Bicyclic inhibitors

Abstract

The present invention provides compounds of formula (I): compositions comprising such compounds; the use of such compounds in therapy (for example in the treatment or prevention of a disease or condition in which plasma kallikrein activity is implicated); and methods of treating patients with such compounds; wherein A, B, W, X and Y are as defined herein. ##STR00001##

Claims

1. A compound as defined by formula (III) or formula (IV): ##STR00035## wherein: B is (i) or (ii): i) a radical of formula II: ##STR00036## or ii) a fused 6,5- or 6,6-heteroaromatic bicyclic ring, containing N and, optionally, one or two additional N, O, or S heteroatoms, which is optionally mono-, di or tri-substituted with a substituent that is alkyl, alkoxy, OH, halo, CN, COOR8, CONR8R9, CF.sub.3 or NR8R9; P is H and Q is —C(R20)(R21)NH.sub.2, or P is —C(R20)(R21)NH.sub.2 and Q is H; U and V are independently C or N such that the aromatic ring containing U and V is phenyl, pyridine or pyrazine; R1 is absent when U is N; R2 is absent when V is N; or, when present, R1 and R2 are independently H, alkyl, alkoxy, CN, halo or CF.sub.3; R3 is H, alkyl, alkoxy, CN, halo or CF.sub.3; A is —(CH.sub.2).sub.0-9-heteroaryl or —(CH.sub.2).sub.0-9-aryl; R8 and R9 are independently H or alkyl; R20 and R21 are independently H and alkyl, or may together form a cycloalkyl ring or a cyclic ether; alkyl is a linear saturated hydrocarbon having up to 10 carbon atoms (C.sub.1-C.sub.10) or a branched saturated hydrocarbon of between 3 and 10 carbon atoms (C.sub.3-C.sub.10); alkyl may optionally be independently substituted with 1 or 2 substituents which are (C.sub.1-C.sub.6)alkoxy, OH, CN, CF.sub.3, COOR10, CONR10R11, fluoro or NR10R11; cycloalkyl is a monocyclic saturated hydrocarbon of between 3 and 7 carbon atoms; a cyclic ether is a monocyclic saturated hydrocarbon of between 4 and 7 carbon atoms, wherein one of the ring carbons is replaced by an oxygen atom; alkoxy is a linear O-linked hydrocarbon of between 1 and 6 carbon atoms (C.sub.1-C.sub.6) or a branched O-linked hydrocarbon of between 3 and 6 carbon atoms (C.sub.3-C.sub.6); alkoxy may optionally be independently substituted with 1 or 2 substituents which are OH, CN, CF.sub.3, COOR10, CONR10R11, fluoro or NR10R11; aryl is phenyl, biphenyl or naphthyl; aryl may be optionally independently substituted with 1, 2 or 3 substituents which are alkyl, alkoxy, methylenedioxy, ethylenedioxy, OH, halo, CN, morpholinyl, piperidinyl, heteroaryl, —(CH.sub.2).sub.0-3—O-heteroaryl, aryl.sup.b, —O-aryl.sup.b, —(CH.sub.2).sub.1-3-aryl.sup.b, —(CH.sub.2).sub.1-3-heteroaryl, —COOR10, —CONR10R11, —(CH.sub.2).sub.1-3—NR14R15, CF.sub.3 or —NR10R11; aryl.sup.b is phenyl, biphenyl or naphthyl, which may be optionally independently substituted with 1, 2 or 3 substituents which are alkyl, alkoxy, OH, halo, CN, morpholinyl, piperidinyl, —COOR10, —CONR10R11, CF.sub.3 or NR10R11; heteroaryl is a 5, 6, 9 or 10 membered mono- or bi-cyclic aromatic ring, containing, where possible, 1, 2 or 3 ring members which are independently N, NR8, S or O; heteroaryl may be optionally independently substituted with 1, 2 or 3 substituents which are alkyl, alkoxy, OH, halo, CN, aryl, morpholinyl, piperidinyl, —(CH.sub.2).sub.1-3-aryl, heteroaryl.sup.b, —COOR10, —CONR10R11, CF.sub.3 or —NR10R11; heteroaryl.sup.b is a 5, 6, 9 or 10 membered mono- or bi-cyclic aromatic ring, containing, where possible, 1, 2 or 3 ring members which are independently N, NR8, S or O; wherein heteroaryl.sup.b may be optionally independently substituted with 1, 2 or 3 substituents which are alkyl, alkoxy, OH, halo, CN, morpholinyl, piperidinyl, aryl, —(CH.sub.2).sub.1-3-aryl, —COOR10, —CONR10R11, CF.sub.3 or NR10R11; R10 and R11 are independently H or alkyl; or R10 and R11 together with the nitrogen to which they are attached form a 4-, 5-, 6- or 7-membered heterocyclic ring which may be saturated or unsaturated with 1 or 2 double bonds; R14 and R15 are independently alkyl, aryl.sup.b or heteroaryl.sup.b; or R14 and R15 together with the nitrogen to which they are attached form a 4-, 5-, 6- or 7-membered heterocyclic ring which may be saturated or unsaturated with 1 or 2 double bonds, and optionally may be oxo substituted; or a tautomer, isomer, stereoisomer, or pharmaceutically acceptable salt or solvate thereof.

