(aza)indolizine derivative and pharmaceutical use thereof

Abstract

(Aza)indolizine derivatives represented by Formula (I) having xanthine oxidase inhibitory activities and useful as agents for the prevention or treatment of a disease associated with abnormality of serum uric acid level, prodrugs thereof, salts thereof or the like. In Formula (I), 0 to 2 of X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are a nitrogen atom and the others are CR.sup.1; one of T.sup.1 and T.sup.2 represents cyano and the other represents a group represented by Formula (A), with the proviso that when T.sup.1 is cyano, at least one of X.sup.1 to X.sup.4 is a nitrogen atom; R.sup.1 independently represents a hydrogen atom, a halogen atom, a hydroxy group, C.sub.1-6 alkyl, C.sub.1-6 alkoxy or the like; ring U represents a benzene ring or the like; m represents integral number from 0 to 2; R.sup.2 independently represents a fluorine atom, a hydroxy group or the like. ##STR00001##

Claims

1. A compound represented by the formula (I): ##STR00039## wherein 0 to 2 of X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are a nitrogen atom and the others are CR.sup.1; one of T.sup.1 and T.sup.2 represents cyano and the other represents a group represented by the formula: ##STR00040## and with the proviso that when T.sup.1 is cyano, at least one of X.sup.1 to X.sup.4 is a nitrogen atom; R.sup.1 independently represents any one of the following (1) to (8): (1) a hydrogen atom; (2) a halogen atom; (3) a hydroxy group; (4) amino; (5) carbamoyl; (6) cyano; (7) carboxy; (8) C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 alkoxy, mono(di) C.sub.1-6 alkylamino, C.sub.2-7 acyl, C.sub.2-7 acylamino, mono(di)C.sub.1-6 alkylcarbamoyl, C.sub.1-6 alkylsulfonyl, C.sub.1-6 alkylsulfonylamino, mono(di)C.sub.1-6 alkylsulfamoyl, C.sub.1-6 alkylthio, C.sub.3-8 cycloalkyl, 3 to 8-membered heterocycloalkyl, C.sub.5-8 cycloalkenyl, 5 to 8-menbered heterocycloalkenyl, C.sub.3-8 cycloalkyloxy, C.sub.3-8 cycloalkylamino, C.sub.6 aryl, 5 or 6-membered heteroary, C.sub.6 aryloxy, C.sub.6 arylamino, C.sub.6 arylcarbonyl or C.sub.6 arylcarbonylamino each of which may have any group selected from substituent group G; substituent group G consists of a fluorine atom, a chlorine atom, a hydroxy group, amino, carboxy, carbamoyl, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy and mono(di)C.sub.1-6 alkylamino; ring U represents a benzene ring; m represents an integral number from 0 to 2; and R.sup.2 represents a fluorine atom, a hydroxy group, amino, methyl or trifluoromethyl, and when m is 2, two R.sup.2 are optionally different from each other, or a pharmaceutically acceptable salt thereof.

2. The compound as claimed in claim 1 wherein the compound, represented by the (I) is a compound, represented by the following formula (Ia) to (Ii): ##STR00041## ##STR00042## wherein R.sup.1a, R.sup.1b and R.sup.1c independently represent any one of the following (a1) to (a4): (a1) a hydrogen atom; (a2) a halogen atom; (a3) a hydroxy group; (a4) C.sub.1-6 alkyl, C.sub.1-6 alkoxy, mono(di)C.sub.1-6 alkylamino, C.sub.2-7 acyl, C.sub.3-8 cycloalkyl, 3 to 8-membered heterocycloalkyl, C.sub.6 aryl or 5 or 6-membered heteroaryl each of which may have any group selected from substituent group G; R.sup.2a represents a hydrogen atom, a fluorine atom, a hydroxy group or amino; and ring U and substituent group G have the same meanings as defined in claim 1, or a pharmaceutically acceptable salt thereof.

3. The compound as claimed in claim 2, wherein the group represented by the formula: ##STR00043## is a group represented by the formula: ##STR00044## and R.sup.2b represents a hydrogen atom or a hydroxy group, or a pharmaceutically acceptable salt thereof.

4. The compound as claimed in claim 2, wherein R.sup.1a, R.sup.1b and R.sup.1c independently represent any one of the following (b1) to (b4): (b1) a hydrogen atom; (b2) a halogen atom; (b3) a hydroxy group; (b4) C.sub.1-6 alkyl, C.sub.1-6 alkoxy, mono(di) C.sub.1-6 alkylamino or hydroxyC.sub.1-6 alkyl each of which may be substituted by a fluorine atom, or a pharmaceutically acceptable salt thereof.

