WO2001002387A1

WO2001002387A1 - N-imidazolylmethyl carboxamides as nitric oxide production inhibitors

Info

Publication number: WO2001002387A1
Application number: PCT/JP2000/004302
Authority: WO
Inventors: Ichiro Shima; Takehiko Ohkawa; Kazuhiko Ohne; Tatsuya Zenkoh; Kentaro Sato
Original assignee: Fujisawa Pharmaceutical Co Ltd
Current assignee: Fujisawa Pharmaceutical Co Ltd
Priority date: 1999-07-05
Filing date: 2000-06-29
Publication date: 2001-01-11
Anticipated expiration: 2002-01-05
Also published as: EP1196407A1; AUPQ142599A0; JP2003503489A

Abstract

The present invention relates to a compound of formula (I) wherein each symbol is as defined in the specification, and pharmaceutically acceptable salts thereof. The compound (I) of the present invention and pharmaceutically acceptable salts thereof possess a strong inhibitory activity on the production of nitric oxide (NO), and are useful for prevention and/or treatment of NO-mediated diseases in human being and animals.

Description

DESCRIPTION

N-IMIDAZOLYLMETHYL CARBOXAMIDES AS NITRIC OXIDE PRODUCTION INHIBITORS

TECHNICAL FIELD

This invention relates to new amide compounds and pharmaceutically acceptable salts thereof which are useful as medicament .

BACKGROUND ART

Some peptide compounds have been known as described in, for example, EP 0 394 989 A2.

DISCLOSURE OF INVENTION

This invention relates to new amide compounds.

One object of this invention is to provide the new and useful amide compounds and pharmaceutically acceptable salts thereof that possess a strong inhibitory activity on the production of nitric oxide (NO).

Another object of this invention is to provide a process for the preparation of the amide compounds and salts thereof.

A further object of this invention is to provide a pharmaceutical composition comprising said amide compound or a pharmaceutically acceptable salt thereof.

Still further object of this invention is to provide a use of said amide compounds or pharmaceutically acceptable salts thereof as a medicament for prophylactic and therapeutic treatment of NO-mediated diseases including respiratory diseases such as adult respiratory distress syndrome (ARDS) and asthma; cardiovascular diseases such as cardiovascular ischemia, myocarditis, heart failure, hypotension and atherosclerosis; endocrine diseases such as diabetes (e.g., insulin-dependent diabetes mellitus , etc . ) , complications of diabetes mellitus (e.g., diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, etc. ) and gout; renal diseases such as glomerulonephritis and renal failure; gastrointestinal diseases such as peptic ulcer and inflammatory bowel disease (e.g., ulcerative colitis , chronic colitis , etc. ) ; pancreatic diseases such as pancreatitis; hepatic diseases such as hepatitis and liver cirrhosis; diseases of bone or joint such as synovitis, arthritis, osteoarthritis , osteoporosis; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and multiple sclerosis; dermal diseases such as dermatitis and eczema; cancer such as solid tumors and metastasis; rejection by organ transplantation; shock (e.g. , septic shock, etc. ) ; and sepsis-induced systemic inflammatory response syndrome.

The object amide compounds of the present invention are novel and can be represented by the following general formula ( I ) :

wherein

R¹ is benzofuranyl substituted by halogen, or styryl substituted by halogen, R² is hydrogen or lower alkyl, R³ is pyridyl, optionally protected hydroxy or lower alkylsulfonylamino, R⁴ is lower alkyl,

R⁵ and R⁶ are the same or different and each is hydrogen or lower alkyl, and

X is CH or N, provided that ( i) R¹ is styryl substituted by halogen when R² is hydrogen, R³ is pyridyl, R⁵ and R⁶ are each hydrogen and X is CH, and (ii) X is N when R¹ is benzofuranyl substituted by halogen, R³ is pyridyl and R⁵ and R⁶ are each hydrogen.

Suitable pharmaceutically acceptable salts of the object compound ( I ) are conventional non-toxic salts and include, for example, a salt with a base or an acid addition salt such as a salt with an inorganic base, for example, an alkali metal salt (e.g. , sodium salt, potassium salt, etc.), an alkaline earth metal salt (e.g., calcium salt, magnesium salt, etc. ) , an ammonium salt; a salt with an organic base, for example, an organic amine salt (e.g., triethylamine salt, pyridinesalt, picolinesalt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc. ) ; an inorganic acid addition salt (e.g., hydrochloride, hydrobromide, sulfate, phosphate, etc.); an organic carboxylic or sulfonic acid addition salt (e.g., formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc. ) ; and a salt with a basic or acidic amino acid (e.g., arginine, aspartic acid, gultamic acid, etc.).

In the above and subsequent descriptions of the present specification, suitable examples and illustration of the various definitions which the present invention intends to include within the scope thereof are explained in detail as follows.

The term "lower" is used to intend a group having 1 to 6, preferably 1 to 4, carbon atom(s), unless otherwise provided.

Suitable "halogen" includes, for example, fluorine, bromine, chlorine and iodine.

Suitable "lower alkyl" and "lower alkyl moiety" in the term "lower alkylsulfonylamino" include straight or branched one having 1 to 6 carbon atom(s) , such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl and hexyl, in which more preferred one is C_j-C^ alkyl.