2. A compound according to claim 1, wherein B is (i) or (ii): i) a radical of formula IIa: ##STR00037## wherein R1 is H or alkyl, R2 is H, R3 is H or alkyl, and P is —CH.sub.2NH.sub.2; or ii) a fused 6,5- or 6,6-heteroaromatic bicyclic ring, containing N and, optionally, one or two additional heteroatoms which are independently N, O or S, which is optionally mono or di-substituted with a substituent that is alkyl, alkoxy, OH, halo, CN, CF.sub.3 or NR8R9.

3. A compound according to claim 1, wherein B is a radical of formula IIa: ##STR00038## wherein R1 is H and alkyl, R2 is H, R3 is H or alkyl, and P is —CH.sub.2NH.sub.2.

4. A compound according claim 1, wherein B is (i) optionally substituted isoquinolinyl, wherein said optional substituent is NH.sub.2, or (ii) 1H-pyrrolo[2,3-b]pyridine.

5. A compound according to claim 1, wherein A is heteroaryl substituted by phenyl; or —(CH.sub.2).sub.0-3phenyl substituted by heteroaryl, or —(CH.sub.2).sub.1-3-heteroaryl, or —(CH.sub.2).sub.1-3—NR14R15.

6. A compound according to claim 1, wherein A is phenyl, ##STR00039##

7. A compound according to claim 1, that is: 6-{[1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-ylamino]-methyl}-isoquinolin-1-ylamine; 6-[(1-Benzyl-1H-pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-isoquinolin-1-ylamine; 1-(4-{4-[(1H-Pyrrolo[2,3-b]pyridin-5-ylmethyl)-amino]-pyrrolo[3,2-c]pyridin-1-ylmethyl}-benzyl)-1H-pyridin-2-one; [1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-(1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-amine; (4-Aminomethyl-benzyl)-[1-(4-phenyl-butyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; (4-Aminomethyl-2-methyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; 1-{4-[4-(4-Aminomethyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one; 1-{4-[4-(4-Aminomethyl-2-methyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one; (4-Aminomethyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; (1-Amino-isoquinolin-6-ylmethyl)-{8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazin-4-yl}-amine; or a pharmaceutically acceptable salt or solvate thereof.

8. A pharmaceutical composition comprising a compound as claimed in claim 1 and a pharmaceutically acceptable carrier, diluent or excipient.

9. The compound of claim 1, wherein the stereoisomer is an enantiomer, diastereoisomer, or racemic or scalemic mixture thereof.

10. The pharmaceutical composition of claim 8, wherein the compound is: 6-{[1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-ylamino]-methyl}-isoquinolin-1-ylamine; 6-[(1-Benzyl-1H-pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-isoquinolin-1-ylamine; 1-(4-{4-[(1H-Pyrrolo[2,3-b]pyridin-5-ylmethyl)-amino]-pyrrolo[3,2-c]pyridin-1-ylmethyl}-benzyl)-1H-pyridin-2-one; [1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-(1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-amine; (4-Aminomethyl-benzyl)-[1-(4-phenyl-butyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; (4-Aminomethyl-2-methyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; 1-{4-[4-(4-Aminomethyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one; 1-{4-[4-(4-Aminomethyl-2-methyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one; (4-Aminomethyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; (1-Amino-isoquinolin-6-ylmethyl)-{8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazin-4-yl}-amine; or a pharmaceutically acceptable salt or solvate thereof.

11. A method of treatment of a disease or condition in which plasma kallikrein activity is implicated, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as claimed in claim 1, wherein the disease or condition in which kallikrein activity is implicated is impaired visual acuity, diabetic retinopathy, diabetic macular edema, hereditary angioedema, diabetes, pancreatitis, cerebral haemorrhage, nephropathy, cardiomyopathy, inflammatory bowel disease, arthritis, septic shock, hypotension, adult respiratory distress syndrome, disseminated intravascular coagulation, blood coagulation during cardiopulmonary bypass surgery, bleeding from post-operative surgery, or retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema.

12. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema.

13. The method of claim 11, wherein said compound is: 6-{[1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-ylamino]-methyl}-isoquinolin-1-ylamine; 6-[(1-Benzyl-1H-pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-isoquinolin-1-ylamine; 1-(4-{4-[(1H-Pyrrolo[2,3-b]pyridin-5-ylmethyl)-amino]-pyrrolo[3,2-c]pyridin-1-ylmethyl}-benzyl)-1H-pyridin-2-one; [1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-(1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-amine; (4-Aminomethyl-benzyl)-[1-(4-phenyl-butyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; (4-Aminomethyl-2-methyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; 1-{4-[4-(4-Aminomethyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one; 1-{4-[4-(4-Aminomethyl-2-methyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one; (4-Aminomethyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine; (1-Amino-isoquinolin-6-ylmethyl)-{8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazin-4-yl}-amine; or a pharmaceutically acceptable salt or solvate thereof.

14. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is cerebral haemorrhage in hyperglycemic patients.

15. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is impaired visual acuity.

16. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is hereditary angioedema.

17. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is diabetes.

18. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is cerebral haemorrhage.

19. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is nephropathy in diabetic patients.

20. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is cardiomyopathy.

21. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is inflammatory bowel disease or arthritis.

22. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is septic shock or hypotension.

23. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is adult respiratory distress syndrome.

24. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is disseminated intravascular coagulation.

25. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is blood coagulation during cardiopulmonary bypass surgery.

26. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is bleeding from post-operative surgery.

27. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is diabetic retinopathy.

28. The method of claim 11, wherein the disease or condition in which plasma kallikrein activity is implicated is diabetic macular edema.

29. A method of treating a disease or condition in which plasma kallikrein activity is implicated comprising administration to a subject in need thereof a therapeutically effective amount of a compound of claim 1, wherein the disease or condition in which plasma kallikrein activity is implicated is impaired visual acuity, diabetic retinopathy, diabetic macular edema, or hereditary angioedema.

Description

EXAMPLES

(1) The invention is illustrated by the following non-limiting examples in which the following abbreviations and definitions are used:

(2) TABLE-US-00001 DCM Dichloromethane DMA N,N-Dimethylacetamide DMF N,N-Dimethylformamide EtOAc Ethyl Acetate HATU 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)- 1,1,3,3-tetramethylisouronium hexafluorophosphate(V) hrs Hours HOBt Hydroxybenzotriazole LCMS Liquid chromatography mass spectrometry Me Methyl MeCN Acetonitrile MeOH Methanol Min Minutes MS Mass spectrum NMR Nuclear magnetic resonance spectrum - NMR spectra were recorded at a frequency of 400 MHz unless otherwise indicated Pet. Ether Petroleum ether fraction boiling at 60-80° C. Ph Phenyl rt room temperature THF Tetrahydrofuran TFA Trifluoroacetic acid

(3) All reactions were carried out under an atmosphere of nitrogen unless specified otherwise.

(4) .sup.1H NMR spectra were recorded on a Bruker (400 MHz) spectrometer with reference to deuterium solvent and at rt.

(5) Molecular ions were obtained using LCMS which was carried out using a Chromolith Speedrod RP-18e column, 50×4.6 mm, with a linear gradient 10% to 90% 0.1% HCO.sub.2H/MeCN into 0.1% HCO.sub.2H/H.sub.2O over 13 min, flow rate 1.5 mL/min, or using Agilent, X-Select, acidic, 5-95% MeCN/water over 4 min. Data was collected using a Thermofinnigan Surveyor MSQ mass spectrometer with electospray ionisation in conjunction with a Thermofinnigan Surveyor LC system.

(6) Chemical names were generated using the Autonom software provided as part of the ISIS draw package from MDL Information Systems, in the IUPAC form using Chemaxon software.

(7) Where products were purified by flash chromatography, ‘silica’ refers to silica gel for chromatography, 0.035 to 0.070 mm (220 to 440 mesh) (e.g. Merck silica gel 60), and an applied pressure of nitrogen up to 10 p.s.i accelerated column elution. Reverse phase preparative HPLC purifications were carried out using a Waters 2525 binary gradient pumping system at flow rates of typically 20 mL/min using a Waters 2996 photodiode array detector.

(8) All solvents and commercial reagents were used as received.

(9) General methods for the preparation of the compounds in Tables below are described here:—

(10) General Method for Alkylation of the Bicyclic Nitrogen

(11) To sodium hydride (2 eq) in DMF at 0° C. was added the free based bicyclic amine (1 eq) and the reaction stirred for 20 mins then benzyl bromide (1.1 eq) added and reaction stirred at room temperature for between 2-16 h. The cooled reaction mixture was quenched with water and extracted with EtOAc (2×) the combined organics were washed with water and brine, dried (MgSO4) and concentrated and purified as necessary.

(12) General Procedures for Chloro Displacement with Primary Amines

(13) A: The aryl chloride (1 eq) and the amine (1-5 eq) in ethanol were heated at 130° C. for between 8-120 h. The reaction mixture was concentrated in vacuo and purified as necessary.

(14) B: The aryl chloride (1 eq) and the amine (1-5 eq) in n-butanol were heated at 130° C. for between 8-120 h. The reaction mixture was concentrated in vacuo and purified as necessary.

(15) C: To the aryl chloride (1 eq) in a microwave tube in dry toluene was added the amine (1-1.4 eq), BINAP (0.8 eq) and sodium tert-butoxide (1.4 eq). A flow of N.sub.2 was passed through reaction mixture for 5 mins. Finally Pd.sub.2dba.sub.3 (0.3 eq) added and reaction stirred for 1 min before placing immediately in the microwave at 170° C. for between 30-90 mins. The reaction mixture was concentrated and purified either by flash chromatography or by reverse phase prep HPLC.
General Method for Nitrile Reduction

(16) To the cooled nitrile (1 eq) in methanol was added nickel (II) chloride hexahydrate (0.1 eq) and di-tert-butyl dicarbonate (2 eq). The sodium borohydride (7 eq) was added portionwise to control the gas evolution. The reaction mixture was stirred at 0° C. to room temperature for 18 hours after which time the MeOH was removed by evaporation. The residue was dissolved in CHCl.sub.3, washed with saturated NaHCO.sub.3, water, brine, dried (Na.sub.2SO.sub.4) and filtered. The filtrate was evaporated and purified as necessary.