5. The compound as claimed in claim 1, represented by the formula (II): ##STR00045## wherein one of T.sup.1 and T.sup.2 represents cyano and the other represents a group represented by the formula: ##STR00046## R.sup.3 independently represents any one of the following (c1) to (c4): (c1) a hydrogen atom; (c2) a halogen atom; (c3) a hydroxy group; (c4) C.sub.1-6 alkyl, C.sub.1-6 alkoxy, mono(di)C.sub.1-6 alkylamino, C.sub.2-7 acyl, C.sub.3-8 cycloalkyl, 3 to 8-membered heterocycloalkyl, C.sub.6 aryl or 5 or 6-membered heteroaryl each of which may have any group selected from substituent group G; R.sup.2a represents a hydrogen atom, a fluorine atom, a hydroxy group or amino; and ring U and substituent group G have the same meanings as defined in claim 1, or a pharmaceutically acceptable salt thereof.

6. The compound as claimed in claim 5, wherein R.sup.3 independently represents any one of the following (d1) to (d4): (d1) a hydrogen atom; (d2) a halogen atom; (d3) a hydroxy group; (d4) C.sub.1-6 alkyl, C.sub.1-6 alkoxy, mono(di)C.sub.1-6 alkylamino or hydroxyC.sub.1-6 alkyl each of which may be substituted by a fluorine atom, or a pharmaceutically acceptable salt thereof.

7. The compound as claimed in claim 6, represented by the formula (IIa): ##STR00047## wherein R.sup.2b represents a hydrogen atom or a hydroxy group; R.sup.3a, R.sup.3b and R.sup.3c independently represent any one of the following (e1) to (e4): (e1) a hydrogen atom; (e2) a halogen atom; (e3) a hydroxy group; (e4) C.sub.1-6 alkyl, C.sub.1-6 alkoxy, mono(di)C.sub.1-6alkylamino or hydroxyC.sub.1-6 alkyl each of which may be substituted by a fluorine atom, or a pharmaceutically acceptable salt thereof.

8. The compound as claimed in claim 6, represented by the formula (IIb): ##STR00048## wherein R.sup.2b represents a hydrogen atom or a hydroxy group; and R.sup.3b and R.sup.3c independently represent any one of the following (f1) to (f4): (f1) a hydrogen atom; (f2) a halogen atom; (f3) a hydroxy group; (f4) C.sub.1-6 alkyl, C.sub.1-6 alkoxy, mono(di)C.sub.1-6 alkylamino or hydroxyC.sub.1-6 alkyl each of which may be substituted by a fluorine atom, or a pharmaceutically acceptable salt thereof.

9. The compound as claimed in claim 7, wherein R.sup.2b is a hydroxy group; R.sup.3a is a hydrogen atom, a fluorine atom or a chlorine atom; R.sup.3b is a hydrogen atom, a fluorine atom, methyl, ethyl or methoxy; and R.sup.3c is a hydrogen atom, a fluorine atom, a chlorine atom, methyl or trifluoromethyl, or a pharmaceutically acceptable salt thereof.

10. The compound as claimed in claim 8, wherein R.sup.2b is a hydroxy group; R.sup.3b is a hydrogen atom, a fluorine atom, methyl, ethyl or methoxy; and R.sup.3c is a hydrogen atom, a fluorine atom, a chlorine atom, methyl or trifluoromethyl, or a pharmaceutically acceptable salt thereof.

11. The compound as claimed in claim 4, represented by the formula (Ij): ##STR00049## wherein R.sup.2b represents a hydrogen atom or a hydroxy group; and R.sup.1a and R.sup.1b have the same meanings as defined in claim 4, or a pharmaceutically acceptable salt thereof.

12. The compound as claimed in claim 11, wherein R.sup.2b is a hydroxy group; R.sup.1a is a hydrogen atom, a fluorine atom, a chlorine atom or methyl; and R.sup.1b is a hydrogen atom, a fluorine atom, methyl, ethyl or methoxy, or a pharmaceutically acceptable salt thereof.

13. 4-(7-Cyanopyrrolo[1,2-b]pyridazine-5-yl)-2-hydroxy benzoic acid, or a pharmaceutically acceptable salt thereof.

14. 4(7-Cyano-2-methyl-pyrrolo[1,2-b]pyridazin-5-yl)-2-hydroxy-benzoic acid, or a pharmaceutically acceptable salt thereof.

15. 4-(3Cyano-indolizin-1-yl)-2hydroxyl-benzoie acid, or a pharmaceutically acceptable salt thereof.

16. 4-(5-Cyano-pyrrolo[1,2-b]pyridazin-7-ul)-2-hydroxy-benzoic acid, or a pharmaceutically acceptable salt thereof.