"Optionally protected hydroxy" includes hydroxy and protected hydroxy. Suitable examples of "hydroxy protective group" in the term "protected hydroxy" include acyl such as lower alkanoyl which may be substituted by one to three halogen atoms (e.g., acetyl, trichloroacetyl, etc.), mono(or di or tri)phenyl(lower)alkyl which may have one or more suitable substituent(s) (e.g., benzyl, 4- methoxybenzyl, trityl, etc.), trisubstituted silyl [e.g., tri( lower)alkylsilyl (e.g., trimethylsilyl, tert-butyldi ethylsilyl, etc.), etc.], tetrahydropyranyl and the like.

"Styryl substituted by halogen" means styryl which has halogen atomas a substituent on the benzene ring. Suitable examples of "styryl substituted by halogen" include 2-(2-chlorophenyl)ethenyl, 2-(3- chlorophenyl)ethenyl, 2-(4-chlorophenyl)ethenyl, 2-(2- bromopheny1)ethenyl, 2-(3-bromopheny1)ethenyl, 2-(4- bromophenyl )ethenyl, 2-( 2-fluorophenyl)ethenyl, 2-(3- fluorophenyl)ethenyl, 2-(4-fluorophenyl)ethenyl, and the like.

The object compound (I) of the present invention can be prepared by the following processes.

Process (1)

derivative group, eof

(II)

(I) or a salt thereof Process (2)

(IV) or a salt thereof

R¹

or a salt thereof

Process ( 3 )

protection of hydroxy group

(I)-2

or a salt thereof

wherein R¹, R², R³, R⁴, R⁵, R⁶ and X are each as defined above, R⁷ is lower alkyl, and R⁸ is hydroxy protective group.

The starting compounds can be prepared by the method of Preparation mentioned below or by a process known in the art for preparing their structurally analogous compounds.

The processes for preparing the object compound are explained in detail in the following.

Process (1)

The compound (I) or a salt thereof can be prepared by reacting the compound (II) or its reactive derivative at the amino group, or a salt thereof with the compound (III) or its reactive derivative at the carboxy group, or a salt thereof.

Suitable reactive derivative of the compound (II) includes Schiff ' s base type imino or its tautomeric enamine type isomer formed by the reaction of the compound (II) with a carbonyl compound such as aldehyde, ketone or the like; a silyl derivative formed by the reaction of the compound (II) with a silyl compound such as N,0- bis(trimethylsilyl)acetamide, N-trimethylsilylacetamide or the like; a derivative formed by the reaction of the compound ( II ) with phosphorus trichloride or phosgene.

Suitable reactive derivative of the compound (III) includes an acid halide, an acid anhydride and an activated ester. The suitable example may be an acid chloride; an acid azide; a mixed acid anhydride with an acid such as substituted phosphoric acid (e.g., dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoric acid, etc.), dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, alkanesulfonic acid (e.g. , methanesulfonic acid, ethanesulfonic acid, etc.), sulfuric acid, alkylcarbonic acid, aliphatic carboxylic acid (e.g., pivalic acid, pentanoic acid, isopentanoic acid, 2- ethylbutyric acid, trichloroacetic acid, etc.); aromatic carboxylic acid (e.g., benzoic acid, etc.); a symmetrical acid anhydride; an activated amide with imidazole, 4-substituted imidazole, dimethylpyrazole, triazole or tetrazole; an activated ester (e.g., cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl [ (CH₃)₂N⁺=CH- ] ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester, etc.); or an ester with an N-hydroxy compound (e.g., N,N- dimethylhydroxylamine, l-hydroxy-2-(lH)-pyridone, N- hydroxysuccinimide, N-hydroxybenzotriazole, N-hydroxyphthalimide, l-hydroxy-6-chloro-lH-benzotriazole, etc.). These reactive derivatives can optionally be selected from them according to the kind of the compound (III) to be used.

The reaction is usually carried out in a conventional solvent such as water, acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N- dimethylformamide, pyridine or any other organic solvents which do not adversely affect the reaction, or the mixture thereof.

When the compound (III) is used in free acid form or its salt form in the reaction, the reaction is preferably carried out in the presence of a conventional condensing agent such as N,N'- dicyclohexylcarbodiimide; N-cyclohexyl-N' - morpholinoethylcarbodiimide; N-cyclohexyl-N' - ( 4- diethylaminocyclohexyl )carbodiimide; N,N' -diisopropylcarbodiimide; N-ethyl-N' - ( 3-dimethylaminopropyl )carbodiimide; N,N-carbonyl-bis- (2-methylimidazole) ; pentamethyleneketene-N-cyclohexylimine; diphenylketene-N-cyclohexylimine; ethoxyacetylene; 1-alkoxy-l- chloroethylene; trialkyl phosphite; isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride); phosphorus trichloride; thionyl chloride; oxalyl chloride; triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt; 2-ethyl-5- (m- sulfophenyl)isoxazolium hydroxide intramolecular salt; l-(p- chlorobenzenesulfonyloxy)-6-chloro-lH-benzotriazole; so-called Vilsmeier reagent prepared by the reaction of N,N-di_methylformamide with thionyl chloride, phosgene, phosphorus oxychloride, etc.; or the like.

The reaction may also be carried out in the presence of an organic or inorganic base such as an alkali metal bicarbonate, tri(lower)alkylamine, pyridine, N-(lower)alkylmorpholine, N,N- di(lower)alkylbenzylamine, or the like.