(17) General Method for Boc Deprotection

(18) To the Boc protected benzylamine was added 4M HCl in dioxane and the reaction stirred at room temperature for between 1-16 h. The solvent was removed in vacuo to afford the target as the HCl salt

Example 7

1-{4-[4-(4-Aminomethyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one

(19) ##STR00014##

A. 1-(4-Hydroxymethyl-benzyl)-1H-pyridin-2-one

(20) 4-(Chloromethyl)benzylalcohol (5 g, 31.93 mmol) was dissolved in acetone (150 ml), and 2-hydroxypyridine (3.6 g, 38.31 mmol) and potassium carbonate (13.2 g, 95.78 mmol) were added. The reaction mixture was stirred at 50° C. for 3 hours. The solvent was removed in vacuo and the residue taken up in chloroform (100 mL). The organic layer was washed with water (30 mL), brine (30 mL), dried (Na.sub.2SO.sub.4), filtered and evaporated. The residue was purified by flash chromatography eluting with 5% MeOH-DCM to give a white solid identified as the title compound (5.4 g, 25.09 mmol, 79% yield).

(21) [M+Na]+=237.8

B. 1-(4-Bromomethyl-benzyl)-1H-pyridin-2-one

(22) 1-(4-Hydroxymethyl-benzyl)-1H-pyridin-2-one (1 g, 4.65 mmol) was dissolved in DCM (75 ml) and phosphorous tribromide (2.5 g, 9.29 mmol) was added. The reaction was stirred at room temperature for 3 hrs. On completion, the reaction mixture was diluted with CHCl.sub.3 (75 mL) and washed with saturated NaHCO.sub.3 (30 mL), water (30 mL) and brine (30 mL). The organic layer was dried (Na.sub.2SO.sub.4), filtered and evaporated to give a white solid identified as the title compound which was used without further purification (1.05 g, 3.78 mmol, 81% yield).

(23) [M+H].sup.+=277.61 and 279.59

C. 4-[(1H-Pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-benzonitrile

(24) 4-(Aminomethyl)benzonitrile. HCl was partitioned between chloroform (50 mL) and saturated NaHCO.sub.3 (10 mL), dried over Na.sub.2SO.sub.4, filtered and evaporated to afford 4-(aminomethyl)benzonitrile free base as a yellow oil. To 4-(aminomethyl)benzonitrile (250 mg, 1.89 mmol) was added 4-chloro-5-azaindole (289 mg, 1.89 mmol) in ethanol (1 mL) and the mixture was heated at 130° C. for 35 hours, adding minimum ethanol when evaporated. The crude residue was purified by flash chromatography eluting with 4% to 12% MeOH-DCM to give a pale yellow gum identified as the title compound (300 mg, 1.21 mmol, 64% yield).

(25) [M+H]+=248.7

D. {4-[(1H-Pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-benzyl}-carbamic acid tert-butyl ester

(26) 4-[(1H-Pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-benzonitrile (300 mg, 1.21 mmol) was dissolved in MeOH (30 ml) and cooled to 0° C. Nickel (II) chloride hexahydrate (29 mg, 0.12 mmol) and di-tertbutyl dicarbonate (527 mg, 2.42 mmol) were added followed by sodium borohydride portionwise (320 mg, 8.46 mmol) over 10 min. The reaction mixture was stirred at 0° C. to rt for 4 hours after which time the MeOH was removed by evaporation. The residue was dissolved in CHCl.sub.3 (100 ml), washed with saturated NaHCO.sub.3 (30 ml), water (30 ml), brine (30 ml), dried (Na.sub.2SO.sub.4), filtered and evaporated. The crude residue was purified by flash chromatography eluting with 12% MeOH-DCM to give an off white solid identified as the title compound (180 mg, 0.51 mmol, 42% yield).

(27) [M+H]+=352.8

E. [4-({1-[4-(2-Oxo-2H-pyridin-1-ylmethyl)-benzyl]-1H-pyrrolo[3,2-c]pyridin-4-ylamino}-methyl)-benzyl]-carbamic acid tert-butyl ester

(28) {4-[(1H-Pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]-benzyl}-carbamic acid tert-butyl ester (70 mg, 0.20 mmol) was dissolved in dry DMF (7 mL), placed under nitrogen and cooled to 0° C. NaH (60% in mineral oil, 16 mg, 0.40 mmol) was added, reaction allowed to warm to rt and stirred for 15 min at rt. During this time the solution turned from a pale yellow colour to a deep red/orange. The reaction mixture was then cooled to 0° C. and 1-(4-bromomethyl-benzyl)-1H-pyridin-2-one (66 mg, 0.24 mmol) in DMF (3 mL) added dropwise, then allowed to warm to rt and stirred for 3 hours at rt. The reaction mixture was quenched with water and diluted with ethyl acetate (50 mL). The organic layer was washed with saturated NaHCO.sub.3 (20 mL), water (3×20 mL), brine (20 mL), dried (Na.sub.2SO.sub.4), and evaporated under vacuum. The crude material was purified by flash chromatography eluting with 8% MeOH-DCM to give a pale yellow gum identified as the title compound (40 mg, 0.073 mmol, 37% yield).

