ANTIBACTERIAL COMPOUNDS

Abstract

The present invention relates to novel antibacterial compounds, pharmaceutical compositions containing them and their use as antimicrobials.

Claims

1: A compound of formula (I)
A-L.sub.1-B-L.sub.2-Y-L.sub.3-C  (I) wherein A is a 10-membered fused bicyclic ring; L.sub.1 is σ bond, —O— or —N(R′)—, wherein R′ is H or (C.sub.1-3)alkyl; B is a divalent residue of a piperazine or piperidine ring; L.sub.2 is σ bond, —CH.sub.2—, —O—, or —N(R′)—, wherein R′ is H or (C.sub.1-3)alkyl; Y is a (C.sub.1-6)alkylenyl, (C.sub.2-6)alkenylenyl, (C.sub.2-6)alkynylenyl, or (C.sub.3-6)cycloalkylenyl group, said group being optionally substituted with one or more groups selected from —OH, (C.sub.1-3)alkyl, (C.sub.3-6)cycloalkyl, 3- to 5-membered oxacycloalkyl, —COOR′, —NR′R″ wherein R′ and R″, identical or different each other, are hydrogen atom or (C.sub.1-3)alkyl, L.sub.3 is —O—, —(CH.sub.2).sub.2—, —N(R′)—, —N(R′)—C(═O)—, —N(R′)—C(═S)—, —N(R′)—(C.sub.1-6)alkylenyl-, —C(═O)—N(R′)—, —C(═O)—N(R′)—(C.sub.1-6)alkylenyl-, —SO.sub.2—N(R′)—, —N(R′)—SO.sub.2—, wherein R′ is hydrogen atom or (C.sub.1-3)alkyl; C is a group having a 5- or 6-membered saturated or unsaturated ring, or a 9- or 10-membered fused bicyclic ring; or an addition salt with a pharmaceutically acceptable organic or inorganic acid or base, an enantiomer, N-oxide, or quaternary ammonium salt of said compound of formula (I).

2: The compound according to claim 1, wherein A is a fused bicyclic ring having the following formula (II): ##STR00250## wherein G1 and G2, identical or different each other, are CH or N; R.sub.1 is H, halogen atom, CN or CF.sub.3; R.sub.2 is H, halogen atom, OH, CN, CF.sub.3, (C.sub.1-6)alkyl or (C.sub.1-6)alkoxy; D is a divalent group selected from ##STR00251## wherein R.sub.3 is H, (C.sub.1-6)alkyl or (C.sub.1-6)alkoxy.

3: The compound according to claim 1, wherein B is a group having the following formula (III) ##STR00252## wherein G3 and G4, identical or different each other, are C(R.sub.4) or N, provided that that at least one of G3 and G4 is N, R.sub.4 is hydrogen atom, —OH, —CN, —COOH, —NR′R″ wherein R′ and R″, identical or different each other, are hydrogen atom or (C.sub.1-3)alkyl; G5 is C(═O) or C(H)(R.sub.5); wherein R.sub.5 is hydrogen atom, CF.sub.3, —(C.sub.1-3)alkyl-CF.sub.3, —COOR′ and —CONR′R″, wherein R′ and R″, identical or different each other, are hydrogen atom or (C.sub.1-3)alkyl.

4: The compound according to claim 1, wherein C has one of the following formulae (IV), (V), (VI), (VII) or (VIII): ##STR00253## wherein n is an integer from 0 to 3 P.sub.1 and P.sub.2, equal or different each other, are CH or N P.sub.3 is O, S or NH; R.sub.6 and R.sub.7 together form a 5- or 6-membered aliphatic or aromatic ring, optionally comprising at least one heteroatom selected from N and O, R.sub.8 and R.sub.9, equal or different each other, are a hydrogen atom or an aryl group, or together form a 5- or 6-membered aliphatic or aromatic ring, optionally comprising at least one heteroatom selected from N and O, R.sub.10 and R.sub.11, equal or different each other, are a hydrogen atom, or a (C.sub.1-3)alkyl group, or together form a 5- or 6-membered aliphatic or aromatic ring, and wherein each hydrogen atom linked to a carbon or nitrogen atom forming the ring of formulae (IV), (V), (VI), (VII) or (VIII) is optionally substituted with at least one substituent selected from the group consisting of halogen atom, OH, (C.sub.1-3)alkyl or (C.sub.1-3)alkoxy.

5: The compound according to claim 4, wherein R.sub.6 and R.sub.7 together form a ring selected from the group consisting of benzene, furan, tetrahydrofuran, dioxolane, piperidine and piperazine.

6: The compound according to claim 4, wherein R.sub.8 and R.sub.9, equal or different each other, are a hydrogen atom or an aryl group, or together form a ring selected from the group consisting of benzene, pyridine, and piperidine.

7: The compound according to claim 1, wherein C is a group having one of the following formulas: ##STR00254## ##STR00255##

8: The compound according to claim 1, wherein said Y is a (C.sub.1-4)alkylenyl, (C.sub.2-4)alkynylenyl, or (C.sub.5-6)cycloalkylenyl group, said group being optionally substituted with one or more groups selected from —OH, methyl, (C.sub.3-4)cycloalkyl, 3- or 4-membered oxacycloalkyl, —COOH, —NR′R″ wherein R′ and R″, identical or different each other, are hydrogen atom or methyl.

9: The compound according to claim 8, wherein said Y is a (C.sub.1-4)alkylenyl, (C.sub.2-4)alkynylenyl, or cyclohexylenyl group, said group being optionally substituted with one or more hydroxy groups.

10: The compound according to claim 1, wherein said L.sub.3 is —O—, —(CH.sub.2).sub.2—, —N(R′)— wherein R′ is H or (C.sub.1-3)alkyl, —NH—C(═O)—, —NH—(C.sub.1-3)alkylenyl-, —C(═O)—NH— or —C(═O)—NH—(C.sub.1-3)alkylenyl-.

11: The compound according to claim 10, wherein said L.sub.3 is —O—, —(CH.sub.2).sub.2—, or —N(R′)— wherein R′ is H or methyl, —NH—C(═O)—, —NH—CH.sub.2—, —C(═O)—NH—, or —C(═O)—NH—CH.sub.2—.

12: A pharmaceutical composition comprising at least one compound of formula (I) according to claim 1, a salt thereof with a pharmaceutically acceptable organic or inorganic acid or base, or an enantiomer thereof, or a quaternary ammonium salt thereof, or a N-oxide thereof, and at least one inert pharmaceutically acceptable excipient.

13: The compound of formula (I) according to claim 1, for use in medicine.

14: The compound of formula (I) according to claim 1, for use in the treatment of bacterial infections.

15: The compound of formula (I) according to claim 14, wherein said bacterial infections is selected from the group consisting of a skin infection, a mucosal infection, a gynaecological infection, a respiratory tract infection (RTI), a CNS infections, a gastro-intestinal infection, a bone infection, a cardiovascular infection, a sexually transmitted infection, or a urinary tract infection.

16: A method for treating a bacterial infection, comprising the administration of a compound of formula (I) according to claim 1 to a patient in need thereof.

Description

EXAMPLES

[0123] List of the abbreviations used in the synthetic pathways described hereinafter: [0124] Boc—tert-butyl carbamate [0125] cHex—cyclohexane [0126] CV—column volume [0127] DCM—dichloromethane [0128] DIPEA—N,N-diisopropylethylamine [0129] DMA—N,N-dimethylacetamide [0130] DMF—N,N-dimethylformamide [0131] DMSO—dimethyl sulfoxide [0132] Et.sub.2O—diethyl ether [0133] EtOAc—ethylacetate [0134] EtOH—ethanol [0135] HATU—1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate [0136] HBTU—N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate [0137] LC—liquid chromatography [0138] MeCN—acetonitrile [0139] MeOH—methanol [0140] MS—mass spectroscopy [0141] TEA—triethylamine [0142] TFA—trifluoroacetic acid [0143] THF—tetrahydrofuran [0144] Pd/C—palladium on activated charcoal [0145] Pd(dba).sub.2—bis(dibenzylideneacetone)palladium(0) [0146] Pd(OAc).sub.2—palladium(II) acetate [0147] UPLC—ultra high performance liquid chromatography

Preparation of compound 2: N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride

[0148] Compound 2 was prepared as described hereinbelow.

##STR00196##

Step 1: N-(3,3-diethoxypropyl)pyridine-3-carboxamide (2a)

[0149] Pyridine-3-carbonyl chloride (10 g, 56.2 mmoles, 1 eq.) was dissolved in DMF (50 mL) and CH.sub.3CN (200 mL) with TEA (19.5 mL, 140.4 mmoles, 2.5 eq.). 3,3-diethoxypropan-1-amine (10.35 g, 70.2 mmoles, 1.25 eq.) in CH.sub.3CN (20 mL) was added at 0° C. The solution was stirred overnight then was concentrated, diluted with DCM and washed with sat. NaHCO.sub.3 and brine. The organic phase was then separated, dried over sodium sulphate and evaporated in vacuum to obtain N-(3,3-diethoxypropyl)pyridine-3-carboxamide 2a (6.8 g, Y=45%). LC-MS (M-H+): 253.0.

Step 2: N-(3-oxopropyl)pyridine-3-carboxamide (2b)

[0150] Intermediate 2a (500 mg, 1.98 mmoles, 1 eq.) was dissolved in DCM (5 mL). TFA (2 mL) was added, and after 3 h the solution was concentrated and washed several times with toluene, diethyl ether and ethyl acetate to afford 900 mg of N-(3-oxopropyl)pyridine-3-carboxamide 2b (Y=quant.). LC-MS (M-H.sup.+): 179.0.

Step 3: 1-(piperazin-1-yl)isoquinoline (2c)

[0151] 1-Chloroisoquinoline (3 g, 18.3 mmoles, 1 eq.) was dissolved in CH.sub.3CN (150 mL) with piperazine (23.7 g, 274.5 mmoles, 15 eq.) and potassium carbonate (3.8 g, 27.5 mmoles, 1.5 eq). The solution was heated to reflux for 48 h then was concentrated, diluted with DCM (250 mL) and washed with sat. NaHCO.sub.3 and brine. The organic phase was separated, dried over sodium sulfate and evaporated in vacuum to obtain 1-(piperazin-1-yl)isoquinoline 2c (3.9 g, Y=94%). LC-MS (M-H.sup.+): 214.1.

Step 4: N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride (2)

[0152] Intermediate 2b (150 mg, 0.51 mmoles, 1 eq.) was dissolved in DCM (5 mL). DIPEA (0.078 mL, 0.45 mmoles, 0.9 eq.), acetic acid (one drop) and intermediate 2c (87 mg, 0.41 mmoles, 0.8 eq.) were added subsequently at room temperature. After ten minutes sodium triacetoxyborohydride (141 mg, 0.66 mmoles, 1.3 eq.) was added and the solution was left stirring overnight. The mixture was diluted with DCM, washed with sat. NaHCO.sub.3 and sat. NaCl, dried over sodium sulphate, filtered and evaporated in vacuum. The crude material was purified with a NH-Column eluting with ethyl acetate to obtain the desired amine (78 mg, 0.21 mmoles). The product was dissolved in DCM (5 mL) and 1M HCl in Et.sub.2O (0.84 mL, 4 eq.) was added dropwise at 0° C. The solution was left stirring for 2 h then concentrated in vacuum and triturated with diethyl ether to obtain title compound N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride 2 (79 mg, Y=39%). LC-MS (M-H.sup.+): 376.2.

[0153] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=11.37 (br. s., 1H), 9.29-9.14 (m, 2H), 8.88 (dd, J=1.5, 5.5 Hz, 1H), 8.60 (td, J=1.8, 8.1 Hz, 1H), 8.21 (d, J=8.4 Hz, 1H), 8.06 (d, J=6.2 Hz, 1H), 8.02 (d, J=7.7 Hz, 1H), 7.93-7.78 (m, 2H), 7.72 (ddd, J=1.4, 7.0, 8.3 Hz, 1H), 7.59 (d, J=6.2 Hz, 1H), 6.71 (br. s., 2H), 4.06 (d, J=13.5 Hz, 2H), 3.88-3.56 (m, 4H), 3.44 (q, J=6.5 Hz, 4H), 3.34-3.20 (m, 2H), 2.09 (quin, J=6.7 Hz, 2H).

Preparation of compound 3: N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}benzamide hydrochloride

[0154] Compound 3 was prepared according to the procedure described for compound 2, using in step 1 benzoyl chloride. LC-MS (M-H.sup.+): 375.2.

[0155] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=11.68 (br. s., 1H), 8.79 (t, J=5.4 Hz, 1H), 8.25 (d, J=8.6 Hz, 1H), 8.06 (d, J=8.6 Hz, 1H), 8.02 (dd, J=1.0, 6.3 Hz, 1H), 7.98-7.86 (m, 3H), 7.75 (ddd, J=1.1, 7.0, 8.5 Hz, 1H), 7.63 (d, J=6.3 Hz, 1H), 7.58-7.41 (m, 3H), 4.16 (d, J=13.5 Hz, 2H), 3.90 (t, J=12.4 Hz, 2H), 3.66 (d, J=13.1 Hz, 2H), 3.56-3.34 (m, 4H), 3.25 (quin, J=5.0 Hz, 2H), 2.08 (quin, J=7.2 Hz, 2H).

Preparation of compound 4: N-{2-[4-(isoquinolin-1-yl)piperazin-1-yl]ethyl}benzamide hydrochloride

[0156] Compound 4 was prepared according to the procedure described for compound 2, using in step 1 benzoyl chloride and 2,2-diethoxyethanamine. LC-MS (M-H.sup.+): 361.2.

[0157] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=8.71 (t, J=5.5 Hz, 1H), 8.15 (d, J=5.7 Hz, 1H), 8.14 (d, J=8.6 Hz, 1H), 7.93 (d, J=7.5 Hz, 1H), 7.91-7.87 (m, 2H), 7.75 (ddd, J=1.5, 7.0, 8.3 Hz, 1H), 7.64 (ddd, J=1.7, 7.0, 8.3 Hz, 1H), 7.61-7.45 (m, 4H), 6.06 (s, 2H), 3.68 (q, J=6.2 Hz, 2H), 3.63-3.19 (m, 10H).

Preparation of Compound 14

[0158] ##STR00197##

[0159] Compound 14 was prepared according to the procedure described for the synthesis of compound 2, using 1,6-dichloroisoquinoline in step 3 (Y=78%). LC-MS (M-H.sup.+)=410.2

[0160] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=11.36 (br. s., 1H), 9.30 (t, J=5.3 Hz, 1H), 9.25 (d, J=1.7 Hz, 1H), 8.90 (dd, J=1.3, 5.3 Hz, 1H), 8.67 (d, J=7.9 Hz, 1H), 8.19 (d, J=8.9 Hz, 1H), 8.14 (d, J=3.3 Hz, 1H), 8.13 (s, 1H), 7.88 (dd, J=5.4, 8.1 Hz, 1H), 7.67 (dd, J=2.1, 9.1 Hz, 1H), 7.51 (d, J=5.9 Hz, 1H), 3.96 (d, J=13.9 Hz, 2H), 3.76-3.54 (m, 4H), 3.50-3.34 (m, 4H), 3.32-3.19 (m, J=4.6 Hz, 2H), 2.19-1.99 (m, 2H).

Preparation of compound 21: N-{3-[4-(3-methylisoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride

[0161] Compound 21 was prepared according to the procedure described for compound 2, using in step 3 1-chloro-3-methylisoquinoline. LC-MS (M-H.sup.+): 390.2.

[0162] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.37 (br. s., 1H), 9.30 (t, J=5.4 Hz, 1H), 9.25 (d, J=1.5 Hz, 1H), 8.90 (dd, J=1.4, 5.1 Hz, 1H), 8.65 (d, J=8.0 Hz, 1H), 8.12 (d, J=8.5 Hz, 1H), 7.92-7.83 (m, 2H), 7.78 (t, J=7.5 Hz, 1H), 7.60 (t, J=7.5 Hz, 1H), 7.37 (s, 1H), 7.30-5.98 (m, 3H), 5.76 (s, 1H), 3.98 (d, J=13.3 Hz, 2H), 3.78-3.54 (m, 4H), 3.51-3.34 (m, 4H), 3.27 (td, J=4.6, 9.6 Hz, 2H), 2.54 (s, 3H), 2.19-1.99 (m, 1H)

Preparation of compound 28: 4-({1-[3-(isoquinolin-1-yl)prop-2-yn-1-yl]piperidin-4-yl}amino)-2H-chromen-2-one

[0163] Compound 28 was prepared as described hereinbelow.

##STR00198##

Step 1: 2-oxo-2H-chromen-4-yl trifluoromethanesulfonate (28a)

[0164] 4-Hydroxycoumarine (1 g, 6.17 mmoles, 1 eq.) was dissolved in DCM (30 mL), triethylamine (1.72 mL, 12.34 mmoles, 2 eq.) was added and the resulting mixture was cooled to −10° C. Trifluoromethanesulfonic anhydride (1.25 mL, 7.4 mmoles, 1.2 eq.) in DCM (5 mL) was added dropwise and the solution was left stirring at −10° C. for 2 h. The resulting reddish brown solution was warmed to room temperature, diluted with cHex/Et.sub.2O 1/1 (50 mL) and filtered through a pad of silica gel using cHex/Et.sub.2O 1/1 as eluent. The solvent was removed in vacuum to obtain 2-oxo-2H-chromen-4-yl trifluoromethanesulfonate 28a (1.76 g, Y=97%). LC-MS (M-H.sup.+): 295.0.

Step 2: tert-butyl 4-[(2-oxo-2H-chromen-4-yl)amino]piperidine-1-carboxylate (28b)

[0165] Triethylamine (1 mL, 7.2 mmoles, 1.2 eq.) in acetonitrile (2 mL) was added dropwise to a stirred solution of intermediate 28a (1.76 g, 5.99 mmoles, 1 eq.) and 1-Boc-4-aminopiperidine (1.2 g, 5.99 mmoles, 1 eq.) in dry acetonitrile (20 mL). The mixture was heated at reflux for 2 h then was cooled to room temperature, diluted with DCM (100 mL) and washed with saturated NaHCO.sub.3 (50 mL) and water (50 mL). The organic phase was separated, dried over sodium sulfate and evaporated in vacuum. The crude material was purified by trituration with methanol to obtain tert-butyl 4-[(2-oxo-2H-chromen-4-yl)amino]piperidine-1-carboxylate 28b (1.51 g, Y=73%). LC-MS (M-H.sup.+): 345.2.