The reaction temperature is not critical, and the reaction is usually carried out under cooling to heating.

Process (?)

The compound (i)-lora salt thereof can be prepared by reacting the compound (IV) or a salt thereof with the compound (V).

This reaction can be carried out in the same manner as in or a manner similar to Example 5. Process (3)

The compound ( I )-3 or a salt thereof can be prepared by subjecting the compound (I)-2 or a salt thereof to protection of hydroxy group.

This reaction can be carried out in the same manner as in or a manner similar to Example 7.

Suitable salts of the starting compounds and their reactive derivatives in Processes ( 1 ) to ( 3 ) can be referred to the ones as exemplified for the compound (I).

The compounds obtained by the above process can be isolated and purified by a conventional method such as pulverization, recrystallization, column chromatography, reprecipitation, or the like.

It is to be noted that the compound ( I ) and the other compounds may include one or more stereoisomer(s ) such as optical isomer(s) and geometrical isomer(s) due to asymmetric carbon atom(s) and double bond(s), and all of such isomers and mixtures thereof are included within the scope of this invention.

The object compounds (I) and pharmaceutically acceptable salts thereof include solvates [e.g., enclosure compounds (e.g., hydrate, etc . ) ] .

The object compounds (I) and pharmaceutically acceptable salts thereof possess a strong inhibitory activity on the production of nitric oxide (NO) .

Accordingly, the object compounds (I) and pharmaceutically acceptable salts thereof are expected to possess a nitric oxide synthase (NOS)-inhibitory activity or a NOS-production inhibitory activity.

Accordingly, the object compounds (I) and pharmaceutically acceptable salts thereof are useful for prevention and/or treatment of NO-mediated diseases including respiratory diseases such as adult respiratory distress syndrome (ARDS) and asthma; cardiovascular diseases such as cardiovascular ischemia, myocarditis, heart failure, hypotension and atherosclerosis; endocrine diseases such as diabetes (e.g., insulin-dependent diabetes mellitus, etc.), complications of diabetes mellitus (e.g., diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, etc.) and gout; renal diseases such as glomerulonephritis and renal failure; gastrointestinal diseases such as peptic ulcer and inflammatory bowel disease (e.g., ulcerative colitis, chronic colitis, etc.); pancreatic diseases such as pancreatitis; hepatic diseases such as hepatitis and liver cirrhosis; diseases of bone or joint such as synovitis , arthritis , osteoarthritis , osteoporosis; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and multiple sclerosis; dermal diseases such as dermatitis and eczema; cancer such as solid tumors and metastasis; rejection by organ transplantation; shock (e.g., septic shock, etc.); and sepsis-induced systemic inflammatory response syndrome.

The object compounds (I) and pharmaceutically acceptable salts thereof are also useful for prevention and/or treatment of NO-mediated nervous diseases including central nervous system diseases such as CNS disorders, cerebrovascular diseases (e.g., cerebral infarction, cerebral ischemia, cerebral hemorrhage, etc . ) , migraine, Alzheimer ' s disease; peripheral nervous system diseases such as neuritis, pain (e.g., postherpetic neuralgia, reflex sympathetic dystrophy (RSD), causalgia, deafferentation pain syndrome, neuropathic pain, etc.), allodynia, hyperalgesia, neurological disorders and neuroprotection; Parkinson's disease; and amyotrophic lateral sclerosis.

Additionally, the object compounds (I) and pharmaceutically acceptable salts thereof are useful for treatment of sexual dysfunction such as male sexual dysfunction including erectile dysfunction, and female sexual dysfunction including orgasmic dysfunction related to clitoral disturbances.

Further, the object compounds (I) and pharmaceutically acceptable salts thereof are useful for prevention and/or treatment of NO-mediated ophthalmic diseases, including conjunctive diseases such as conjunctivitis (e.g., allergic conjunctivitis, vernal conjunctivitis, keratoconjunctivitis sicca, viral conjunctivitis, bacterial conjunctivitis, etc. ) ; uveal diseases such as uveitis (e.g. , Behcet disease, Harada disease, sympathetic ophthalmia, sarcoidosis, diabetic iritis , etc . ) ; scleral diseases such as scleritis ; corneal diseases such as corneal neovascularization, keratitis, corneal edema, corneal opacity, corneal dystrophy, keratoconus and neuroparalytic keratitis; retinal, vitreous diseases such as diabetic retinopathy, retinal artery occlusion, retinal vein occlusion, central serous chorioretinopathy, central hemorrhagic chorioretinitis , macular degeneration (e.g., age-related macular degeneration, etc.), retinal detachment, retinal pigmentary degeneration, macular neovascularization, macular hole, proliferative vitreoretinopathy, vitreous hemorrhage and vitreous opacity; lens diseases such as cataract (e.g., senile cataract, traumatic cataract, diabetic cataract, atopic cataract, etc. ) ; glaucoma such as primary open-angle glaucome, primary angle-closure glaucoma, normal tension glaucoma and neovascular glaucoma; ocular hypertension; vision disorders such as amblyopia, color vision defect and night blindness; refractive errors such as astigmatism, hyperopia, myopia and presbyopia; and lacrimal apparatus diseases such as dry eye syndromes , lacrimal duct obstruction and dacryocystitis .