(29) [M+H]+=550.0

F. 1-{4-[4-(4-Aminomethyl-benzylamino)-pyrrolo[3,2-c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one

(30) [4-({1-[4-(2-Oxo-2H-pyridin-1-ylmethyl)-benzyl]-1H-pyrrolo[3,2-c]pyridin-4-ylamino}-methyl)-benzyl]-carbamic acid tert-butyl ester (40 mg, 0.07 mmol) was dissolved in MeOH (1 ml) and treated with 4N HCl in dioxan (4 ml). After three hours at rt the solvent was removed in vacuo and the residue azeotroped with toluene (10 ml). The crude reaction mixture was purified by preparative HPLC to afford an off white solid identified as the title compound as a bis trifluoroacetic acid salt (24 mg, 0.035 mmol, 49% yield).

(31) [M+H]+=449.8

(32) NMR (d6-DMSO) δ 4.03 (2H, d, J=5.8 Hz), 4.76 (2H, d, J=6.2 Hz), 5.05 (2H, s), 5.48 (2H, s), 6.21-6.24 (1H, m), 6.38 (1H, d, J=8.9 Hz), 7.12 (1H, d, J=3.2 Hz), 7.21-7.28 (5H, m), 7.39-7.46 (5H, m), 7.53 (1H, t, J=6.2 Hz), 7.64 (1H, d, J=3.3 Hz), 7.77 (1H, dd, J=6.6, 1.7 Hz), 8.17 (3H, s), 9.39 (1H, d, J=5.8 Hz), 12.63 (1H, d, J=5.0 Hz) ppm.

Example 10

(1-Amino-isoquinolin-6-ylmethyl)-{8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazin-4-yl}-amine

(33) ##STR00015##

A. 2-((E)-2-Dimethylamino-vinyl)-terephthalonitrile ester

(34) Methylterephthalonitrile (1.42 g, 9.99 mmol) and Bredereck's reagent (3.48 g, 19.98 mmol) were dissolved in DMF (15 mL). The reaction mixture was heated at 75° C. under nitrogen for 72 hrs after which time the solvent was removed in vacuo. Trituration with Pet Ether gave a bright yellow solid identified as 2-((E)-2-dimethylamino-vinyl)-terephthalonitrile ester (1.88 g, 0.95 mmol, 95%).

(35) .sup.1H NMR (CD.sub.3OD) δ: 3.20 (6H, s), 5.34 (1H, d, J=13.4 Hz), 7.21 (1H, dd, J=8.0 Hz, 1.4 Hz), 7.9 (1H, d, 13.4 Hz), 7.61 (1H, d, J=8.0 Hz), 7.94 (1H, d, J=1.2 Hz)

B. 1-Amino-2-(2,4-dimethoxy-benzyl)-1,2-dihydro-isoquinoline-6-carbonitrile

(36) 2-((E)-2-Dimethylamino-vinyl)-terephthalonitrile ester (1.85 g, 9.38 mmol) was dissolved in 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (5 mL) and 2,4-dimethoxybenzylamine (2.35 g, 14.07 mmol) was added. The reaction mixture was heated at 75° C. under nitrogen. After 3 hrs the reaction mixture was cooled and diethyl ether/Pet Ether (15:85) was added. The yellow solid was filtered off, dried in vacuo, and identified as 1-amino-2-(2,4-dimethoxy-benzyl)-1,2-dihydro-isoquinoline-6-carbonitrile (2.65 g, 8.38 mmol, 89%).

(37) [M+H]+=320.0

(38) .sup.1H NMR (CD.sub.3OD) δ: 3.85 (3H, s), 3.92 (3H, s), 5.02 (2H, s), 6.39 (1H, d, J=7.4 Hz), 6.57 (1H, dd, J=8.4 Hz, 2.4 Hz), 6.66 (1H, d, 2.4 Hz), 7.18 (1H, d, 8.4 Hz), 7.24 (1H, d, 7.4 Hz), 7.72 (1H, dd, J=8.5 Hz, 1.4 Hz), 7.93 (1H, s), 8.45 (1H, d, J=8.5 Hz)

C. 1-Amino-isoquinoline-6-carbonitrile

(39) 1-Amino-2-(2,4-dimethoxy-benzyl)-1,2-dihydro-isoquinoline-6-carbonitrile (1.6 g, 5.0 mmol) was dissolved in anisole (17 mL) and trifluoroacetic acid (20 mL). The reaction mixture was heated at 105° C. under nitrogen for 12 hrs after which time the reaction mixture was cooled, diethyl ether/Pet Ether (3:7) was added, the resultant solid was filtered off, dried in vacuo and identified as 1-amino-isoquinoline-6-carbonitrile (770 mg, 4.54 mmol, 91%).

(40) [M+H]+=170.0

(41) .sup.1H NMR (CD.sub.3OD) δ: 7.23-7.25 (1H, d, J=6.9 Hz), 7.65 (1H, d, J=6.8 Hz), 8.11 (1H, dd, J=8.7 Hz, 1.6 Hz), 8.33 (1H, s), 8.45 (1H, d, J=8.7 Hz).

D. (1-Amino-isoquinolin-6-ylmethyl)-carbamic acid tert-butyl ester

(42) 1-Amino-isoquinoline-6-carbonitrile (200 mg, 1.18 mmol) was dissolved in methanol (20 mL). This solution was cooled to 0° C. Nickel (II) chloride hexahydrate (28 mg, 0.12 mmol) and di-tertbutyl dicarbonate (516 g, 2.36 mmol) were added followed by sodium borohydride (313 g, 8.22 mmol) portionwise. The reaction mixture was stirred at 0° C. to room temp for 3 days. The MeOH was removed by evaporation. The residue was dissolved in CHCl.sub.3 (70 ml), washed with sat NaHCO.sub.3 (1×30 mL), water (1×30 mL), brine (1×30 mL), dried (Na.sub.2SO.sub.4) and evaporated in vacuo to give a yellow oil identified as (1-amino-isoquinolin-6-ylmethyl)-carbamic acid tert-butyl ester (110 mg, 0.4 mmol, 34%).