Step 3: 4-(piperidin-4-ylamino)-2H-chromen-2-one trifluoroacetic acid salt (28c)

[0166] Intermediate 28b (1 g, 2.90 mmoles, 1 eq.) was dissolved in DCM (10 ml) and TFA (3 mL) was added dropwise at 0° C. The solution was left stirring for 2 h then was concentrated in vacuum and washed with toluene (25 ml) and diethyl ether (25 ml) to obtain 4-(piperidin-4-ylamino)-2H-chromen-2-one trifluoroacetic acid salt 28c, which was used without any further purification (1.22 g, Y=quant.). LC-MS (M-H.sup.+): 245.1.

Step 4: 4-{[1-(prop-2-yn-1-yl)piperidin-4-yl]amino}-2H-chromen-2-one (28d)

[0167] To a solution of intermediate 28c (50 mg, 0.14 mmoles, 1 eq.) and potassium carbonate (38.7 mg, 0.28 mmoles, 2 eq.) in DMF (2 mL) 3-bromo-1-propyne (20.8 mg, 0.14 mmoles, 1 eq.) was added at room temperature. The reaction was left stirring overnight then was filtered and the filtrate concentrated. The residue was purified by Si-column eluting with eluting with EtOAc to EtOAc/Methanol 8:2 to give 4-{[1-(prop-2-yn-1-yl)piperidin-4-yl]amino}-2H-chromen-2-one 28d (25.6 mg, Y=64%). LC-MS (M-H.sup.+): 283.1.

Step 5: 4-({1-[3-(isoquinolin-1-yl)prop-2-yn-1-yl]piperidin-4-yl}amino)-2H-chromen-2-one (28)

[0168] Intermediate 28d (186 mg, 0.66 mmoles, 1 eq.), 1-chloroisoquinoline (120 mg, 0.72 mmoles, 1.1 eq.) and CuI (12 mg, 0.066 mmoles, 0.1 eq.) were dissolved in dry DMF (3.6 ml). DIPEA (0.473 ml, 2.64 mmoles, 4 eq.) was added. The mixture was degassed by alternatively applying vacuum and nitrogen, then bis(triphenylphosphine)palladium(II) dichloride (45 mg, 0.066 mmoles, 0.1 eq.) was added and the mixture was heated at 60° C. After 3 h water (10 mL) was added and the mixture was extracted with ethyl acetate (3×10 mL). The organic phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give 283 mg of crude material. After purification by Si-column eluting with EtOAc to EtOAc/MeOH 8:2 compound 4-({1-[3-(isoquinolin-1-yl)prop-2-yn-1-yl]piperidin-4-yl}amino)-2H-chromen-2-one 28 (87 mg, Y=32%) was obtained. LC-MS (M-H+): 410.2.

[0169] .sup.1H NMR (400 MHz, METHANOL-d.sub.4) δ=8.42-8.34 (m, 2H), 7.97-7.93 (m, 1H), 7.89 (d, J=7.8 Hz, 1H), 7.79-7.66 (m, 3H), 7.58-7.49 (m, 1H), 7.28-7.20 (m, 2H), 5.30 (s, 1H), 3.78 (s, 2H), 3.60-3.47 (m, 1H), 3.13 (d, J=12.0 Hz, 2H), 2.59 (dt, J=1.8, 11.8 Hz, 2H), 2.11 (d, J=11.3 Hz, 2H), 1.79 (dq, J=3.0, 12.0 Hz, 2H).

Preparation of compound 31: N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}furo[3,2-b]pyridine-6-carboxamide hydrochloride

[0170] Compound 31 was prepared as described hereinbelow.

##STR00199##

Step 1: tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}carbamate (31a)

[0171] A mixture of 1-(piperazin-1-yl)isoquinoline 2c (1.40 g, 6.54 mmoles. 1 eq.), tert-butyl N-(3-bromopropyl)carbamate (1.48 g, 6.22 mmoles, 0.95 eq.), potassium iodide (0.54 g, 3.27 mmoles, 0.50 eq.) and potassium carbonate (1.81 g, 13.1 mmoles, 2 eq.) in DMF (20 mL) was stirred at room temperature for 2 days. The reaction was partitioned between EtOAc (200 mL) and brine (250 mL). The organic phase was separated then was washed with brine (3×200 mL), dried over sodium sulfate and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 100) eluting with a gradient of 20-100% of A in cyclohexane, where A is MeOH/EtOAc (5:95), to give 2.10 g (Y=87%) of tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}carbamate 31a. LC-MS (M-H+)=371.2.

Step 2: 3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-amine TFA salt (31b)

[0172] Intermediate 31a (6.35 g, 8.74 mmoles) was dissolved in DCM (25 mL) and TFA (1 mL) was added dropwise at 0° C. The solution was stirred for 2 h then was concentrated in vacuum, washed with toluene and diethyl ether to obtain 3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-amine TFA salt 31b (6.35 g, Y=93%). LC-MS (M-H.sup.+)=271.2.

Step 3: N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}furo[3,2-b]pyridine-6-carboxamide hydrochloride (31)

[0173] Intermediate 31b (250 mg, 0.34 mmoles, 1 eq.) and commercially available furo[3,2-b]pyridine-6-carboxylic acid (56 mg, 0.34 mmoles, 1 eq.) were dissolved in dry DMF (2 mL). TEA (0.25 mL, 1.7 mmoles, 5 eq.) was added, the mixture was cooled to 0° C. and HATU (136 mg, 0.36 mmoles, 1.05 eq.) was added. After stirring at room temperature for 2 h DCM was added, the organic phase was washed with sat. NaHCO.sub.3 and brine and evaporated in vacuum. The crude material was purified by Si column eluting with EtOAc to EtOAc/MeOH 8:2 to obtain 100 mg of the desired compound. The product was dissolved in DCM (5 mL) and HCl (1M solution in diethyl ether, 0.70 mL, 4 eq.) was added dropwise at 0° C. The solution was left stirring for 30 min then concentrated in vacuum and triturated with diethyl ether to obtain N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}furo[3,2-b]pyridine-6-carboxamide hydrochloride 31 (120 mg, 0.23 mmoles, Y=68%). LC-MS (M-H.sup.+)=416.2.

[0174] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=11.42 (br. s., 1H), 9.09 (d, J=1.7 Hz, 1H), 9.05 (t, J=5.6 Hz, 1H), 8.57 (s, 1H), 8.51 (d, J=2.4 Hz, 1H), 8.23 (d, J=8.7 Hz, 1H), 8.10-8.00 (m, 2H), 7.91 (t, J=7.5 Hz, 1H), 7.74 (ddd, J=1.0, 7.0, 8.4 Hz, 1H), 7.61 (d, J=6.3 Hz, 1H), 7.25 (dd, J=1.0, 2.4 Hz, 1H), 4.11 (d, J=13.2 Hz, 2H), 3.81 (t, J=12.7 Hz, 2H), 3.67 (d, J=11.5 Hz, 2H), 3.55-3.36 (m, 4H), 3.35-3.20 (m, 2H), 2.11 (quin, J=7.0 Hz, 2H).

Preparation of compound 35: N-{3-[4-(6-methylisoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide

[0175] Compound 35 was prepared according to the procedure described for compound 2, using in step 3, 1-chloro-6-methylisoquinoline. LC-MS (M-H.sup.+): 390.2.

[0176] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.44 (br. s., 1H), 9.26 (br. s., 1H), 9.22 (s, 1H), 8.88 (d, J=4.8 Hz, 1H), 8.59 (br. s., 1H), 8.14 (d, J=8.5 Hz, 1H), 7.98 (d, J=6.3 Hz, 1H), 7.84 (s, 2H), 7.59 (d, J=8.5 Hz, 1H), 7.53 (d, J=6.3 Hz, 1H), 4.12 (d, J=14.3 Hz, 2H), 3.82 (t, J=11.7 Hz, 2H), 3.66 (d, J=12.3 Hz, 2H), 3.50-3.35 (m, 4H), 3.32-3.21 (m, 2H), 2.54 (s, 3H), 2.08 (quin, J=7.3 Hz, 2H)

Preparation of compound 36: N-{3-[4-(6-methoxyisoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride

[0177] Compound 36 was prepared according to the procedure described for compound 2, using in step 3 1-chloro-6-methoxyisoquinoline. LC-MS (M-H.sup.+): 406.2.

[0178] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.03 (br. s., 1H), 9.13 (d, J=1.3 Hz, 1H), 9.07 (t, J=5.0 Hz, 1H), 8.80 (dd, J=1.0, 5.0 Hz, 1H), 8.40 (d, J=7.5 Hz, 1H), 8.14 (d, J=9.3 Hz, 1H), 7.98 (d, J=6.3 Hz, 1H), 7.72-7.64 (m, 1H), 7.52 (d, J=6.5 Hz, 1H), 7.48 (s, 1H), 7.32 (dd, J=2.3, 9.3 Hz, 1H), 4.39-3.53 (m, 8H), 3.49-3.33 (m, 4H), 3.31-3.19 (m, 2H), 2.16-1.95 (m, 2H)

Preparation of compound 38: N-(2,5-difluorobenzyl)-3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-amine hydrochloride

[0179] Intermediate 31b (0.21 mmoles, 1 eq.) was dissolved in dry DCE (3 mL) and 2,5-difluorobenzaldehyde (32.8 mg, 0.23 mmoles, 1.1 eq), DIPEA (0.146 mL, 0.84, 4 eq.) and acetic acid (one drop) were added subsequently at room temperature. The mixture was stirred for 4 h at 60° C. Sodium borohydride (12 mg, 0.315 mmoles, 1.5 eq.) was added. The mixture was stirred at room temperature for 12 h then DCM was added and the mixture was washed with sat. NaHCO.sub.3. The collected organic phases were evaporated in vacuum and the crude material was purified by Si-column eluting with ethyl acetate to ethyl acetate/MeOH 6:4 to afford the desired compound as free base. The product was dissolved in DCM then HCl (1M solution in diethyl ether, 0.156 mL, 3 eq.) was added dropwise at 0° C. The solution was left stirring for 2 h then was concentrated in vacuum and triturated with diethyl ether to obtain N-(2,5-difluorobenzyl)-3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-amine hydrochloride 38 (26.6 mg, Y=24%). LC-MS (M-H+): 397.2.

[0180] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.00 (br. s., 1H), 9.47 (br. s., 2H), 8.21-8.08 (m, 2H), 7.95 (d, J=8.0 Hz, 1H), 7.77 (t, J=7.0 Hz, 1H), 7.70-7.59 (m, 2H), 7.51 (d, J=5.8 Hz, 1H), 7.45-7.32 (m, 2H), 5.75 (s, 1H), 4.23 (t, J=5.4 Hz, 2H), 3.90 (d, J=12.8 Hz, 2H), 3.58 (br. s., 4H), 3.43-3.22 (m, 4H), 3.18-3.07 (m, 2H), 2.22 (quin, J=7.4 Hz, 2H)

Preparation of compound 39: 2,5-difluoro-N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}benzamide hydrochloride

[0181] Compound 39 was prepared according to the procedure described for compound 31, using in step 3, 2,5-difluorobenzoic acid. LC-MS (M-H.sup.+): 411.2.

[0182] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.04 (br. s., 1H), 8.63-8.53 (m, 1H), 8.15 (d, J=5.8 Hz, 2H), 7.95 (d, J=8.0 Hz, 1H), 7.76 (t, J=7.0 Hz, 1H), 7.69-7.60 (m, 1H), 7.55-7.45 (m, 2H), 7.44-7.32 (m, 2H), 3.88 (d, J=10.0 Hz, 2H), 3.63 (br. s., 3H), 3.46-3.32 (m, 6H), 3.26 (td, J=5.3, 10.7 Hz, 2H), 2.00 (quin, J=7.2 Hz, 2H)

Preparation of compound 42: N-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}furo[2,3-c]pyridine-5-carboxamide hydrochloride

[0183] The compound was prepared according to the procedure described for compound 31, using in step 3 furo[2,3-c]pyridine-5-carboxylic acid. LC-MS (M-H.sup.+): 489.2.

[0184] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.24 (br. s., 1H), 9.07 (t, J=5.9 Hz, 1H), 9.03 (s, 1H), 8.43 (d, J=0.8 Hz, 1H), 8.36 (d, J=2.3 Hz, 1H), 8.15 (d, J=8.3 Hz, 1H), 8.12 (d, J=5.8 Hz, 1H), 7.96 (d, J=8.3 Hz, 1H), 7.79 (t, J=7.4 Hz, 1H), 7.66 (t, J=7.4 Hz, 1H), 7.52 (d, J=5.8 Hz, 1H), 7.23 (d, J=1.5 Hz, 1H), 3.91 (d, J=11.5 Hz, 2H), 3.60 (br. s., 4H), 3.50-3.33 (m, 5H), 3.31-3.16 (m, 2H), 2.16-1.95 (m, 2H)

Preparation of compound 43: N-{3-[4-(1,6-naphthyridin-5-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride

[0185] Compound 43 was prepared according to the procedure described for compound 2, using in step 3 5-chloro-1,6-naphthyridine. LC-MS (M-H.sup.+): 377.2.

[0186] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=(CHLOROFORM-d) 9.04 (d, J=2.0 Hz, 1H), 9.00 (dd, J=1.6, 4.1 Hz, 1H), 8.71 (dd, J=1.8, 4.8 Hz, 1H), 8.39-8.33 (m, 2H), 8.22 (br. s., 1H), 8.17 (td, J=1.9, 8.0 Hz, 1H), 7.52 (d, J=5.8 Hz, 1H), 7.43 (dd, J=4.3, 8.5 Hz, 1H), 7.39 (dd, J=4.9, 7.9 Hz, 1H), 3.65 (q, J=5.8 Hz, 2H), 3.56-3.41 (m, 4H), 2.81 (br. s., 4H), 2.73 (t, J=5.8 Hz, 2H), 1.91 (quin, J=6.0 Hz, 2H)

Preparation of compound 45: N-{3-[4-(6-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}pyridine-3-carboxamide hydrochloride

[0187] Compound 45 was prepared according to the procedure described for compound 2, using in step 3 1-chloro-6-fluoroisoquinoline. LC-MS (M-H.sup.+): 394.2.

[0188] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=9.40-9.32 (m, 1H), 9.28 (s, 1H), 8.93 (d, J=5.3 Hz, 1H), 8.72 (d, J=7.6 Hz, 1H), 8.30 (dd, J=5.6, 9.1 Hz, 1H), 8.09 (d, J=6.1 Hz, 1H), 7.99-7.89 (m, 1H), 7.84 (dd, J=2.2, 9.5 Hz, 1H), 7.63-7.52 (m, 2H), 4.02 (d, J=13.4 Hz, 2H), 3.75 (t, J=12.6 Hz, 2H), 3.64 (d, J=12.0 Hz, 2H), 3.43 (ddd, J=5.7, 5.8, 11.8 Hz, 4H), 3.34-3.20 (m, 2H), 2.20-1.97 (m, 2H).

Preparation of Compound 50

[0189] Compound 50 was prepared as described herein below.

Step 1—Synthesis of tert-butyl N-{3-[4-(naphthalen-1-yl)piperazin-1-yl]propyl}carbamate

[0190] ##STR00200##

[0191] A mixture of commercial 1-(naphthalen-1-yl)piperazine hydrochloride (200 mg, 0.80 mmol), tert-butyl N-(3-bromopropyl)carbamate (182 mg, 0.76 mmol), potassium iodide (66 mg, 016 mmol) and potassium carbonate (221 mg, 1.6 mmol) in DMF (3 mL) was stirred 18 h at room temperature. The reaction mixture was partitioned between EtOAc (50 mL) and brine (50 mL), the organic phase was washed with brine (50 mL), dried over sodium sulfate and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 25) eluting with a gradient of 30-100% EtOAc in cyclohexane to give 263 mg (0.71 mmol, 94% yield) of the title compound. LC-MS (M-H.sup.+)=370.4

Step 2—Synthesis of 3-[4-(naphthalen-1-yl)piperazin-1-yl]propan-1-amine

[0192] ##STR00201##

[0193] TFA (2 mL) was added to a solution of tert-butyl N-{3-[4-(naphthalen-1-yl)piperazin-1-yl]propyl}carbamate (261 mg, 071 mmol) in dichloromethane (6 mL). The resulting mixture was stirred 1 h at room temperature then the volatiles were evaporated under reduced pressure. The residue was dissolved in dichloromethane (10 mL) and evaporated under reduced pressure twice. The resulting residue was dissolved in MeOH (4 mL) and loaded onto a preconditioned SCX cartridge (5 g) eluting with MeOH and with a 2M solution of ammonia in methanol. The basic fractions were dried under reduced pressure to give 193 mg (0.71 mmol, quant.) of the title compound as a white solid. LC-MS (M-H.sup.+)=270.3

Step 3—Synthesis of 4-({3-[4-(naphthalen-1-yl)piperazin-1-yl]propyl}amino)-2H-chromen-2-one hydrochloride (compound 50)

[0194] ##STR00202##

[0195] Compound 50 was prepared according to the procedure described for the synthesis of 53 (step 3, Y=21%). LC-MS (M-H+)=414.2

[0196] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.79 (br. s., 1H), 8.19-8.08 (m, 2H), 7.98-7.87 (m, 2H), 7.67 (d, J=8.0 Hz, 1H), 7.61 (t, J=7.3 Hz, 1H), 7.57-7.50 (m, 2H), 7.46 (t, J=7.9 Hz, 1H), 7.34 (t, J=7.7 Hz, 1H), 7.32 (d, J=8.3 Hz, 1H), 7.18 (d, J=7.3 Hz, 1H), 5.28 (s, 1H), 3.64 (d, J=10.0 Hz, 2H), 3.53-3.19 (m, 10H), 2.16 (quin, J=7.0 Hz, 2H).

Preparation of compound 53: 4-[{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}(methyl)amino]-2H-chromen-2-one hydrochloride

[0197] Compound 53 was prepared as described hereinbelow.

