by one or more R.sub.4; n is 0, 1, 2, 3, 4 or 5; R.sub.4 is selected from the group consisting of halogen, cyano, —R, —OR, ═O, —SR, —NR.sub.2, ═NR, —C(halogen).sub.3, —CR(halogen).sub.2, —CR.sub.2(halogen), —OCN, —SCN, —N═C═O, —NCS, —NO, —NO.sub.2, —NRC(═O)R, —NRC(═O)OR, —NRC(═O)NRR, —C(═O)NRR, —C(═O)OR, —OC(═O)NRR, —OC(═O)OR, —C(═O)R, —S(═O).sub.2OR, —S(═O).sub.2R, —OS(═O).sub.2OR, —S(═O).sub.2NRR, —S(═O)R, —NRS(═O).sub.2R, —NRS(═O).sub.2NRR, —NRS(═O).sub.2OR, —OP(═O)(OR).sub.2, —P(═O)(OR).sub.2, —C(═O)R, —C(═S)R, —C(═O)OR, —C(═S)OR, —C(═O)SR, —C(═S)SR, —C(═O)NRR, —C(═S)NRR, —C(═NR)NRR and —NRC(═NR)NRR; R is independently selected from the group consisting of H, C.sub.1-8 alkyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl; and when L.sub.1 is —CH.sub.2— or —NH—, R.sub.3 is not H.

2. The compound according to claim 1, characterized in that, L.sub.1 and L.sub.2 are each independently selected from the group consisting of —O—, —CH.sub.2—, —S—, —NH—, —C(═O)—, —S(═O)— and —S(═O).sub.2—, wherein the above —CH.sub.2— and —NH— are optionally substituted by one or more R.sub.4.

3. The compound according to claim 2, characterized in that, L.sub.1 and L.sub.2 are each independently selected from the group consisting of —O— and —CH.sub.2—, wherein the above —CH.sub.2— is optionally substituted by one or more R.sub.4.

4. The compound according to claim 1, characterized in that, R.sub.1 is selected from the group consisting of hydrogen, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cyclohydrocarbyl, 3-6 membered heterocyclohydrocarbyl, aryl and heteroaryl, wherein the above C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cyclohydrocarbyl, 3-6 membered heterocyclohydrocarbyl, aryl and heteroaryl are optionally substituted by one or more R.sub.4.

5. The compound according to claim 4, characterized in that, R.sub.1 is selected from the group consisting of C.sub.1-6 alkyl, wherein the above C.sub.1-6 alkyl is optionally substituted by one or more R.sub.4.

6. The compound according to claim 1, characterized in that, R.sub.2 is selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, amino, —CONH.sub.2 and C.sub.1-6 alkyl, wherein the above hydroxyl, amino, —CONH.sub.2 and C.sub.1-6 alkyl are optionally substituted by one or more R.sub.4.

7. The compound according to claim 6, characterized in that, R.sub.2 is selected from the group consisting of hydrogen, cyano and —CONH.sub.2, wherein the above —CONH.sub.2 is optionally substituted by one or more R.sub.4.

8. The compound according to claim 1, characterized in that, B is selected from the group consisting of aryl and heteroaryl.

9. The compound according to claim 8, characterized in that, B is selected from the group consisting of phenyl and pyridyl.

10. The compound according to claim 1, characterized in that, L.sub.3 is selected from the group consisting of C.sub.0-6 alkylene, wherein the above C.sub.0-6 alkylene is optionally substituted by one or more R.sub.4.

11. The compound according to claim 1, characterized in that, R.sub.3 is selected from the group consisting of hydrogen, amino, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, amyl and heteroaryl, wherein the above amino, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, aryl and heteroaryl are optionally substituted by one or more R.sub.4; or R.sub.3 and L.sub.3 together with the adjacent atom at the ring B form a saturated or unsaturated 5-8 membered ring, the 5-8 membered ring is optionally substituted by one or more R.sub.4.

12. The compound according to claim 1, characterized in that, R.sub.4 is selected from the group consisting of halogen, cyano, —R, —OR, ═O, —SR, —NR.sub.2, ═NR, —C(halogen).sub.3, —CR(halogen).sub.2, —CR.sub.2(halogen), —OCN, —SCN, —N═C═O, —NCS, —NO, —NO.sub.2, —NRC(═O)R, —C(═O)NRR, —C(═O)OR, —OC(═O)NRR, —C(═O)R, —S(═O).sub.2OR, —S(═O).sub.2R, —OS(═O).sub.2OR, —S(═O).sub.2NRR, —S(═O)R, —NRS(═O).sub.2R, —C(═O)R, —C(═O)OR and —C(═O)NRR.

13. The compound according to claim 12, characterized in that, R.sub.4 is selected from the group consisting of halogen, cyano, —R, —OR, ═O, —NR.sub.2, ═NR, —C(halogen).sub.3, —CR(halogen).sub.2 and —CR.sub.2(halogen).

14. The compound according to claim 1, selected from: ##STR00076## ##STR00077## ##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082## or the pharmaceutically acceptable salt thereof.

15. A method for treating viral infection, comprising administering the compound according to claim 1 or the pharmaceutically acceptable salt thereof to a subject in need thereof.

16. A pharmaceutical composition, comprising the compound according to claim 1 or the pharmaceutically acceptable salt thereof in a therapeutically effective amount and one or more pharmaceutically acceptable carriers or excipients.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0072] FIG. 1: in vivo pharmacodynamics in HDI mouse model infected with hepatitis b virus (plasma).

[0073] FIG. 2: in vivo pharmacodynamics in HDI mouse model infected with hepatitis b virus (liver).

EXAMPLES

[0074] The following Examples are intended to illustrate the invention and should not be understood as a limitation to the scope thereof.

Example 1

2-butoxy-7-(3-((4-methylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0075] ##STR00015##

##STR00016## ##STR00017##

Example 1 Procedures

[0076] Step A: 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (4 g, 21.4 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), to which was added sodium hydride (1.03 g, 60% mineral oil mixture, 25.6 mmol) in portions at 0° C. The reaction liquid was stirred at room temperature for 30 min and (2-(chloromethoxyl)ethyl)trimethylsilane (3.9 g, 23.5 mmol) was added dropwise. The mixture was further stirred at room temperature for 2 h and was diluted with water (120 mL) and extracted with ethyl acetate (100 mL×2). The combined organic layer was washed with saturated aqueous sodium carbonate solution and saline, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: ethyl acetate/petroleum ether 5% to 10%) to give 2,4-dichloro-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine (5.8 g, 85%) as yellow solid.

[0077] MS(ESI) M/Z: 318[M+H.sup.+].

[0078] Step B: In 1000 mL high pressure reactor, 2,4-dichloro-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine (5 g, 15.8 mmol), isopropanol (15 mL) and aqueous ammonia (250 mL) were mixed and the mixture was stirred at 100-110° C. for 3 h. After the mixture was cooled to room temperature, it was diluted with water (250 mL) and filtered to give 2-chloro-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (4 g, 85%), which was not further purified.

[0079] MS(ESI) M/Z: 299[M+H.sup.+].

[0080] Step C: 2-chloro-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (4 g, 13.4 mmol) and sodium butoxide (5.15 g, 53.6 mmol) were dissolved in n-butanol (55 mL). The mixture was heated to 100° C. under nitrogen atmosphere and stirred for 8 h. After the mixture was cooled to room temperature, it was diluted with water (200 mL), extracted with ethyl acetate (100 mL×3). The combined organic layer was washed with saline, dried with anhydrous sodium sulfate, and concentrated under 560 reduced pressure. The residue was purified with silica gel column chromatography (eluent: ethyl acetate/petroleum ether 15% to 25%) to give 2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (4.1 g, 91%) as yellow solid.

[0081] MS(ESI) M/Z: 337[M+H.sup.+].

[0082] Step D: 2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (4 g, 12 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). NBS (2.2 g, 12.5 mmol) was formulated as saturated solution in anhydrous tetrahydrofuran, which was added into the above solution over 20 min at a temperature below 0° C. After addition, the reaction mixture was stirred for 30 min at 0° C., and diluted with saline (150 mL), and extracted with ethyl acetate (100 mL×3). The combined organic layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: ethyl acetate/petroleum ether 5% to 15%) to give 7-bromo-2-butoxy-5-(2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (3.85 g, 78%) as white solid.

[0083] MS(ESI) M/Z: 415, 417[M+H.sup.+].

[0084] Step E: At −78° C., n-butyllithium (2.5 M, 12 mL, 30 mmol) was added into a solution of 7-bromo-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (3 g, 7.25 mmol) in anhydrous tetrahydrofuran (40 mL) under nitrogen atmosphere with stirring. The reaction mixture was stirred at −78° C. for 1 h and then a solution of 1,3-benzenedialdehyde (1.26 g, 9 mmol) in anhydrous tetrahydrofuran (5 mL) was added slowly. The mixture was further stirred for 30 min at −78° C., then poured into saturated ammonium chloride aqueous solution (15 mL) and was extracted with ethyl acetate (60 mL×2). The combined organic layer was concentrated under reduced pressure and the residue was purified with preparative HPLC to give 1.1 g of 3-((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(hydroxyl)methyl)benzaldehyde salt.

[0085] MS(ESI) M/Z: 471[M+H.sup.+].

[0086] Step F: At 0° C., to a solution of 3-((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(hydroxyl)methyl)benzaldehyde (200 mg, 0.43 mmol) and 1-methylpiperazine (87 mg, 0.87 mmol) in ethanol (2.5 mL) was added sodium cyanoborohydride (40 mg, 0.64 mmol) in portions with stirring. The reaction mixture was stirred at room temperature for 2 h, diluted with water (10 ml) and extracted with ethyl acetate (15 mL×2). The combined organic layer was dried with anhydrous sodium sulfate and concentrated under 595 reduced pressure to give crude (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(3-((4-methylpiperazine-1-yl)methyl)phenyl)methanol, which was used for the next step directly.

[0087] MS(ESI) M/Z: 555 [M+H.sup.+].

[0088] Step G: To a solution of (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(3-((4-methylpiperazine-1-yl)methyl)phenyl)methanol (100 mg) in trifluoroacetic acid (2 mL) was added triethylsilane (0.4 mL) in portions with stirring. The reaction mixture was stirred at 55° C. for 1 h under nitrogen atmosphere and concentrated under reduced pressure. The residue was dissolved in an anhydrous solution of potassium carbonate (100 mg) in methanol (5 mL). The mixture was further stirred at 50° C. for 30 min and filtered. The filtrate was concentrated under reduced pressure and the residue was purified with preparative HPLC to give 36 mg of 2-butoxy-7-(3-((4-methylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine trifluoroacetate.

[0089] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.33-7.21 (m, 4H), 4.55 (t, J=6.8 Hz, 2H), 4.01 (s, 2H), 3.67 (s, 2H), 3.29-3.24 (m, 4H), 2.87-2.80 (m, 7H), 1.87-1.80 (m, 2H), 1.56-1.49 (m, 2H), 1.02 (t, J=6.8 Hz, 3H).

[0090] MS(ESI) m/z: 409 [M+H.sup.+].

Example 2

2-butoxy-7-(3-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0091] ##STR00018##

[0092] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(3-(morpholino methyl)phenyl)methanol was prepared according to Example 1, wherein morpholine was used instead of 1-methylpiperazine in Step F.

[0093] LCMS(ESI) m/z: 542 [M+H.sup.+].

[0094] Step B: 2-butoxy-7-(3-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step G according to Example 1.

[0095] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.41 (s, 2H), 7.35-7.24 (m, 5H), 4.49 (t, J=6.8 Hz, 2H), 4.03 (s, 2H), 3.82 (s, 2H), 3.77-3.75 (m, 4H), 2.77-2.73 (m, 4H), 1.83-1.79 (m, 2H), 1.55-1.49 (m, 2H), 1.01 (t, J=6.8 Hz, 3H). MS(ESI) m/z: 396 [M+H.sup.+].