17. A xanthine oxidase inhibitor comprising a compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof as an active ingredient.

18. A pharmaceutical composition comprising a compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof as an active ingredient.

19. A method of treatment of a diease selected from the group consisting of hyperuricemia, gouty tophus, gouty arthritis, tenal disorder associeated with hyperuricemia and Urinary calculi comprising administering an effective amount of the pharmaceutical composition as Claimed in claim 18 to a patient in need thereof.

20. The method of treatment of claim 19, wherein the disease is hyperuricemia.

21. A method for lowering serum uric acid level comprising administering an effective amount of the pharmaceutical composition as claimed in claim 18 to a patient in need thereof.

22. A method of inhibiting the production of uric acid comprising administering an effective amount of the pharmaceutical composition as claimed in claim 18 to a patient in need thereof.

Description

REFERENCE EXAMPLE 1

2-Methoxymethoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolane-2-yl)benzoic acid methyl ester

(1) To a solution of 4-iodo-2-methoxymethoxy benzoic acid methyl ester (1.29 g) in N,N-dimethylformamide (10 mL) were added bis(pinacolato)diboron (1.12 g), palladium acetate (II) (0.05 g) and potassium acetate (1.18 g) at room temperature, and the mixture was stirred at 80 C. for 8 hours. After cooling to ambient temperature, to the reaction mixture was added water, and the resulting mixture was extracted with toluene. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give the title compound (0.52 g).

REFERENCE EXAMPLE 2

5-Bromopyrrolo[1,2-b]pyridazine-7-carbonitrile

(2) To a solution of pyridazine (1.10 g) in tetrahydrofuran (30 mL) was added tetracyanoethylene oxide (2.00 g) at 0 C., and the mixture was stirred at the same temperature overnight. The mixture was concentrated under reduced pressure, and the obtained solid was washed with dichloromethane and dried under reduced pressure at 50 C. To a suspension of the obtained solid (1.90 g) in toluene (30 mL) was added bis(trimethylsilyl)acetylene (22.9 g) at room temperature, and the mixture was heated under reflux for 2 days. The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give a mixture (1.60 g) of 5,6-bistrimethylsilanylpyrrolo[1,2-b]pyridazine-7-carbonitrile and 5-trimethylsilanylpyrrolo[1,2-b]pyridazine-7-carbonitrile. To a solution of the obtained mixture (1.60 g) in tetrahydrofuran (25 mL) was added 1 mol/L solution of tetra-n-butyl ammonium fluoride in tetrahydrofuran (5.7 mL) under ice-cooling, and the mixture was stirred at room temperature overnight. To the reaction mixture was added water, and the resulting mixture was extracted with dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to give pyrrolo[1,2-b]pyridazine-7-carbonitrile (1.10 g). To a solution of the obtained compound (1.10 g) in dichloromethane (30 mL) was added N-bromosuccinimide (1.50 g) at room temperature, and the mixture was stirred at the same temperature for 3 hours. To the mixture was added 1 mol/L sodium thiosulfate aqueous solution. The resulting mixture was extracted with dichloromethane, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give the title compound (1.23 g).

REFERENCE EXAMPLE 3

5-Bromo-2-methylpyrrolo[1,2-b]pyridazine-7-carbonitrile

(3) The title compound was prepared in a similar manner to that described in Reference Example 2 using the corresponding starting materials.

REFERENCE EXAMPLE 4

5-Bromo-2-chloro-3-methylpyrrolo[1,2-b]pyridazine-7-carbonitrile

(4) The title compound was prepared in a similar manner to that described in Reference Example 2 using the corresponding starting materials.

REFERENCE EXAMPLE 5

5-Bromo-2-chloropyrrolo[1,2-b]pyridazine-7-carbonitrile

(5) The title compound was prepared in a similar manner to that described in Reference Example 2 using the corresponding starting materials.

REFERENCE EXAMPLE 6

5-Bromo-2-methoxypyrrolo[1,2-b]pyridazine-7-carbonitrile

(6) The title compound was prepared in a similar manner to that described in Reference Example 2 using the corresponding starting materials.