Step 1: tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl} methylcarbamate (53a)

[0198] ##STR00203##

[0199] Sodium hydride (24 mg, 60% dispersion in mineral oil, 0.61 mmoles, 1.5 eq.) was added to a stirred solution of intermediate 31a (150 mg, 0.41 mmoles, 1 eq.) in DMF (2 mL) at room temperature. The mixture was stirred for 10 minutes then iodomethane (38 μL, 0.61 mmoles, 1.5 eq.) was added. Stirring was continued for 2 hours then the reaction was quenched with water (50 mL) and extracted with EtOAc (50 mL). The organic phase was washed with brine (50 mL), dried over sodium sulfate and evaporated under reduced pressure. The residue was chromatographed on NH-modified silica gel (2× SNAP 11 in series) eluting with a gradient of 10-50% EtOAc in cyclohexane to give 78 mg of tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}methyl carbamate 53a (Y=50%). LC-MS (M-H+)=385.4.

Step 2: 3-[4-(isoquinolin-1-yl)piperazin-1-yl]-N-methylpropan-1-amine (53b)

[0200] TFA (1 mL) was added to a solution of intermediate 53a (78 mg, 0.2 mmoles, 1 eq.) in dichloromethane (3 mL) at room temperature and the resulting mixture was stirred for 60 minutes. The volatiles were evaporated under reduced pressure then the residue was dissolved in MeOH (2 mL) and loaded onto a preconditioned SCX cartridge (1 g). The SCX was eluted with MeOH and then a 2M solution of ammonia in methanol. The basic fractions were evaporated under reduced pressure to give 56 mg of 3-[4-(isoquinolin-111)piperazin-1-yl]-N-methylpropan-1-amine 53b (Y=quant.). LC-MS (M-H+)=285.3.

Step 3: 4-[{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}(methyl)amino]-2H-chromen-2-one hydrochloride (53)

[0201] A solution of intermediate 53b (56 mg, 0.2 mmoles, 1.1 eq.), triethylamine (37 μL, 0.27 mmoles, 1.5 eq.) and 2-oxo-2H-chromen-4-yl trifluoromethanesulfonate 28a (53 mg, 0.18 mmoles, 1 eq.) in acetonitrile (2 mL) was heated to 70° C. for 1 hour. The reaction mixture was concentrated under reduced pressure and the residue was partitioned between dichloromethane (10 mL) and a brine/sodium bicarbonate mixture (1:1, 10 mL). The mixture was filtered through a hydrophobic frit (Phase Separator) washing with dichloromethane (10 mL). The organic phase was evaporated under reduced pressure and the residue was chromatographed on NH-modified silica gel (2× SNAP 11 in series) eluting with a gradient of 20-100% EtOAc in cyclohexane to give 79 mg of a colorless sticky gum. The product was dissolved in dichloromethane (3 mL) and treated with 1M HCl solution in diethyl ether (0.46 mL) causing precipitation. The resulting mixture was evaporated under reduced pressure and the residue was triturated with diethyl ether. The solids were dried to give 85 mg of 4-[{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}(methyl)amino]-2H-chromen-2-one hydrochloride 53 as a white solid (Y=95%). LC-MS (M-H+)=429.2.

[0202] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.19 (d, J=8.3 Hz, 1H), 8.12 (d, J=5.8 Hz, 1H), 8.00 (d, J=8.0 Hz, 1H), 7.88-7.80 (m, 1H), 7.71 (t, J=7.5 Hz, 1H), 7.63 (t, J=7.5 Hz, 1H), 7.57 (d, J=5.8 Hz, 1H), 7.43-7.34 (m, 1H), 5.66 (s, 1H), 3.98 (d, J=13.8 Hz, 2H), 3.73-3.53 (m, 4H), 3.51-3.35 (m, 4H), 3.31-3.14 (m, 2H), 3.08 (s, 3H), 2.24 (br. quin, J=6.5 Hz, 2H).

Preparation of compound 57: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]-3-oxopropyl}amino)-2H-chromen-2-one hydrochloride

[0203] Compound 57 was prepared as described hereinbelow.

##STR00204##

Step 1: tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]-3-oxopropyl}carbamate (57a)

[0204] 1-(piperazin-1-yl)isoquinoline 2c (100 mg, 0.46 mmoles, 1 eq.) was added to a stirred solution of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (104 mg, 0.54 mmoles, 1.2 eq.), 1-hydroxybenzotriazole hydrate (72 mg, 0.54 mmoles, 1.2 eq.), 3-{[(tert-butoxy)carbonyl]amino}propanoic acid (86 mg, 0.46 mmoles, 1 eq.) and TEA (190 μl, 0.138 mmoles, 3 eq.) in DCM (5 mL) at room temperature. The resulting mixture was stirred overnight then the organic phase was washed with sodium bicarbonate solution, filtered through a hydrophobic frit (Phase Separator) and evaporated under reduce pressure. The residue was chromatographed on silica gel (SNAP 10) eluting with a gradient of 10-100% EtOAc in cyclohexane to give 148 mg of tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]-3-oxopropyl} carbamate 57a (Y=85%). LC-MS (M-H+)=385.4.

Step 2: 3-amino-1-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-one (57b)

[0205] Intermediate 57b was prepared according to the procedure described for the synthesis of intermediate 53b in the step 2 (Y=92%). LC-MS (M-H+)=285.3.

Step 3: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]-3-oxopropyl}amino)-2H-chromen-2-one hydrochloride (57)

[0206] Compound 57 was prepared according to the procedure described for the synthesis of 53 in step 3 (Y=87%). LC-MS (M-H+)=429.3.

[0207] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.26 (d, J=8.5 Hz, 1H), 8.11-8.00 (m, 2H), 7.98-7.87 (m, 2H), 7.80-7.68 (m, 2H), 7.63-7.56 (m, 1H), 7.54 (d, J=6.5 Hz, 1H), 7.38-7.25 (m, 2H), 5.19 (s, 1H), 4.15 (br. s., 1H), 3.87-3.62 (m, 8H), 3.55 (q, J=6.4 Hz, 2H), 2.82 (t, J=6.8 Hz, 2H)

Preparation of compound 58: 4,4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)quinolin-2(1H)-one hydrochloride

[0208] Compound 58 was prepared as described hereinbelow.

##STR00205##

Step 1: 2-oxo-1,2-dihydroquinolin-4-yl trifluoromethanesulfonate (58a)

[0209] 1,1,1-Trifluoro-N-phenyl-N-(trifluoromethane)sulfonylmethane sulfonamide (1.06 g, 2.98 mmoles, 1.2 eq.) was added to a stirred suspension of 4-hydroxy-1,2-dihydroquinolin-2-one (0.4 g, 2.48 mmoles, 1 eq.) and triethylamine (1.04 mL, 7.44 mmoles, 3 eq.) in DMF (15 mL) at room temperature. The reaction was stirred for 2 hours then was poured into water (100 mL) and extracted with EtOAc (60 mL). The organic phase was washed with brine (50 mL), dried over sodium sulphate and evaporated under reduced pressure. The residue was absorbed onto silica gel (10 g) and placed on top of a silica gel column (SNAP 50) which was eluted with 50-100% EtOAc in cyclohexane to give 537 mg (1.83 mmoles, Y=74%) of 2-oxo-1,2-dihydroquinolin-4-yl trifluoromethanesulfonate 58a as a white solid. LC-MS (M-H+)=294.1.

Step 2: 4,4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino) quinolin-2(1H)-one hydrochloride (58)

[0210] Compound 58 was prepared according to the procedure described for the synthesis of compound 53 (Y=28%), using in step 3 intermediates 58a and 31b. LC-MS (M-H+)=414.4.

[0211] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=12.16 (br. s., 1H), 11.06 (br. s., 1H), 8.27 (br. s., 1H), 8.18 (d, J=8.3 Hz, 1H), 8.10 (d, J=5.3 Hz, 1H), 7.99 (d, J=7.8 Hz, 1H), 7.89-7.78 (m, 1H), 7.74-7.61 (m, 2H), 7.60-7.45 (m, 2H), 7.35 (d, J=5.8 Hz, 1H), 5.82 (br. s., 1H), 5.02 (br. s., 2H), 3.97 (d, J=11.5 Hz, 2H), 3.79-3.55 (m, 4H), 3.53-3.23 (m, 6H), 2.19 (br. s., 2H)

Preparation of compound 60: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-6-phenyl-2H-pyran-2-one hydrochloride

[0212] Compound 60 was prepared as described hereinbelow.

##STR00206##

Step 1: 4-hydroxy-6-phenyl-2H-pyran-2-one (60a)

[0213] Malonyl dichloride (0.25 mL, 2.6 mmoles, 1 eq.) was added dropwise to a stirred solution of trimethyl[(1-phenylethenyl)oxy]silane (1.07 mL, 5.2 mmoles, 2 eq.) in diethyl ether (16 mL) at −78° C. The reaction mixture was stirred overnight allowing the reaction to gradually warm to room temperature. The precipitated solid was collected, washed with diethyl ether (20 mL) and dried under vacuum to give 0.34 g of 4-hydroxy-6-phenyl-2H-pyran-2-one 60a (1.8 mmoles, Y=70%), which was progressed without any further purification. LC-MS (M-H+)=189.1.

Step 2: 2-oxo-6-phenyl-2H-pyran-4-yl trifluoromethanesulfonate (60b)

[0214] Intermediate 60b was prepared according to the procedure described for the synthesis of 28a, using in step 1 intermediate 60a (Y=77%). LC-MS (M-H+)=321.1.

Step 3: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-6-phenyl-2H-pyran-2-one hydrochloride (60)

[0215] Compound 60 was prepared according to the procedure described for the synthesis of 53, using in step 3 intermediates 60b and 31b (Y=37%). LC-MS (M-H+)=441.4.

[0216] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.05 (br. s., 1H), 8.19 (d, J=8.3 Hz, 1H), 8.10 (d, J=5.8 Hz, 1H), 7.99 (d, J=7.8 Hz, 1H), 7.83 (t, J=7.2 Hz, 1H), 7.70 (dd, J=6.7, 14.7 Hz, 4H), 7.61-7.46 (m, 4H), 6.60 (br. s., 1H), 5.62 (br. s., 1H), 5.03 (d, J=1.3 Hz, 1H), 3.98 (d, J=11.8 Hz, 2H), 3.64 (d, J=11.5 Hz, 4H), 3.49-3.35 (m, 2H), 3.28 (br. s., 4H), 2.15-2.00 (m, 2H)

Preparation of compound 68: 4-(4-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazin-1-yl)quinolin-2(1H)-one hydrochloride

[0217] Compound 68 was prepared as described hereinbelow.

##STR00207## ##STR00208##

Step 1: tert-butyl 4-[3-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)propyl]piperazine-1-carboxylate (68a)

[0218] N-(3-Bromopropyl)phthalimide (2.1 g, 7.66 mmoles, 1 eq.), 1-Boc-piperazine (1.5 g, 8 mmoles, 1 eq.), potassium iodide (2.54 g, 15.3 mmoles, 2 eq.) and potassium carbonate (1.76 g, 12.7 mmoles, 1.7 eq.) were dissolved in DMA (15 mL) and stirred at room temperature for 16 h. The solvent was evaporated and the residue was dissolved in DCM. The salts were removed by filtration and the filtrate, after solvent evaporation, was purified by silica column (cHex: EtOAc 8:2 to cHex: EtOAc 1:1). The desired product tert-butyl 4-[3-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)propyl]piperazine-1-carboxylate 68a was obtained as a pale yellow crystalline solid (2.8 g, Y=quant.). LC-MS (M-H+)=374.1.

Step 2: tert-butyl 4-(3-aminopropyl)piperazine-1-carboxylate (68b)

[0219] Intermediate 68a (1.9 g, 5.08 mmoles) was dissolved in methylamine (33% in absolute ethanol, 20 mL) and heated at 40° C. for 4 h. The solvent was evaporated, the residue was dissolved in diethyl ether and filtered and the filtrate was evaporated to give tert-butyl 4-(3-aminopropyl)piperazine-1-carboxylate (68b) as a colourless oil (1.1 g, Y=89%). LC-MS (M-H.sup.+)=244.3.

Step 3: tert-butyl 4-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl} piperazine-1-carboxylate (68c)

[0220] A solution of intermediate 68b (1.1 g, 4.5 mmoles, 1.1 eq.), triethylamine (0.674 mL, 4.8 mmoles, 1.2 eq.) and 2-oxochromen-4-yl trifluoromethanesulfonate 28a (1.2 g, 4.1 mmoles, 1 eq.) in acetonitrile (20 mL) was heated to 70° C. for 1 hour. The reaction mixture was concentrated under reduced pressure and the residue was partitioned between dichloromethane and a brine/sodium bicarbonate mixture (1:1). The mixture was filtered through a hydrophobic frit (Phase Separator) washing with dichloromethane. The organic phase was evaporated under reduced pressure and the residue was chromatographed on silica gel (SNAP50) eluting with a gradient of EtOAc in cyclohexane to give tert-butyl 4-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazine-1-carboxylate 68c (700 mg, Y=44%). LC-MS (M-H.sup.+)=388.3.

Step 4: 4-{[3-(piperazin-1-yl)propyl]amino}chromen-2-one (68d)

[0221] TFA (2 mL) was added to a solution of intermediate 68c (700 mg, 1.8 mmoles) in dichloromethane (6 mL) at room temperature and the resulting mixture stirred for 20 minutes. The residue was evaporated under reduced pressure, dissolved in MeOH and loaded onto a preconditioned SCX cartridge. The SCX was eluted with MeOH and then a 2M solution of ammonia in methanol. The basic fractions were evaporated under reduced pressure to give 4-{[3-(piperazin-1-yl)propyl]amino}chromen-2-one 68d (456 mg, Y=88%). LC-MS (M-H.sup.+)=288.3.

Step 5: 4-(4-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazin-1-yl)quinolin-2(1H)-one hydrochloride (68)

[0222] A solution of intermediate 68d (70 mg, 024 mmoles, 1.1 eq.), triethylamine (35 uL, 0.26 mmoles, 1.2 eq.) and intermediate 58a (71 mg, 0.22 mmoles, 1 eq.) in DMSO (5 mL) was heated to 80° C. for 1 hour. The mixture was treated with water, the resulting solid was filtered, dissolved in dichloromethane (5 mL) and treated with HCl (1 M solution in diethyl ether, 0.630 mL) causing precipitation. The resulting mixture was evaporated under reduced pressure and the residue was triturated with diethyl ether. The solid was dried to give 113 mg of 4-(4-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazin-1-yl)quinolin-2(1H)-one hydrochloride 68 as a white solid (Y=quant.). LC-MS (M-H.sup.+)=431.3.

[0223] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.46 (br. s., 1H), 11.20 (br. s., 1H), 8.18 (d, J=7.5 Hz, 1H), 7.98 (br. s., 1H), 7.69 (d, J=8.0 Hz, 1H), 7.60 (t, J=7.3 Hz, 1H), 7.49 (t, J=7.4 Hz, 1H), 7.32 (d, J=7.5 Hz, 3H), 7.16 (t, J=7.3 Hz, 1H), 5.95 (s, 1H), 5.26 (s, 1H), 3.78-3.19 (m, 13H), 2.15 (br. s., 2H)

Preparation of compound 70: 4-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propoxy}-2H-chromen-2-one hydrochloride

[0224] Compound 70 was prepared as described hereinbelow.

##STR00209##

Step 1: 3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-ol (70a)

[0225] A mixture of potassium carbonate (634 mg, 4.6 mmoles, 1.5 eq.), 1-chloroisoquinoline (500 mg, 3.1 mmoles, 1 eq.) and 3-(piperazin-1-yl)propan-1-ol (881 mg, 6.1 mmoles, 2 eq.) in DMSO (4 mL) was heated to 120° C. under microwave irradiation for 5 hours. The reaction was allowed to cool to room temperature. The solid was filtered, washed with water then dried under reduced pressure to give 690 mg of 3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-ol 70a (Y=81%). LC-MS (M-H.sup.+)=272.2.

Step 2: 4-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propoxy}-2H-chromen-2-one hydrochloride (70)

[0226] Methanesulfonyl chloride (57 μL, 0.74 mmoles, 2 eq.) was added to a stirred solution of intermediate 70a (100 mg, 0.37 mmoles, 1 eq.) and TEA (206 μL, 1.48 mmoles, 4 eq.) in dichloromethane (10 mL) at 0° C. The reaction was stirred for 30 minutes then was left to warm to room temperature. The reaction mixture was washed with saturated aqueous NaHCO.sub.3 solution and water. The organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The intermediate was used without any further purification for the following reaction.

[0227] Sodium hydride (60% dispersion in mineral oil, 15 mg, 0.37 mmoles, 1 eq.) was added to a stirred solution of 4-hydroxy-2H-chromen-2-one (60 mg, 0.37 mmoles, 1 eq.) in acetonitrile (4 mL). The suspension was stirred at room temperature for 30 minutes then was added to a solution of the previously obtained intermediate in acetonitrile (4 mL). K.sub.2CO.sub.3 (76 mg, 0.55 mmoles, 1.5 eq.) was added and the mixture was heated to 80° C. for 2 h. The volatiles were removed under reduced pressure, the residue was dissolved in dichloromethane and washed with water and brine. The organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel eluting with a gradient of mixture A in dichloromethane, where A is MeOH/NH.sub.4OH 95/5, to give 72 mg of a white foam. The product was dissolved in dichloromethane (4 mL) and the minimum amount of MeOH then was treated with HCl (1 M solution in diethyl ether, 0.44 mL) causing precipitation. The solvents were evaporated under reduced pressure and the residue was triturated with diethyl ether. The resulting solid was dried to give 92 mg of 4-{3-[4-(isoquinolin-1-yl)piperazin-1-yl]propoxy}-2H-chromen-2-one hydrochloride 70 (Y=51%). LC-MS (M-H.sup.+)=416.3.

[0228] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.41 (br. s., 1H), 8.20 (d, J=8.5 Hz, 1H), 8.10 (d, J=6.0 Hz, 1H), 8.00 (d, J=8.3 Hz, 1H), 7.95 (dd, J=1.4, 7.9 Hz, 1H), 7.85 (t, J=7.4 Hz, 1H), 7.74-7.66 (m, 2H), 7.57 (d, J=6.0 Hz, 1H), 7.45-7.37 (m, 2H), 5.95 (s, 1H), 5.36 (br. s., 1H), 4.38 (t, J=5.6 Hz, 2H), 4.01 (d, J=13.1 Hz, 2H), 3.79-3.59 (m, 4H), 3.55-3.36 (m, 4H), 2.45-2.35 (m, 2H)

Preparation of compound 71: 4-(isoquinolin-1-yl)-1-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazin-2-one hydrochloride

[0229] Compound 71 was prepared as described hereinbelow.