In order to illustrate the usefulness of the object compound ( I ) , the pharmacological test result of the compound ( I ) is shown in the following.

Test Compound

Compound (a): (2E)-3-(4-chlorophenyl)-N-[ (lS)-l-[ l-methyl-5-[4-

(morpholin-4-yl )phenyl ] imidazol-2-yl ]-2-(2-pyridyl ) ethyl]-2- propenamide

Test 1 : Assay for inhibitory activity on the production of nitric oxide The murine macrophage cell line RAW264.7 (American Type Culture Collection, No. TIB71) was used in this study. RAW264.7 cells were grown on F75 plastic culture flasks at 37°C, 5% in Dulbecco's modified Eagle's medium (DMEM) supplemented with L-glutamine, penicillin, streptomycin and 10% heat-inactivated fetal bovine serum. They were removed from culture flasks by rubber cell scraper and were centrifuged and resuspended in DMEMwithout phenol red. Theywere plated in 96-well microtiter plates (10⁵ cells per well) and allowed to adhere over 2 hours. The test samples were added and the cells were preincubated for 1 hour. Thereafter the cells were activated with both of lipopolysaccharide (LPS) ( ljLLgl l ) and interferon γ (INF y ) (3 u/ml) for 18-24 hours. An equal volume of Griess reagent (1% sulfanilamide/0.1% N-naphthylethylenediamine dihydrochloride/2.5% H₃P0₄) was added and the cells were incubated at room temperature for 10 minutes. The absorbance was read at 570 nm using microplate reader and N0₂ ^~ was measured using NaN0₂ as a standard.

Test result:

Table 1

Test compound (10^"6M) Inhibition ( % )

(a) 100

Test 2: Protective effect of the compound (I) combined with FK 506 on rat cardiac allograft Method:

Experiments were performed on male Lewis and ACI rats weighing 175-200 g. Rats were anesthetizedwith sodium pentobarbital (50 mg/kg, i.p.), and underwent allogeneic (Lewis donor to ACI recipient) heterotopic intra-abdominal cardiac transplantation. Experimental groups were divided into single-drug group and combined-drug group. Single-drug dose of FK506, which was prepared in a manner similar to that disclosed in EP-0184162, was 0.32 mg/kg. Combined-drug dose was FK506 (0.32 mg/kg) + the compound (I) (10 mg/kg). The grafted hearts were monitored by daily palpation where complete rejection was defined as the cessation of palpable contractile activity. Each drug was suspended in a solution of 0.5% methylcellulose, and administered by daily gastric intubation in a volume of 5 ml/kg of body weight for 14 days.

The combination of the compound (I) and FK506 dramatically prolonged the graft survival.

The above experimental results indicate that the activity and/or efficacy of an immunosuppressant in rejection of transplantation can be remarkably and synergistically increased by administering compound (I) in combination, which has a strong inhibitory activity on the production of nitric oxide.

For therapeutic administration, the object compound (I) of the present invention and pharmaceutically acceptable salts thereof are used in the form of a conventional pharmaceutical preparation in admixture with a conventional pharmaceutically acceptable carrier such as an organic or inorganic solid or liquid excipient which is suitable for oral, parenteral or external administration. The pharmaceutical preparation may be compounded in a solid form such as granule, capsule, tablet, dragee, suppository or ointment, or in a liquid form such as solution, suspension or emulsion for injection, intravenous drip, ingestion, eye drop, etc. If needed, there may be included in the above preparation auxiliary substance such as stabilizing agent, wetting or emulsifying agent, buffer or any other commonly used additives.

The effective ingredient may usually be administered in a unit dose of 0.001 mg/kg to 500 mg/kg, preferably 0.01 mg/kg to 10 mg/kg, 1 to 4 times a day. However, the above dosage may be increased or decreased according to age, body weight and conditions of the patient or administering method.

The preferred embodiments of the amide compound of the present invention represented by the general formula (I) are as follows.

1) The compound of the formula (I) wherein

R¹ is benzofuranyl substituted by halogen, and

R³ is optionally protected hydroxy or lower alkylsulfonylamino, or a pharmaceutically acceptable salt thereof.

2) The compound of the formula (I) wherein R¹ is styryl substituted by halogen, and

R³ is pyridyl, optionally protected hydroxy or lower alkylsulfonylamino, or a pharmaceutically acceptable salt thereof.

3) The compound of the formula (I) wherein R¹ is styryl substituted by halogen, and

R³ is pyridyl or optionally protected hydroxy, or a pharmaceutically acceptable salt thereof. 4 ) The compound of the formula ( I ) wherein R¹ is benzofuranyl substituted by halogen, R³ is pyridyl, and X is N, or a pharmaceutically acceptable salt thereof.

The following Preparations and Examples are given for the purpose of illustrating the present invention in detail. Preparation 1 l-[4-(Morpholin-4-yl)phenyl]ethan-l-one

A mixture of l-(4-fluorophenyl)ethan-l-one (100 g), morpholine (126ml) and potassium carbonate (95 g) in N,N-dimethylformamide (DMF) (1 L) was heated at 150°C (bath temperature) for 48 hours. Thin-layer chromatography showed the exhaustion of the starting material. The reaction mixture was poured into water (2 L) , and the precipitate was collected and washed with water. Then, the residue was dissolved in ethyl acetate, dried over magnesium sulfate, and filtered. After removal of the solvent, l-[4-(morpholin-4-yl)phenyl]ethan-l-one (81.5 g) was obtained as a yellow-brown powder.