REFERENCE EXAMPLE 7

1-Bromoindolizine-3-carbonitrile

(7) To a solution of pyridine (2.00 g) in ethyl acetate (20 mL) was added bromoacetonitrile (3.64 g) at room temperature, and the mixture was heated under reflux overnight. The precipitated solid was collected by filtration, washed with ethyl acetate and dried under reduced pressure at 50 C. To a suspension of the obtained compound (4.9 g) in chlorobenzene (30 mL) were added benzyl acrylate (7.99 g), manganese dioxide (10.70 g) and triethylamine (2.49 g) at room temperature, and the mixture was stirred at 80 C. for 5 hours. The mixture was filtered through a Celite pad, and the filtrate was concentrated. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give 3-cyanoindolizine-1-carboxylic acid benzyl ester (4.90 g). To a mixed solution of the obtained compound (2.50 g) in methanol (15 mL) and tetrahydrofuran (15 mL) was added palladium 10% on carbon (wetted with ca. 50% water) (0.25 g), and the mixture was stirred under a hydrogen atmosphere at room temperature overnight. To the mixture was added 1 mol/L sodium hydroxide aqueous solution, and the resulting mixture was extracted with ether. To the aqueous layer was added 2 mol/L hydrochloric acid, and the precipitated solid was collected by filtration. The obtained solid was dried under reduced pressure at 50 C. to give 3-cyanoindolizine-1-carboxylic acid (0.63 g). To a suspension of the obtained compound (0.63 g) in quinoline (5 mL) was added copper (0.05 g), and the mixture was stirred at 220 C. for 45 minutes. To the mixture was added 2 mol/L hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic layer was washed with 1 mol/L hydrochloric acid and brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give indolizine-3-carbonitrile (0.23 g). To a solution of the obtained compound (0.23 g) in dichloromethane (5 mL) was added N-bromosuccinimide (0.37 g) at room temperature, and the mixture was stirred at same temperature for 2 hours. To the reaction mixture was added 1 mol/L sodium thiosulfate aqueous solution, and the resulting mixture was extracted with dichloromethane. The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give the title compound (0.30 g).

REFERENCE EXAMPLE 8

1-Bromo-6-fluoroindolizine-3-carbonitrile

(8) The title compound was prepared in a similar manner to that described in Reference Example 7 using the corresponding starting materials.

REFERENCE EXAMPLE 9

7-Bromopyrrolo[1,2-b]pyridazine-5-carbonitrile

(9) To a solution of pyridazine (25 g) in ethyl acetate (200 ml) was added bromoacetic acid (52 g) at room temperature, and the mixture was stirred at 80 C. overnight. The mixture was concentrated under reduced pressure. The precipitated solid was collected by filtration, and dried under reduced pressure. To a suspension of the obtained compound (17 g) in chlorobenzene (160 mL) were added acrylonitrile (8.28 g), manganese dioxide (33 g) and triethylamine (7.89 g), and the mixture was stirred at 80 C. for 5 hours. The reaction mixture was filtered through a Celite pad, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give pyrrolo[1,2-b]pyridazine-5-carbonitrile (0.30 g). To a solution of the obtained compound (0.30 g) in dichloromethane (10 mL) was added N-bromosuccinimide (0.41 g) at room temperature, and the mixture was stirred at same temperature for 3 hours. To the mixture was added 1 mol/L sodium thiosulfate aqueous solution, and the resulting mixture was extracted with dichloromethane. The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give the title compound (0.20 g).

REFERENCE EXAMPLE 10

4-(5-Cyano-4-methyl pyrrolo[1,2-b]pyridazine-7-yl)benzoic acid methyl ester

(10) To a solution of 4-methyl pyridazine (1.00 g) in ethyl acetate (30 ml) was added 4-bromomethyl benzoic acid methyl ester (2.92 g) at room temperature, and the mixture was stirred at 80 C. overnight. The precipitated solid was collected by filtration, washed with ethyl acetate and dried under reduced pressure. To a solution of the obtained compound (4.50 g) in 1,2-dimethoxyethane (26 mL) were added acrylonitrile (3.69 g), manganese dioxide (6.05 g) and triethylamine (4.22 g) at room temperature, and the mixture was stirred at 80 C. for 7 hours. The mixture was filtered through a Celite pad, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give the title compound (0.330 g).

REFERENCE EXAMPLE 11

4-(8-Cyanopyrrolo[1,2-a]pyrazine-6-yl)benzoic acid methyl ester

(11) The title compound was prepared in a similar manner to that described in Reference Example 10 using the corresponding starting materials.

REFERENCE EXAMPLE 12

8-Bromopyrrolo[1,2-a]pyrazine-6-carbonitrile

(12) The title compound was prepared in a similar manner to that described in Reference Example 2 using the corresponding starting materials.