##STR00210##

Step 1: 4-(isoquinolin-1-yl)piperazin-2-one (71a)

[0230] Intermediate 71a was prepared according to the procedure described in the step 1 of the synthesis of compound 70, using piperazin-2-one (Y=77%). LC-MS (M-H.sup.+)=228.1.

Step 2: tert-butyl {3-[4-(isoquinolin-1-yl)-2-oxopiperazin-1-yl] propyl}carbamate (71b)

[0231] NaH (60% dispersion in mineral oil, 81 mg, 2 mmoles, 1.1 eq.) and tert-butyl N-(3-bromopropyl)carbamate (330 mg, 1.4 mmoles, 0.75 eq.) were added to a stirred solution of intermediate 71a (420 mg, 1.8 mmoles, 1 eq.) in anhydrous DMF (6 mL). The mixture was stirred for 4 hours then was quenched with water and extracted with ethyl acetate. The organic phase was dried over sodium sulphate and chromatographed on silica eluting with a gradient of 50-100% EtOAc in petroleum ether to give 273 mg of tert-butyl {3-[4-(isoquinolin-1-yl)-2-oxopiperazin-1-yl]propyl}carbamate 71b (Y=51%). LC-MS (M-H+)=385.2.

Step 3: 1-(3-aminopropyl)-4-(isoquinolin-1-yl)piperazin-2-one (71c)

[0232] Intermediate 71c was prepared according to the procedure described in step 2 of the synthesis of compound 53 (Y=quant.). LC-MS (M-H.sup.+)=285.3.

Step 4: 4-(isoquinolin-1-yl)-1-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazin-2-one hydrochloride (71)

[0233] Compound 71 was prepared according to the procedure described in step 3 of the synthesis of compound 53 (Y=80%). LC-MS (M-H.sup.+)=429.3.

[0234] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.23 (d, J=8.0 Hz, 1H), 8.05 (d, J=8.0 Hz, 1H), 8.02-7.93 (m, 2H), 7.87 (t, J=6.9 Hz, 1H), 7.76-7.63 (m, 2H), 7.61-7.53 (m, 1H), 7.50 (d, J=6.3 Hz, 1H), 7.35-7.23 (m, 2H), 5.19 (s, 1H), 5.04-4.38 (m, 1H), 4.24 (br. s., 2H), 3.92 (br. s., 2H), 3.60 (br. s., 2H), 3.51 (t, J=7.0 Hz, 2H), 3.30 (q, J=7.0 Hz, 2H), 1.93 (quin, J=7.0 Hz, 2H)

Preparation of compound 74: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]cyclohexyl}amino)-2H-chromen-2-one hydrochloride

[0235] Compound 74 was prepared as described hereinbelow.

##STR00211##

Step 1: tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]cyclohexyl} carbamate (74a)

[0236] Sodium triacetoxyborohydride (591 mg, 2.8 mmoles, 3 eq.) was added to a stirred solution of 1-(piperazin-1-yl)isoquinoline 2c (200 mg, 0.93 mmoles, 1 eq.) and tert-butyl N-(3-oxocyclohexyl)carbamate (200 mg, 0.93 mmoles, 1 eq.) in dichloromethane (5 mL). The reaction was stirred at room temperature for 2 hours then was diluted with DCM and quenched with water. The mixture was filtered through a hydrophobic frit (Phase Separator). The organic phase was washed with a mixture of brine and sodium bicarbonate solution (1:1) and filtered through a hydrophobic frit (Phase Separator). The organic phase was evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 10) eluting with a gradient of EtOAc in cyclohexane to give 245 mg of tert-butyl {3-[4-(isoquinolin-1-yl)piperazin-1-yl]cyclohexyl}carbamate 74a (Y=63%). LC-MS (M-H+)=411.4.

Step 2: 3-[4-(isoquinolin-1-yl)piperazin-1-yl]cyclohexanamine (74b)

[0237] Intermediate 74b was prepared according to the procedure described in step 2 of the synthesis of compound 53 (Y=quant.). LC-MS (M-H.sup.+)=311.3.

Step 3: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]cyclohexyl}amino)-2H-chromen-2-one hydrochloride (74)

[0238] Compound 74 was prepared according to the procedure described in step 3 of the synthesis of compound 53 (Y=87%). LC-MS (M-H+)=455.4.

[0239] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ ppm 1.60-1.76 (m, 2H), 1.76-1.89 (m, 3H), 1.92-2.01 (m, 1H), 2.13-2.22 (m, 1H), 2.42 (d, J=12.72 Hz, 1H), 3.36-4.06 (m, 8H), 3.76-3.84 (m, 1H), 4.24 (br. s., 1H), 5.31 (s, 1H), 6.97 (d, J=6.36 Hz, 1H), 7.28-7.38 (m, 2H), 7.55 (d, J=5.90 Hz, 1H), 7.58-7.64 (m, 1H), 7.69 (t, J=7.58 Hz, 1H), 7.83 (t, J=7.60 Hz, 1H), 7.99 (d, J=7.60 Hz, 1H), 8.08 (d, J=5.87 Hz, 1H), 8.21 (d, J=7.60 Hz, 1H), 8.31 (d, J=7.34 Hz, 1H), 10.88 (br. s., 1H)

Preparation of compound 75: N-{2-[4-(isoquinolin-1-yl)piperazin-1-yl]ethyl}-2-oxo-2H-chromene-4-carboxamide hydrochloride

[0240] Compound 75 was prepared as described hereinbelow.

##STR00212##

Step 1: 2-oxo-2H-chromene-4-carboxylic acid (75a)

[0241] Palladium dichloride (80 mg, 0.45 mmoles, 0.1 eq.) was added to a stirred mixture of 2-iodophenol (1 g, 4.5 mmoles, 1 eq.), dimethyl maleate (1.71 mL, 13.6 mmoles, 3 eq.) and triethylamine (1.90 mL, 13.6 mmoles, 3 eq.) in water (40 mL). The reaction mixture was heated to 85° C. for 24 hours. The reaction mixture was acidified with 1M HCl solution and extracted repeatedly with dichloromethane. The combined organic phases were filtered through a hydrophobic frit (Phase Separator) and evaporated under reduced pressure. The residue was chromatographed on RP-18 (SNAP 60) eluting with a gradient of 2-100% MeCN in water. The MeCN was removed under reduced pressure then the aqueous phase was basified with 1 M NaOH solution and washed with diethyl ether (2×50 mL). The aqueous phase was acidified with 1M HCl solution and extracted repeatedly with dichloromethane. The combined organic phases were filtered through a hydrophobic frit (Phase Separator) and evaporated under reduced pressure to give 228 mg of 2-oxo-2H-chromene-4-carboxylic acid 75a as a pale yellow solid (Y=26%). LC-MS (M-H+)=191.0.

Step 2: 2-[4-(isoquinolin-1-yl)piperazin-1-yl]ethanamine (75b)

[0242] Intermediate 75b was prepared as described in step 2 of the synthesis of compound 68 using N-(2-bromoethyl)phthalimide (Y=47%). LC-MS (M-H.sup.+)=257.2.

Step 3: N-{2-[4-(isoquinolin-1-yl)piperazin-1-yl]ethyl}-2-oxo-2H-chromene-4-carboxamide hydrochloride (75)

[0243] Compound 75 (59 mg, Y=35%) was prepared following the conditions described in step 1 of the synthesis of compound 57. LC-MS (M-H.sup.+)=429.3.

[0244] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.25 (br. s., 1H), 9.33 (br. s., 1H), 8.20 (d, J=8.3 Hz, 1H), 8.11 (d, J=5.8 Hz, 1H), 8.00 (d, J=8.0 Hz, 1H), 7.90-7.79 (m, 2H), 7.69 (q, J=8.1 Hz, 2H), 7.56 (d, J=5.8 Hz, 1H), 7.47 (d, J=8.3 Hz, 1H), 7.39 (t, J=7.5 Hz, 1H), 6.86 (s, 1H), 4.61-3.04 (m, 13H)

Preparation of compound 81: N-{2-[4-(isoquinolin-1-yl)piperazin-1-yl]ethyl}-2-oxo-2H-chromene-4-carboxamide hydrochloride

[0245] Compound 81 was prepared as described hereinbelow.

##STR00213## ##STR00214##

[0246] Compound 81 was prepared as described for compound 71 starting from methyl piperazine-2-carboxylate. LC-MS (M-H.sup.+)=473.3.

[0247] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.14 (d, J=5.8 Hz, 1H), 8.08 (d, J=6.8 Hz, 2H), 7.85-7.75 (m, 2H), 7.71-7.64 (m, 1H), 7.63-7.57 (m, 1H), 7.54 (d, J=5.8 Hz, 1H), 7.38-7.28 (m, 2H), 5.27 (s, 1H), 4.64 (br. s., 7H), 4.12-3.56 (m, 6H), 3.38 (d, J=6.8 Hz, 4H), 2.24-1.92 (m, 2H)

Preparation of compound 82: 4-(isoquinolin-1-yl)-1-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazine-2-carboxylic acid hydrochloride

[0248] ##STR00215##

[0249] To a solution of methyl 4-(isoquinolin-1-yl)-1-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazine-2-carboxylate hydrochloride 81 (40 mg, 0.085 mmoles, 1 eq.) in THF/water 9:1 (1 mL) LiOH (4 mg, 0.093 mmoles, 1.1 eq.) was added. The mixture was stirred at 60° C. for 5 hours then was concentrated under reduced pressure. The residue was dissolved in water and 1 M HCl was added (6 eq.). The solvent was evaporated and the crude product was purified by C18 reversed chromatography (acetonitrile/water from 5:95 to 100:0) to obtain 4-(isoquinolin-1-yl)-1-{3-[(2-oxo-2H-chromen-4-yl)amino]propyl}piperazine-2-carboxylic acid hydrochloride 82 (38 mg, Y=84%). LC-MS (M-H+)=459.3.

[0250] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.18-8.10 (m, 2H), 8.05 (d, J=7.5 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.77-7.66 (m, 2H), 7.59 (t, J=7.8 Hz, 2H), 7.42 (d, J=5.8 Hz, 1H), 7.36-7.28 (m, 2H), 5.21 (s, 1H), 3.71-3.62 (m, 1H), 3.61-3.54 (m, 1H), 3.52-3.08 (m, 9H), 2.96 (td, J=6.7, 13.2 Hz, 1H), 2.82-2.70 (m, 2H), 1.86 (quin, J=6.7 Hz, 2H)

Preparation of compound 83: 4-({3-[4-(isoquinolin-1-yl)-2-(2,2,2-trifluoroethyl)piperazin-1-yl]propyl}amino)-2H-chromen-2-one hydrochloride

[0251] Compound 83 was prepared as described hereinbelow.

[0252] Compound 83 was prepared as described for compound 71 starting from 2-(2,2,2-trifluoroethyl)piperazine. LC-MS (M-H.sup.+)=497.3.

[0253] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.51 (br. s., 1H), 8.25-8.08 (m, 3H), 7.97 (d, J=8.3 Hz, 1H), 7.88 (br. s., 1H), 7.79 (t, J=7.4 Hz, 1H), 7.70-7.63 (m, 1H), 7.63-7.57 (m, 1H), 7.54 (d, J=5.5 Hz, 1H), 7.40-7.26 (m, 1H), 5.30 (s, 1H), 5.42 (br. s., 2H), 4.46-2.74 (m, 13H), 2.14 (quin, J=7.0 Hz, 2H)

Preparation of compound 85: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-7-methoxy-2H-chromen-2-one hydrochloride

[0254] Compound 85 was prepared as described hereinbelow.

##STR00216##

Step 1: 7-methoxy-2-oxo-2H-chromen-4-yl trifluoromethanesulfonate (85a)

[0255] Intermediate 85a was prepared as described in step 1 of the synthesis of compound 28, starting from 4-hydroxy-7-methoxy-2H-chromen-2-one (Y=quant.). LC-MS (M-H.sup.+)=325.1.

Step 2: 4-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-7-methoxy-2H-chromen-2-one hydrochloride (85)

[0256] Compound 85 was prepared as described in step 4 of the synthesis of compound 71, using intermediates 85a and 31b (Y=22%). LC-MS (M-H.sup.+)=445.4.

[0257] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.07 (br. s., 1H), 8.18 (d, J=8.3 Hz, 1H), 8.09 (d, J=6.0 Hz, 1H), 8.06 (d, J=8.8 Hz, 1H), 7.99 (d, J=8.0 Hz, 1H), 7.83 (t, J=7.2 Hz, 2H), 7.69 (t, J=7.5 Hz, 1H), 7.55 (d, J=6.0 Hz, 1H), 6.93 (dd, J=2.3, 8.8 Hz, 1H), 6.88 (d, J=2.5 Hz, 1H), 5.14 (s, 1H), 4.10 (br. s., 1H), 3.97 (d, J=12.5 Hz, 2H), 3.89-3.77 (m, 3H), 3.71-3.52 (m, 4H), 3.39 (d, J=5.8 Hz, 4H), 3.29 (br. s., 2H), 2.14 (quin, J=7.0 Hz, 2H)

Preparation of compound 87: 3-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-2H-chromen-2-one hydrochloride

[0258] ##STR00217##

[0259] Compound 87 was prepared as described hereinbelow.

Step 1: 3-bromo-2H-chromen-2-one (87a)

[0260] Bromine (0.35 mL, 6.84 mmoles, 1 eq.) was added dropwise to a stirred solution of coumarin (1 g, 6.84 mmoles, 1 eq.) in chloroform (10 mL) at 0° C. The reaction mixture was allowed to gradually warm to room temperature overnight. Triethylamine (2 mL) was added and the reaction mixture was washed with water (2×10 mL). The organic phase was dried over Na.sub.2SO.sub.4 and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 50) eluting with 5% EtOAc in petroleum ether to give 504 mg of 3-bromo-2H-chromen-2-one 87a as a white solid (Y=33%). LC-MS (M-H+)=225.0.

Step 2: 3-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-2H-chromen-2-one hydrochloride (87)

[0261] A mixture of intermediates 31b (50 mg, 0.185 mmoles, 1 eq.) and 87a (50 mg, 0.222 mmoles, 1.2 eq.), Pd(OAc).sub.2 (5 mg, 0.019 mmoles, 0.1 eq.), Xantphos (11 mg, 0.019 mmoles, 0.1 eq.) and Cs.sub.2CO.sub.3 (90 mg, 0.28 mmoles, 1.5 eq.) in 1,4-dioxane (2.5 mL) was heated to 130° C. in a sealed tube under microwave irradiation for 2 hours. The reaction mixture was concentrated under reduced pressure and the residue was chromatographed on silica gel (SNAP25) eluting with a gradient of 0-2% mixture A in dichloromethane, where mixture A is MeOH/NH.sub.4OH 10:1. The partially pure product was further purified by preparative LC-MS to give 20 mg of a yellow gum. The product was dissolved in dichloromethane (4 mL) and treated with 1M HCl solution in diethyl ether (0.16 mL) causing precipitation. The resulting mixture was evaporated under reduced pressure and the residue was triturated with diethyl ether. The solids were dried to give 24 mg of 3-({3-[4-(isoquinolin-1-yl)piperazin-1-yl]propyl}amino)-2H-chromen-2-one hydrochloride 87 (Y=27%). LC-MS (M-H+)=415.3.

[0262] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.16 (br. s., 1H), 8.19 (d, J=7.5 Hz, 1H), 8.08 (d, J=5.8 Hz, 1H), 8.00 (d, J=7.5 Hz, 1H), 7.85 (t, J=6.5 Hz, 1H), 7.70 (t, J=7.3 Hz, 1H), 7.57 (d, J=5.3 Hz, 1H), 7.47 (br. s., 1H), 7.37-7.10 (m, 3H), 6.68 (s, 1H), 6.27 (br. s., 1H), 4.00 (d, J=12.3 Hz, 2H), 4.37-3.81 (m, 1H), 3.64 (d, J=11.0 Hz, 4H), 3.40 (d, J=9.3 Hz, 2H), 3.28 (d, J=5.8 Hz, 4H), 2.11 (br. quin, J=7.0 Hz, 2H)

Preparation of compound 96: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-5-methyl-2H-chromen-2-one hydrochloride

[0263] Compound 96 was prepared as described hereinbelow.

##STR00218##

Step 1: 1-(2-hydroxy-6-methylphenyl)ethanone (96a)

[0264] A solution of 2-hydroxy-6-methylbenzoic acid (2 g, 12.7 mmoles, 1 eq.) in THF (13 mL) was cooled to −78° C. MeLi (1.6 M in Et.sub.2O, 38.1 mmoles, 3 eq.) was added dropwise. The mixture was stirred at room temperature overnight then was cooled to 0° C. and quenched with sat. NH.sub.4Cl. Ethyl acetate was added and the organic phase was washed with brine. The solvent was removed in vacuum and the crude material was purified by Si-column eluting with cHex to cHex/ethyl acetate 7:3 to obtain 980 mg of 1-(2-hydroxy-6-methylphenyl)ethanone 96a (Y=51%). LC-MS (M-H+)=151.0.

Step 2: 4-hydroxy-5-methyl-2H-chromen-2-one (96b)

[0265] A solution of intermediate 96a (0.98 g, 6.5 mmoles, 1 eq.) in toluene (10 mL) was added dropwise to a stirred suspension of NaH (60% dispersion in mineral oil, 0.784 g, 19.6 mmoles, 3.0) in toluene (20 mL)

[0266] at room temperature. Stirring was continued for 10 minutes, then a solution of diethyl carbonate (1.58 mL, 13.1 mmoles, 2 eq.) in toluene (10 mL) was added dropwise and the mixture was heated to 115° C. for 48 h. The solvent was evaporated under reduced pressure, the solid residue was added portion-wise to ice-cold water (100 mL) to give a turbid solution. The aqueous phase was washed with diethyl ether (50 mL) then acidified with 1M HCl solution (60 mL) causing precipitation of a white solid. The solid was collected by filtration and washed with water. The wet filter cake was suspended in dichloromethane (100 mL) and evaporated under reduced pressure to give 949 mg of 4-hydroxy-5-methyl-2H-chromen-2-one 96b (Y=82%). LC-MS (M-H+)=177.0.