^-NMR (DMS0-d₆) δ 2.46(3H,s), 3.30(4H,dd,J=4Hz,J=5Hz ) , 3.72(4H,dd,J=4Hz,J=5Hz), 6.98(2H,d,J=9Hz) , 7.83(2H,d,J=9Hz). Preparation 2

2-Bromo-l-[ 4-(morpholin-4-yl )phenyl]ethan-1-one

To a solution of l-[4-(morpholin-4-yl)phenyl]ethan-l-one (170 g) in 48% hydrobromic acid (340 ml) was added a solution of bromine (43 ml) in 48% hydrobromic acid (60 ml) dropwise at 65-75°C during the period of 0.5 hour. The mixture was stirred at 65°C for an additional 1 hour. The reaction mixture was cooled to 10°C and the resulting precipitate was collected by filtration. The resulting solid (hydrobromide salt) was basified with saturated aqueous sodium hydrogencarbonate solution carefully, and extracted with chloroform (500 ml). The organic layer was dried over magnesium sulfate, and filtered. After removal of the solvent, 2-bromo-l-[4-(morpholin- 4-yl)phenyl]ethan-l-one (200 g) was obtained as a greenish yellow powder. Further purification was not attempted.

^-NMR (DMSO-d₆) δ 3.30-3.34(4H,dd,J=4Hz, =5Hz) , 3.34- 3.43(4H,m), 6.99(2H,d,J=9Hz) , 7.86(2H,d,J=9Hz) . Preparation 3

2- [ 2-[ 4-(Morpholin-4-yl )phenyl ]-2-oxoethyl ] isoindoline-1 , 3- dione

A mixture of 2-bromo-l-[4-(morpholin-4-yl)phenyl]ethan-l-one (200 g) and potassium phthalimide (137 g) in DMF (1500 ml) was stirred at 90°C for 1 hour. After cooling to 10°C, the precipitate was collected, and washed with cold DMF (150 ml) and water (250 mlX2), successively. 2-[ 2- [ 4-(Morpholin-4-yl )phenyl ] -2- oxoethyl] isoindoline-1, 3-dione (186 g) was obtained as a pale yellow wet solid. This compound was used for next step without further dry-up or purification.

^-N R (DMSO-d₆) δ 3.34 (4H,dd,J=4Hz,J=5Hz ) ,

3.72(4H,dd,J=4Hz,J=5Hz), 5.10(2H,s), 7.04 (2H,d,J=9Hz) , 7.86-7.97(6H,m). Preparation 4

2-Amino-l-[ 4-(morpholin-4-yl )phenyl ]ethan-1-one dihydrochloride

A slurry of 2-[2-[4-(morpholin-4-yl)phenyl]-2- oxoethyl] isoindoline-1, 3-dione (185 g) in concentrated hydrochloric acid (1900 ml) was heated under reflux for 8 hours. As a result, the slurry gradually dissolved and a clear pale yellow solution was obtained. After removal of the solvent in vacuo, the residual oily solid was triturated with methanol (400 ml). The resulting precipitate was collected by filtration, and washed with methanol (100 ml) to give 2-amino-l-[4- (morpholin-4-yl)phenyl]ethan-l-one dihydrochloride (125 g) as a white powder.

^-NMR (DMSO-d₆) δ 3.35(4H,dd,J=4Hz, J=5Hz ) ,

3.73(4H,dd,J=4Hz,J=5Hz), 4.45(2H,d,J=4.5Hz ) , 7.03(2H,d,J=9Hz), 7.87(2H,d, J=9Hz ) . Preparation 5

( 2S )-2- (tert-Butoxycarbonylamino) -N- [ 2-[4-(morpholin-4- yl )phenyl ] -2-oxoethyl]-3-( 2-pyridyl )propanamide

To a solution of 2-amino-l-[ 4-(morpholin-4-yl)phenyl ]ethan- 1-one dihydrochloride (3.71 g), (2S)-2-(tert- butoxycarbonylamino)-3-(2-pyridyl)propanoic acid (5.73 g) and diphenylphosphoryl azide (3.48 g) in DMF (70 ml) was added dropwise N,N-diisopropylethylamine (4.41 ml) at 0°C and the mixture was stirred for 20 minutes. The mixture was heated to ambient temperature and stirred for 8 hours. The resulting mixture was diluted with ethyl acetate (200 ml) and washed successively with water, saturated aqueous sodium hydrogencarbonate solution and brine. The organic layer was dried over magnesium sulfate and concentrated in vacuo. The residual solid was triturated with ethyl acetate-diisopropyl ether (1:2) to give ( 2S)-2- (tert-butoxycarbonylamino)-N- [ 2-[4- (morpholin-4- yl)phenyl]-2-oxoethyl] -3- (2-pyridyl )propanamide (2.06 g) as off- white crystals.