REFERENCE EXAMPLE 13

4-(3-Cyanopyrrolo[3,2-b]pyridine-1-yl)-2-methoxymethoxy benzoic acid ethyl ester

(13) To a solution of 1H-pyrrolo[3,2-b]pyridine (0.4 g) in N,N-dimethylformamide (6 mL) were added 4-fluoro-2-methoxymethoxy benzoic acid ethyl ester (0.85 g) and cesium carbonate (2.21 g) at room temperature, and the mixture was stirred at 80 C. overnight. To the mixture was added water, and the resulting mixture was extracted with dichloromethane. The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give 2-methoxymethoxy-4-pyrrolo[3,2-b]pyridine-1-yl-benzoic acid ethyl ester (0.60 g). To a solution of the obtained compound (0.60 g) in dichloromethane (30 mL) was added N-bromosuccinimide (0.33 g) under ice-cooling, and the mixture was stirred at the same temperature for 2 hours. To the mixture was added 1 mol/L sodium thiosulfate aqueous solution. The resulting mixture was extracted with dichloromethane, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give 4-(3-bromopyrrolo[3,2-b]pyridine-1-yl)-2-methoxymethoxy benzoic acid ethyl ester (0.49 g). To a solution of the obtained compound (0.49 g) in N-methylpyrrolidone (10 mL) were added zinc cyanide and tetrakis (triphenylphosphine) palladium (0) (0.28 g) at room temperature, and the mixture was stirred at 150 C. for 1 hour using microwave reactor. To the reaction mixture was added water, and the resulting mixture was extracted with ethyl acetate. The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate) to give the title compound (0.25 g).

EXAMPLE 1

4-(7-Cyanopyrrolo[1,2-b]pyridazine-5-yl)benzoic acid

(14) To a mixed solution of 5-bromopyrrolo[1,2-b]pyridazine-7-carbonitrile (0.022 g) in 1,2-dimethoxyethane (2 mL) and water (0.3 mL) were added 4-methoxycarbonyl phenyl boronic acid (0.020 g), tetrakis(triphenylphosphine)palladium (0) (0.006 g) and cesium carbonate (0.048 g) at room temperature, and the mixture was stirred at 80 C. under an argon atmosphere for 3 hour. To the mixture was added water, and the resulting mixture was extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate=95/5-10/90) to give 4-(7-cyanopyrrolo[1,2-b]pyridazine-5-yl)benzoic acid methyl ester (0.018 g). To a mixed solution of the obtained compound (0.018 g) in tetrahydrofuran (2 mL), ethanol (1 mL) and water (1 mL) was added lithium hydroxide monohydrate (0.008 g) at room temperature, and the mixture was stirred at same temperature overnight. To the reaction mixture was added 1 mol/L hydrochloric acid, and the precipitated solid was collected by filtration, washed with water and dried under reduced pressure at 50 C. to give the title compound (0.002 g).

EXAMPLE 2

4-(7-Cyanopyrrolo[1,2-b]pyridazine-5-yl)-2-hydroxy benzoic acid

(15) To a mixed solution of 5-bromopyrrolo[1,2-b]pyridazine-7-carbonitrile (0.38 g) in N,N-dimethylformamide (5 mL) and water (0.5 mL) were added 2-methoxymethoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolane-2-yl) benzoic acid methyl ester (0.55 g), tetrakis (triphenylphosphine) palladium (0) (0.10 g) and cesium carbonate (0.83 g) at room temperature, and the mixture was stirred at 80 C. for 7 hours. After cooling to room temperature, to the reaction mixture was added water, and the precipitated solid was collected by filtration and purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate=95/5-10/90) to give 4-(7-cyanopyrrolo[1,2-b]pyridazine-5-yl)-2-methoxymethoxy benzoic acid methyl ester (0.41 g). To a mixed solution of the obtained compound (0.41 g) in tetrahydrofuran (12 mL), ethanol (6 mL) and water (6 mL) was added lithium hydroxide monohydrate (0.24 g), and the mixture was stirred at room temperature for 5 hours. To the reaction mixture was added 2 mol/L hydrochloric acid, and the mixture was stirred at 50 C. overnight. After cooling to room temperature, to the reaction mixture was added water. The precipitated solid was collected by filtration, washed with water and dried under reduced pressure at 50 C. to give the title compound (0.30 g).

EXAMPLES 3 TO 6

(16) The compounds of Examples 3 to 6 were prepared in a similar manner to that described in Example 2 using the corresponding starting materials.

EXAMPLE 7

(17) The compound of Example 7 was prepared in a similar manner to that described in Example 1 using the corresponding starting materials.

EXAMPLES 8 TO 9

(18) The compounds of Examples 8 to 9 were prepared in a similar manner to that described in Example 2 using the corresponding starting materials.

EXAMPLE 10

(19) The compound of Example 10 was prepared in a similar manner to that described in Example 1 using the corresponding starting materials.