Step 3: 5-methyl-2-oxo-2H-chromen-4-yl trifluoromethanesulfonate (96c)

[0267] Intermediate 96c was prepared as described in step 1 of the synthesis of compound 28, using intermediate 96b (Y=58%). LC-MS (M-H.sup.+)=309.0.

Step 4: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-5-methyl-2H-chromen-2-one hydrochloride (96)

[0268] Compound 96 was prepared as described in step 4 of the synthesis of compound 71, using intermediate 96d. Intermediate 96d was prepared according to the procedure described in step 2 of the synthesis of compound 31, starting from 1-chloro-7-fluoroisoquinoline (Y=31%). LC-MS (M-H.sup.+)=447.3.

[0269] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=11.17 (br. s., 1H), 8.13 (d, J=5.8 Hz, 1H), 8.09 (dd, J=5.5, 8.8 Hz, 1H), 7.87 (d, J=9.8 Hz, 1H), 7.74 (t, J=8.5 Hz, 1H), 7.59 (d, J=5.8 Hz, 1H), 7.44 (t, J=7.9 Hz, 1H), 7.16 (d, J=8.0 Hz, 1H), 7.12 (d, J=7.5 Hz, 1H), 6.65 (br. s., 1H), 5.59 (br. s., 1H), 5.27 (s, 1H), 3.87 (d, J=12.8 Hz, 2H), 3.63 (d, J=11.3 Hz, 2H), 3.55 (t, J=12.8 Hz, 2H), 3.44 (d, J=5.5 Hz, 4H), 3.28 (br. s., 2H), 2.84 (s, 3H), 2.27-2.06 (m, 2H)

Preparation of compound 97: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-3-methyl-2H-chromen-2-one hydrochloride

[0270] Compound 97 was prepared as described hereinbelow.

##STR00219##

Step 1: 4-hydroxy-3-methyl-2H-chromen-2-one (97a)

[0271] Intermediate 97a was prepared according to the procedure described in step 2 of the synthesis of compound 96, using 1-(2-hydroxyphenyl)propan-1-one (2.1 g, Y=20%). LC-MS (M-H.sup.+)=177.0.

Step 2: 3-methyl-2-oxo-2H-chromen-4-yl trifluoromethanesulfonate (97b)

[0272] Intermediate 97b was prepared according to the procedure described in step 1 of the synthesis of compound 28, using intermediate 97a (Y=58%). LC-MS (M-H.sup.+)=309.0.

Step 3: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-3-methyl-2H-chromen-2-one hydrochloride (97)

[0273] Compound 97 was prepared as described in step 4 of the synthesis of compound 71, using intermediate 96d. Intermediate 96d was prepared according to the procedure described in step 2 of the synthesis of compound 31, starting from 1-chloro-7-fluoroisoquinoline (Y=38%). LC-MS (M-H.sup.+)=447.3.

[0274] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.84 (br. s., 1H), 8.20-8.11 (m, 2H), 8.07 (dd, J=5.5, 9.0 Hz, 1H), 7.84 (dd, J=2.4, 10.2 Hz, 1H), 7.71 (dt, J=2.5, 8.8 Hz, 1H), 7.61-7.48 (m, 2H), 7.38-7.24 (m, 2H), 6.72 (br. s., 1H), 4.48 (br. s., 1H), 3.81 (d, J=11.8 Hz, 2H), 3.65 (t, J=6.7 Hz, 2H), 3.59 (d, J=11.0 Hz, 2H), 3.53-3.32 (m, 4H), 3.22 (br. s., 2H), 2.16 (s, 3H), 2.14-2.04 (m, 2H)

Preparation of compound 101: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-3-methyl-2H-chromen-2-one hydrochloride

[0275] Compound 101 was prepared as described hereinbelow.

##STR00220## ##STR00221##

Step 1: tert-butyl 4-(3-methoxy-3-oxopropyl)piperazine-1-carboxylate (101a)

[0276] Methyl 3-bromopropanoate (0.607 mL, 5.56 mmoles, 1.05 eq.) was added to a solution of N-Boc-piperazine (1 g, 5.3 mmoles, 1 eq.) and TEA (0.870 mL, 6.36 mmoles, 1.2 eq.) in anhydrous THF (8 mL), and the reaction was stirred overnight at room temperature. The mixture was diluted with EtOAc, the organic phase was washed with sodium bicarbonate solution and brine, dried over sodium sulphate and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 25) eluting with a gradient of 20-100% of EtOAc in ciclohexane to give 1.3 g of tert-butyl 4-(3-methoxy-3-oxopropyl)piperazine-1-carboxylate 101a (Y=90%). LC-MS (M-H+)=273.2.

Step 2: 3-[4-(tert-butoxycarbonyl)piperazin-1-yl]propanoic acid (101b)

[0277] To a solution of intermediate 101a (1.3 g, 4.7 mmoles, 1 eq.) in THF/water (9:1) LiOH (123 mg, 5.17 mmoles, 1.1 eq.) was added. The mixture was stirred at 60° C. for 1 h then was concentrated under reduced pressure. The crude 3-[4-(tert-butoxycarbonyl)piperazin-1-yl]propanoic acid 101b (607 mg) was progressed without any further purification. LC-MS (M-H.sup.+)=259.2.

Step 3: tert-butyl 4-(3-amino-3-oxopropyl)piperazine-1-carboxylate (101c)

[0278] Hexamethyldisilazane (0.488 mL, 2.35 mmoles, 1 eq.) was added to a stirred solution of N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (540 mg, 2.82 mmoles, 1.2 eq.), 1-hydroxybenzotriazole hydrate (381 mg, 2.82 mmoles, 1.2 eq.), intermediate 101b (607 mg, 2.35 mmoles, 1 eq.) and TEA (977 μl, 7.05 mmoles, 3 eq.) in DCM (8 mL). The resulting mixture was stirred overnight at room temperature. The organic phase was washed with sodium bicarbonate solution, filtered through a hydrophobic frit (Phase Separator) and evaporated under reduce pressure. The residue was chromatographed on modified NH silica gel (SNAP 26) eluting with a gradient of 10-100% A in EtOAc, where A is MeOH/AcOEt (5:95), to give 454 mg of tert-butyl 4-(3-amino-3-oxopropyl)piperazine-1-carboxylate 101c as a white solid (Y=75%).

[0279] LC-MS (M-H+)=258.2.

Step 4: 3-(piperazin-1-yl)propanamide (101d)

[0280] Intermediate 101d was prepared according to the procedure described in step 4 of the synthesis of compound 68 (Y=quant.). LC-MS (M-H.sup.+)=158.1.

Step 5: 3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propanamide (101e)

[0281] Intermediate 101e was prepared as described in step 1 of the synthesis of compound 73, using 1-chloro-7-fluoroisoquinoline (Y=84%). LC-MS (M-H.sup.+)=303.2.

Step 6: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-3-methyl-2H-chromen-2-one hydrochloride (101)

[0282] 4-(trifluoromethane)sulfonylchromen-2-one 28a (149 mg, 0.51 mmoles, 1.3 eq.), 3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propanamide 101e (117 mg, 0.39 mmoles, 1 eq.), K.sub.3PO.sub.4 (165 mg, 0.78 mmoles, 2 eq.), Pd(dba).sub.2 (11 mg, 0.019 mmoles, 0.05 eq.) and Xantphos (11 mg, 0.019 mmoles, 0.05 eq.) were loaded into a Schlenk tube, which was filled with N.sub.2. Dry dioxane (4 mL) was added and the resulting mixture was stirred at 100° C. for 30 min. The organic phase was washed with H.sub.2O, filtered through a hydrophobic frit (Phase Separator) and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 10) eluting with a gradient of 50-100% of EtOAc in ciclohexane. The resultant solid was dissolved in DCM and cooled to 0° C. 3 eq. of 1M HCl in diethyl ether was added, after 1 h the mixture was evaporated in vacuo and the residue was triturated with diethyl ether to obtain 58 mg of 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-3-methyl-2H-chromen-2-one hydrochloride 101 (Y=29%). LC-MS (M-H+)=447.3.

[0283] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.81 (br. s., 1H), 10.44 (s, 1H), 8.34 (dd, J=1.4, 8.4 Hz, 1H), 8.16 (d, J=5.8 Hz, 1H), 8.08 (dd, J=5.8, 9.0 Hz, 1H), 7.86 (dd, J=2.5, 10.3 Hz, 1H), 7.76-7.66 (m, 2H), 7.58 (d, J=5.8 Hz, 1H), 7.48-7.41 (m, 2H), 7.24 (s, 1H), 4.99 (br. s., 1H), 3.85 (d, J=10.5 Hz, 2H), 3.66-3.59 (m, 2H), 3.59-3.52 (m, 2H), 3.50-3.41 (m, 4H), 3.31 (t, J=7.0 Hz, 2H)

Preparation of compound 107: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-7-hydroxy-2H-chromen-2-one hydrochloride

[0284] Compound 107 was prepared as described hereinbelow.

##STR00222## ##STR00223##

Step 1: 7-(benzyloxy)-4-hydroxy-2H-chromen-2-one (107a)

[0285] Intermediate 107a was prepared according to the procedure described in step 2 of the synthesis of compound 96, using 1-[4-(benzyloxy)-2-hydroxyphenyl]ethanone (Y=86%). LC-MS (M-H.sup.+)=269.2.

Step 2: 7-(benzyloxy)-2-oxo-2H-chromen-4-yl trifluoromethanesulfonate (107b)

[0286] Intermediate 107b was prepared according to the procedure described in step 1 of the synthesis of compound 28 (Y=quant.). LC-MS (M-H.sup.+)=401.1.

Step 3: 7-(benzyloxy)-4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-2H-chromen-2-one (107c)

[0287] Intermediate 107c was prepared as described in step 4 of the synthesis of compound 71, using intermediate 96d. Intermediate 96d was prepared according to the procedure described in step 2 of the synthesis of compound 31, starting from 1-chloro-7-fluoroisoquinoline (Y=37%). LC-MS (M-H.sup.+)=539.4.

Step 4: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-7-hydroxy-2H-chromen-2-one hydrochloride (107)

[0288] To a suspension of intermediate 107c (49 mg, 0.09 mmoles, 1 eq.) in EtOH (4 mL) Pd/C 10% (0.05 eq.) was added and the mixture was stirred at room temperature under hydrogen (1 atm). After 30 h the mixture was filtered and concentrated in vacuum. The residue was loaded onto a SCX cartridge (1 g), which was eluted with MeOH and with a 1M NH.sub.3 solution in MeOH. The basic solution was concentrated in vacuum to give a colorless oil (34 mg). This was purified by flash chromatography (Biotage KP-Sil 10 g SNAP cartridge, eluent A: EtOAc, B: EtOAc/MeOH 80/20, gradient A/B from 90:10 to 0:100) to give 18.2 mg of a white solid. The product was dissolved in MeOH and a 1 M HCl solution in diethyl ether (0.1 mL) was added. The mixture was concentrated under nitrogen and the residue was triturated with diethyl ether (2×) to give 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-7-hydroxy-2H-chromen-2-one hydrochloride 107 as a white solid (18.7 mg, Y=40%). LC-MS (M-H+)=449.3.

[0289] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.51 (br. s., 1H), 10.42 (br. s., 1H), 8.16 (d, J=5.8 Hz, 1H), 8.07 (dd, J=5.5, 9.0 Hz, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.85 (dd, J=2.3, 10.3 Hz, 1H), 7.75-7.66 (m, 2H), 7.57 (d, J=5.8 Hz, 1H), 6.75 (dd, J=2.3, 8.8 Hz, 1H), 6.63 (d, J=2.3 Hz, 1H), 5.09 (s, 1H), 3.82 (d, J=8.8 Hz, 2H), 3.73 (br. s., 1H), 3.61 (d, J=6.3 Hz, 2H), 3.47-3.33 (m, 6H), 3.28 (td, J=5.0, 10.0 Hz, 2H), 2.10 (quin, J=7.2 Hz, 2H)

Preparation of compound 111: 4-({1-[(isoquinolin-1-yloxy)acetyl] piperidin-4-yl}amino)-2H-chromen-2-one

[0290] Compound 111 was prepared as described hereinbelow.

##STR00224##

Step 1: Ethyl (isoquinolin-1-yloxy)acetate (111a)

[0291] To a suspension of ethyl hydroxyacetate (3.45 mL, 36.4 mmoles, 3 eq.) in THF (15 mL) NaH (60% dispersion in mineral oil, 1.47 g, 36.6 mmoles, 3 eq.) was added portion-wise. The mixture was heated to 70° C. and stirred for 1 h then a solution of 1-chloroisoquinoline (2 g, 12.3 mmoles, 1 eq.) in THF (15 mL) was added. The reaction mixture was stirred at 70° C. overnight then was cooled to room temperature, poured into a saturated solution of NaCl (15 mL) and extracted with EtOAc (3×10 mL). The collected organic phases were dried over Na.sub.2SO.sub.4 and concentrated under vacuum. The residue was purified via flash chromatography on silica gel eluting with hexane/EtOAc 9:1 to give 1.85 g of ethyl (isoquinolin-1-yloxy)acetate 111a (Y=65%). GC/MS: 231.

Step 2: (isoquinolin-1-yloxy)acetic acid (111b)

[0292] To a solution of intermediate 111a (1.2 g, 5.2 mmoles) in a mixture of THF/EtOH 1:1 (10 mL) NaOH (1 M solution, 20.4 mL, 4 eq.) was added. The solution was stirred for 3 h at room temperature then the solvents were removed under vacuum, water (10 mL) was added to the residue followed by 1 M HCl (neutralization). The mixture was stirred for 15 min. then the resulting solid was filtered, washed with water and dried to give 0.83 g of (isoquinolin-1-yloxy)acetic acid 111b (Y=78%). LC-MS (M-H.sup.+)=204.1.

Step 3: 4-({1-[(isoquinolin-1-yloxy)acetyl]piperidin-4-yl}amino)-2H-chromen-2-one (111)

[0293] To a solution of intermediate 111b (0.3 g, 1.48 mmoles, 1 eq.) in DMF (10 mL) TEA (0.46 mL, 3.25 mmoles, 2.2 eq.), intermediate 28c (0.5 g, 1.63 mmoles, 1.1 eq.) and HBTU (616 mg, 1.63 mmoles, 1.1 eq.) were added. The mixture was stirred at room temperature overnight then was diluted with EtOAc and filtered. The solution was washed with water and concentrated. The residue was diluted with EtOAc, a white solid separated that was filtered and dried to give 182 mg of 4-({1-[(isoquinolin-1-yloxy)acetyl]piperidin-4-yl}amino)-2H-chromen-2-one 111 (Y=29%). LC-MS (M-H.sup.+)=430.2.

[0294] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=8.25 (dd, J=0.7, 7.7 Hz, 1H), 8.15 (d, J=7.9 Hz, 1H), 7.97 (d, J=5.8 Hz, 1H), 7.92 (d, J=8.2 Hz, 1H), 7.79 (ddd, J=1.5, 7.0, 8.3 Hz, 1H), 7.71-7.55 (m, 2H), 7.41 (d, J=5.3 Hz, 1H), 7.38-7.25 (m, 3H), 5.40 (s, 1H), 5.34 (s, 2H), 4.35 (d, J=12.9 Hz, 1H), 4.07-3.94 (m, 1H), 3.92-3.70 (m, 1H), 3.27 (d, J=11.8 Hz, 1H), 2.81 (tdd, J=11.9, 12.1, 12.3 Hz, 1H), 2.19-1.81 (m, 2H), 1.66 (dq, J=3.0, 11.8 Hz, 1H), 1.49 (dq, J=3.0, 11.8 Hz, 1H).

Preparation of compound 117: N-(isoquinolin-1-yl)-2-{4-[(2-oxo-2H-chromen-4-yl)amino]piperidin-1-yl}acetamide

[0295] Compound 117 was prepared as described hereinbelow.

##STR00225##

Step 1: Ethyl {4-[(2-oxo-2H-chromen-4-yl)amino]piperidin-1-yl}acetate (117a)

[0296] To a solution of intermediate 28c (0.64 g, 1.79 mmoles, 1 eq.) in MeCN (10 mL) K.sub.2CO.sub.3 (0.74 g, 5.34 mmoles, 3 eq.) was added. The mixture was stirred under reflux for 1 h then a solution of ethyl bromoacetate (0.2 mL, 1.78 mmoles, 1 eq.) in MeCN (5 mL) was added dropwise. After stirring 2 h under reflux the mixture was cooled to room temperature, diluted with EtOAc, filtered and concentrated under vacuum. The residue was purified via flash chromatography on silica gel eluting with CHCl.sub.3/MeOH 9:1 to give 0.45 g of ethyl {4-[(2-oxo-2H-chromen-4-yl)amino]piperidin-1-yl}acetate 117a (Y=76%). LC-MS (M-H.sup.+)=331.2.

Step 2: {4-[(2-oxo-2H-chromen-4-yl)amino]piperidin-1-yl}acetic acid (117b)

[0297] Intermediate 117b was prepared according to the procedure described in step 2 of the synthesis of compound 111, starting from intermediate 117a (Y=quant.). LC-MS (M-H.sup.+)=303.1.

Step 3: N-(isoquinolin-1-yl)-2-{4-[(2-oxo-2H-chromen-4-yl)amino]piperidin-1-yl}acetamide (117)

[0298] Compound 117 was prepared according to the procedure described in step 3 of the synthesis of compound 111, using isoquinolin-1-amine (Y=43%). LC-MS (M-H.sup.+)=429.2. .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=11.47-10.96 (m, 1H), 10.12-9.65 (m, 1H), 8.34 (br. s., 1H), 8.17 (d, J=7.7 Hz, 2H), 8.08-7.76 (m, 3H), 7.72 (t, J=7.8 Hz, 1H), 7.66-7.54 (m, 1H), 7.48-7.23 (m, 3H), 5.38 (s, 1H), 4.81-4.09 (m, 2H), 4.05-3.41 (m, 4H), 2.33-1.83 (m, 4H).

Preparation of compound 118: N-(isoquinolin-1-ylmethyl)-2-{4-[(2-oxo-2H-chromen-4-yl)amino]piperidin-1-yl}acetamide

[0299] Compound 118 was prepared according to the procedure described in step 3 of the synthesis of compound 111, using intermediate 117b and 1-(isoquinolin-1-yl)methanamine (Y=26%). LC-MS (M-H.sup.+)=443.2.