Example 3

7-(3-(aminomethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0096] ##STR00019##

[0097] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(3-(aminomethyl)phenyl)methanol was prepared according to Example 1, wherein ammonium acetate was used instead of 1-methylpiperazine in Step F.

[0098] LCMS(ESI) m/z: 472 [M+H.sup.+].

[0099] Step B: 7-(3-(aminomethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step G according to Example 1.

[0100] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.31-7.15 (m, 4H), 7.06 (s, 1H), 4.32 (t, J=6.6 Hz, 2H), 4.00 (s, 2H), 3.80 (s, 2H), 1.79-1.73 (m, 2H), 1.56-1.50 (m, 2H), 1.01 (t, J=7.4 Hz, 3H).

[0101] MS(ESI) m/z: 326 [M+H.sup.+].

Example 4

2-butoxy-7-(3-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0102] ##STR00020##

[0103] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(3-(pyrrolidine-1-ylmethyl)phenyl)methanol was prepared according to Example 1, wherein pyrrolidine was used instead of 1-methylpiperazine in Step F.

[0104] Step B: 2-butoxy-7-(3-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step G according to Example 1.

[0105] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.50 (s, 2H), 7.41-7.28 (m, 5H), 4.45 (t, J=6.8 Hz, 2H), 4.31 (s, 2H), 4.06 (s, 2H), 3.31-3.29 (m, 4H), 2.10-2.07 (m, 4H), 1.81-1.76 (m, 2H), 1.54-1.49 (m, 2H), 1.01 (t, J=6.8 Hz, 3H).

[0106] MS(ESI) m/z: 380 [M+H.sup.+].

Example 5

2-butoxy-7-(4-((3,3-difluoropyrrolidine-1-yl)methyl)benzyl-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0107] ##STR00021##

[0108] Step A: 4-((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(hydroxyl)methyl)benzaldehyde was prepared according to Example 1, wherein 1,4-benzenedialdehyde was used instead of 1,3-benzenedialdehyde in Step E.

[0109] LCMS(ESI) m/z: 471 [M+H.sup.+].

[0110] Step B: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((3,3-difluoropyrrolidine-1-yl)methyl)phenyl)methanol was prepared according to Example 1, wherein 3,3-difluoropyrrolidine was used instead of 1-methylpiperazine in Step F.

[0111] LCMS(ESI) m/z: 562 [M+H.sup.+].

[0112] Step C: 2-butoxy-7-(4-((3,3-difluoropyrrolidine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step G according to Example 1.

[0113] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.28-7.15 (m, 4H), 7.04 (s, 1H), 4.30 (t, J=6.4 Hz, 2H), 3.97 (s, 2H), 3.59 (s, 2H), 2.88-2.71 (m, 4H), 2.30-2.19 (m, 2H), 1.78-1.71 (m, 2H), 1.55-1.46 (m, 2H), 0.98 (t, J=7.2 Hz, 3H).

[0114] MS(ESI) m/z: 416 [M+H.sup.+].

Example 6

2-butoxy-7-(4-((3-fluoropyrrolidine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0115] ##STR00022##

[0116] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((3-fluoropyrrolidine-1-yl)methyl)phenyl)methanol was prepared according to Example 5, wherein 3-fluoropyrrolidine was used instead of 3,3-difluoropyrrolidine in Step B.

[0117] LCMS(ESI) m/z: 544 [M+H.sup.+].

[0118] Step B: 2-butoxy-7-(4-((3-fluoropyrrolidine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0119] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.30-7.24 (m, 4H), 7.06 (s, 1H), 5.24-5.08 (m, 1H), 4.32 (t, J=6.4 Hz, 2H), 3.99 (s, 2H), 3.69-3.57 (m, 2H), 2.88-2.65 (m, 4H), 2.45-2.43 (m, 1H), 2.25-2.11 (m, 1H), 2.02-1.91 (m, 1H), 1.78-1.73 (m, 2H), 1.57-1.50 (m, 2H), 1.01 (t, J=7.2 Hz, 3H).

[0120] MS(ESI) m/z: 398[M+H.sup.+].

Example 7

1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-7-1)methyl)benzyl)pyrrolidine-3-ol

[0121] ##STR00023##

[0122] Step A: 1-(4-((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(hydroxyl)methyl)benzyl)pyrrolidine-3-ol was prepared according to Example 5, wherein pyrrolidine-3-ol was used instead of 3,3-difluoropyrrolidine in Step B.

[0123] LCMS(ESI) m/z: 542 [M+H.sup.+].

[0124] Step B: 1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)benzyl)pyrrolidine-3-ol formate was prepared with the procedures of Step C according to Example 5.

[0125] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.43 (s, 2H), 7.45-7.39 (m, 4H), 7.25 (s, 1H), 4.53 (m, 1H), 4.44-4.27 (m, 2H), 4.04 (s, 2H), 3.54-3.47 (m, 1H), 3.38-3.36 (m, 4H), 3.22-3.19 (m, 1H), 2.28-2.24 (m, 1H), 2.05-2.01 (m, 1H), 1.82-1.76 (m, 2H), 1.56-1.50 (m, 2H), 1.01 (t, J=7.2 Hz, 3H).

[0126] MS(ESI) m/z: 396 [M+H.sup.+].

Example 8

2-butoxy-7-(4-(piperidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0127] ##STR00024##

[0128] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-(piperidine-1-ylmethyl)phenyl)methanol was prepared according to Example 5, wherein piperidine was used instead of 3,3-difluoropyrrolidine in Step B.

[0129] LCMS(ESI) m/z: 540 [M+H.sup.+].

[0130] Step B: 2-butoxy-7-(4-(piperidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0131] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.28 (d, J=8.0 Hz, 2H), 7.22 (d, J=8.0 Hz, 2H), 7.04 (s, 1H), 4.30 (t, J=6.6 Hz, 2H), 3.98 (s, 2H), 3.47 (s, 2H), 2.42 (s, 4H), 1.77-1.73 (m, 2H), 1.60-1.57 (m, 4H), 1.52-1.46 (m, 4H), 0.99 (t, J=7.4 Hz, 3H).

[0132] MS(ESI) m/z: 394 [M+H.sup.+].

Example 9

2-butoxy-7-(4-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0133] ##STR00025##

[0134] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-(morpholino methyl)phenyl)methanol was prepared according to Example 5, wherein morpholine is used instead of 3,3-difluoropyrrolidine in Step B.

[0135] LCMS(ESI) m/z: 542 [M+H.sup.+].

[0136] Step B: 2-butoxy-7-(4-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0137] .sup.1HNMR (Methanol-d4, 400 MHz): 7.28 (d, J=8.0 Hz, 2H), 7.22 (d, J=8.0 Hz, 2H), 7.03 (s, 1H), 4.29 (t, J=6.6 Hz, 2H), 3.96 (s, 2H), 3.67-3.64 (m, 4H), 3.46 (s, 2H), 2.43 (s, 4H), 1.77-1.72 (m, 2H), 1.55-1.45 (m, 2H), 0.98 (t, J=7.4 Hz, 3H).

[0138] MS(ESI) m/z: 396 [M+H.sup.+].

Example 10

2-butoxy-7-(4-((4-methylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0139] ##STR00026##

[0140] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((4-methylpiperazine-1-yl)methyl)phenyl)methanol was prepared according to Example 5, wherein 1-methylpiperazine was used instead of 3,3-difluoropyrrolidine in Step B.

[0141] LCMS(ESI) m/z: 555 [M+H.sup.+].

[0142] Step B: 2-butoxy-7-(4-((4-methylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0143] .sup.1HNMR (Methanol-d4, 400 MHz): 7.29 (d, J=8.0 Hz, 2H), 7.22 (d, J=8.0 Hz, 2H), 7.04 (s, 1H), 4.31 (t, J=6.6 Hz, 2H), 3.97 (s, 2H), 3.50 (s, 2H), 2.49-2.26 (m, 11H), 1.79-1.72 (m, 2H), 1.56-1.47 (m, 2H), 0.99 (t, J=7.4 Hz, 3H).

[0144] MS(ESI) m/z: 409 [M+H.sup.+].

Example 11

2-butoxy-7-(4-((dimethylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0145] ##STR00027##

[0146] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((dimethylamino)methyl)phenyl)methanol was prepared according to Example 5, wherein dimethylamine was used 775 instead of 3,3-difluoropyrrolidine in Step B.

[0147] LCMS(ESI) m/z: 500 [M+H.sup.+].

[0148] Step B: 2-butoxy-7-(4-((dimethylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step C according to Example 5.

[0149] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.48 (s, 2H), 7.41 (s, 4H), 7.26 (s, 1H), 4.43 (t, J=6.8 Hz, 2H), 4.22 (s, 2H), 4.06 (s, 2H), 2.79 (s, 6H), 1.79 (m, J=6.8 Hz, 2H), 1.55-1.49 (m, 2H), 1.01 (t, J=6.8 Hz, 3H).

[0150] MS(ESI) m/z: 354 [M+H.sup.+].

Example 12

2-butoxy-7-(4-((diethylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0151] ##STR00028##

[0152] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((diethylamino)methyl)phenyl)methanol was prepared according to Example 5, wherein diethylamine was used instead of 3,3-difluoropyrrolidine in Step B.

[0153] LCMS(ESI) m/z: 528 [M+H.sup.+].

[0154] Step B: 2-butoxy-7-(4-((diethylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step C according to Example 5.

[0155] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.48 (s, 2H), 7.42 (s, 4H), 7.25 (s, 1H), 4.41 (t, J=6.8 Hz, 2H), 4.28 (s, 2H), 4.06 (s, 2H), 3.20-3.15 (m, 4H), 1.82-1.77 (m, 2H), 1.55-1.49 (m, 2H), 1.34 (t, J=6.8 Hz, 6H), 1.01 (t, J=6.8 Hz, 3H).

[0156] MS(ESI) m/z: 382 [M+H.sup.+].

Example 13

2-butoxy-7-(4-((dipropylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0157] ##STR00029##

[0158] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxy)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((dipropylamino)methyl)phenyl)methanol was prepared according to Example 5, wherein dipropylamine was used instead of 3,3-difluoropyrrolidine in Step B.

[0159] LCMS(ESI) m/z: 556 [M+H.sup.+].

[0160] Step B: 2-butoxy-7-(4-((dipropylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0161] .sup.1HNMR (Methanol-d4, 400 MHz): 7.29-7.19 (m, 4H), 7.04 (s, 1H), 4.32 (t, J=6.5 Hz, 1H), 3.99 (s, 2H), 3.55 (s, 2H), 2.41-2.37 (m, 4H), 1.78-1.74 (m, 2H), 1.57-1.47 (m, 6H), 1.00 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.4 Hz, 6H).

[0162] MS(ESI) m/z: 410 [M+H.sup.+].

Example 14

7-(4-(azetidin-1-ylmethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0163] ##STR00030##

[0164] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-(azetidin-1-ylmethyl)phenyl)methanol was prepared according to Example 5, wherein azetidin was used instead of 3,3-difluoropyrrolidine in Step B.

[0165] LCMS(ESI) m/z: 512 [M+H.sup.+].

[0166] Step B: 7-(4-(azetidin-1-ylmethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0167] .sup.1HNMR (Methanol-d4, 400 MHz): 7.28 (d, J=8.0 Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 7.04 (s, 1H), 4.31 (t, J=6.8 Hz, 2H), 3.98 (s, 2H), 3.59 (s, 2H), 3.30-3.27 (m, 4H), 2.15-2.10 (m, 2H), 1.78-1.73 (m, 2H), 1.56-1.52 (m, 2H), 1.01 (t, J=6.8 Hz, 3H).

[0168] MS(ESI) m/z: 366 [M+H.sup.+].

Example 15

2-butoxy-7-(4-((3-methoxylazetidin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0169] ##STR00031##

[0170] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((3-methoxylazetidin-1-yl)methyl)phenyl)methanol was prepared according to Example 5, wherein 3-methoxylazetidin 840 was used instead of 3,3-difluoropyrrolidine in Step B.

[0171] LCMS(ESI) m/z: 542 [M+H.sup.+].