EXAMPLE 11

4-(5-Cyano-4-methyl pyrrolo[1,2-b]pyridazine-7-yl)benzoic acid

(20) To a mixed solution of 4-(5-cyano-4-methyl pyrrolo[1,2-b]pyridazine-7-yl)benzoic acid methyl ester (0.35 g) in tetrahydrofuran (7 mL), ethanol (3.5 mL) and water (3.5 mL) was added lithium hydroxide monohydrate (0.14 g) at room temperature, and the mixture was stirred at same temperature overnight. To the mixture was added 1 mol/L hydrochloric acid. The precipitated solid was collected by filtration, washed with water and methanol and dried under reduced pressure at 50 C. to give the title compound (0.088 g).

EXAMPLE 12

(21) The compound of Example 12 was prepared in a similar manner to that described in Example 2 using the corresponding starting materials.

EXAMPLE 13

(22) The compound of Example 13 was prepared in a similar manner to that described in Example 11 using the corresponding starting materials.

EXAMPLE 14

4-(6-Cyanopyrrolo[1,2-a]pyrazine-8-yl)-2-hydroxy benzoic acid

(23) To a mixed solution of 8-bromopyrrolo[1,2-a]pyrazine-6-carbonitrile (0.038 g) in N,N-dimethylformamide (2 mL) and water (0.2 mL) were added 2-methoxymethoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolane-2-yl)benzoic acid methyl ester (0.066 g), tetrakis (triphenylphosphine) palladium (0) (0.01 g) and cesium carbonate (0.084 g) at room temperature, and the mixture was stirred at 80 C. for 6 hours. After cooling to room temperature, to the reaction mixture was added water, and the resulting mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: n-hexane/ethyl acetate=95/5-10/90) to give 4-(6-cyanopyrrolo[1,2-a]pyazine-8-yl)-2-methoxymethoxy benzoic acid methyl ester (0.016 g). To a mixed solution of the obtained compound (0.016 g) in tetrahydrofuran (2 mL), ethanol (1 mL) and water (1 mL) was added lithium hydroxide monohydrate (0.01 g), and the mixture was stirred at room temperature for 4 hours. To the reaction mixture was added 2 mol/L hydrochloric acid, and the mixture was stirred at 50 C. overnight. After cooling to room temperature, to the reaction mixture was added water. The precipitated solid was collected by filtration, washed with water and dried under reduced pressure at 50 C. to give 4-(6-carbamoyl pyrrolo[1,2-a]pyrazine-8-yl)-2-hydroxy benzoic acid (0.013 g). To a solution of the obtained solid (0.01 g) in dichloromethane were added trifluoroacetic anhydride (0.035 g) and triethylamine (0.027 g) under ice-cooling, and the mixture was stirred at room temperature overnight. To the mixture was added methanol, and the mixture was stirred for 30 minutes. The solvent was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (eluent: methanol/dichloromethane=30/70-50/50). To the obtained solid were added ethyl acetate and 1 mol/L hydrochloric acid, and the organic layer was concentrated under reduced pressure to give the title compound (0.001 g).

EXAMPLE 15

4-(3-Cyanopyrrolo[3,2-b]pyridine-1-yl)-2-hydroxy benzoic acid

(24) To a mixed solution of 4-(3-cyanopyrrolo[3,2-b]pyridine-1-yl)-2-methoxymethoxy benzoic acid ethyl ester (0.25 g) in tetrahydrofuran (7.2 mL), ethanol (3.6 mL) and water (3.6 mL) was added lithium hydroxide monohydrate (0.15 g), and the mixture was stirred at room temperature for 5 hours. To the reaction mixture was added 2 mol/L hydrochloric acid, and the mixture was stirred at 50 C. overnight. After cooling to room temperature, to the reaction mixture was added water. The precipitated solid was collected by filtration, washed with water and dried under reduced pressure at 50 C. to give the title compound (0.15 g).

EXAMPLE 16

(25) The compound of Example 16 was prepared in a similar manner to that described in Example 2 using the corresponding starting materials.

(26) Tables 1 to 3 show the chemical structures and .sup.1H-NMR data of the above compounds of Examples 1 to 16.

(27) The abbreviations in these Tables: Ex No. and Str., represent Example number and chemical structure, respectively.