[0300] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=8.53 (t, J=5.1 Hz, 1H), 8.47 (d, J=5.8 Hz, 1H), 8.26 (qd, J=1.0, 8.3 Hz, 1H), 8.18 (dd, J=1.3, 8.0 Hz, 1H), 8.01 (d, J=7.9 Hz, 1H), 7.86-7.76 (m, 2H), 7.72 (ddd, J=1.5, 7.0, 8.0 Hz, 1H), 7.59 (dt, J=1.5, 7.7 Hz, 1H), 7.39-7.15 (m, 3H), 5.25 (s, 1H), 5.01 (d, J=5.3 Hz, 2H), 3.63-3.39 (m, 1H), 3.07 (s, 2H), 2.95 (d, J=11.7 Hz, 2H), 2.38-2.22 (m, 2H), 2.02-1.88 (m, 2H), 1.76 (dq, J=3.6, 11.7 Hz, 2H).

Preparation of compound 119: N-(1-benzothiophen-2-ylmethyl)-3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-amine

[0301] Compound 119 was prepared according to the procedure described in step 4 of the synthesis of compound 2 using intermediate 31b and 1-benzothiophene-2-carbaldehyde (Y=20%). LC-MS (M-H.sup.+)=417.2.

[0302] .sup.1H NMR (300 MHz, CHLOROFORM-d) δ=8.13 (d, J=5.8 Hz, 1H), 8.07 (d, J=8.6 Hz, 1H), 7.83-7.66 (m, 3H), 7.60 (ddd, J=1.3, 7.0, 8.3 Hz, 1H), 7.49 (ddd, J=1.5, 7.0, 8.3 Hz, 1H), 7.36-7.27 (m, 2H), 7.24 (d, J=5.8 Hz, 1H), 7.20 (d, J=0.7 Hz, 1H), 4.14 (d, J=0.9 Hz, 2H), 3.47 (t, J=5.0 Hz, 4H), 2.85 (t, J=6.5 Hz, 3H), 2.77 (t, J=5.0 Hz, 4H), 2.59 (t, J=6.9 Hz, 2H), 1.84 (quin, J=6.7 Hz, 2H).

Preparation of compound 120: N-(1,3-benzodioxol-5-ylmethyl)-3-[4-(isoquinolin-1-yl)piperazin-1-yl]propan-1-amine

[0303] Compound 120 was prepared according to the procedure described in step 4 of the synthesis of compound 2 using intermediate 31b and 1,3-benzodioxole-5-carbaldehyde (Y=10%). LC-MS (M-H.sup.+)=405.2.

[0304] .sup.1H NMR (300 MHz, CHLOROFORM-d) δ=8.13 (d, J=5.8 Hz, 1H), 8.02 (d, J=8.5 Hz, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.61 (ddd, J=1.1, 7.0, 8.3 Hz, 1H), 7.50 (ddd, J=1.7, 7.0, 8.3 Hz, 1H), 7.26 (dd, J=0.6, 5.8 Hz, 1H), 6.93-6.85 (m, 2H), 6.81 (d, J=7.8 Hz, 1H), 5.94 (s, 2H), 3.99 (s, 2H), 3.47-3.23 (m, 4H), 3.11 (t, J=5.9 Hz, 2H), 2.87-2.72 (m, 4H), 2.65 (t, J=5.8 Hz, 2H), 1.96 (quin, J=5.8 Hz, 2H).

Preparation of compound 122: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-5,6,7,8-tetrahydro-2H-pyrano[3,2-c]pyridin-2-one hydrochloride

[0305] Compound 122 was prepared as described hereinbelow.

##STR00226##

Step 1: tert-butyl 4-(pyrrolidin-1-yl)-3,6-dihydropyridine-1(2H)-carboxylate (122a)

[0306] Pyrrolidine (3.5 g, 50.2 mmoles, 2 eq.) was added to a solution of 1-Boc-4-piperidone (5 g, 25 mmoles, 1 eq.) in dry toluene (30 mL). The mixture was heated at 90° C. with a Dean-Stark apparatus. After 3 h the mixture was concentrated in vacuum to give tert-butyl 4-(pyrrolidin-1-yl)-3,6-dihydropyridine-1(2H)-carboxylate 122a as yellow oil (6.3 g, Y=quant.). The compound was used without any further purification and characterization.

Step 2: tert-butyl 3-acetyl-4-oxopiperidine-1-carboxylate (122b)

[0307] Intermediate 122a (6.3 g, 25 mmoles, 1 eq.) was dissolved in 1,4-dioxane (25 mL). Acetic anhydride (5.21 mL, 55.2 mmol, 2.2 eq.) was added and the resulting mixture was allowed to stand at room temperature under nitrogen for 2 days. Water (6 mL) was added and the resulting mixture was refluxed for 1 hour then cooled to room temperature and concentrated in vacuum. Water (20 mL) was added and the aqueous phase was extracted twice with EtOAc (20 mL). The combined organics extracts were washed with a 5% w/w HCl aqueous solution (20 mL), dried over MgSO.sub.4 and concentrated in vacuum. The resultant orange oil was purified by Si-column eluting with cyclohexane to cyclohexane/ethyl acetate 9:1 to afford 2.98 g of tert-butyl 3-acetyl-4-oxopiperidine-1-carboxylate 122b (Y=49%). LC-MS (M-H+)=242.2.

Step 3: tert-butyl 4-hydroxy-2-oxo-7,8-dihydro-2H-pyrano[3,2-c]pyridine-6(5H)-carboxylate (122c)

[0308] A solution of intermediate 122b (1 g, 4.14 mmoles, 1 eq.) in THF (20 mL) was cooled to −78° C. Lithium bis(trimethylsilyl)amide (1 M solution in THF, 12.4 mL, 3 eq.) was added dropwise. After stirring for 1 h at −78° C., dimethylcarbonate (0.4 mL, 4.6 mmoles, 1.1 eq.) was added. The resulting mixture was allowed to warm slowly to −10° C. After 1 h the reaction was cooled to 0° C. and quenched with 1 M HCl until pH=6. Ethyl acetate was added, the organic phase was separated, washed with brine and evaporated in vacuum to obtain crude tert-butyl 3-(3-methoxy-3-oxopropanoyl)-4-oxopiperidine-1-carboxylate, that was used without any further purification.

[0309] The crude material was dissolved in toluene (8 mL), 1,8-diazabicyclo[5.4.0]undec-7-ene (0.62 mL, 4.14 mmoles, 1 eq.) was added and the mixture was stirred under reflux. After 3 hours the reaction was cooled to 0° C. and quenched with 1 M HCl. Ethyl acetate was added, the organic phase was separated, washed with brine and evaporated in vacuum. The crude material was purified by Si-column eluting with EtOAc to EtOAc/MeOH 7:3 to obtain 600 mg of tert-butyl 4-hydroxy-2-oxo-7,8-dihydro-2H-pyrano[3,2-c]pyridine-6(5H)-carboxylate 122c (Y=54%). LC-MS (M-H+)=268.2.

Step 4: tert-butyl 2-oxo-4-{[(trifluoromethyl)sulfonyl]oxy}-7,8-dihydro-2H-pyrano[3,2-c]pyridine-6(5H)-carboxylate (122d)

[0310] Intermediate 122d was prepared according to the procedure described in step 1 of the synthesis of compound 28 (Y=quant.). LC-MS (M-H.sup.+)=400.1.

Step 5: tert-butyl 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-2-oxo-7,8-dihydro-2H-pyrano[3,2-c]pyridine-6(5H)-carboxylate (122e)

[0311] Intermediate 122e (Y=17%) was prepared as described in step 4 of the synthesis of compound 71, using intermediate 96d. Intermediate 96d was prepared according to the procedure described in step 2 of the synthesis of compound 31, starting from 1-chloro-7-fluoroisoquinoline. LC-MS (M-H.sup.+)=538.4.

Step 6: 4-({3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]propyl}amino)-5,6,7,8-tetrahydro-2H-pyrano[3,2-c]pyridin-2-one hydrochloride (122)

[0312] Compound 122 was prepared as described in step 4 of the synthesis of compound 68 (Y=72%). LC-MS (M-H.sup.+)=438.2.

[0313] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=10.74 (br. s., 1H), 9.61 (br. s., 2H), 8.16 (d, J=5.8 Hz, 1H), 8.07 (dd, J=5.8, 9.0 Hz, 1H), 7.84 (dd, J=2.5, 10.3 Hz, 1H), 7.71 (dt, J=2.5, 8.8 Hz, 1H), 7.57 (d, J=5.8 Hz, 1H), 7.16 (t, J=5.3 Hz, 1H), 5.06 (s, 1H), 4.16-3.86 (m, 3H), 3.81 (d, J=10.8 Hz, 2H), 3.59 (d, J=9.0 Hz, 2H), 3.50-3.32 (m, 6H), 3.31-3.16 (m, 4H), 2.73 (t, J=5.8 Hz, 2H), 2.14-1.93 (m, J=7.0, 7.0, 7.0, 7.0 Hz, 2H).

Preparation of Compound 123

[0314] Compound 123 was prepared as described herein below.

##STR00227##

Step 1: 2-oxo-2H-chromen-4-yl trifluoromethanesulfonate (123a)

[0315] 4-Hydroxycoumarine (1 g, 6.17 mmoles, 1 eq.) was dissolved in DCM (30 mL), triethylamine (1.72 mL, 12.34 mmoles, 2 eq.) was added and the resulting mixture was cooled to −10° C. Trifluoromethanesulfonic anhydride (1.25 mL, 7.4 mmoles, 1.2 eq.) in DCM (5 mL) was added dropwise and the solution was left stirring at −10° C. for 2 h. The resulting reddish brown solution was warmed to room temperature, diluted with cHex/Et2O 1/1 (50 mL) and filtered through a pad of silica gel using cHex/Et2O 1/1 as eluent. The solvent was removed in vacuum to obtain 2-oxo-2H-chromen-4-yl trifluoromethanesulfonate 123a (1.76 g, Y=97%). LC-MS (M-H+): 295.0.

Step 2: tert-butyl 4-[(2-oxo-2H-chromen-4-yl)amino]piperidine-1-carboxylate (123b)

[0316] Triethylamine (1 mL, 7.2 mmoles, 1.2 eq.) in acetonitrile (2 mL) was added dropwise to a stirred solution of intermediate 123a (1.76 g, 5.99 mmoles, 1 eq.) and 1-Boc-4-aminopiperidine (1.2 g, 5.99 mmoles, 1 eq.) in dry acetonitrile (20 mL). The mixture was heated at reflux for 2 h then was cooled to room temperature, diluted with DCM (100 mL) and washed with saturated NaHCO3 (50 mL) and water (50 mL). The organic phase was separated, dried over sodium sulfate and evaporated in vacuum. The crude material was purified by trituration with methanol to obtain tert-butyl 4-[(2-oxo-2H-chromen-4-yl)amino]piperidine-1-carboxylate 123b (1.51 g, Y=73%). LC-MS (M-H+): 345.2.

Step 3: 4-(piperidin-4-ylamino)-2H-chromen-2-one trifluoroacetic acid salt (123c)

[0317] Intermediate 123b (1 g, 2.90 mmoles, 1 eq.) was dissolved in DCM (10 ml) and TFA (3 mL) was added dropwise at 0° C. The solution was left stirring for 2 h then was concentrated in vacuum and washed with toluene (25 ml) and diethyl ether (25 ml) to obtain 4-(piperidin-4-ylamino)-2H-chromen-2-one trifluoroacetic acid salt 123c, which was used without any further purification (1.22 g, Y=quant.). LC-MS (M-H+): 245.1.

Step 4: 4-{[1-(prop-2-yn-1-yl)piperidin-4-yl]amino}-2H-chromen-2-one (123d)

[0318] To a solution of intermediate 123c (50 mg, 0.14 mmoles, 1 eq.) and potassium carbonate (38.7 mg, 0.28 mmoles, 2 eq.) in DMF (2 mL) 3-bromo-1-propyne (20.8 mg, 0.14 mmoles, 1 eq.) was added at room temperature. The reaction was left stirring overnight then was filtered and the filtrate concentrated. The residue was purified by Si-column eluting with eluting with EtOAc to EtOAc/Methanol 8:2 to give 4-{[1-(prop-2-yn-1-yl)piperidin-4-yl]amino}-2H-chromen-2-one 123d (25.6 mg, Y=64%). LC-MS (M-H+): 283.1.

Step 5: 4-({1-[3-(isoquinolin-1-yl)prop-2-yn-1-yl]piperidin-4-yl}amino)-2H-chromen-2-one (123e)

[0319] Intermediate 123d (186 mg, 0.66 mmoles, 1 eq.), 1-chloroisoquinoline (120 mg, 0.72 mmoles, 1.1 eq.) and CuI (12 mg, 0.066 mmoles, 0.1 eq.) were dissolved in dry DMF (3.6 ml). DIPEA (0.473 ml, 2.64 mmoles, 4 eq.) was added. The mixture was degassed by alternatively applying vacuum and nitrogen, then bis(triphenylphosphine)palladium(II) dichloride (45 mg, 0.066 mmoles, 0.1 eq.) was added and the mixture was heated at 60° C. After 3 h water (10 mL) was added and the mixture was extracted with ethyl acetate (3×10 mL). The organic phase was washed with brine, dried over Na2SO4, filtered and concentrated in vacuum to give 283 mg of crude material. After purification by Si-column eluting with EtOAc to EtOAc/MeOH 8:2 compound 4-({1-[3-(isoquinolin-1-yl)prop-2-yn-1-yl]piperidin-4-yl}amino)-2H-chromen-2-one 123e (87 mg, Y=32%) was obtained. LC-MS (M-H+): 410.2.

Step 6—Synthesis of 4-({1-[3-(isoquinolin-1-yl)propyl]piperidin-4-yl}amino)-2H-chromen-2-one (123f)

[0320] To a solution of intermediate 123e 4-({1-[3-(isoquinolin-1-yl)prop-2-yn-1-yl]piperidin-4-yl}amino)-2H-chromen-2-one (82 mg, 0.2 mmoles, 1 eq.) in EtOAc (5 ml) Pd/C 10% (0.05 eq.) was added and the mixture was stirred under hydrogen (3.5 atm) at room temperature. After 3 hours DCM was added, the mixture was then filtered and concentrated in vacuum to give crude material, which was purified by Si-column eluting with DCM to DCM/Methanol 8:2 to give 30 mg (0.072 mmol, 36% yield) of intermediate 123f. LC-MS (M-H.sup.+): 414.1

Step 7—Synthesis of 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propan-1-ol (compound 123)

[0321] To a solution of 4-({1-[3-(isoquinolin-1-yl)propyl]piperidin-4-yl}amino)-2H-chromen-2-one (123f, 25 mg, 0.06 mmol) in acetone (5 mL) K2CO3 (43 mg, 0.31 mmol) was added. After stirring 1 h the mixture was cooled to 0° C. then methyl iodide (3.7 μL, 0.06 mmol) was added. The mixture was left to return to room temperature then was stirred overnight, filtered and concentrated in vacuo. The residue was purified via preparative HPLC (C18, acetonitrile from 10% to 30% in water) to give 13.3 mg (0.024 mmol, 40% yield) of the title compound. LC-MS (M-H.sup.+): 428.2

[0322] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=8.44 (d, J=5.6 Hz, 1H), 8.32 (d, J=8.5 Hz, 1H), 8.16-8.09 (m, 1H), 7.99 (d, J=7.6 Hz, 1H), 7.83-7.76 (m, 1H), 7.76-7.68 (m, 2H), 7.67-7.58 (m, 1H), 7.54 (d, J=7.3 Hz, 1H), 7.40-7.30 (m, 2H), 5.36 (s, 1H), 3.96-3.79 (m, 1H), 3.75-3.59 (m, 4H), 3.57-3.36 (m, 4H), 3.13 (s, 3H), 2.39-2.05 (m, 6H).

Preparation of compound 128: (2S,3S) and (2R,3R) 4-({3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]-2,3-dihydroxypropyl}amino)-2H-chromen-2-one hydrochloride

[0323] Compound 128 was prepared as described hereinbelow.

##STR00228## ##STR00229##

Step 1: [1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]methanol (128a)

[0324] Intermediate 128a was prepared according to the procedure described in step 1 of the synthesis of compound 70, using piperidin-4-ylmethanol (Y=82%). LC-MS (M-H.sup.+)=261.2.

Step 2: 1-(7-fluoroisoquinolin-1-yl)piperidine-4-carbaldehyde (128b)

[0325] Intermediate 128a (675 mg, 2.6 mmoles, 1 eq.), sodium bicarbonate (1.09 g, 13 mmoles, 5 eq.) and Dess-Martin periodinane (1.65 g, 3.9 mmoles, 1.5 eq.) were suspended in DCM (4 mL). The reaction mixture was stirred at r.t. for 3 hours then was quenched with sat. NaHCO.sub.3/2N Na.sub.2S.sub.2O.sub.3 in a 1:1 ratio. After 15 minutes water and DCM were added, the organic phase was separated, washed with brine, treated with Na.sub.2SO.sub.4 and concentrated under reduced pressure to give 1-(7-fluoroisoquinolin-1-yl)piperidine-4-carbaldehyde 128b, which was progressed without any further purification (671 mg, Y=quant.). LC-MS (M-H+)=259.1.

Step 3: methyl (2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-enoate (128c)

[0326] Methyl 2-(dimethoxyphosphoryl)acetate (224 mg, 1.23 mmoles, 1.6 eq.) was added to a solution of potassium tert-butoxide (138 mg, 1.23 mmoles, 1.6 eq.) in THF (3 mL) at 0° C. The mixture was stirred at room temperature for 30 min. then a solution of intermediate 128b (199 mg, 0.77 mmoles, 1 eq.) in THF (2 mL) was added dropwise at 0° C. The reaction was stirred at room temperature overnight. The reaction was quenched with water and extracted with EtOAc (2×). The organic phase was dried over sodium sulphate and evaporated under reduced pressure. The residue was chromatographed on silica gel (SNAP 10) eluting with a gradient of 0-30% EtOAc in cyclohexane to give 125 mg of methyl (2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-enoate 128c (Y=52%). LC-MS (M-H+)=315.1.