[0172] Step B: 2-butoxy-7-(4-((3-methoxylazetidin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0173] .sup.1HNMR (Methanol-d4, 400 MHz): 7.28 (d, J=8.0 Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 7.04 (s, 1H), 4.31 (t, J=6.8 Hz, 2H), 4.06-4.04 (m, 1H), 3.98 (s, 2H), 3.60 (s, 2H), 3.54-3.52 (m, 2H), 3.24 (s, 3H), 3.04-3.02 (m, 2H), 1.78-1.73 (m, 2H), 1.56-1.52 (m, 2H), 1.01 (t, J=6.8 Hz, 3H).

[0174] MS(ESI) m/z: 396 [M+H.sup.+].

Example 16

2-butoxy-7-(4-((4-methyl-1,4-diazepan-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0175] ##STR00032##

[0176] Step A: ((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((4-methyl-1,4-diazepan-1-yl)methyl)phenyl)methanol was prepared according to Example 5, wherein 1-methyl-1,4-diazepane was used instead of 3,3-difluoropyrrolidine in Step B.

[0177] LCMS(ESI) m/z: 569 [M+H.sup.+].

[0178] Step B: 2-butoxy-7-(4-((4-methyl-1,4-diazepan-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step C according to Example 5.

[0179] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.41 (s, 3H), 7.34-7.24 (m, 5H), 4.52 (t, J=6.8 Hz, 2H), 3.99 (s, 2H), 3.76 (s, 2H), 3.38-3.36 (m, 2H), 3.29-3.27 (m, 2H), 2.95 (s, 2H), 2.87-2.84 (m, 5H),

2.07-2.05 (m, 2H), 1.84-1.80 (m, 2H), 1.55-1.49 (m, 2H), 1.03-0.99 (t, J=8.0 Hz, 3H).

[0180] MS(ESI) m/z: 423 [M+H.sup.+].

Example 17

2-butoxy-7-(4-((2,6-dimethylmorpholinyl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0181] ##STR00033##

[0182] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((2,6-dimethylmorpholinyl)methyl)phenyl)methanol was prepared according to Example 5, wherein 2,6-dimethylmorpholine was used instead of 3,3-difluoropyrrolidine in Step B.

[0183] LCMS(ESI) m/z: 570 [M+H.sup.+].

[0184] Step B: 2-butoxy-7-(4-((2,6-dimethylmorpholinyl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step C according to Example 5.

[0185] .sup.1HNMR (Methanol-d4, 400 MHz): 7.30-7.28 (d, J=8.0 Hz, 2H), 7.23-7.21 (d, J=8.0 Hz, 2H), 7.06 (s, 1H), 4.34-4.30 (t, J=8.0 Hz, 2H), 3.99 (s, 2H), 3.69-3.64 (m, 2H), 3.47 (s, 2H), 2.73 (d, J=12.0 Hz, 2H), 1.77-1.70 (m, 4H), 1.54-1.51 (m, 2H), 1.11 (d, J=10.4 Hz, 6H), 1.00 (t, J=8.0 Hz, 3H).

[0186] MS(ESI) m/z: 424 [M+H.sup.+].

Example 18

7-(4-((1 S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-ylmethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0187] ##STR00034##

[0188] Step A: (4-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-ylmethyl)phenyl)(4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methanol was prepared according to Example 5, wherein (1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane was used instead of 3,3-difluoropyrrolidine in Step B.

[0189] LCMS(ESI) m/z: 554 [M+H.sup.+].

[0190] Step B: 7-(4-((1 S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-ylmethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-D]pyrimidine-4-amine formate was prepared with the procedures of Step C according to Example 5.

[0191] .sup.1HNMR (Methanol-d4, 400 MHz): δ:8.38 (brs, 2H), 7.45 (d, J=8.4 Hz, 2H), 7.37 (d, J=8.4 Hz, 2H), 7.29 (s, 1H), 4.66 (s, 1H), 4.47 (t, J=6.8 Hz, 2H), 4.36-4.27 (m, 1H), 4.24-4.23 (m, 2H), 4.16-4.13 (m, 1H), 4.04 (s, 2H), 3.82-3.81 (m, 1H), 3.33-3.31 (m, 2H), 2.33-2.29 (m, 1H), 2.14-2.11 (m, 1H), 1.83-1.76 (m, 2H), 1.56-1.48 (m, 2H), 1.01 (t, J=7.2 Hz, 3H).

[0192] MS(ESI) m/z: 408 [M+H.sup.+].

Example 19

2-butoxy-7-(4-((4-methoxylpiperidine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0193] ##STR00035##

[0194] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((4-methoxylpiperidine-1-yl)methyl)phenyl)methanol was prepared according to Example 5, wherein 4-methoxylpiperidine was used instead of 3,3-difluoropyrrolidine in Step B.

[0195] LCMS(ESI) m/z: 570 [M+H.sup.+].

[0196] Step B: 2-butoxy-7-(4-((4-methoxylpiperidine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate with the procedures of Step C according to Example 5.

[0197] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.45 (s, 2H), 7.43-7.38 (m, 4H), 7.28 (s, 1H), 4.45 (t, J=6.4 Hz, 2H), 4.21 (s, 2H), 4.05 (s, 2H), 3.52-3.53 (m, 1H), 3.33-3.39 (m, 3H), 3.26-3.24 (m, 2H), 3.13-3.10 (m, 2H), 1.99-1.92 (m, 4H), 1.84-1.77 (m, 2H), 1.56-1.50 (m, 2H), 1.01 (t, J=7.2 Hz, 3H).

[0198] MS(ESI) m/z: 424[M+H.sup.+].

Example 20

2-butoxy-7-(4-((4-isopropylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0199] ##STR00036##

[0200] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-((4-isopropylpiperazine-1-yl)methyl)phenyl)methanol was prepared according to Example 5, wherein 1-isopropylpiperazine was used instead of 3,3-difluoropyrrolidine in Step B.

[0201] LCMS(ESI) m/z: 583 [M+H.sup.+].

[0202] Step B: 2-butoxy-7-(4-((4-isopropylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step C according to Example 5.

[0203] .sup.1HNMR (Methanol-d4, 300 MHz): δ8.45 (s, 2H), 7.31-7.25 (m, 5H), 4.49 (t, J=8.4 Hz, 2H), 3.99 (s, 2H), 3.64 (s, 2H), 3.42-3.40 (m, 1H), 3.21-3.25 (m, 4H), 2.66-2.82 (m, 4H), 1.84-1.79 (m, 2H), 1.56-1.51 (m, 2H), 1.35 (d, J=8.8 Hz, 6H), 1.04-0.99 (t, J=10.0 Hz, 3H).

[0204] MS(ESI) m/z: 437 [M+H.sup.+].

Example 21

2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0205] ##STR00037##

[0206] Step A: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(4-(pyrrolidine-1-ylmethyl)phenyl)methanol was prepared according to Example 5, wherein pyrrole was used instead of 3,3-difluoropyrrolidine in Step B.

[0207] LCMS(ESI) m/z: 526 [M+H.sup.+].

[0208] Step B: 2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step C according to Example 5.

[0209] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.41 (s, 2H), 7.46 (d, J=8.0 Hz, 2H), 7.40 (d, J=8.0 Hz, 2H), 7.30 (s, 1H), 4.48 (t, J=6.8 Hz, 2H), 4.33 (s, 2H), 4.05 (s, 2H), 3.32-3.30 (m, 4H), 2.10-2.06 (m, 4H), 1.83-1.89 (m, 2H), 1.55-1.48 (m, 2H), 1.02 (t, J=7.2 Hz, 3H).

[0210] MS(ESI) m/z: 380 [M+H.sup.+].

Example 22

2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0211] ##STR00038##

[0212] Scheme for preparing 6-(pyrrolidine-1-ylmethyl)nicotinaldehyde:

##STR00039##

[0213] Step A: At room temperature, to a solution of methyl 6-methylnicotinate (10 g, 0.0662 mol) in CCl.sub.4 (100 mL) was added NBS (13.0 g, 0.0728 mol) and BPO (1.6 g, 0.0066 mol). The reaction mixture was heated to 75° C. and stirred for 12 h. After cooling, water was added (80 mL) and the mixture was extracted with ethyl acetate (200 mL×2). The organic layer was washed with saturated sodium thiosulfate aqueous solution (80 mL), dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=20/1) to give methyl 6-(bromomethyl)nicotinate (5.2 g, yield 34%) as brown solid.

[0214] .sup.1HNMR (CDCl.sub.3, 400 MHz): 9.18 (d, J=1.6 Hz, 1H), 8.32 (dd, J=8.0 Hz, J.sub.2=2.0 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 4.60 (s, 2H), 3.97 (s, 3H).

[0215] MS(ESI) m/z: 230, 232 [M+H.sup.+].

[0216] Step B: At 0° C., to a solution of pyrrolidine (3.09 g, 43.47 mmol) and triethylamine (3 mL, 21.73 mmol) in anhydrous tetrahydrofuran (100 mL) was added methyl 6-(bromomethyl)nicotinate (5.0 g, 21.73 mmol) in portions. After addition, the reaction mixture was stirred at room temperature for 16 h, diluted with water (80 mL) and extracted with ethyl acetate (100 mL). The organic layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=10/1) to give methyl 6-(pyrrolidine-1-ylmethyl)nicotinate (4.1 g, yield 86%) as brown solid.

[0217] .sup.1HNMR (CDCl.sub.3, 400 MHz): 9.11 (d, J=2.0 Hz, 1H), 8.22 (dd, J=8.0 Hz, J=2.0 Hz, 1H), 7.48 (d, J=8.0 Hz, 1H), 3.91 (s, 3H), 3.81 (s, 2H), 2.58-2.53 (m, 4H), 1.81-1.77 (m, 4H).

[0218] MS(ESI) m/z: 221 [M+H.sup.+].

[0219] Step C: At a temperature below 0° C., to a solution of methyl 6-(pyrrolidine-1-ylmethyl)nicotinate (3.0 g, 13.62 mmol) in anhydrous tetrahydrofuran (70 mL) was added lithium aluminum hydride (1.03 g, 27.24 mmol) in portions with stirring. The reaction was performed at about 0° C. for 2 h and at room temperature for a further 30 min. TLC showed disappearance of reactants. The mixture was cooled to 0° C. and water (1 mL) was added very slowly. Then 15% sodium hydroxide aqueous solution (1 mL) and water (3 mL) were added with vigor stirring. The resultant mixture was filtered. The filtrate was dried with anhydrous Mg.sub.2SO.sub.4 and concentrated to dryness under reduced pressure to give (6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methanol (2.5 g).

[0220] .sup.1HNMR (CDCl.sub.3, 400 MHz): δ8.41 (d, J=1.6 Hz, 1H), 7.67 (dd, J.sub.1=8.0 Hz, J.sub.2=20.0 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 4.67 (s, 2H), 3.75 (s, 2H), 2.57-2.543 (m, 4H), 1.81-1.76 (m, 4H).

[0221] Step D: (6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methanol (2.5 g, 13 mmol) was dissolved in anhydrous dichloromethane (50 mL). At 0° C., manganese dioxide (5.0 g, 58 mmol) was added in portions. The reaction mixture was stirred at room temperature for 24 h and filtered. The filtrate was concentrated under vacuum and the residue was purified with silica gel column chromatography (eluent: 15% ethyl acetate in 1000 petroleum ether) to give 6-(pyrrolidine-1-ylmethyl)nicotinaldehyde (2.2 g, crude) as yellow oil.

[0222] LCMS(ESI) m/z: 191 [M+H.sup.+].

[0223] Scheme for preparing 2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine:

##STR00040##

Example 22 Procedure

[0224] Step E: (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methanol was prepared according to Example 1, wherein 6-(pyrrolidine-1-ylmethyl)nicotinaldehyde was used instead of 1,3-benzenedialdehyde in Step E.

[0225] LCMS(ESI) m/z: 527 [M+H.sup.+].

[0226] Step F: 2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared as white solid with the procedures of Step G according to Example 1.