(43) [M+H].sup.+=274.1.

E. 6-Aminomethyl-isoquinolin-1-ylamine Hydrochloride

(44) (1-Amino-isoquinolin-6-ylmethyl)-carbamic acid tert-butyl ester (110 mg, 0.40 mmol) was dissolved in 4M HCl in dioxan (40 mL). After 18 hrs at room temperature the solvent was removed in vacuo to give a pale brown solid identified as 6-aminomethyl-isoquinolin-1-ylamine hydrochloride (67 mg, 0.39 mmol, 96%).

(45) [M+H]+=174.3

F. (4-((4-Methyl-1H-pyrazol-1-yl)methyl)phenyl)methanol

(46) To a round bottom flask under N.sub.2 was added: (4-(chloromethyl)phenyl)methanol (10.04 g, 60.9 mmol), 4-methyl-1H-pyrazole (5.05 ml, 60.9 mmol) and dry MeCN (100 mL). Next, potassium carbonate (9.26 g, 67.0 mmol) was added and the white suspension was heated to 60° C. for 18 h. The volatiles were removed in vacuo. The residue was partitioned between EtOAc (100 mL) and water (150 mL). Aqueous layer was neutralised to pH 7 with 1 N HCl and extracted with EtOAc (2×100 mL). The combined organic layers were washed with water (100 mL), brine (50 mL) then dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude product was purified by chromatography (10-80% EtOAc in iso-hexanes) to afford (4-((4-methyl-1H-pyrazol-1-yl)methyl)phenyl)methanol (2.9 g, 14.05 mmol, 23.07% yield) as a free-flowing oil that solidified on standing.

(47) [M+H].sup.+=203.2

G. 1-(4-(Bromomethyl)benzyl)-4-methyl-1H-pyrazole

(48) To a flask under N.sub.2 was added: (4-((4-methyl-1H-pyrazol-1-yl)methyl)phenyl)methanol (250 mg, 1.236 mmol), triphenylphosphine (373 mg, 1.421 mmol) and dry DCM (5.0 mL). Cooled in an ice bath before perbromomethane (451 mg, 1.360 mmol) was added. Stirred at rt for 1 h. Concentrated in vacuo and purified by column chromatography (0-20% EtOAc in iso-hexanes) to afford 1-(4-(bromomethyl)benzyl)-4-methyl-1H-pyrazole (0.33 g, 1.182 mmol, 96% yield) as an oil that solidified on standing to a white solid.

(49) [M+H].sup.+=265.1/267.1

H. 2-[(6-Chloro-5-methoxy-4-pyrimidinyl)amino]-ethanol

(50) To a solution of 4,6-dichloro-5-methoxypyrimidine (1.00 g, 5.59 mmol) in dioxane (15 mL) was added 2-aminoethanol (348 mg, 5.70 mmol) and potassium carbonate (926 mg, 6.70 mmol). The reaction was refluxed at 125° C. On completion, the reaction mixture was cooled to room temperature, the resulting suspension was filtered and the filtrate concentrated in vacuo. Both the filtered solid and the solid obtained from concentration of the filtrate were identified as the title compound and combined to afford 1.2 g (5.89 mmol, quantitative yield) of the title compound.

(51) [M+H].sup.+=203.9

I. 4-Chloro-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine

(52) 2-[(6-Chloro-5-methoxy-4-pyrimidinyl)amino]-ethanol (1.2 g, 5.89 mmol) was dissolved in a solution of boron tribromide (1.0 M in DCM, 40 mL) and the resulting reaction was heated to reflux and stirred for 3 hrs. The reaction mixture was cooled to rt, treated with ice-water (30 ml) and extracted with EtOAc (3×50 ml). The organic layer was dried (MgSO.sub.4), filtered and concentrated in vacuo to afford the title compound as the HBr salt (0.96 g, 3.81 mmol, 65% yield).

(53) [M+H].sup.+=172.0

J. 4-Chloro-8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine

(54) 4-Chloro-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (109 mg, 0.64 mmol) was dissolved in DMF (2 ml) to which was added diisopropylethylamine (332 μl, 1.91 mmol) followed by 1-(4-(bromomethyl)benzyl)-4-methyl-1H-pyrazole (168 mg, 0.64 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with CHCl.sub.3 (40 ml) and washed sequentially with water (5×40 ml) and brine (40 ml). The organic layer was dried (MgSO.sub.4), filtered and concentrated to afford a yellow solid. The crude material was purified by flash chromatography (8% MeOH/DCM-10% MeOH/DCM/1% NH.sub.4OH). Fractions containing the title compound were concentrated to afford the title compound as a pale yellow oil (119 mg, 0.33 mmol, 52.6% yield).