ESI-MS: 469 (M+H)

'H-NMR (300MHz,CDCl₃) δ 1.46(9H,s), 3.20-3.38(6H,m) , 3.82- 3.88(4H,m), 4.60(2H,d,J=5Hz), 4.64-4.74 (lH,br) , 6.37- 6.45(lH,br), 6.86(2H,d,J=9Hz ) , 7.14 ( lH,dd, J=5Hz,J=8Hz) , 7.21(lH,d,J=8Hz), 7.59( lH,t,J=8Hz) , 7.82-7.90(3H,m) , 8.56(lH,d,J=5Hz).

Preparation 6 l-(tert-Butoxy)-N-[(lS)-l-[l-methyl-5-[4-(morpholin-4- yl )phenyl] imidazol-2-yl] -2- ( 2-pyridyl )ethyl] formamide

To a solution of ( 2S)-2- (tert-butoxycarbonylamino )-N-[ 2-[4- (morpholin-4-yl )phenyl]-2-oxoethyl]-3- ( 2-pyridyl )propanamide (2.0 g) in acetic acid (4.0 ml) and xylene (60 ml) was added methylamine (40% in water, 4.0 ml) and the mixture was refluxed for 3 hours in a round-bottomed flask equipped with a Dean-Stark apparatus. The mixture was cooled to ambient temperature and a mixture of acetic acid (4.0ml) andmethylamine (40% in water, 4.0 ml) was added to the solution. The solutionwas refluxed for 2 hours and cooled to ambient temperature. The solution was extracted with IN hydrochloric acid (100 ml) and the aqueous layer was washed with ethyl acetate ( 50 ml ) . The aqueous layer was basifiedwith saturated aqueous sodium hydrogencarbonate solution and extractedwith ethyl acetate ( 100 ml ) . The organic layer was washed successivelywith aqueous sodium hydrogencarbonate solution and brine, dried over magnesium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent; 2% methanol in chloroform) to give 1- (tert-butoxy) -N- [ (lS)-l-[l-methyl-5-[4- (morpholin-4-yl )phenyl ] imidazol-2-yl ] -2-( 2- pyridyl )ethyl ] formamide (1.45 g) as yellow crystals.

ESI-MS: 464 (M+H)

^-NMR (300MHz,CDCl₃) δ 1.37(9H,s), 3.17-3.23 (4H,m) ,

3.40(3H,s), 3.41-3.47(2H,m), 3.83-3.92(4H,m) , 5.33- 5.47(2H,m), 6.93(1H,S), 6.94 (2H,d,J=9Hz) , 7.08-7.16(2H,m) , 7.21(2H,d,J=9Hz), 7.56( lH,t, J=8Hz) , 8.55(lH,d,J=5Hz) . Preparation 7

( IS ) -1-[ l-Methyl-5- [ 4- (morpholin-4-yl)phenyl] imidazol-2- yl ]-2- (2-pyridyl )ethylamine

To a solution of l-(tert-butoxy)-N-[ (lS)-l-[l-methyl-5-[4- (morpholin-4-yl )phenyl] imidazol-2-yl ]-2-( 2- pyridyl )ethyl]formamide (1.43 g) in dichloromethane (25 ml) was added trifluoroacetic acid (5.0 ml) at 0°C and the mixture was stirred at ambient temperature for 2.5 hours. The resulting mixture was concentrated in vacuo and the residue was dissolved in water (20 ml) . The aqueous layer was basified with saturated aqueous sodium hydrogencarbonate solution and extracted with chloroform ( 80 ml ) . The organic layer was dried over magnesium sulfate and concentrated in vacuo. The residue was triturated with ethyl acetate-diisopropyl ether (1:2) to give (lS)-l-[l-methyl-5-[4-(morpholin-4- yl)phenyl]imidazol-2-yl] -2- (2-pyridyl) ethylamine (1.02 g) as yellow crystals .

ESI-MS: 364 (M+H)

'H-NMR (300MHz,CDCl₃) δ 3.17-3.23(4H,m), 3.27-3.47 (2H,m) , 3.49(3H,s), 3.84-3.91(4H,m), 4.58( lH,dd,J=5Hz,J=8Hz ) , 6.95(2H,d,J=9Hz), 6.97(lH,s), 7.11-7.18(2H,m) , 7.23(2H,d,J=9Hz), 7.59( lH,t, J=8Hz ) , 8.58(lH,d, J=5Hz ) . Example 1

(2E)-3-(4-Chlorophenyl )-N-[ ( IS)-1-[ l-methyl-5-[4- (morpholin-4-yl )phenyl] imidazol-2-yl ]-2- ( 2-pyridyl ) ethyl]-2- propenamide

To a solution of 4-chlorocinnamic acid (9.04 g) in dichloromethane (90 ml) were added oxalyl chloride (6.48 ml) and 1 drop of DMF, and the mixture was stirred at ambient temperature for 2 hours. After removal of the solvent by evaporation, the residual acid chloride was dissolved in dichloromethane (50 ml) . This solution was added to a solution of ( lS)-l-[l-methyl-5-[4-(morpholin-4- yl)phenyl]imidazol-2-yl] -2- (2-pyridyl) ethylamine (18 g) in dichloromethane (150 ml) at 0°C. After the mixture was stirred at ambient temperature for 4 hours, the mixture was washed successively with saturated aqueous sodium hydrogencarbonate solution and brine and dried over sodium sulfate. The solvent was removed by evaporation to give (2E)-3-(4-chlorophenyl)-N-[ ( lS)-l-[l-methyl-5-[4- (morpholin-4-yl )phenyl ] imidazol-2-yl ]-2- (2-pyridyl )ethyl]-2- propenamide (26 g) .