[0227] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.62 (s, 1H), 8.40 (brs, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.40 (d, J=8.0 Hz, 1H), 7.35 (s, 1H), 4.48 (s, 2H), 4.45 (t, J=6.4 Hz, 2H), 4.08 (s, 2H), 3.42-3.38 (m, 4H), 2.13-2.10 (m, 4H), 1.83-1.76 (m, 2H), 1.55-10.49 (m, 2H), 1.01 (t, J=7.2 Hz, 3H).

[0228] MS(ESI) m/z: 381[M+H.sup.+].

Example 23

2-butoxy-7-(3-(2-(pyrrolidine-1-yl)ethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0229] ##STR00041##

[0230] Scheme for preparing 3-(2-(pyrrolidine-1-yl)ethyl)benzaldehyde:

##STR00042##

[0231] Step A: Under nitrogen atmosphere, a solution of methyl 3-bromobenzoate (17.0 g, 79.0 mmol), tributylvinyltin (33 g, 102 mmol) and Pd(PPh.sub.3).sub.4 (4.5 g, 4 mmol) in dioxan (200 mL) was stirred at 110° C. for 6 h and the reaction was quenched with addition of 10% potassium fluoride aqueous solution (100 mL). The resultant mixture was stirred at room temperature for a further 10 min and extracted with ethyl acetate (150 mL×3). The combined organic layer was washed with saline, dried with anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: 25% ethyl acetate in petroleum ether) to give 15 g of crude methyl 3-vinylbenzoate as yellow oil.

[0232] MS(ESI) m/z: 163 [M+H.sup.+].

[0233] Step B: Under nitrogen atmosphere, to a solution of methyl 3-vinylbenzoate in anhydrous tetrahydrofuran (100 mL) was added 9-BBN (0.5M, 166 mL, 83 mmol) through a dropping funnel with stirring and the temperature was kept below −30° C. After addition, the reaction mixture was warmed to room temperature and stirred for 16 h. Then the mixture was cooled to −30° C., to which was added H.sub.2O.sub.2 aqueous solution (30 mass %, 19 mL) dropwise and 15% sodium hydroxide aqueous solution (40 mL) dropwise slowly. The resultant mixture was stirred for a further 1 h at ambient temperature, diluted with water (200 mL) and extracted with ethyl acetate (200 mL×2). The combined organic layer was washed with saline, dried with anhydrous sodium sulfate and concentrated under reduced pressure to give 9 g of crude methyl 3-(2-hydroxylethyl)benzoate as yellowy oil, which was used for the next step directly.

[0234] .sup.1HNMR (CDCl.sub.3, 400 MHz): 7.92-7.90 (m, 2H), 7.45-7.37 (m, 2H), 3.92 (s, 3H), 3.89 (t, J=6.5 Hz, 2H), 2.93 (t, J=6.5 Hz, 2H).

[0235] MS(ESI) m/z: 181 [M+H.sup.+].

[0236] Step C: At about 0° C., to a solution of methyl 3-(2-hydroxylethyl)benzoate (10 g) in anhydrous dichloromethane (90 mL) were added methanesulfonyl chloride (34 g, 299 mmol) and triethylamine (12 g, 118 mmol) with stirring. The reactants were stirred at 0° C. for 1 h, quenched with water (50 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: 10% ethyl acetate in petroleum ether) to give 2.7 g of methyl 3-(2-((methylsulfonyl)oxy)ethyl)benzoate as colorless oil.

[0237] MS(ESI) m/z: 259 [M+H.sup.+].

[0238] Step D: pyrrolidine (2.3 g, 31.3 mmol) and potassium carbonate (2.2 g, 16 mmol) were dissolved in anhydrous acetonitrile (20 mL), to which was added a solution of methyl 3-(2-((methylsulfonyl)oxy)ethyl)benzoate (2.7 g, 10.4 mmol) in acetonitrile (5 mL) over 10 min. The reaction liquid was stirred at 70° C. for 16 h, which after being cooled to room temperature was diluted with water (20 mL) and extracted with ethyl acetate (20 mL×3). The combined organic layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified with silica gel 1065 column chromatography (eluent: methanol/dichloromethane is 2%-5%) to give methyl 3-(2-(pyrrolidine-1-yl)ethyl)benzoate (1.7 g, 71%) as yellow oil.

[0239] MS(ESI) m/z: 234 [M+H.sup.+].

[0240] Step E: 3-(2-(pyrrolidine-1-yl)ethyl)benzaldehyde was prepared with the procedures of Step C, D according to Example 22.

[0241] MS(ESI) m/z: 204 [M+H.sup.+].

[0242] Step F: 2-butoxy-7-(3-(2-(pyrrolidine-1-yl)ethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step E, F according to Example 22.

[0243] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.42 (s, 2H), 7.30-7.13 (m, 5H), 4.38 (t, J=6.4 Hz, 2H), 4.01 (s, 1H), 3.41 (t, J=7.6 Hz, 2H), 3.35-3.32 (m, 4H), 3.01 (t, J=7.6 Hz, 2H), 2.09-2.05 (m, 4H), 1.81-1.74 (m, 2H), 1.57-1.48 (m, 2H), 1.01 (t, J=7.6 Hz, 3H).

[0244] MS(ESI) m/z: 394 [M+H.sup.+].

Example 24

2-butoxy-7-(4-(1-(pyrrolidine-1-yl)ethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0245] ##STR00043##

[0246] Scheme for preparing 4-(1-(pyrrolidine-1-yl)ethyl)benzaldehyde:

##STR00044##

[0247] Step A: To a solution of 4-cyanoacetophenone (4 g, 27.56 mmol) and pyrrolidine (2.94 g, 41.33 mmol) in methanol (100 mL) were added acetic acid (0.5 mL) and sodium cyanoborohydride (5.2 g, 82.67 mmol) with stirring and the temperature was kept below 0° C. The reactants were stirred at room temperature for 16 h and concentrated under reduce pressure. The resultant oil was purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/3) to give 2.8 g of 4-(1-(pyrrolidine-1-yl)ethyl)benzonitrile as colorless oil.

[0248] MS(ESI) m/z: 201 [M+H.sup.+].

[0249] Step B: At −20 to −10° C., to a solution of 4-(1-(pyrrolidine-1-yl)ethyl)benzonitrile (2 g, 10 mmol) in anhydrous toluene (100 mL) was added a solution of DIBAL-H (1 M, 20 mL, 20 mmol) over 1 h. The reaction liquid was stirred for a further 3 h, quenched with saturated ammonium chloride aqueous solution and extracted with ethyl acetate. The organic layer was washed with saline, dried with anhydrous sodium sulfate and concentrated under reduced pressure. The resultant solid was purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=50/1-10/1) to give 4-(1-(pyrrolidine-1-yl)ethyl)benzaldehyde (680 mg, 33.5%) as colorless oil.

[0250] (ESI) m/z: 204 [M+H.sup.+].

[0251] Step C: 2-butoxy-7-(4-(1-(pyrrolidine-1-yl)ethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step E, F according to Example 22.

[0252] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.50 (s, 2H), 7.44-7.38 (m, 4H), 7.27 (s, 1H), 4.45 (t, J=6.4, 2H), 4.33-4.28 (m, 1H), 4.04 (s, 2H), 3.37-3.33 (m, 2H), 3.14-3.11 (m, 2H), 2.04-2.02 (m, 4H), 1.83-1.78 (m, 2H), 1.72-1.70 (m, 3H), 1.55-1.49 (m, 2H), 1.01 (t, J=7.4, 3H).

[0253] MS(ESI) m/z: 394 [M+H.sup.+].

Example 25

2-butoxy-7-(4-(1-methylpiperidine-4-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0254] ##STR00045##

[0255] Scheme for preparing tert-butyl 4-(4-formylphenyl)piperidine-1-formate:

##STR00046##

[0256] Step A: Under nitrogen atmosphere, a mixture of 4-bromopyridine (3.0 g, 19.0 mmol), (4-(methoxycarbonyl)phenyl)boric acid (2.63 g, 14.6 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (0.35 g, 0.5 mmol) and sodium carbonate (6.91 g, 65.2 mmol) in 1,2-dimethoxylethane (40 mL) was heated to 90° C. and stirred for 10 h. The resultant mixture was concentrated under reduced pressure and the residue was purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=6/1-2/1) to give methyl 4-(pyridine-4-yl)benzoate (2.7 g, yield: 86.8%) as white solid.

[0257] MS(ESI) m/z: 214[M+H.sup.+].

[0258] Step B: To a solution of methyl 4-(pyridine-4-yl)benzoate (3.8 g, 17.8 mmol) and PtO.sub.2 (0.2 g) in methanol (40 mL) was added 2 mL hydrochloric acid and the mixture was heated to about 50° C. and stirred under hydrogen atmosphere (50 psi) for 16 h. The resultant mixture was filtered and the filtrate was concentrated under reduced pressure to give crude methyl 4-(piperidine-4-yl)benzoate (4.0 g) as hydrochloride without further purification.

[0259] MS(ESI) m/z: 220 [M+H.sup.+].

[0260] Step C: To a mixed solution of methyl 4-(piperidine-4-yl)benzoate (5.0 g, 22.8 mmol) and potassium carbonate (25.0 g, 182.2 mmol) in tetrahydrofuran (50 mL)/water (50 mL) was added di-tert-butyl dicarbonate (10.0 g, 45.8 mmol) in portions with stirring and the temperature was kept below 10° C. After addition, the reaction mixture was stirred at room temperature for a further 0.5 h, diluted with water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic layer was washed with saline, dried with anhydrous sodium sulfate and concentrated under vacuum. The residue was purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=6/1-1/1) to give tert-butyl 4-(4-(methoxycarbonyl)phenyl)piperidine-1-formate (1.9 g, yield: 26.4%) as white solid.

[0261] .sup.1HNMR (CDC.sub.3, 400 MHz): δ7.98 (d, J=8.4 Hz, 2H), 7.28 (d, J=7.6 Hz, 2H), 4.27 (s, 1H), 3.91 (s, 3H), 2.84-2.68 (m, 3H), 1.85 (d, J=12.8 Hz, 2H), 1.66-1.59 (m, 2H), 1.49 (s, 9H).

[0262] MS(ESI) m/z: 320 [M+H.sup.+].

[0263] Step D: tert-butyl 4-(4-formylphenyl)piperidine-1-formate was prepared with the procedures of Step C, D according to Example 22.

[0264] MS(ESI) m/z: 312.1 [M+Na.sup.+].

[0265] Step F: 2-butoxy-7-(4-(piperidine-4-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step E, F according to Example 22.

[0266] MS(ESI) m/z: 380.2 [M+H.sup.+].

Preparation of 2-butoxy-7-(4-(1-methylpiperidine-4-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0267] ##STR00047##

[0268] Step G: After stirring for 5 min, to a solution of 2-butoxy-7-(4-(piperidine-4-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (100 mg, 0.264 mmol) and HCHO (20 mg, 0.666 mmol) in methanol (5 mL) was added sodium cyanoborohydride (50 mg, 0.796 mmol). The reactants were stirred at room temperature for 0.5 h, diluted with water and extracted with ethyl acetate. The organic layer was concentrated under vacuum and the residue was purified with preparative HPLC to give 7.48 mg of 2-butoxy-7-(4-(1-methylpiperidine-4-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine.

[0269] .sup.1HNMR (Methanol, 400 MHz): 7.21 (d, J=8.0 Hz, 2H), 7.11 (d, J=8.0 Hz, 2H), 7.00 (s, 1H), 4.32-4.28 (m, 2H), 3.94 (s, 2H), 3.00-2.97 (m, 2H), 2.52-2.47 (m, 1H), 2.32 (s, 3H), 2.19-2.15 (m, 2H), 1.80-1.72 (m, 6H), 1.53-1.48 (m, 2H), 0.98 (t, J=7.4 Hz, 3H).

[0270] MS(ESI) m/z: 394[M+H.sup.+].