(55) LCMS: 355.9 @ 6.29 mins

(56) .sup.1H NMR: (CDCl.sub.3) 2.10 (3H, s), 4.29 (2H, t, J=9.4 Hz), 4.64 (2H, br. s), 5.21 (2H, s), 5.25 (2H, s), 7.19 (2H, d, J=8.1 Hz), 7.26 (1H, br. s), 7.34-7.37 (2H, m), 7.69 (2H, d, J=7.9 Hz).

K. (1-Amino-isoquinolin-6-ylmethyl)-{8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazin-4-yl}-amine

(57) 4-Chloro-8-[4-(4-methyl-pyrazol-1-ylmethyl)-benzyl]-7,8-dihydro-6H-pyrimido[5,4-b][1,4]oxazine (100 mg, 0.28 mmol) and 6-(aminomethyl)isoquinolin-1-ylamine (48.7 mg, 0.28 mmol) were suspended in ethanol (0.5 ml) and heated under microwave irradiation (CEM focussed microwave, Power 300W, 120° C. for 90 mins). The reaction was filtered and the solid washed with ethanol. The filtrate was concentrated under reduced pressure. The crude material was purified by preparative HPLC 0. Fractions containing product were combined and freeze dried to afford an off-white solid identified as the title compound (19.5 mg, 0.027 mmol, 10% yield).

(58) [M+H].sup.+=493.0

(59) HPLC: 99% purity

(60) 1H NMR d6-DMSO: 1.98 (3H, s), 3.84 (2H, t, J=8.9 Hz), 4.45 (2H, t, J=8.9 Hz), 4.76 (2H, d, J=5.9 Hz), 4.92 (2H, s), 5.24 (2H, s), 7.21-7.17 (3H, m), 7.25 (1H, s), 7.50 (1H, s), 7.52 (2H, d, J=2.3 Hz), 7.56 (1H, dd, J=8.7, 1.6 Hz), 7.66 (1H, d, J=6.8 Hz), 7.69 (1H, s), 8.36 (1H, s), 8.52-8.46 (2H, m), 8.98 (2H, s).

(61) The compounds in the following tables were synthesised as described in the general methods above and as described in Examples 7 and 10 above.

(62) TABLE-US-00002 TABLE 1 Example Free base no. A B MW [M + H].sup.+ 1 462.57 462.9 2 0 379.46 380.0 3 460.53 461.0 4 436.53 436.8 5 384.52 385.1 6 439.58 440.0 7 0 449.55 449.8 8 463.57 464.0 9 425.55 423.0

(63) TABLE-US-00003 TABLE 3 Example No Name 1 6-{[1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]- pyridin-4-ylamino]-methyl}-isoquinolin-1-ylamine 2 6-[(1-Benzyl-1H-pyrrolo[3,2-c]pyridin-4-ylamino)-methyl]- isoquinolin-1-ylamine 3 1-(4-{4-[(1H-Pyrrolo[2,3-b]pyridin-5-ylmethyl)-amino]- pyrrolo[3,2-c]pyridin-1-ylmethyl}-benzyl)-1H-pyridin-2-one 4 [1-(2-Phenyl-thiazol-4-ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4- yl]-(1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-amine 5 (4-Aminomethyl-benzyl)-[1-(4-phenyl-butyl)-1H-pyrrolo[3,2- c]-pyridin-4-yl]-amine 6 (4-Aminomethyl-2-methyl-benzyl)-[1-(2-phenyl-thiazol-4- ylmethyl)-1H-pyrrolo[3,2-c]pyridin-4-yl]-amine 8 1-{4-[4-(4-Aminomethyl-2-methyl-benzylamino)-pyrrolo[3,2- c]pyridin-1-ylmethyl]-benzyl}-1H-pyridin-2-one 9 (4-Aminomethyl-benzyl)-[1-(2-phenyl-thiazol-4-ylmethyl)- 1H-pyrrolo[3,2-c]pyridin-4-yl]-amine