Step 4: (2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-en-1-ol (128d)

[0327] Diisobutylaluminum hydride (1 M solution in cyclohexane, 13 mL, 3.7 eq.) was added dropwise to a stirred solution of 128d (1.1 g, 3.5 mmoles, 1 eq.) in THF (25 mL) at −78° C. The mixture was stirred for 1 hour at the same temperature then was quenched with MeOH (4 mL) and was allowed to warm to room temperature. EtOAc (100 mL) and Rochelle saturated solution (150 mL) were added, the resulting mixture was stirred vigorously for 15 minutes, the organic phase was separated and dried over sodium sulphate. The volatiles were removed under reduced pressure to give 0.9 g of (2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-en-1-ol 128d (Y=90%). LC-MS (M-H+)=287.4.

Step 5: di-tert-butyl {(2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-en-1-yl}imidodicarbonate (128e)

[0328] To a solution of intermediate 128d (800 mg, 2.8 mmoles, 1 eq.) in DCM (4.5 mL) TEA (0.97 mL, 7 mmoles, 2.5 eq.) was added. The mixture was cooled to 0° C. then methanesulfanonyl chloride (0.26 mL, 3.35 mmoles, 1.2 eq.) was added under N.sub.2 atmosphere. The mixture was allowed to warm to room temperature and stirred for 1 h. DCM was added and the solution was washed with sat. NaHCO.sub.3. The organic phase was separated and evaporated in vacuum to give (2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-en-1-yl methanesulfonate (Y=quant.), that was used without any further purification.

[0329] The intermediate (2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-en-1-yl methanesulfonate (2.8 mmoles, 1 eq.) was added dropwise to a stirred mixture of di-tert-butyl-iminodicarboxylate (1 g, 4.75 mmoles, 1.7 eq.), K.sub.2CO.sub.3 (0.7 g, 5 mmoles, 1.8 eq.) and LII (12 mg, 0.092 mmoles, 0.03 eq.) in acetonitrile (20 mL). The reaction was refluxed overnight then sat. NaHCO.sub.3 and EtOAc were added. The organic phase was washed with brine and dried over sodium sulphate. The residue was chromatographed on silica gel (SNAP 50) eluting with a gradient of 0-15% solution A in cHex, where A is 20% MeOH in EtOAc, to give 580 mg of di-tert-butyl {(2E)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-en-1-yl}imidodicarbonate 128e (Y=48%). LC-MS (M-H+)=486.4.

Step 6: (2S,3S) and (2R,3R) 3-amino-1-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]propane-1,2-diol hydrochloride (128f)

[0330] To a stirred solution of intermediate 128e (580 mg, 1.2 mmoles, 1 eq.) in t-butanol/water 1:1 (8 mL) 4-methylmorpholine-N-oxide (167 mg, 1.4 mmoles, 1.2 eq.) and one drop of osmium tetroxide (4% water solution) were added. After stirring 18 hours at room temperature, EtOAc was added and the organic phase was washed with water and brine, separated and evaporated in vacuum. 300 mg of the residue was dissolved in DCM/MeOH (5 mL) and cooled to 0° C. 3 eq. of 1 M HCl in diethyl ether were added and the mixture was stirred at room temperature overnight and then at 60° C. for 5 h. The solvent was evaporated in vacuum to obtain 199 mg of (1S,2S)-3-amino-1-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]propane-1,2-diol hydrochloride 128f as a racemic mixture (Y=95%). LC-MS (M-H+)=320.1.

Step 7: (2S,3S) and (2R,3R) 4-({3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]-2,3-dihydroxypropyl}amino)-2H-chromen-2-one hydrochloride (128)

[0331] Compound 128, as a racemic mixture, was prepared as described in step 4 of the synthesis of compound 71, using intermediates 128f and 28a (Y=86%). LC-MS (M-H.sup.+)=464.3.

[0332] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ ppm 1.46-1.61 (m, 2H), 1.73-1.84 (m, 2H), 2.06-2.13 (m, 1H), 2.77-2.88 (m, 1H), 3.25 (t, J=7.83 Hz, 1H), 3.29-3.36 (m, 1H), 3.40-3.48 (m, 1H), 3.64-3.77 (m, 2H), 3.84-3.98 (m, 1H), 4.45 (d, J=7.83 Hz, 1H), 4.83 (d, J=6.36 Hz, 1H), 5.24 (s, 1H), 7.29-7.36 (m, 2H), 7.40 (d, J=5.87 Hz, 1H), 7.56-7.71 (m, 4H), 7.98 (dd, J=8.80, 5.87 Hz, 1H), 8.07 (d, J=7.83 Hz, 1H), 8.09 (d, J=5.87 Hz, 1H).

Preparation of compound 129: (2R,3S) and (2S,3R) 4-({3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]-2,3-dihydroxypropyl}amino)-2H-chromen-2-one hydrochloride

[0333] Compound 129 was prepared as described hereinbelow.

##STR00230##

[0334] Compound 129, as a racemic mixture, was prepared as described for the synthesis of compound 128, using methyl (2Z)-3-[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]prop-2-enoate as key intermediate. LC-MS (M-H.sup.+)=464.3.

[0335] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ ppm 1.58-1.96 (m, 4H), 2.05 (br. s., 1H), 3.11-3.27 (m, 3H), 3.33 (dd, J=7.83, 2.93 Hz, 1H), 3.61-3.69 (m, 1H), 3.71-3.79 (m, 1H), 4.00 (br. s., 2H), 5.23 (s, 1H), 7.25-7.38 (m, 2H), 7.53 (d, J=5.87 Hz, 1H), 7.59 (t, J=7.58 Hz, 1H), 7.72-7.77 (m, 1H), 7.82 (d, J=9.78 Hz, 2H), 7.93 (d, J=4.40 Hz, 1H), 8.13 (d, J=7.83 Hz, 2H)

Preparation of compound 130: N-(1,3-benzodioxol-5-ylmethyl)-2-[4-(isoquinolin-1-yl)piperazin-1-yl]ethanamine

[0336] Compound 130 was prepared according to the procedure described in step 4 of the synthesis of compound 2, using intermediate 75b and 1,3-benzodioxole-5-carbaldehyde (Y=25%). LC-MS (M-H.sup.+)=391.2.

[0337] .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ=8.09 (d, J=5.7 Hz, 2H), 8.08 (d, J=8.5 Hz, 1H), 7.89 (dd, J=0.9, 8.3 Hz, 1H), 7.71 (ddd, J=1.2, 6.8, 8.0 Hz, 1H), 7.61 (ddd, J=1.2, 6.8, 8.4 Hz, 1H), 7.40 (d, J=5.4 Hz, 1H), 7.05 (t, J=1.0 Hz, 1H), 6.95 (d, J=1.0 Hz, 2H), 6.02 (s, 2H), 3.97 (s, 2H), 3.34 (br. s., 4H), 2.94 (t, J=6.2 Hz, 2H), 2.76-2.58 (m, 6H).

Preparation of Compound 136

[0338] Compound 136 was prepared as described herein below.

Step 1—Synthesis of 3-cyclohexyl-3-({(2R)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)propanoic acid (formate salt, compound 136)

[0339] ##STR00231##

[0340] Compound 3-cyclohexyl-3-({(2R)-3-[4-(7-fluoroisoquinolin-1-yl) piperazin-1-yl]-2-hydroxypropyl}amino)propanoic acid (formate salt) as a mixture of diastereoisomers was prepared according to the synthesis described for compound 139 using racemic 3-amino-3-cyclohexylpropanoic acid (Y=44%).

[0341] LC-MS (M-H.sup.+)=459.5. .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 0.86-1.29 (m, 5H), 1.49-1.79 (m, 6H), 2.05-2.17 (m, 1H), 2.25 (dt, J=16.14, 4.16 Hz, 1H), 2.48 (d, J=7.34 Hz, 2H), 2.62-2.79 (m, 5H), 2.80-2.95 (m, 2H), 3.28 (br. s., 4H), 3.84-3.94 (m, 1H), 7.45 (d, J=5.38 Hz, 1H), 7.65 (td, J=8.68, 2.69 Hz, 1H), 7.69 (d, J=9.78 Hz, 1H), 8.01 (dd, J=9.05, 5.62 Hz, 1H), 8.12 (d, J=5.87 Hz, 1H), 8.15 (s, 1H).

Preparation of Compound 137

[0342] Compound 137 was prepared as described herein below.

Step 1—Synthesis of 3-cyclohexyl-3-({(2S)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)propanoic acid (formate salt, compound 137)

[0343] ##STR00232##

[0344] Compound 3-cyclohexyl-3-({(2S)-3-[4-(7-fluoroisoquinolin-1-yl) piperazin-1-yl]-2-hydroxypropyl}amino)propanoic acid (formate salt) as a mixture of diastereoisomers was prepared according to the synthesis described for compound 138 using racemic 3-amino-3-cyclohexylpropanoic acid. LC-MS (M-H.sup.+)=459.5. .sup.1H NMR (500 MHz, DMSO-d6) δ 1H NMR (500 MHz, DMSO-d6) d ppm 0.82-1.32 (m, 5H), 1.47-1.80 (m, 6H), 2.04-2.16 (m, 1H), 2.20-2.28 (m, 1H), 2.48 (d, J=6.85 Hz, 2H), 2.72 (br. s., 5H), 2.80-2.94 (m, 2H), 3.29 (br. s., 4H), 3.88 (d, J=5.87 Hz, 1H), 5.02 (br. s., 1H), 7.45 (d, J=5.38 Hz, 1H), 7.65 (td, J=8.56, 2.45 Hz, 1H), 7.69 (d, J=9.78 Hz, 1H), 8.01 (dd, J=9.05, 5.62 Hz, 1H), 8.12 (d, J=5.87 Hz, 1H), 8.15 (s, 1H).

Preparation of Compound 138

[0345] Compound 138 was prepared as described herein below.

Step 1—Synthesis of 7-fluoro-1-(4-{[(2R)-oxiran-2-yl]methyl}piperazin-1-yl)isoquinoline

[0346] ##STR00233##

[0347] The title intermediate was prepared according to the procedure described for the synthesis of 7-fluoro-1-(4-{[(2R)-oxiran-2-yl]methyl}piperazin-1-yl)isoquinoline (see compound 139) using (R)-(+)-epichlorohydrin (Y=92%). LC-MS (M-H.sup.+)=288.3

Step 2—Synthesis of (2S)-cyclohexyl({(2S)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)acetic acid (formate salt, compound 138)

[0348] ##STR00234##

[0349] Compound 138 was prepared according to the procedure described for compound 139 using 7-fluoro-1-(4-{[(2R)-oxiran-2-yl]methyl}piperazin-1-yl)isoquinoline and (2S)-amino(cyclohexyl)acetic acid (Y=27%). LC-MS (M-H.sup.+)=445.2. .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 1.09-1.37 (m, 5H), 1.62-1.85 (m, 6H), 2.57-2.60 (m, 4H), 2.73-2.89 (m, 5H), 2.94 (dd, J=11.98, 4.16 Hz, 1H), 3.11 (d, J=4.40 Hz, 1H), 3.34 (d, J=7.34 Hz, 4H), 3.96-4.09 (m, 1H), 7.52 (d, J=5.38 Hz, 1H), 7.71 (td, J=8.68, 2.69 Hz, 1H), 7.76 (dd, J=10.03, 2.20 Hz, 1H), 8.07 (dd, J=9.29, 5.87 Hz, 1H), 8.18 (d, J=5.87 Hz, 1H), 8.21 (s, 1H).

Preparation of Compound 139

[0350] Compound 139 was prepared as described herein below.

Step 1—Synthesis of tert-butyl 4-(7-fluoroisoquinolin-1-yl)piperazine-1-carboxylate

[0351] ##STR00235##

[0352] The title intermediate was prepared as described for the synthesis of intermediate 2c using 1-chloro-7-fluoroisoquinoline and N-Boc-piperazine (Y=94%). LC-MS (M-H.sup.+)=332.3

Step 2—Synthesis of 7-fluoro-1-(piperazin-1-yl)isoquinoline (trifluoroacetic acid salt)

[0353] ##STR00236##

[0354] The title intermediate was prepared as described for the synthesis of intermediate 28c using tert-butyl 4-(7-fluoroisoquinolin-1-yl)piperazine-1-carboxylate (Y=quant.). LC-MS (M-H.sup.+)=232.2

Step 3—Synthesis of 7-fluoro-1-(4-{[(2S)-oxiran-2-yl]methyl} piperazin-1-yl)isoquinoline

[0355] ##STR00237##

[0356] 7-fluoro-1-(piperazin-1-yl)isoquinoline (trifluoroacetic acid salt, 448 mg, 1.3 mmol) was dissolved in MeOH (12 mL), NaOH (156 mg, 3.9 mmol) was added followed by (S)-(+)-epichlorohydrin (0.92 mL, 11.7 mmol) at −10° C. The reaction mixture was stirred at the same temperature for 30 h then was placed at 4° C. overnight. The mixture was filtered, the solid was washed with acetone and the organic phase was evaporated in vacuum. The residue was chromatographed on silica gel (SNAP 50, Cy:EtOAc:MeOH 6:3:1) to give 370 mg (98% yield) of 7-fluoro-1-(4-{[(2S)-oxiran-2-yl]methyl}piperazin-1-yl)isoquinoline as a yellow oil. LC-MS (M-H.sup.+)=288.3

Step 4—Synthesis of (2R)-cyclohexyl({(2R)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)acetic acid (formate salt, compound 139)

[0357] ##STR00238##

[0358] 7-fluoro-1-(4-{[(2S)-oxiran-2-yl]methyl}piperazin-1-yl)isoquinoline (60 mg, 0.21 mmol), (2R)-amino(cyclohexyl)acetic acid (98 mg, 0.63 mmol) and DIPEA (150 μL, 0.84 mmol) were dissolved in a mixture of isopropanol/water 1:1 and stirred at 90° C. for 3 h. The solvent was evaporated in vacuo, the residue was treated with EtOAc and chromatographed on C18 (SNAP 30, gradient: acetonitrile+0.1% formic acid 0-23% in water+0.1% formic acid) to give 35 mg (0.07 mmol, 33% yield) of (2R)-cyclohexyl({(2R)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)acetic acid (formate salt). LC-MS (M-H.sup.+)=445.2.

[0359] .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 0.99-1.36 (m, 5H), 1.53-1.78 (m, 6H), 2.47 (d, J=6.36 Hz, 2H), 2.65-2.81 (m, 5H), 2.86 (dd, J=11.98, 4.16 Hz, 1H), 3.02 (d, J=4.40 Hz, 1H), 3.28 (br. s., 6H), 3.88-4.00 (m, 1H), 7.45 (d, J=5.87 Hz, 1H), 7.65 (td, J=8.68, 2.69 Hz, 1H), 7.70 (dd, J=10.27, 1.96 Hz, 1H), 8.01 (dd, J=9.05, 5.62 Hz, 1H), 8.12 (d, J=5.87 Hz, 1H), 8.17 (s, 1H).

Preparation of Compound 140

[0360] Compound 140 was prepared as described herein below.

Step 1—Synthesis of (2S)-cyclohexyl({(2R)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)acetic acid (formate salt, compound 140)

[0361] ##STR00239##

[0362] Compound (2S)-cyclohexyl({(2R)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)acetic acid (formate salt) was prepared according to the synthesis described for compound 139 using (2S)-amino(cyclohexyl)acetic acid (Y=15%). LC-MS (M-H.sup.+)=445.3.

[0363] .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 0.99-1.39 (m, 5H), 1.54-1.78 (m, 6H), 2.52-2.54 (m, 2H), 2.67-2.90 (m, 6H), 3.06 (d, J=4.40 Hz, 1H), 3.27 (br. s., 4H), 3.88-3.99 (m, 1H), 7.45 (d, J=5.38 Hz, 1H), 7.65 (td, J=8.80, 2.45 Hz, 1H), 7.70 (dd, J=10.03, 2.20 Hz, 1H), 8.01 (dd, J=9.05, 5.62 Hz, 1H), 8.11 (d, J=5.38 Hz, 1H), 8.16 (s, 1H).

Preparation of Compound 141

[0364] Compound 141 was prepared as described herein below.

Step 1—Synthesis of (3R)-3-({(2R)-3-[4-(7-fluoroisoquinolin-1-yl) piperazin-1-yl]-2-hydroxypropyl}amino)-3-phenylpropanoic acid (formate salt, compound 141)

[0365] ##STR00240##

[0366] Compound (3R)-3-({(2R)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)-3-phenylpropanoic acid (formate salt) was prepared according to the synthesis described for compound 139 using (3R)-3-amino-3-phenylpropanoic acid (Y=19%). LC-MS (M-H+)=453.4.

[0367] .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 2.33-2.46 (m, 4H), 2.55-2.73 (m, 6H), 3.23 (br. s., 4H), 3.78-3.85 (m, 1H), 4.11 (dd, J=9.78, 4.89 Hz, 1H), 7.25-7.31 (m, 1H), 7.36 (t, J=7.58 Hz, 2H), 7.39-7.47 (m, 3H), 7.61-7.70 (m, 2H), 8.00 (dd, J=8.80, 5.87 Hz, 1H), 8.11 (d, J=5.87 Hz, 1H).

Preparation of Compound 142

[0368] Compound 142 was prepared as described herein below.

Step 1—Synthesis of (3R)-3-({(2S)-3-[4-(7-fluoroisoquinolin-1-yl) piperazin-1-yl]-2-hydroxypropyl}amino)-3-phenylpropanoic acid (formate salt, compound 142)

[0369] ##STR00241##

[0370] Compound (3R)-3-({(2S)-3-[4-(7-fluoroisoquinolin-1-yl)piperazin-1-yl]-2-hydroxypropyl}amino)-3-phenylpropanoic acid (formate salt) was prepared according to the synthesis described for compound 138 using (3R)-3-amino-3-phenylpropanoic acid (Y=19%). LC-MS (M-H+)=453.4.

[0371] .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 2.33-2.56 (m, 5H), 2.57-2.72 (m, 5H), 3.23 (br. s., 4H), 3.74-3.81 (m, 1H), 4.08 (dd, J=10.03, 4.65 Hz, 1H), 7.26-7.31 (m, 1H), 7.36 (t, J=7.58 Hz, 2H), 7.40-7.46 (m, 3H), 7.61-7.70 (m, 2H), 8.00 (dd, J=9.05, 5.62 Hz, 1H), 8.11 (d, J=5.87 Hz, 1H).

Preparation of Compound 167

[0372] Compound 167 was prepared as described herein below.