Example 26

2-butoxy-7-(4-(1-methylpyrrolidine-2-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0271] ##STR00048##

[0272] Scheme for preparing tert-butyl 2-(4-formylphenyl)pyrrolidine-1-formate:

##STR00049##

[0273] Step A: At 0° C. under N.sub.2 atmosphere, to a mixture of NaH (446 mg, 18.6 mmol) in anhydrous tetrahydrofuran (20 mL) was added 1-allyl-pyrrole-2-one (1.14 g, 9.11 mmol) and then a solution of methyl 4-bromobenzoate in anhydrous tetrahydrofuran (10 mL) slowly. The mixture was stirred at 90° C. for 2 h, then cooled to room temperature, and diluted with 6N hydrochloric acid. The resultant mixture was stirred at 10° C. for 12 h and the aqueous phase was washed with ethyl acetate (50 mL). The mixture was basified with 1N sodium hydroxide until pH was about 9 and then extracted with ethyl acetate (50 mL×2). The combined organic layer was concentrated to dryness under vacuum to give 2.0 g of 5-(4-bromophenyl)-3,4-dihydro-2H-pyrrole as yellow solid, which was used for the next step directly.

[0274] Step B: At 0° C. to a solution of 5-(4-bromophenyl)-3,4-dihydro-2H-pyrrole (2.0 g, 9.0 mmol) in methanol (20 mL) was slowly added sodium borohydride (684 mg, 18.1 mmol) with stirring. After addition, the reaction mixture was stirred at room temperature for 1 h. TLC (petroleum ether/ethyl acetate=2:1) showed depletion of starting materials. The resultant mixture was diluted with water (30 mL). To the mixture of the above step was added potassium carbonate (1.51 g, 10.9 mmol) and Boc.sub.2O (2.3 g, 10.5 mmol). The mixture was stirred at 20° C. for 2 h and thin-layer chromatography plate (developing agent: petroleum ether/ethyl acetate=2/1) showed depletion of starting materials. The mixture was then extracted with ethyl acetate (50 mL×2) and the extract was concentrated under reduced pressure. The residue was purified with silica gel column chromatography to give tert-butyl 2-(4-bromophenyl)pyrrolidine-1-formate (1.5 g, yield: 51.1%) as yellow solid.

[0275] Step C: At −78° C. under nitrogen atmosphere, to a solution of tert-butyl 2-(4-bromophenyl)pyrrolidine-1-formate (0.6 g, 1.839 mmol) in anhydrous tetrahydrofuran (20 mL) was added n-BuLi (1.5 mL, 2.76 mmol) with stirring. The reaction mixture was stirred at −78° C. for 30 min, to which was slowly added N,N-dimethylformamide (192 mg, 2.63 mmol). The resultant mixture was warmed to room temperature, stirred for a further 30 min and quenched with 3 mL sodium bicarbonate aqueous solution. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (25 mL×3). The combined organic layer was washed with saline, dried with sodium sulfate, filtered and distilled to dryness. The residue was purified with silica gel column chromatography (petroleum ether: ethyl acetate=15:1-10:1) to give tert-butyl 2-(4-formylphenyl)pyrrolidine-1-formate (0.4 g, yield: 79.1%) as colorless oil.

[0276] MS(ESI) m/z: 276.0 [M+1.sup.+].

Preparation of 2-butoxy-7-(4-(pyrrolidine-2-yl)benzyl)-5H-pyrrolo[3,2-d]2pyrimidine-4-amine

[0277] Step D: 2-butoxy-7-(4-(pyrrolidine-2-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step E, F according to Example 22.

[0278] MS(ESI) m/z: 366.2 [M+1.sup.+].

Preparation of 2-butoxy-7-(4-(1-methylpyrrolidine-2-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0279] Step E: 2-butoxy-7-(4-(1-methylpyrrolidine-2-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step G according to Example 25.

[0280] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.27 (d, J=8.0 Hz, 2H), 7.22 (d, J=8.0 Hz, 2H), 7.03 (s, 1H), 4.30 (t, J=7.4 Hz, 2H), 3.97 (s, 2H), 3.31-3.19 (m, 1H), 3.07-3.03 (m, 1H), 2.31-2.87 (m, 1H), 2.18-2.15 (m, 1H), 2.13 (s, 3H), 1.89-1.72 (m, 5H), 1.54-1.48 (m, 2H), 0.98 (t, J=7.4 Hz, 3H).

[0281] MS(ESI) m/z: 380[M+1.sup.+].

Example 27

1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)phenyl)-4-methylpiperazine-2-one

[0282] ##STR00050##

Preparation of 4-(4-methyl-2-oxopiperazine-1-yl)benzaldehyde

[0283] ##STR00051##

[0284] Step A: To a solution of 4-bromo-benzaldehyde (1.8 g, 9.73 mmol), 4-methylpiperazine-2-one (1.44 g, 12.6 mmol), Pd.sub.2(dba).sub.3 (768 mg, 0.84 mmol), Xantphos (435 mg, 0.75 mmol) and cesium carbonate (5.48 g, 16.8 mmol) in dioxan (30 mL) was added water (1 drop). The mixture was stirred under nitrogen atmosphere at 90° C. for 1.5 h. After cooling, the mixture was filtered. The filtrate was concentrated to dryness under vacuum. The residue was purified with silica gel chromatography to give 4-(4-methyl-2-oxopiperazine-1-yl)benzaldehyde (1.8 g, 84.80%) as white solid.

[0285] MS(ESI) m/z: 219[M+H.sup.+].

Preparation of 1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)phenyl)-4-methylpiperazine-2-one

[0286] Step B: 1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)phenyl)-4-methylpiperazine-2-one was prepared with the procedures of Step E, F according to Example 22.

[0287] .sup.1HNMR (Methanol-d4, 400 MHz) δ7.36 (s, 1H), 7.30 (d, J=8.4 Hz, 2H), 7.22 (d, J=8.4 Hz, 2H), 4.52 (t, J=6.4 Hz, 2H), 4.02 (s, 2H), 3.72-3.69 (m, 2H), 3.27 (s, 2H), 2.89-2.86 (m, 2H), 2.44 (s, 3H), 1.83-1.79 (m, 2H), 1.54-1.48 (m, 2H), 1.00 (t, J=7.4 Hz, 3H).

[0288] MS(ESI) m/z: 409 [M+H.sup.+].

Example 28

2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0289] ##STR00052##

[0290] Scheme for preparing tert-butyl 7-formyl-3,4-dihydroisoquinoline-2(1H)-carboxylate:

##STR00053##

[0291] Step A: Under nitrogen atmosphere at 0° C., to a solution of 2-(4-bromophenyl)ethylamine (27 g, 0.13 mol) and triethylamine (16.4 g, 0.16 mol) in anhydrous dichloromethane (300 mL) was added trifluoroacetic acid anhydride (34 g, 0.16 mol) dropwise. The reaction mixture was stirred at room temperature for 1 h and then diluted with water. The organic layer was isolated and concentrated to dryness under vacuum to give N-(4-bromophenethyl)-trifluoroacetamide (37 g, 96.10%) as white solid.

[0292] MS(ESI) m/z: 296, 298 [M+H.sup.+].

[0293] Step B: To a suspension of N-(4-bromophenethyl)-trifluoroacetamide (37 g, 0.12 mmol) in concentrated sulfuric acid (200 mL)/acetic acid (300 mL) was added paraformaldehyde (10.2 g, 0.34 mol) in portions with stirring. After addition, the mixture was stirred at room temperature for 12 h, then poured into ice water (1 L) and extracted with ethyl acetate (400 mL×2). The combined organic layer was successively washed with saturated sodium bicarbonate aqueous solution and saline, dried with anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified with silica gel column chromatography (eluent: 5% ethyl acetate in petroleum ether) to give 1-(7-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-trifluoroethyl ketone (33 g, 89.30/0).

[0294] MS(ESI) m/z: 308, 310[M+H.sup.+].

[0295] Step C: To a solution of 1-(7-bromo-3,4-dihydroisoquinoline-2(1H)-yl)-trifluoroethyl ketone (30 g, 0.1 mol) in anhydrous methylpyrrolidine-2-one (300 mL) was added cuprous cyanide (18 g, 0.2 mol). The reaction mixture was stirred at 180° C. under nitrogen atmosphere for 4 h. After being cooled to room temperature, the mixture was slowly poured into ice water (500 mL) and extracted with ethyl acetate (200 mL×2). The combined organic layer was washed with water, dried with anhydrous sodium sulfate and concentrated under vacuum to give 25 g of crude 2-trifluoroacetyl-tetrahydroisoquinoline-7-carbonitrile, which was used for the next step directly.

[0296] MS(ESI) m/z: 255 [M+H.sup.+].

[0297] Step D: 2-trifluoroacetyl-tetrahydroisoquinoline-7-carbonitrile (25 g, 0.1 mol) and potassium carbonate (25 g, 0.18 mol) were dissolved in mix solvents of methanol (300 mL) and water (60 mL) and the mixture was stirred at room temperature for 2 h. di-tert-butyl dicarbonate (26 g, 0.12 mol) was added in portions over 10 min. The reaction mixture was stirred for a further 4 h, diluted with water (200 mL) and extracted with ethyl acetate (200 mL×2). The combined organic layer was washed with saline, dried with anhydrous sodium sulfate and concentrated under vacuum. The residue was purified with silica gel column chromatography (eluent: 5% ethyl acetate in petroleum ether) to give tert-butyl 7-cyano-3,4-dihydroisoquinoline-2(1H)-carboxylate (14 g, 54%) as white solid.

[0298] MS(ESI) m/z: 259[M+H.sup.+].

[0299] Step E: Under nitrogen atmosphere at −10° C., to a solution of tert-butyl 7-cyano-3,4-dihydroisoquinoline-2(1H)-carboxylate (1 g, 3.9 mmol) in anhydrous tetrahydrofuran (20 mL) was added diisobutyl aluminium hydride (1 M, 6 mL, 6.0 mmol) dropwise. After addition, the reaction mixture was stirred at 0° C. for 5 h and quenched with water (0.24 mL). Then 15% sodium hydroxide aqueous solution (0.24 mL) was added followed by 0.6 mL water. The resultant mixture was stirred at room temperature for a further 15 min, dried with anhydrous magnesium sulfate and filtered. The filtrate was concentrated under vacuum and the residue was purified with silica gel column chromatography (eluent: 10% ethyl acetate in petroleum ether) to give tert-butyl 7-formyl-3,4-dihydroisoquinoline-2(1H)-carboxylate (700 mg, 70%) as yellow oil.

[0300] MS(ESI) m/z: 262 [M+H.sup.+].

Preparation of 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0301] Step F: 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step E, F according to Example 22.

[0302] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.49 (s, 2H), 7.23-7.15 (m, 3H), 7.10 (s, 1H), 4.44 (t, J=6.5 Hz, 2H), 4.30 (s, 2H), 3.98 (s, 2H), 3.47 (t, J=6.1 Hz, 2H), 3.08 (t, J=6.1 Hz, 2H), 1.83-1.76 (m, 2H), 1.55-1.49 (m, 2H), 1.01 (t, J=7.4 Hz, 3H).

[0303] MS(ESI) m/z: 352 [M+H.sup.+].

Example 29

2-butoxy-7-((2-methyl-1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0304] ##STR00054##

[0305] Using 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as starting material, with the procedures of Step G according to Example 25, 2-butoxy-7-((2-methyl-1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared.

[0306] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.11-7.09 (m, 1H), 7.03-7.00 (m, 3H), 4.32 (t, J=6.4 Hz, 2H), 3.92 (s, 2H), 3.55 (s, 2H), 2.91-2.88 (m, 2H), 2.73-2.71 (m, 2H), 2.43 (s, 3H), 1.80-1.73 (m, 2H), 1.56-1.52 (m, 2H), 1.01 (t, J=7.6 Hz, 3H)

[0307] MS(ESI) m/z: 366 [M+H.sup.+].

Example 30

2-butoxy-7-((2-ethyl-1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0308] ##STR00055##

[0309] Using 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as starting material, with the procedures of Step G according to Example 25, 2-butoxy-7-((2-ethyl-1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared.