(64) TABLE-US-00004 TABLE 4 NMR data of examples (d6-DMSO) Example No Chemical Shift (ppm) 1 4.75 (2H, d, J = 6.3 Hz), 5.48 (2H, s), 6.37 (1H, d, J = 6.3 Hz), 6.77 (1H, s), 6.91 (1H, s), 7.33 (1H, s), 7.41 (2H, d, J = 6.3 Hz), 7.47-7.52 (4H, m), 7.64 (1H, d, J = 4.4 Hz), 7.89-7.92 (3H, m), 8.26 (1H, d, J = 6.3 Hz), s), 11.52 (1H, s) 2 4.98 (2H, d, J = 6.3 Hz), 5.53 (2H, s), 6.37 (1H, d, J = 6.3 Hz), 6.77 (1H, s), 6.91 (1H, s), 7.33 (1H, s), 7.41 (2H, d, J = 4.4 Hz), 7.47-7.52 (4H, m), 7.64 (1H, d, J = 4.4 Hz), 7.89-7.92 (3H, m), 8.26 (1H, d, J = 6.3 Hz), s), 11.52 (1H, s) 3 4.74 (2H, d, J = 6.3 Hz), 5.04 (2H, s), 5.30 (2H, s), 6.19-6.22 (1H, m), 6.38 (2H, d, J = 6.3 Hz), 6.74 (2H, d, J = 6.77 Hz), 7.13 (2H, d, J = 6.8 Hz), 7.21 (2H, d, J = 6.8 Hz), 7.25-7.26 (1H, m), 7.33-7.42 (2H, m), 7.59 (1H, d, J = 4.4 Hz), 7.72-7.74 (1H, m), 7.91 (1H, d, J = 6.3 Hz), 8.15 (1H, s), 8.25 (1H, d, J = 6.3 Hz), 11.52 (1H, s) 4 4.75 (2H, d, J = 6.3 Hz), 5.48 (2H, s), 6.37 (1H, d, J = 6.3 Hz), 6.77 (1H, s), 6.91 (1H, s), 7.33 (1H, s), 7.41 (2H, d, J = 6.3 Hz), 7.47-7.52 (4H, m), 7.64 (1H, d, J = 4.4 Hz), 7.89-7.92 (3H, m), 8.26 (1H, d, J = 6.3 Hz), s), 11.52 (1H, s) 5 1.50 (2H, quintet, J = 7.2 Hz), 1.72 (2H, quintet, J = 7.2 Hz), 1.85 (2H, br.s), 2.56 (2H, t, J = 8.0 Hz), 3.65 (2H, s), 4.08 (2H, t, J = 6.9 Hz), 4.63 (2H, d, J = 6.0 Hz), 6.65-6.68 (2H, m), 7.08 (1H, t, J = 6.2 Hz), 7.12-7.17 (4H, m), 7.21-7.27 (6H, m), 7.57 (1H, d, J = 5.9 Hz) 6 4.50 (2H, d, J = 5.6 Hz), 5.05 (2H, s), 5.11 (2H, s), 6.21 (1H, dt, J = 1.4, 6.7 Hz), 6.37-6.41 (2H, m), 6.43 (1H, d, 9.5 Hz), 7.23- 7.27 (4H, m), 7.40 (1H, dq, J = 2.1, 9.2 Hz), 7.44 (1H, t, J = 2.8 Hz), 7.74 (1H, dd, J = 1.6, 6.8 Hz), 7.87 (1H, d, J = 1.6 Hz), 7.90 (1H, dd, J = 2.6, 9.5 Hz), 8.19 (1H, d, J = 2.0 Hz), 8.44 (1H, d, J = 2.5 Hz), 8.76 (1H, t, J = 5.6 Hz), 11.58 (1H, s). 8 2.51 (3H, s), 3.98 (2H, s), 4.74 (2H, s), 5.06 (2H, s), 5.49 (2H, s), 6.23 (1H, s), 6.39 (1H, s), 7.25-7.78 (13H, m), 8.27 (2H, s), 9.35 (1H, s), 12.74 (1H, s). 9 4.01 (2H, q, J = 5.6 Hz), 4.77 (2H, d, J = 6.3 Hz), 5.65 (2H, s), 7.132 (1H, s), 7.43-7.51 (8H, m), 7.60 (1H, d, J = 4.4 Hz), 7.60- 7.68 (2H, m), 7.87-7.90 (2H, m), 8.12 (2H, br.s + 1HCl salt), 9.38 (1H, s), 12.59 (1H, br.s)
Biological Methods

(65) The ability of the compounds of formula (I) to inhibit plasma kallikrein may be determined using the following biological assays:

(66) Determination of the IC.sub.50 for Plasma Kallikrein

(67) Plasma kallikrein inhibitory activity in vitro was determined using standard published methods (see e.g. Johansen et al., Int. J. Tiss. Reac. 1986, 8, 185; Shori et al., Biochem. Pharmacol., 1992, 43, 1209; Stürzebecher et al., Biol. Chem. Hoppe-Seyler, 1992, 373, 1025). Human plasma kallikrein (Protogen) was incubated at 37° C. with the fluorogenic substrate H-DPro-Phe-Arg-AFC and various concentrations of the test compound. Residual enzyme activity (initial rate of reaction) was determined by measuring the change in optical absorbance at 410 nm and the IC.sub.50 value for the test compound was determined.

(68) Data acquired from these assays are shown in Table 6 below:

(69) TABLE-US-00005 TABLE 5 Example No IC.sub.50 (human PKal) nM 1 6540 2 28900 3 5460 4 5320 5 29000 6 8590 7 5370 8 6300 9 8100 10 20500

(70) Selected compounds were further screened for inhibitory activity against the related enzyme KLK1. The ability of the compounds of formula (I) to inhibit KLK1 may be determined using the following biological assay:

(71) Determination of the IC.sub.50 for KLK1

(72) KLK1 inhibitory activity in vitro was determined using standard published methods (see e.g. Johansen et al., Int. J. Tiss. Reac. 1986, 8, 185; Shori et al., Biochem. Pharmacol., 1992, 43, 1209; Stürzebecher et al., Biol. Chem. Hoppe-Seyler, 1992, 373, 1025). Human KLK1 (Callbiochem) was incubated at 37° C. with the fluorogenic substrate H-DVal-Leu-Arg-AFC and various concentrations of the test compound. Residual enzyme activity (initial rate of reaction) was determined by measuring the change in optical absorbance at 410 nm and the IC.sub.50 value for the test compound was determined.

(73) Data acquired from this assay are shown in Table 7 below:

(74) TABLE-US-00006 TABLE 7 (KLK1 Activity) Example No IC.sub.50 (human KLK1) nM 1 5340 2 3740 3 5690 4 10300 5 38600 6 8140 7 29900 8 5300 9 8260 10 5040