(28) TABLE-US-00001 TABLE1 Ex. No. Str. .sup.1H-NMR ppm (DMSO-d6) 1 7.10-7.30 (1H, m), 7.70-8.05 (4H, m), 8.08 (1H, s), 8.45-8.65 (2H, m) 2 7.15-7.95 (4H, m), 8.09 (1H, s), 8.45-8.70 (2H, m) 3 2.56 (3H, s), 7.12 (1H, d, J = 9.5 Hz), 7.20-7.35 (2H, m), 7.80-7.95 (1H, m), 7.99 (1H, s), 8.40 (1H, d, J = 9.5 Hz) 4 2.20-2.35 (3H, m), 7.05-7.25 (2H, m), 7.89 (1H, s), 7.90-8.20 (2H, m) 5 7.55-7.70 (2H, m), 7.75-8.05 (3H, m), 8.15-8.30 (1H, m)

(29) TABLE-US-00002 TABLE 2 Ex. No. Str. .sup.1H-NMR ppm (DMSO-d6) 6 4.00 (3H, s), 6.80-7.30 (3H, m), 7.75-8.45 (3H, m) 7 7.00-7.40 (2H, m), 7.60-8.20 (6H, m), 8.45-8.65 (1H, m) 8 0 7.00-7.45 (4H, m), 7.75-8.15 (3H, m), 8.45-8.65 (1H, m) 9 7.10-7.45 (3H, m), 7.75-8.15 (3H, m), 8.70-8.85 (1H, m) 10 7.15-7.35 (1H, m), 7.99 (1H, s), 8.00-8.45 (5H, m), 8.55-8.70 (1H, m), 13.05 (1H, brs)

(30) TABLE-US-00003 TABLE 3 Ex. No. Str. .sup.1H-NMR ppm (DMSO-d6) 11 2.74 (3H, s), 7.08 (1H, d, J = 4.6 Hz), 7.93 (1H, s), 7.95-8.30 (4H, m), 8.51 (1H, d, J = 4.6 Hz), 13.03 (1H, brs) 12 7.20-7.90 (4H, m), 8.03 (1H, s), 8.30-8.70 (2H, m) 13 7.78 (1H, s), 7.80-8.20 (5H, m), 8.50-8.65 (1H, m), 9.15-9.30 (1H, m), 13.18 (1H, brs) 14 7.25-7.45 (2H, m), 7.80-8.20 (3H, m), 8.45-8.65 (1H, m), 9.30-9.50 (1H, m) 15 7.20-7.50 (3H, m), 7.95-8.25 (2H, m), 8.55-8.70 (1H, m), 8.90-9.05 (1H, m) 16 2.34 (3H, s), 2.90 (3H, s), 7.05-7.30 (3H, m), 7.75-7.90 (2H, m), 7.95 (1H, s)

TEST EXAMPLE 1

(31) Xanthine Oxidase Inhibitory Activity

(32) (1) Preparation of Test Compounds

(33) Test compounds were dissolved in dimethyl sulfoxide (DMSO) (manufactured by Wako pure chemical) at 40 mM concentration and then diluted to intended concentrations with phosphate-buffered saline (PBS).

(34) (2) Method of Measurement

(35) Xanthine oxidase (from bovine milk, manufactured by Sigma) was prepared with phosphate-buffered saline (PBS) at 0.02 units/mL, and then the solution was added to 96-well plates at 50 L/well. In addition, test compounds diluted with PBS were added at 50 L/well. Xanthine (manufactured by Wako pure chemical) at 200 M prepared with PBS was added at 100 L/well, and the reaction was conducted for 10 minutes at room temperature. Absorbance at 290 nm was measured by using a microplate reader SpectraMax Plus 384 (manufactured by Molecular device). The absorbance under a condition without xanthine is 0%, and control without test compounds is 100%. Fifty % inhibitory concentration of a test compound (IC.sub.50) was calculated (Table 4). Ex. No in the table indicates Example number.

(36) TABLE-US-00004 TABLE 4 Ex. No. IC.sub.50 (nM) 1 7 2 5 3 5 6 3 7 8 8 12 9 4 10 12 11 10 12 4 15 10 16 13

TEST EXAMPLE 2

(37) Inhibitory Activity of Uric Acid Transport with Human URAT1-Expressing Cells

(38) (1) Cells Used in the Test

(39) URAT1-expressing HEK293 cells (HEK293 cells transfected with vector containing human URAT1 cDNA) and control cells (HEK293 cells transfected with vector alone) were used.

(40) URAT1-expressing cells and control cells were seeded in collagen I-coated 24-well plates (manufactured by BD FALCON) at 1 to 410.sup.5 cells/well, and cultured in CO.sub.2 incubator (37 C., CO.sub.2: 5%) for 1 to 3 days, and then the following measurement of uric acid transport was conducted. In addition, Dulbecco's Modified Eagle Medium (manufactured by Invitrogen) containing 9% fetal bovine serum (manufactured by Invitrogen), antibiotic-antimycotic (manufactured by Invitrogen) and 2 mmol/L L-glutamine were used for the culture.