[0310] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.43 (s, 2H), 7.25-7.18 (m, 3H), 7.10 (s, 1H), 4.45 (t, J=6.4 Hz, 2H), 4.34 (s, 2H), 3.99 (s, 2H), 3.51 (t, J=6.0 Hz, 2H), 3.32-3.26 (m, 2H), 3.15 (t, J=6.0 Hz, 2H), 1.84-1.77 (m, 2H), 1.58-1.48 (m, 2H), 1.42 (t, J=8.0 Hz, 3H), 1.01 (t, J=6.0 Hz, 3H).

[0311] MS(ESI) m/z: 380 [M+H.sup.+].

Example 31

2-butoxy-7-((2-isopropyl-1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0312] ##STR00056##

[0313] Using 2-butoxy-7-((1,2,3,4-tetrahydroisoquinyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as starting material, with the procedures of Step G according to Example 25, 2-butoxy-7-((2-isopropyl-1,2,3,4-tetrahydroisoquinoline-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as prepared.

[0314] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.10-7.08 (m, 1H), 7.03-7.00 (m, 3H), 4.32 (t, J=6.4 Hz, 2H), 3.93 (s, 2H), 3.70 (s, 2H), 2.90-2.86 (m, 3H), 2.83-2.80 (m, 2H), 1.80-1.73 (m, 2H), 1.56-1.50 (m, 2H), 1.17 (d, J=6.4 Hz, 6H), 1.01 (t, J=7.6 Hz, 3H).

[0315] MS(ESI) m/z: 394[M+H.sup.+].

Example 32

2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0316] ##STR00057##

[0317] Scheme for preparing N-t-butoxycarbonyl 1,2,3,4-tetrahydroisoquinoline-6-formaldehyde:

##STR00058##

[0318] Step A: To a mixed solution of 6-bromoisoquinoline (10 g, 48 mmol) in N,N-dimethylformamide/methanol (V/V=1/1) (200 mL) were added sodium acetate (5.0 g, 61 mmol), triphenylphosphine (3.0 g, 11.4 mmol) and palladium acetate (2.8 g, 12 mmol). The mixture was place in a clave with CO at 300 kPa and heated to 100° C. After stirring for 15 h, completion of the reaction was determined by LC-MS and the reactants were filtered with diatomaceous earth (elution with ethyl acetate). The resultant mixture was concentrated under reduced pressure and purified with silica gel column chromatography (eluent: petroleum ether/ethyl acetate=5/1) to give methyl isoquinoline-6-carboxylate (8.9 g, yield: 98%).

[0319] MS(ESI) m/z: 188[M+H.sup.+].

[0320] Step B: Under nitrogen atmosphere, to a solution of methyl isoquinoline-6-carboxylate (10 g, 53.5 mmol) in methanol (100 mL) were added acetic acid (2 mL) and PtO.sub.2 (200 mg) with stirring. Under hydrogen atmosphere, the mixture was stirred at 40° C. for 3 h and the catalyst was filtered off with diatomaceous earth. The mixture was concentrated under vacuum to give methyl 1,2,3,4-tetrahydroisoquinoline-6-carboxylate (9 g, yield: 88%) without further purification.

[0321] MS(ESI) m/z: 192 [M+H.sup.+].

[0322] Step C: methyl N-t-butoxycarbonyl 1,2,3,4-tetrahydroisoquinoline-6-carboxylate was prepared with the procedures of Step C according to Example 25.

[0323] MS(ESI) m/z: 292 [M+H.sup.+].

[0324] Step D: N-t-butoxycarbonyl 1,2,3,4-tetrahydroisoquinoline-6-formaldehyde was prepared with the procedures of Step C, D according to Example 22.

[0325] MS(ESI) m/z: 262 [M+H.sup.+].

Preparation of 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0326] Step E: 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared with the procedures of Step E, F according to Example 22.

[0327] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.12-7.09 (m, 1H), 7.08 (s, 1H), 7.04 (s, 1H), 6.96 (d, J=7.6 Hz, 1H), 4.32 (t, J=7.4 Hz, 2H), 3.98 (s, 2H), 3.93 (s, 2H), 3.13 (t, J=6.2 Hz, 2H), 2.85-2.82 (m, 2H), 1.79-1.73 (m, 2H), 1.58-1.48 (m, 2H), 1.01 (s, 3H).

[0328] MS(ESI) m/z: 352 [M+H.sup.+].

Example 33

2-butoxy-7-((2-methyl-1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0329] ##STR00059##

[0330] Using 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as starting material, with the procedures of Step G according to Example 25, 2-butoxy-7-((2-methyl-1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared.

[0331] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.10-7.09 (m, 2H), 7.03 (s, 1H), 6.96 (d, J=8.4 Hz, 1H), 4.32 (t, J=6.6 Hz, 2H), 3.93 (s, 2H), 3.60 (s, 2H), 2.92-2.89 (m, 2H), 2.77-2.74 (m, 2H), 2.46 (s, 3H), 1.81-1.73 (m, 2H), 1.58-1.48 (m, 2H), 1.01 (t, J=7.4 Hz, 3H).

[0332] MS(ESI) m/z: 366 [M+H.sup.+].

Example 34

2-butoxy-7-((2-ethyl-1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0333] ##STR00060##

[0334] Using 2-butoxy-7-((1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as starting material, with the procedures of Step G according to Example 25, 2-butoxy-7-((2-ethyl-1,2,3,4-tetrahydroisoquinoline-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine was prepared.

[0335] .sup.1HNMR (Methanol-d4, 400 MHz): 7.11-7.08 (m, 2H), 7.03 (s, 1H), 6.97 (d, J=8.0 Hz, 1H), 4.32 (t, J=6.6 Hz, 2H), 3.94 (s, 2H), 3.63 (s, 2H), 2.93-2.88 (m, 2H), 2.79-2.76 (m, 2H), 2.65-2.60 (m, 2H), 1.79-1.75 (m, 2H), 1.56-1.52 (m, 2H), 1.21 (t, J=7.2 Hz, 3H), 1.01 (t, J=7.2 Hz, 3H).

[0336] MS(ESI) m/z: 380[M+H.sup.+].

Example 35

7-benzyl-2-(2-methoxylethoxyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0337] ##STR00061##

[0338] Step A: (4-amino-2-(2-methoxylethoxyl)-5-((2-(trimethylsilylethyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(phenyl)methanol was prepared with the procedures of Step C, D, E according to Example 1.

[0339] MS(ESI) m/z: 445 [M+H.sup.+].

[0340] Step B: 7-benzyl-2-(2-methoxylethoxyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step G according to Example 1.

[0341] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.39 (s, 1H), 7.29-7.19 (m, 6H), 4.61-4.58 (m, 2H), 4.00 (s, 1H), 3.79-3.76 (m, 2H), 3.42 (s, 3H).

[0342] MS(ESI) m/z: 299[M+H.sup.+].

Example 36

2-(2-methoxylethoxyl)-7-((6-methylpyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0343] ##STR00062##

[0344] 2-(2-methoxylethoxyl)-7-((6-methylpyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step A, B according to Example 35.

[0345] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.34 (s, 3H), 7.66 (dd, J=2.4 Hz/J=8.0 Hz, 1H), 7.31 (s, 1H), 7.24 (d, J=8.0 Hz, 1H), 4.57-4.55 (m, 2H), 4.01 (s, 2H), 3.77-3.75 (m, 2H), 3.41 (s, 3H), 2.51 (s, 3H).

[0346] MS(ESI) m/z: 314[M+H.sup.+].

Example 37

7-((5-chloropyridine-2-yl)methyl)-2-(2-methoxylethoxyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0347] ##STR00063##

[0348] 7-((5-chloropyridine-2-yl)methyl)-2-(2-methoxylethoxyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step A, B according to Example 35.

[0349] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.45 (s, 1H), 8.40 (s, 1H), 7.77 (dd, J=2.4 Hz/J=8.0 Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.32 (s, 1H), 4.52 (t, J=4.0 Hz, 2H), 4.17 (s, 2H), 3.75 (t, J=4.0 Hz, 2H), 3.42 (s, 3H).

[0350] MS(ESI) m/z: 334[M+H.sup.+].

Example 38

2-(2-methoxylethoxyl-)-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0351] ##STR00064##

[0352] 2-(2-methoxylethoxyl)-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared with the procedures of Step A, B according to Example 35.

[0353] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.62 (s, 1H), 8.41 (s, 2H), 7.79-7.76 (m, 1H), 7.36 (d, J=8.4 Hz, 1H), 7.28 (s, 1H), 4.49-4.44 (m, 4H), 4.05 (s, 2H), 3.74-3.72 (m, 2H), 3.39 (s, 3H), 3.33-3.30 (m, 4H), 2.10-2.07 (m, 4H).

[0354] MS(ESI) m/z: 383 [M+H.sup.+].

Example 39

1-(4-((4-amino-2-(2-methoxylethoxyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)phenyl)-4-methylpiperazine-2-one

[0355] ##STR00065##

[0356] 1-(4-((4-amino-2-(2-methoxylethoxyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)phenyl)-4-methylpiperazine-2-one was prepared with the procedures of Step A, B according to Example 35.

[0357] .sup.1HNMR (Methanol-d4, 400 MHz): 7.35 (s, 1H), 7.31 (d, J=8.4 Hz, 2H), 7.22 (d, J=8.4 Hz, 2H), 4.65-4.62 (m, 2H), 4.01 (s, 2H), 3.77-3.76 (m, 2H), 3.70-3.67 (m, 2H), 3.35 (s, 3H), 3.32-3.28 (m, 2H), 2.90-2.88 (m, 2H), 2.45 (s, 3H).

[0358] MS(ESI) m/z: 411 [M+H.sup.+].

Example 40

2-butoxy-7-((5-(pyrrolidine-1-ylmethyl)pyridine-2-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine

[0359] ##STR00066##

[0360] 2-butoxy-7-((5-(pyrrolidine-1-ylmethyl)pyridine-2-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine formate was prepared according the procedures of Example 22.

[0361] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.61 (s, 1H), 8.46 (brs, 2H), 7.91 (d, J=8.0 Hz, 1H), 7.47 (d, J=7.6 Hz, 1H), 7.37 (s, 1H), 4.44 (t, J=6.4 Hz, 2H), 4.35 (s, 2H), 4.22 (s, 2H), 3.33-3.27 (m, 4H), 2.09-2.06 (m, 4H), 1.83-1.76 (m, 2H), 1.57-1.50 (m, 2H), 1.01 (t, J=7.6 Hz, 3H).

[0362] MS(ESI) m/z: 381 [M+H.sup.+].

Example 41

4-amino-2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile

[0363] ##STR00067##

Example 41 Procedures

[0364] ##STR00068## ##STR00069##

Example 41 Procedures

[0365] Step A: Under nitrogen atmosphere at −78° C., to a solution of 7-bromo-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (10.00 g, 24.07 mmol) in anhydrous tetrahydrofuran (200 mL) was added n-BuLi (6.17 g, 96.28 mmol). The mixture was stirred at −78° C. for 1 h, to which was added a solution of 6-chloronicotinaldehyde (10.22 g, 72.21 mmol) in tetrahydrofuran (200 mL) dropwise. The reaction mixture was stirred at −78° C. for a further 1 h, slowly poured into water (150 mL), stirred at room temperature for 20 min and extracted with ethyl acetate (100 mL×3). The combined organic phase was washed with saturated saline (50 mL×2), dried with anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified with silica gel chromatography (eluent: petroleum ether/ethyl acetate=5/1-1/3) to give (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(6-chloropyridine-3-yl)methanol (5.00 g, 43.45%) as yellow solid.

[0366] .sup.1HNMR (400 MHz, CHLOROFORM-d) δ8.52 (d, J=2.3 Hz, 1H), 7.87 (dd, J=2.4, 8.2 Hz, 1H), 7.34 (d, J=8.0 Hz, 1H), 6.65 (s, 1H), 6.14 (s, 1H), 5.97 (br.s., 2H), 5.39-5.26 (m, 2H), 4.31 (t, J=6.7 Hz, 2H), 3.62-3.49 (m, 2H), 1.86-1.71 (m, 2H), 1.51 (qd, J=7.5, 14.9 Hz, 2H), 1.28 (t, J=7.2 Hz, 1H), 1.06-0.87 (m, 5H), 0.00 (s, 9H).