^XH-NMR (300MHz, CDC1₃) δ 3.21(4H,t, J=7Hz ) , 3.46-3.64. (5H,m) , 3.88(4H,t,J=7Hz), 3.83 ( lH,q, J=7Hz) , 6.31 ( lH,d, J=15Hz) , 6.89-6.97(3H,m), 7.08-7.22(4H,m) , 7.23-7.33 (3H,m) , 7.43(lH,d,J=15Hz), 7.55(lH,m), 7.68( lH,d,J=8Hz ) , 8.54(lH,m). Preparation 8

( 2R^*, 6S^*)-4 ' - ( 2 , 6-Dimethylmorpholin-4-yl )acetophenone

A suspension of 4-fluoroacetophenone (40 g), cis-2,6- dimethylmorpholine (66.7 g) and potassium carbonate (38 g) in DMF (400 ml) was stirred at 120°C for 65 hours. The resulting mixture was diluted with water (800 ml) and extracted with ethyl acetate (800 ml) . The organic layer was washed successively with saturated aqueous sodium hydrogencarbonate solution and brine, dried over magnesium sulfate and concentrated in vacuo. The residual solid was triturated with n-hexane (200 ml) to give (2R^*,6S*)-4 '-(2,6-dimethylmorpholin-4- yl)acetophenone (45.5 g) as brown crystals.

ESI-MS: 234.2 (M+H)

'H-NMR (300MHz, CDC1₃) δ 1.28(6H,d,J=6Hz) , 2.47-2.52 (2H,m) , 2.53(3H,s), 3.58-3.66(2H,m), 3.70-3.83 (2H,m) , 6.86(2H,d,J=9Hz), 7.88(2H,d, J=9Hz) . Preparation 9

2-Bromo-[ (2R^*, 6S*)-4 '- ( 2 , 6-dimethylmorpholin-4- yl ) ]acetophenone

To a solution of (2R*,6S*)-4 '-(2,6-dimethylmorpholin-4- yl )acetophenone (20.8 g) in 47% hydrobromic acid (40 ml) was added dropwise a solution of bromine in 47% hydrobromic acid (5.0 ml) at 55°C over 45 minutes and the mixture was stirred at 65°C for 1.5 hours. The resulting mixture was cooled to 10°C and the solid was collected by filtration. The solid was dissolved in water and the aqueous solution was basified with saturated aqueous sodium hydrogencarbonate solution. The aqueous layer was extracted with chloroform (100 ml) and the organic layer was washed with brine, then dried over magnesium sulfate and concentrated in vacuo . The residual solid was treated with cold ethyl acetate-diisopropyl ether (1:5, 18 ml), collected by filtration, and then washed successively with ethyl acetate- diisopropyl ether (1:10, 11 ml) and diisopropyl ether (5.0 ml) to give 2-bromo-[ ( 2R^*, 6S*) -4 ' - ( 2 , 6-dimethylmorpholin-4-yl ) ]acetophenone (9.54 g) as slightly green crystals.

ESI-MS: 312.0(M+)

^-NMR (300MHz, CDCl₃) δ 1.28( 6H,d,J=6Hz ) , 2.52-2.63(2H,m) , 3.61-3.69(2H,m), 3.70-3.82 (2H,m) , 4.37(2H,s), 6.86(2H,d,J=9Hz), 7.90(2H,d,J=9Hz) . Preparation 10

[ ( 2R*, 6S*) -4 ' - ( 2 , 6-Dimethylmorpholin-4-yl) ]-2- phthalimidoacetophenone

A mixture of 2-bromo-[ (2R^*,6S*)-4 '-(2,6-di_methylmorpholin-4- yl )]acetophenone (13.8 g) and phthalimide potassium salt (8.6 g) in DMF ( 140 ml ) was stirred at 100°C for 30 minutes . The resulting mixture was cooled to 5°C and diluted with water (300 ml). The solid was collected by filtration and washed with water (40 mlX2) to give [ ( 2R^*, 6S*) -4 ' - (2 , 6-dimethylmorpholin-4-yl) ]-2- phthalimidoacetophenone (17.5 g) as yellow crystals. ESI-MS: 379.2 (M+H) 'H-NMR (300MHz, CDC1₃) δ 1.29(6H,d,J=6Hz) ,

2.57(2H,dd,J=10Hz,J=12Hz), 3.65(2H,dd,J=2Hz,J=10Hz) , 3.72-3.84(2H,m), 5.07(2H,s), 6.88(2H,d,J=9Hz) , 7.70- 7.79(2H,m), 7.85-7.94(4H,m). Preparation 11

2-Aπtino-[ (2R*, 6S^*)-4 '-( 2 , 6-dimethylmorpholin-4- yl) ]acetophenone dihydrochloride A suspension of [ (2R*,6S*)-4 '-(2,6-dimethylmorpholin-4-yl) ]- 2-phthalimidoacetophenone (17.5 g) in concentrated hydrochloric acid (170 ml) was stirred at 120°C for 20 hours. The resulting clear solution was concentrated in vacuo and the residue was diluted with water (80 ml). The mixture was filtered and the filtrate was concentrated in vacuo. The residual solid was treated with cold methanol (15 ml) and the mixture was stirred at 0°C for 45 minutes. The solid was collected by filtration and washed with cold methanol-ethyl acetate (1:2, 24 ml) to give 2-amino-[ (2R^*,6S*)-4'~ ( 2, 6-dimethylmorpholin-4-yl )]acetophenone dihydrochloride (10.13 g) as colorless crystals.