Step 1—Synthesis of 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl) piperidin-4-yl]amino}ethyl)amino]propan-1-ol (compound 167)

[0373] ##STR00242##

[0374] To a solution of methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate (compound 189, 45 mg, 0.1 mmol) in THF (10 mL) NaBH.sub.4 (19 mg, 0.5 mmol) was added. The mixture refluxed for 1 h then MeOH (0.13 mL, 3.3 mmol) was added. After stirring 18 h at reflux the mixture was cooled to rt, concentrated under vacuum and partitioned between water and DCM. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated under vacuum. The residue was purified by preparative HPLC (C18, CH.sub.3CN from 30% to 65% in H.sub.2O) to afford 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propan-1-ol (14 mg, 0.03 mmol, 30% yield over two steps). LC-MS (M-H.sup.+)=429.3.

[0375] .sup.1H NMR (300 MHz, CHLOROFORM-d) δ=8.12 (d, J=5.9 Hz, 1H), 7.76 (dd, J=5.6, 8.9 Hz, 1H), 7.67 (dd, J=2.6, 10.2 Hz, 1H), 7.40 (dt, J=2.6, 8.6 Hz, 1H), 7.27 (d, J=5.5 Hz, 1H), 4.01 (ddd, J=3.8, 5.8, 11.6 Hz, 1H), 3.86-3.63 (m, 2H), 3.49-2.72 (m, 13H), 2.20 (d, J=11.2 Hz, 2H), 2.09-1.54 (m, 8H), 1.45-0.87 (m, 4H).

Preparation of Compound 189

[0376] Compound 189 was prepared as described herein below.

Step 1—Synthesis of tert-butyl [1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]carbamate

[0377] ##STR00243##

[0378] Potassium carbonate (4.6 g, 33 mmol) was added to a stirred solution of 1-chloro-7-fluoroisoquinoline (5 g, 27.5 mmol) and tert-butyl piperidin-4-ylcarbamate (13.8 g, 68.8 mmol) in methyl isopropyl ketone (50 mL) at room temperature. The resulting mixture was heated to reflux overnight. UPLC check showed the reaction was complete. The mixture was then allowed to cool to room temperature then water was added. The mixture was extracted with EtOAc, the combined organic phases were washed with brine and dried over sodium sulfate. The solvents were evaporated under reduced pressure and the resulting residue was purified by flash chromatography on silica gel (from 100% hexane to hexane/Ethyl acetate 8:2) to obtain tert-butyl [1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]carbamate (7.1 g, 20.6 mmol, 75% yield). LC-MS (M-H.sup.+)=346.2

Step 2—Synthesis of 1-(7-fluoroisoquinolin-1-yl)-N-[(4-methoxyphenyl)methyl]piperidin-4-amine hydrochloride

[0379] ##STR00244##

[0380] To a solution of tert-butyl [1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]carbamate (5.5 g, 15.9 mmol) in dry DMF (50 mL) NaH (60% mineral oil, 1.5 g, 31.8 mmol) was added. After stirring 1 h at room temperature p-methoxybenzyl chloride (5 mL, 31.8 mmol) was added. The reaction mixture was stirred at room temperature overnight then was poured into cold water (300 mL). The solution was extracted with EtOAc, the combined organic layers were washed with brine, dried over sodium sulfate and concentrated to dryness. The dried residue was dissolved in a solution of HCl in MeOH (1.25 M, 200 mL) and the mixture was refluxed for 2 h. The solvent was removed under vacuum and the resulting residue was treated with isopropanol and dried to give 1-(7-fluoroisoquinolin-1-yl)-N-[(4-methoxyphenyl)methyl]piperidin-4-amine hydrochloride (5.6 g, 15.3 mmol, 96% yield). LC-MS (M-H.sup.+)=366.2

Step 3—Synthesis of N-(2-chloroethyl)-1-(7-fluoroisoquinolin-1-yl)-N-[(4-methoxyphenyl)methyl]piperidin-4-amine

[0381] ##STR00245##

[0382] To a solution of 1-(7-fluoroisoquinolin-1-yl)-N-[(4-methoxyphenyl)methyl]piperidin-4-amine hydrochloride (2 g, 5 mmol) and DIPEA (0.9 mL, 5 mmol) in acetonitrile (10 mL) chloroacetaldehyde (50 wt % solution in H.sub.2O, 2.5 mL, 19.9 mmol) was added. After stirring 15′ at rt, acetic acid (0.57 ml) and NaBH(OAc).sub.3 (2.1 g, 10 mmol) were added. The mixture was stirred 3 h at rt adjusting pH to 5-6 by addition of 0.1 M HCl. After addition of NaHCO.sub.3 the solution was extracted with DCM, the organic phase was washed with water, treated with Na.sub.2SO.sub.4 and concentrated under vacuum. EtOAc was added to the residue, the resulting solid was filtered off and the solvent was evaporated under reduced pressure. A purification by flash chromatography (silica gel, CHCl.sub.3/MeOH 9:1) afforded N-(2-chloroethyl)-1-(7-fluoroisoquinolin-1-yl)-N-[(4-methoxyphenyl)methyl]piperidin-4-amine (650 mg, 1.5 mmol, 30% yield). LC-MS (M-H.sup.+)=428.1

Step 4—Synthesis of methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl][(4-methoxyphenyl)methyl] amino}ethyl) amino]propanoate

[0383] ##STR00246##

[0384] A solution of sodium iodide (39 mg, 0.26 mmol) and N-(2-chloroethyl)-1-(7-fluoroisoquinolin-1-yl)-N-[(4-methoxyphenyl)methyl] piperidin-4-amine (110 mg, 0.26 mmol) in DMF (5 mL) was stirred 15′ then was cooled to 0° C. Potassium carbonate (107 mg, 0.77 mmol) was added followed by one-pot addition of methyl 3-amino-3-cyclohexylpropanoate hydrochloride (114 mg, 0.51 mmol) in DMF (5 mL). The mixture was stirred at 90° C. overnight then was cooled and poured into cold water. The solution was extracted with DCM, the organic phase was dried over Na.sub.2SO.sub.4 and concentrated under vacuum to give methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl][(4-methoxyphenyl)methyl]amino}ethyl)amino]propanoate (110 mg, 0.20 mmol, 77% yield), that was progressed without further purification. LC-MS (M-H.sup.+)=577.3

Step 5—Synthesis of methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate (compound 189)

[0385] ##STR00247##

[0386] A mixture of methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl][(4-methoxyphenyl)methyl]amino}ethyl)amino]propanoate (110 mg, 0.20 mmol) and TFA (1 mL) in DCM (10 mL) was refluxed overnight. After cooling to rt sat. NaHCO.sub.3 was added, the resulting solution was extracted with EtOAc, the organic phase was dried over Na.sub.2SO.sub.4 and concentrated under vacuum. Purification by preparative HPLC (C-18, CH.sub.3CN from 10% to 45% in H.sub.2O) provided 58 mg (0.13 mmol, 65% yield) of methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate. LC-MS (M-H.sup.+)=457.3.

[0387] .sup.1H NMR (500 MHz, DMSO-d6/D.sub.2O) δ ppm 0.94-1.33 (m, 5H), 1.53-2.03 (m, 8H), 2.22 (d, J=11.11 Hz, 2H), 2.77-2.90 (m, 2H), 3.04-3.18 (m, 2H), 3.32-3.52 (m, 6H), 3.64-3.71 (m, 3H), 3.72-4.22 (m, 2H), 7.53 (d, J=5.90 Hz, 1H), 7.66-7.80 (m, 2H), 7.98-8.12 (m, 2H).

Preparation of Compound 190

[0388] Compound 190 was prepared as described herein below.

Step 1—Synthesis of lithium 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate (compound 190)

[0389] ##STR00248##

[0390] To a solution of methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate (compound 189, 60 mg, 0.13 mmol) in a mixture of THF (0.4 mL), MeOH (0.2 mL) and water (0.2 mL) LiOH×H2O (11 mg, 0.26 mmol) was added. The reaction mixture was stirred at 55° C. for 20 minutes then the solvent was evaporated and the crude was purified by reverse chromatography (C-18, from 100% water to 7/3 water/acetonitrile) to obtain lithium 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate (32 mg, 0.07 mmol, 56% yield). LC-MS (M-H.sup.+)=443.3.

[0391] .sup.1H NMR (500 MHz, DMSO-d6) δ ppm 0.82-1.29 (m, 5H), 1.41-1.80 (m, 8H), 1.86-2.04 (m, 3H), 2.09 (dd, J=15.30, 3.91 Hz, 1H), 2.58-2.79 (m, 6H), 2.93 (t, J=12.50 Hz, 2H), 3.64 (d, J=12.49 Hz, 2H), 7.41 (d, J=5.76 Hz, 1H), 7.59-7.69 (m, 2H), 7.99 (dd, J=8.71, 5.69 Hz, 1H), 8.09 (d, J=5.80 Hz, 1H).

Preparation of Compound 191

[0392] Compound 191 was prepared as described herein below.

Step 1—Synthesis of 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]-N-hydroxypropanamide (compound 191)

[0393] ##STR00249##

[0394] To a stirred solution of hydroxylamine hydrochloride (2.6 g, 40 mmol) in MeOH (12 mL) a solution of potassium hydroxide (3.3 g, 59 mmol) in MeOH (7 mL) was added slowly at 0° C. After addition, the mixture was stirred for 30 minutes at 0° C. then was allowed to reach room temperature. The resulting precipitate was filtered and the solution of free hydroxylamine was placed in 100 mL flask. Methyl 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]propanoate (compound 189, 87 mg, 0.19 mmol) in MeOH (5 mL) was added to this solution. The resulting solution was degassed at 0° C. for 30 minutes then was concentrated to dryness. The residue was dissolved in water, the aqueous phase was acidified to pH 5 by adding 1M HCl and extracted with EtOAc. The organic layer was washed with brine, dried over sodium sulfate and concentrated. The residue was purified by reverse chromatography (C-18, from 100% water to 8:2 water/acetonitrile) to give 3-cyclohexyl-3-[(2-{[1-(7-fluoroisoquinolin-1-yl)piperidin-4-yl]amino}ethyl)amino]-N-hydroxypropanamide (13 mg, 0.028 mmol, 15% yield). LC-MS (M-H.sup.+)=458.3.

[0395] .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 0.92-1.81 (m, 12H), 1.31-1.47 (m, 1H), 1.90-2.16 (m, 4H), 2.65-2.72 (m, 3H), 2.74-2.81 (m, 2H), 2.81-2.89 (m, 1H), 2.88-2.99 (m, 2H), 3.69 (d, J=12.72 Hz, 2H), 7.43 (d, J=5.26 Hz, 1H), 7.60-7.70 (m, 2H), 8.00 (dd, J=9.87, 5.92 Hz, 1H), 8.07-8.12 (m, 1H).

EXAMPLES

Example 1

Inhibition of DNA Gyrase or Topo IV in E. coli and S. aureus

[0396] The above compounds were tested for the inhibition of the enzyme DNA gyrase in a gyrase supercoiling assay and for the inhibition of the enzyme topoisomerase IV in a decatenation assay, in both Gram positive and Gram negative bacteria, according to the following methods.

[0397] Both the assays were carried out according to a set-up method modified from the article to Blanche F, et al. “Differential Behaviors of Staphylococcus aureus and Escherichia coli Type II DNA Topoisomerases”, Antimicrob. Agents Chemother., 1996, Vol. 40, No. 12 p. 2714-2720.

[0398] The compounds were screened at single concentration (200, 100 or 50 μM), in duplicate.

[0399] Ciprofloxacin and novobiocin were used as reference compounds, at single concentration of 200 and 50 μM, respectively.

[0400] DNA Gyrase Supercoiling Assay.

[0401] Reagents from S. aureus and E. coli Gyrase Supercoiling Assay kits (Inspiralis, UK) were used. A master mix with a total volume sufficient for the number of reactions to perform was prepared with the following reagents: 5× assay buffer, relaxed pBR322 substrate (0.5 μg/reaction), RNase-DNase free water. Aliquotes of this mix were dispensed in each tube, then 10× compound stock solutions or vehicle control (DMSO), were added to each reaction tube.

[0402] Reaction was started with E. Coli (2 U/reaction) or S. aureus (1 U/reaction) gyrase enzyme addition.

[0403] A sample added with an equal volume of dilution buffer was used as negative control (without enzyme).

[0404] The reaction tubes were gentle vortexed and incubated 30 minutes at 37° C. Each reaction was stopped by adding 30 μl of Stop Buffer and 30 μl chloroform/isoamyl alcohol (24/1), briefly vortexed for 5-10 seconds and centrifuged at 20000×g for 2 minutes. Samples were loaded onto 1% agarose gel and subjected to electrophoresis for 1 hour at 80V constant voltage in TAE (40 mM Tris-acetate, 2 mM EDTA).

[0405] Data acquisition and analysis. Treatment of relaxed pBR322 with DNA gyrase converted the relaxed topoisomers (DNAs of different linking number) to the supercoiled form of the plasmid, which migrates faster on an agarose gel. An upper band might also be visible, which consists of open-circular (nicked) DNA which is present in the relaxed substrate but co-migrates with some of the relaxed topoisomers.

[0406] Bands were visualized by ethidium bromide staining (dil 1:20000) for 30 minutes followed by destaining in distilled water for 10 minutes.

[0407] In order to evaluate the compounds activity on the enzyme, the bands of supercoiled DNAs in the gel were photographed by a digital imaging system ImageQuant LAS 4000 (GE Healthcare) according to manufacturer's instructions.

[0408] The fluorescent intensity of each band was analysed by ImageQuant TL software and it was expressed as volume (volume of the uncalibrated quantity of material in the image feature after subtraction of the background intensity by using rolling ball method).

[0409] Each band intensity was compared, as percentage, to vehicle sample band intensity, which served as positive control, on the same gel.

[0410] Inhibitory activity was expressed as percent of inhibition versus the positive control.

[0411] The results are summarized in the following Table 2.

[0412] Topoisomerase IV Decatenation Assay

[0413] S. aureus and E. coli Topoisomerase IV decatenation kits (Inspiralis, UK) were used. A master mix with a total volume sufficient for the number of reactions to perform was prepared with the following reagents: 5× assay buffer (50 mM HEPES-KOH (pH 7.6), 100 mM potassium glutamate, 10 mM magnesium acetate, 10 mM DTT, 1 mM ATP, 50 μg/ml albumin), kDNA substrate (200 ng/reaction), RNase-DNase free water. Aliquots of this mix were dispensed in each tube, then 10× compound stock solutions or vehicle control (DMSO), were added in each reaction tube.

[0414] Reaction was started with Topoisomerase IV enzyme (0.5 U/reaction) addition.

[0415] A sample added with an equal volume of dilution buffer was used as negative control (without enzyme).

[0416] The reaction tubes were gentle vortexed and incubated 30 minutes at 37° C. Each reaction was stopped by adding 30 μl of Stop Buffer and 30 μl of chloroform/isoamyl alcohol (24/1), briefly vortexed for 5-10 seconds and centrifuged at 20000×g for 2 minutes. Samples taken from the upper phase were loaded into 1% agarose gel and subjected to electrophoresis for 1 hour at 80V constant voltage in TAE (40 mM Tris-acetate, 2 mM EDTA).

[0417] Data acquisition and analysis. Due to the high molecular mass, kDNA could not enter an agarose gel under normal electrophoresis conditions, but remained in the wells. In the presence of Topo IV topoisomerase mini-circles (2.5 Kb) were released from kDNA by decatenation and were quickly and easily resolved in the gel at relatively high voltages.

[0418] Bands were visualized by ethidium bromide staining (dil 1:20000) for 30 minutes followed by destaining in distilled water for 10 minutes.

[0419] For single concentration screening assay, in order to evaluate the compounds activity on the enzymes, the bands of decatenated DNAs in the gel were photographed by a digital imaging system ImageQuant LAS 4000 (GE Healthcare) according to manufacturer's instructions.

[0420] The fluorescent intensity of each band was analyzed by ImageQuant TL software and it was expressed as volume (volume of the uncalibrated quantity of material in the image feature after subtraction of the background intensity by using rolling ball method).

[0421] Each band intensity was compared, as percentage, to vehicle sample band intensity, which served as positive control, on the same gel.

[0422] Inhibitory activity was expressed as percent of inhibition versus the positive control.

[0423] The results are summarized in the following Table 2.

TABLE-US-00002 TABLE 2 E. coli S. aureus Compound % inhibition % inhibition % inhibition % inhibition No DNA gyrase Topo IV DNA gyrase Topo IV 2 n/a 65 n/a n/a 3 59 n/a n/a n/a 4 n/a 53 84 n/a 21 n/a n/a 67 n/a 31 n/a n/a 65 n/a 35 n/a n/a 78 n/a 36 n/a n/a 90 n/a 38 n/a n/a 63 n/a 39 50 n/a 74 n/a 42 n/a n/a 84 n/a 43 n/a n/a 100  n/a 45 n/a n/a 81 n/a 50 92 n/a n/a n/a 53 64 n/a 57 n/a 58 100  n/a n/a n/a 60 100  n/a 89 n/a 68 100  n/a n/a n/a 70 86 n/a 75 n/a 71 56 n/a n/a n/a 74 65 n/a 61 n/a 75 69 n/a 61 n/a 81 80 n/a n/a n/a 82 85 n/a 82 n/a 83 69 n/a n/a n/a 85 78 n/a 86 n/a 87 79 n/a 96 n/a 96 88 n/a 91 n/a 97 85 n/a 86 n/a 101 100  n/a 100  n/a 107 92 n/a 74 n/a 117 75 n/a n/a n/a 118 78 n/a n/a n/a 119 n/a n/a 60 n/a 120 n/a n/a 76 n/a 122 58 n/a n/a n/a 123 52 n/a n/a n/a 128 n/a n/a 66 n/a 129 69 n/a n/a n/a 130 n/a n/a 74 n/a 132 n/a n/a 53 n/a 133 53 n/a 58 n/a 136 57 n/a 75 n/a 137 57 n/a 75 n/a 138 n/a n/a 89 n/a 139 n/a n/a 54 n/a 140 68 n/a 92 n/a 141 n/a n/a 65 n/a 142 n/a n/a 52 n/a 167 n/a n/a 70 n/a 189 n/a n/a 84 n/a 190 61 n/a 73 n/a 191 n/a n/a 81 n/a n/a = not active

[0424] The above results showed that the exemplified compounds effectively inhibited DNA gyrase or topoisomerase IV in E. coli, which is a Gram positive bacterium, and/or S. aureus, which is a Gram negative bacterium.