[0367] MS(ESI) m/z: 478 [M+H.sup.+].

[0368] Step B: At room temperature, to a solution of (4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)(6-chloropyridinepyridine-3-yl)methanol (5.00 g, 10.46 mmol) in trifluoroacetic acid (50 mL) was added triethylsilane (6.08 g, 52.30 mmol) in portions. The reaction mixture was stirred at ambient temperature for 24 h, poured into sodium bicarbonate saturated aqueous solution (150 mL) and further stirred for 20 min followed by extraction with ethyl acetate (100 mL×3). The combined organic phase was washed with saline (20 mL×2), dried with anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified with silica gel chromatography (eluent: petroleum ether/ethyl acetate=3/1) to give 2-butoxy-7-((6-chloropyridine-3-yl)methyl)-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (2.30 g, 47.59%) as yellow solid.

[0369] .sup.1HNMR (300 MHz, CHLOROFORM-d) δ8.52 (d, J=2.3 Hz, 1H), 7.88 (dd, J=2.4, 8.1 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 6.64 (s, 1H), 6.14 (s, 1H), 5.89 (br.s., 2H), 5.40-5.23 (m, 2H), 4.31 (t, J=6.6 Hz, 2H), 3.66-3.47 (m, 2H), 1.88-1.70 (m, 2H), 1.60-1.46 (m, 2H), 1.07-0.82 (m, 5H), 0.00 (s, 9H).

[0370] MS(ESI) m/z: 462 [M+H.sup.+].

[0371] Step C: To a solution of 2-butoxy-7-((6-chloropyridine-3-yl)methyl)-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-D]pyrimidine-4-amine (2.30 g, 4.98 mmol) in N,N-dimethylformamide (15 mL) was added palladium acetate (111.75 mg, 0.5 mmol), 1,3-bis(diphenylphosphino)propane (205.30 mg, 0.5 mmol), triethylamine (1.51 g, 14.93 mmol) and methanol (797.43 mg, 24.89 mmol). The suspension was vacuumized and aerated with CO several times. The mixture was heated to 100° C. and stirred under CO atmosphere (3 M Pa) for 24 h. Thin-layer chromatography plate (developing agent: petroleum ether/ethyl acetate=1/1) showed depletion of starting materials. Insolubles were filtered off and concentration was performed. The crude product was purified with silica gel chromatography (eluent: petroleum ether/ethyl acetate=1/1) to give methyl 5-((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)picolinate (1.10 g, 45.48%) as yellow solid.

[0372] .sup.1HNMR (400 MHz, CHLOROFORM-d) δ8.76 (d, J=1.8 Hz, 1H), 8.06 (d, J=8.0 Hz, 1H), 7.85 (dd, J=2.0, 8.0 Hz, 1H), 6.82 (s, 1H), 5.71 (br.s., 2H), 5.35 (s, 2H), 4.33 (t, J=6.5 Hz, 2H), 4.19-4.08 (m, 3H), 4.00 (s, 3H), 3.60-3.51 (m, 2H), 1.85-1.74 (m, 2H), 1.53 (qd, J=7.4, 15.0 Hz, 2H), 1.28 (t, J=7.2 Hz, 2H), 1.02-0.90 (m, 5H), 0.00 (s, 9H).

[0373] MS(ESI) m/z: 486 [M+H.sup.+].

[0374] Step D: At a temperature below 0° C., to a solution of methyl 5-((4-amino-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)picolinate (800.00 mg, 1.65 mmol) in tetrahydrofuran (10 mL) was added bromosuccinamide (293.18 mg, 1.65 mmol) in portions. The reaction mixture was stirred at 0° C. for 1 h, diluted with water (30 mL) and extracted with dichloromethane (20 mL×2). The combined organic phase was dried with magnesium sulfate and concentrated under vacuum. The residue was purified with thin-layer chromatography plate to give methyl 5-((4-amino-6-bromo-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)picolinate (160.00 mg, 17.18%) as yellow solid.

[0375] .sup.1HNMR (400 MHz, CHLOROFORM-d) δ8.83 (s, 1H), 8.03 (d, J=8.0 Hz, 1H), 7.86 (d, J=8.0 Hz, 1H), 5.85 (br.s., 2H), 5.55 (s, 2H), 4.34 (t, J=6.5 Hz, 2H), 4.10 (s, 2H), 4.00 (s, 3H), 3.71-3.60 (m, 2H), 1.84-1.72 (m, 4H), 1.59-1.47 (m, 2H), 0.98 (q, J=7.8 Hz, 5H), 0.01 (s, 9H).

[0376] MS(ESI) m/z: 565, 567[M+H.sup.+].

[0377] Step E: Under nitrogen atmosphere at −78° C., to a solution of methyl 5-((4-amino-6-bromo-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)picolinate (150.00 mg, 0.266 mmol) in anhydrous tetrahydrofuran (8 mL) was added diisobutyl aluminum hydride (56.28 mg, 0.396 mmol) dropwise with stirring. After addition, the reaction mixture was stirred at −78° C. for 1 h. Then the reaction mixture was quenched with methanol (5 mL), diluted with water (20 mL) and extracted with ethyl acetate (30 mL×2). The combined organic layer was concentrated to dryness under vacuum to give about 150 mg of crude 5-((4-amino-6-bromo-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7-yl)methyl)pyridinealdehyde without further purification.

[0378] .sup.1HNMR (400 MHz, CHLOROFORM-d) δ10.05 (s, 1H), 8.87 (s, 1H), 7.96-7.80 (m, 2H), 5.72 (br. s, 2H), 5.56 (s, 2H), 4.34 (t, J=6.5 Hz, 2H), 4.12 (s, 2H), 3.71-3.62 (m, 2H), 1.84-1.72 (m, 2H), 1.56-1.48 (m, 2H), 1.06-0.81 (m, 5H), 0.01 (s, 9H).

[0379] MS(ESI) m/z: 535, 537[M+H.sup.+].

[0380] Step F: To a solution of 5-((4-amino-6-bromo-2-butoxy-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-7yl)methyl)pyridinealdehyde (150.00 mg, 0.281 mmol), pyrrolidine (29.94 mg, 0.421 mmol), acetic acid (0.2 mL) 1565 in tetrahydrofuran (5 mL) was added sodium cyanoborohydride (35.27 mg, 0.561 mmol) and the mixture was stirred at room temperature for 12 h. The mixture was poured into ice/water mixture (volume ratio=1/1, 15 mL), stirred for 20 min, and extracted with ethyl acetate (40 mL×3). The combined organic phase was washed with saline (20 mL×2), dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified with preparative HPLC to give 150 mg of 6-bromo-2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine as yellow solid.

[0381] MS(ESI) m/z: 589, 591 [M+H.sup.+].

[0382] Step G: To anhydrous N,N-dimethylformamide (2 mL) were added 6-bromo-2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-4-amine (150.00 mg, 254.39 mol), Pd.sub.2(dba).sub.3 (23.30 mg, 25.44 mol), 1,1′-bis(diphenylphosphino)ferrocene (14.10 mg, 25.44 mol), zinc cyanide (59.74 mg, 508.78 mol) and Zn (33.27 mg, 508.78 mol), and the mixture was replaced with nitrogen and heated under nitrogen atmosphere to 110° C. for 3 h. After cooling, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (25 mL×3). The combined organic phase was washed with saline (30 mL), dried with anhydrous sodium sulfate and concentrated under vacuum. The residue was purified with preparative TLC to give 4-amino-2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile (120 mg, 88.05%).

[0383] MS(ESI) m/z: 536 [M+H.sup.+].

[0384] Step H: At 20° C. a solution of 4-amino-2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5-((2-(trimethylsilyl)ethoxyl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile (120 mg, 0.224 mmol) in trifluoroacetic acid (5 mL) was stirred at 20° C. for 12 h and concentrated to dryness under vacuum. The residue was purified with preparative HPLC to give 8.7 mg of 4-amino-2-butoxy-7-((6-(pyrrolidine-1-ylmethyl)pyridine-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile.

[0385] .sup.1HNMR (Methanol-d4, 400 MHz): δ8.52 (s, 1H), 7.79 (d, J=8.0 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H), 4.33 (t, J=6.8 Hz, 2H), 4.17 (s, 2H), 3.76 (s, 2H), 2.61 (s, 4H), 1.82-1.72 (m, 6H), 1.54-1.49 (m, 2H), 1.02-0.99 (t, J=7.2 Hz, 3H).

[0386] MS(ESI) m/z: 406 [M+H.sup.+].

Example 42

4-amino-2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile

[0387] ##STR00070##

[0388] 4-amino-2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile was prepared according to the procedures of Example 41 and Step A, B, C, D, E, F, G, H of Example 41 were followed.

[0389] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.34-7.32 (d, J=8.4 Hz, 2H), 7.26-7.24 (d, J=8.4 Hz, 2H), 4.36-4.33 (t, J=6.8 Hz, 2H), 4.13 (s, 2H), 3.62 (s, 2H), 2.57 (brs, 4H), 1.82-1.77 (m, 6H), 1.52-1.49 (m, 2H), 1.00 (t, J=7.2 Hz, 3H).

[0390] MS(ESI) m/z: 405 [M+H.sup.+].

Example 43

4-amino-2-butoxy-7-(4-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile

[0391] ##STR00071##

[0392] 4-amino-2-butoxy-7-(4-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile hydrochloride was prepared according to the procedures of Example 41 and Step A, B, C, D, E, F, G H of Example 41 were followed.

[0393] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.55 (d, J=7.8 Hz, 2H), 7.43 (d, J=7.8 Hz, 2H), 4.60 (t, J=6.5 Hz, 2H), 4.38 (s, 2H), 4.23 (s, 2H), 4.06-4.02 (m, 2H), 3.80-3.73 (m, 2H), 3.47-3.35 (m, 2H), 3.28-3.14 (m, 2H), 1.89-1.82 (m, 2H), 1.59-1.51 (m, 2H), 1.03 (t, J=7.4 Hz, 3H).

[0394] LCMS(ESI) m/z: 421 [M+H.sup.+].

Example 44

4-amino-2-butoxy-7-(4-((4-methylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile

[0395] ##STR00072##

[0396] 4-amino-2-butoxy-7-(4-((4-methylpiperazine-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile hydrochloride was prepared according to the procedures of Example 41 and Step A, B, C, D, E, F, G H of Example 41 were followed.

[0397] .sup.1HNMR (Methanol-d4, 400 MHz): δ:7.61 (d, J=7.8 Hz, 2H), 7.42 (d, J=7.8 Hz, 2H), 4.60 (t, J=6.5 Hz, 2H), 4.47 (s, 2H), 4.23 (s, 2H), 3.89-3.45 (m, 8H), 3.02 (s, 3H), 1.92-1.80 (m, 2H), 1.61-1.44 (m, 2H), 1.03 (t, J=7.3 Hz, 3H).

[0398] LCMS(ESI) m/z: 434 [M+H.sup.+].

Example 45

4-amino-2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-formamide

[0399] ##STR00073##

Example 45 Procedures

[0400] ##STR00074##

[0401] Step A: 4-amino-2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-carbonitrile (90 mg, 0.22 mmol) and sodium hydroxide (34 mg, 0.85 mmol) were dissolved in mixed solvents of methanol (10 mL) and water (10 mL) and the mixture was stirred at 80° C. for 12 h. After cooling, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (15 mL×2). The combined organic layer was concentrated to dryness under vacuum and was purified with preparative HPLC to give 10 mg of 4-amino-2-butoxy-7-(4-(pyrrolidine-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidine-6-formamide.

[0402] .sup.1HNMR (Methanol-d4, 400 MHz): δ7.46 (d, J=8.0 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 4.58 (t, J=6.4 Hz, 2H), 4.39 (s, 2H), 4.34 (s, 2H), 3.34-3.32 (m, 2H), 3.18-3.16 (m, 2H) 2.17-2.16 (m, 2H), 2.03-2.00 (m, 2H), 1.86-1.82 (m, 2H), 1.56-1.50 (m, 2H), 1.02 (t, J=7.2 Hz, 3H).