(41) (2) Preparation of Test Compounds

(42) .sup.14C-labeled uric acid (.sup.14C uric acid) (manufactured by American Radiolabeled Chemicals, Inc.) was dissolved in Hanks' balanced salt solution (HBSS) (manufactured by Invitrogen) to prepare HBSS containing .sup.14C uric acid at 50 M. Test compounds were dissolved in DMSO and then diluted to 1000-fold with the prepared HBSS containing .sup.14C uric acid above to prepare .sup.14C uric acid solution containing the indicated concentration of test compounds (final concentration of DMSO: 0.1%). .sup.14C uric acid solution containing 0.1% DMSO was prepared as a control.

(43) (3) Measurement of Uric Acid Transport

(44) After the medium was removed from the plates in which cells were seeded, and 1 mL of HBSS was added to the cells. After the HBSS was removed, 0.3 mL of HBSS was newly added to the cells, which were incubated at 37 C. for 15 minutes. After the HBSS was removed, 0.3 mL of .sup.14C uric acid solution containing 0.1% DMSO or test compounds was added to the cells, which were then incubated at 37 C. for 2 minutes. After the incubation, the solution was removed, and the cells were washed once with 1 mL of ice-cold phosphate-buffered saline containing 0.2% BSA, and washed twice with 1 mL of ice-cold PBS. After the PBS was removed, the cells were lysed by addition of 0.5 mL of 0.1 mol/L NaOH aqueous solution for each well. The cell lysates (0.3 mL/well) were transferred into glass vials, and mixed with 10 mL of a scintillator (Hionic-Fluor, manufactured by Perkin Elmer). The radioactivity was measured by means of a liquid scintillation counter.

(45) (4) Protein Determination

(46) Protein concentration in the cell lysates was determined by BCA Protein Assay Kit (manufactured by Pierce) and then amount of protein (mg/well) was calculated.

(47) (5) Calculation of the Percent Inhibition of Uric Acid Uptake for Each Compound

(48) Uric acid uptake activity in each well was calculated by the following formula.
Uric acid uptake activity (p mol/mg protein)=radioactivity (dpm/well)/[amount of protein (mg/well)concentration of radioactivity in HBSS containing .sup.14C uric acid (dpm/p mol)]

(49) Percent inhibition was calculated according to the following formula.
Percent inhibition (%)=[1(BC)/(AC)]100 A: Uric acid uptake activity in URAT1-expressing HEK293 cells in the presence of 0.1% DMSO B: Uric acid uptake activity in URAT1-expressing HEK293 cells in the presence of test compounds C: Uric acid uptake activity in control cells in the presence of 0.1% DMSO
(6) Results

(50) The compounds of Examples 2, 3, 6, 8, 9, 12 and 16 showed not less than 50 percent inhibition in a concentration of 10 M.

TEST EXAMPLE 3

(51) Hypouricemic Effect

(52) (1) Method of Measurement

(53) Test compounds (1 mg/kg) suspended in 0.5% methylcellulose solution were orally administered to overnight fasted male CD (SD) IGS rats (5-week-old, Charles River Japan). At 2 hours after administration, blood was collected under ether anesthesia from abdominal aorta, and serum was separated according to a general method. Serum uric acid levels were determined by use of uric acid measurement kit (Uric acid C-Test Wako: manufactured by Wako pure chemical), and percent decrease in uric acid was calculated according to the formula described below.
Percent decrease in uric acid (%)=(Serum uric acid levels in control animalsSerum uric acid levels in animals administered test compounds)100/Serum uric acid levels in control animals
(2) Results

(54) Oral administration of test compounds of Examples 2 and 12 at 3 mg/kg showed not less than 60 percent decrease in uric acid.

(55) Oral administration of a test compound of Example 3 at 1 mg/kg showed not less than 60 percent decrease in uric acid.

TEST EXAMPLE 4

(56) Acute Toxicity Test

(57) Test compounds of Examples 2 and 12 (300 mg/kg) suspended in 0.5% methylcellulose solution were orally administered to male ICR mice (7-week-old, 5 mice per group), and the general condition during 24 hours after administration was observed. As a result, there were no deaths, and no abnormalities in general conditions were observed.

(58) Industrial Applicability

(59) The (aza)indolizine derivatives represented by the formula (I) of the present invention or prodrugs thereof, or pharmaceutically acceptable salts thereof exert an excellent xanthine oxidase inhibitory activity, and therefore, can exert an inhibitory activity of uric acid production and lower serum uric acid level. Therefore, the present invention can provide an agent for the prevention or treatment of hyperuricemia, gouty tophus, gouty arthritis, renal disorder associated with hyperuricemia, urinary calculi or the like.