[0403] MS(ESI) m/z: 423 [M+H.sup.+].

Experimental Example 1: Toll-Like Receptor 7 and Toll-Like Receptor 8 In Vitro Receptor Binding Activity Screen

Reagents:

[0404] HEK-blue hTLR7 cell and HEK-blue hTLR8 cell (available from InvivoGen)
DMEM medium
heat inactivated fetal bovine serum
Anti Mycoplasma reagent Normocin™
bleomycin
blasticidin

Scheme:

[0405] 1. Preparation of 96-well compound plate:

[0406] The compounds were gradient diluted with DMSO in 3-fold using liquid work station POD starting at a concentration of 10 mmol/L and 10 points were diluted (2nd column to 11th column, and each point was duplicated). At 12th column, 1 μL of 5 mg/mL positive compound R848 was added as positive control; and at 1st column, 1 μL of DMSO was added as negative control. Each well contained 1 μL of DMSO.

2. The cells in culture flask were collected and the cell density was diluted to 250,000 cells/mL.
3. 200 μL (50,000 cells/well) of cell suspension was added into prepared compound plate and the final concentration of DMSO in each well was 0.5%.
4. The culture plates containing cells and the compounds were incubated in CO.sub.2 incubator for 24 h at 37° C., 5% CO.sub.2.
5. After 24 h incubation, 20 μL of supernatant was removed from each well to a 96-well transparent assay plate. To each well of the assay plate was added 180 μL of Quanti-Blue reagent and the plate was incubated in an incubator at 37° C., 5% CO.sub.2 for 1 h.
6. After 1 h, the content of alkaline phosphatase in 20 μL of supernatant was determined using Microplate Reader OD650.
7. EC.sub.50 of each compound was obtained with Prism software.

[0407] Results were shown in Table 1:

TABLE-US-00001 TABLE 1 compound TLR7 EC.sub.50 Example 1 C Example 2 C Example 3 C Example 4 B Example 5 C Example 6 B Example 7 B Example 8 B Example 9 C Example 10 C Example 11 B Example 12 B Example 13 B Example 14 B Example 15 B Example 16 B Example 17 B Example 18 B Example 19 B Example 20 B Example 21 B Example 22 B Example 23 C Example 24 B Example 25 A Example 26 B Example 27 B Example 28 B Example 29 B Example 30 B Example 31 B Example 32 B Example 33 B Example 34 B Example 35 C Example 36 C Example 37 C Example 38 B Example 39 B Example 40 B Example 41 A Example 42 A Example 43 A Example 44 A Example 45 B Note: 1 nM ≦ A ≦ 100 nM; 100 nM < B ≦ 1000 nM; 1000 nM < C ≦ 50 μM.

[0408] The head-to-head test results of Example 21 compound and control Toll-like receptor 7 agonist GS-9620 were shown in table 2:

TABLE-US-00002 TABLE 2 Sample (title compound) TLR7 EC.sub.50 (nM) TLR8 EC.sub.50 (nM) GS-9620 517  7867 Example 21 160 11632

[0409] Results: Example 21 compound according to the invention showed higher in vitro receptor binding activity to Toll-like receptor 7 than the control Toll-like receptor 7 agonist GS-9620 and lower in vitro receptor binding activity to Toll-like receptor 8 than the control Toll-like receptor 7 agonist GS-9620.

Experimental Example 2: Peripheral Blood Mononuclear Cell Assay

[0410] The purpose of this example is to determine the expression level of cytokines 24 h after stimulation to human peripheral blood mononuclear cells (PBMC) with the compounds. The cell supernatant was assayed without dilution and the levels of IFN-α and TNF-α were directly determined. The compound was firstly formulated into 20 mM DMSO stock solution and was gradient diluted with cell medium in 10-fold with the total number of 11 diluting points. The compounds in 9 diluting points (the highest concentration was 200 μmol/L) were added into 96-well plate with 50 μL in each well. Fresh human peripheral blood mononuclear cells were inoculated, with 150 μL in each well containing 450,000 cells. The cell culture plate was incubated in an incubator at 37° C., 5% CO.sub.2 for 24 h. After incubation, the culture plate was centrifuged at 1200 rpm for 5 min and the supernatant was collected and stored at −20° C. for determination. The determination of cytokine was performed using Cytometric Bead Array (CBA) of BD-Pharmingen on flow cytometer. Using the above determining method, the lowest drug concentration stimulating cytokine level which is over 3 times greater than the lowest detectable limit was designated as the MEC (Minimal Effective Concentration) value in the cytokine stimulating test.

[0411] The results were shown in Table 3:

TABLE-US-00003 TABLE 3 Example INF-α MEC  4 C 21 A 22 B 28 B 29 A 30 B 31 B 42 A Note: 0.01 nM ≦ A ≦ 1 nM; 1 nM < B ≦ 10 nM; 10 nM < C ≦ 100 μM.

[0412] The head-to-head test results of Example 21 compound and control Toll-like receptor 7 agonist GS-9620 were shown in table 4:

TABLE-US-00004 TABLE 4 Sample (title compound) INF-α MEC (nM) TNF-α MEC (nM) GS-9620 50 500 Example 21 compound  5 500

[0413] Results: Example 21 compound according to the invention showed higher in vitro IFN-α inducing activity than the control Toll-like receptor 7 agonist GS-9620 and comparable TNF-α inducing activity as GS-9620 in PBMC.

Experimental Example 3: Pharmacokinetics in Rat

[0414] 12 male SD rats were divided into 4 groups with 3 SD rats in each group. 2 groups of animals were administered by intravenous injection 1 mg/kg of the control Toll-like receptor 7 agonist GS-9620 and Example 21 compound according to the invention as 10% hydroxypropyl-β-cyclodextrin aqueous solution (concentration is 0.5 mg/mL), respectively. The other 2 groups were administered orally 5 mg/kg of GS-9620 and Example 21 compound as 0.5% methylcellulose/0.2% Tween 80 pure water suspension (concentration is 1 mg/mL). Each rat with intravenous injection was collected for whole blood samples which were prepared into plasma 2, 15, 30 min and 1, 2, 4, 8, 24 h continuously after administration. Each rat with oral administration was collected for whole blood samples which were prepared into plasma 15, 30 min and 1, 2, 4, 8, 24 h continuously after administration. The plasma concentrations of GS-9620 and Example 21 compound were determined with LC-MS/MS. The results were shown in Table 5.

TABLE-US-00005 TABLE 5 Mean plasma drug concentration compound name GS-9620 Example 21 compound IV1 PO1 IV2 PO2 Time (h) (1 mpk) (5 mpk) (1 mpk) (5 mpk) 0.083 170 — 318 — 0.25 102 56.3 141 69.4 0.5 65.4 33.2 109 41.6 1 48.1 83.4 74.3 36.4 2 21.6 136 48.9 186 4 13 16.7 37.7 51.2 8 4.17 9.49 31.6 23.9 24 ND ND 3.94 5.25 C0 or Cmax(nM) 220 164 478 186 T½ (hr) 2.57 2.24 5.76 6.24 Vdss (L/kg) 32.8 — 29 — C1 (mL/min/kg) 205 — 65.8 — AUC0-last (nM .Math. hr) 185 316 641 699 AUC0-inf (nM .Math. hr) 201 359 676 749

[0415] Results: Under the same condition, Example 21 compound according to the invention, as compared to the control Toll-like receptor 7 agonist GS-9620, showed longer half-life and higher exposure in rat.

Experimental Example 4: In Vivo Pharmacodynamics in Duckling Model Infected with Hepatitis B Virus

[0416] Experimental design and procedures: Beijing ducks of 1 day old were intravenously administered duck hepatitis b virus positive duck serum. After 7 days, the animals were administered according to grouping, 6 ducks in each group. Control group: normal saline. Test sample: GS-9620 and Example 21 compound, two dosing groups for each sample: 20 mg/kg and 5 mg/kg. The samples were administered intragastricly: 20 mg/kg groups were administered once every third day (one administration every 3 days) and 5 mg/kg groups were administered once every day for 16 days. The positive control drug lamivudine is manufactured by GlaxoSmithKline, as 50 mg/kg for intragastric administration, which was administered twice a day for 16 days. For control group infected with duck hepatitis b virus, solvent was used instead of drug. 7 days after infection, the blood was collected before administration (TO), 8 days after administration (T8), 16 days after administration (T16) and 3 day after ceasing administration (P3), and the duck serum was separated and frozen for storage. Duck serum was used in the determination of duck hepatitis b virus DNA (DHBV-DNA) and the efficacies of GS-9620, Example 21 compound and positive control lamivudine for duck hepatitis b virus were compared. Duck serum DNA (DHBV-DNA) determination: different duck sera in a batch were determined for duck blood DHBV-DNA level with real time fluorescent quantitative PCR. Statistics analysis: paired and grouped analysis was used to calculate the significance of inhibition of drug on duck serum DHBV-DNA for assessment. The efficacies were shown in Table 6.

TABLE-US-00006 TABLE 6 Duck serum HBV-DNA inhibition % before and after administration Group T8 T16 P3 Control group: normal saline 32.01 ± 44.57 35.96 ± 56.40    65.2 ± 16.7 GS-9620 20 mg/kg 99.13 ± 1.83** 98.26 ± 1.50** −132.97 ± 352.35 Example 21 compound 20 mg/kg 100.0 ± 0** 98.80 ± 1.84*    92.81 ± 13.79** GS-9620 5 mg/kg 98.66 ± 2.75** 78.02 ± 51.69    70.60 ± 47.66 Example 21 compound 5 mg/kg 99.96 ± 0.06** 99.36 ± 1.07**    95.55 ± 3.56** lamivudine 50 mg/kg 99.76 ± 0.28** 99.44 ± 0.99**    95.26 ± 11.20** Grouped t-test, as compared to virus control group at the same time point. *p < 0.05, **p < 0.01.

[0417] Results: As compared to the control Toll-like receptor 7 agonist GS-9620, Example 21 compound according to the invention, under the same condition, showed better efficacies in duckling model infected with hepatitis b virus: for 20 mg/kg (one administration every third day), the inhibition rates are roughly comparable; for 5 mg/kg (one administration everyday), the inhibition rate of Example 21 compound showed significant advantage; 3 days after ceasing administration, GS-9620 20 mg/kg group (one administration every third day) showed rebound of HBV-DNA replication while no rebound was found in the corresponding Example 21 compound group.

Experimental Example 5: In Vivo Pharmacodynamics in HDI (Hydrodynamic Injection) Mouse Model Infected with Hepatitis b Virus

Experimental Design and Procedures:

[0418] Route: intragastric administration
Administration time: day 1 to day 7, 7 days in total
Administration groups: group 1: vehicle, 10% HP-β-CD; group 2: GS-9620, 20 mg/kg; groups 3: Example 21 compound, 20 mg/kg

[0419] At day 1, 3, 5 and 7, plasma samples were collected 4 h after administration; and at day 7, liver sample was collected 4 h after administration. The details were shown in Table 7.

TABLE-US-00007 TABLE 7 Number Plasmid injection of mice Plasmid Administration Time for Time for in each Plasmid injected dosage volume Administration collecting collecting Group group (μg/animal) and time compound (mg/kg) (ml/kg) route blood liver 1 7 ~20 HDI Vehicle / 10 intragastric day 1, day 7, 2 pAAV2-HBV GS-9620 20 administration, 3, 5, 7, 4 h after 3 1.3 mer, Example 20 day 1 to day 7, 4 h after administration (21) once a day administation

[0420] The detailed results of in vivo pharmacodynamics in HDI (hydrodynamic injection) mouse model infected with hepatitis b virus were shown in FIGS. 1 and 2. Results: The data of HBV copy numbers in plasma and liver showed, Example 21 compound, under the same condition had a better efficacy than the control Toll-like receptor 7 agonist GS-9620.