ESI-MS: 249.1 (M+H)

^-NMR (300MHz, DMSO-d₆) δ 1.17 ( 6H,d,J=6Hz) ,

2.44(2H,dd,J=llHz,J=13Hz), 3.58-3.72 (2H,m) , 3.87(2H,dd,J=2Hz,J=13Hz), 4.42 (2H,d, J=4Hz) , 7.05(2H,d,J=9Hz), 7.85(2H,d, J=9Hz ) , 8.27-8.41(2H,br s) . Preparation 12

(2S)-2-[ (tert-Butoxy)carbonylamino]-N-[2-[ (2R*,6S^*)-4-(2,6- dimethylmorpholin-4-yl)phenyl]-2-oxoethyl] -3- ( 2- pyridy1)propanamide

To a solution of 2-amino-[ (2R*,6S^*)-4 '-(2,6- dimethylmorpholin-4-yl ) ]acetophenone dihydrochloride (1.50 g) , (2S)-2-[ (tert-butoxy)carbonylamino]-3- ( 2-pyridyl)propanoic acid (1.31 g) and diphenylphosphoryl azide (1.11 ml) in DMF (30 ml) was added dropwise N,N-diisopropylethylamine (2.44 ml) at 0°C and the mixture was stirred for 30 minutes. The temperature of the reaction mixture was gradually raised to ambient temperature and mixture was stirred for 15 hours under a nitrogen atmosphere. The resulting mixture was diluted with ethyl acetate ( 75 ml) and washedwith saturated aqueous sodium hydrogencarbonate solution (40 ml) . The organic layer was extracted with IN hydrochloric acid (60 ml) and the aqueous layer was basifiedwith saturated aqueous sodium hydrogencarbonate solution, and then extracted with chloroform (100 ml). The organic layer was dried over magnesium sulfate and concentrated in vacuo. The residual solid was triturated with ethyl acetate-diisopropyl ether (1:3, 20 ml) to give (2S)-2-[ (tert-butoxy)carbonylamino ]-N-[2- [ (2R*,6S*)-4- ( 2 , 6-dimethylmorpholin-4-yl )phenyl ] -2-oxoethyl ]-3- (2- pyridyl)propanamide (1.66 g) as off-white crystals. ESI-MS: 497.4 (M+H)

'H-NMR (300MHZ, CDC1₃) δ 1.28(6H,d,J=6Hz) , 1.46(9H,s), 2.55(2H,dd, J=llHz,J=13Hz) , 3.26 ( lH,dd,J=7Hz, J=15Hz) , 3.31-3.43(lH,m), 3.63 (2H,dd, J=2Hz,J=13Hz) , 3.69- 3.82(2H,m), 4.59(2H,d,J=4Hz) , 4.63-4.74( lH,m) , 6.34(lH,m), 6.85(2H,d,J=9Hz), 7.14 (lH,dd,J=5Hz,J=8Hz) , 7.21(lH,d,J=8Hz), 7.59( lH,dt,J=2Hz,J=8Hz) , 7.79- 7.88(lH,m), 7.84(2H,d,J=9Hz), 8.54 ( lH,d,J=5Hz) . Preparation 13

1- (tert-Butoxy) -N-[ ( 1S )-1- [ 1-methyl-5- [ ( 2R^*, 6S*)-4- (2 , 6- dimethylmorpholin-4-yl )phenyl] imidazol-2-yl ] -2- ( 2- pyridyl)ethyl ] formamide

To a solution of (2S )-2- [ (tert-butoxy)carbonylamino ] -N-[2- [ ( 2R^*, 6S*) -4-( 2 , 6-dimethylmorpholin-4-yl)phenyl ]-2-oxoethyl ]-3- ( 2- pyridy1)propanamide (2.80 g) in acetic acid (9.7 ml) and xylene (70 ml) was added methylamine (40% in water, 4.1 ml) and the mixture was refluxed for 1.5 hours in a round-bottomed flask equipped with a

Dean-Stark apparatus . The mixture was cooled to 50°C under a nitrogen atmosphere and then acetic acid (9.7 ml) and methylamine (40% in water, 4.1 ml) were added to the mixture. The mixture was refluxed for 1.5 hours. The mixture was extracted with IN hydrochloric acid (85 ml) and the aqueous layer was basified with saturated aqueous sodium hydrogencarbonate solution, then extracted with ethyl acetate (100 ml 2). The organic layer was washed successively with saturated aqueous sodium hydrogencarbonate solution and brine, dried over magnesium sulfate and concentrated in vacuo . The residue was purified by silica gel column chromatography (eluent; 2-5% methanol in chloroform) to give l-(tert-butoxy)-N-[ (lS)-l-[l-methyl-5- [ ( 2R^*, 6S*) -4- ( 2 , 6-dimethylmorpholin-4-yl)phenyl] imidazol-2-yl] -2- ( 2-pyridyl) ethyl]formamide (2.37 g) as a brown amorphous solid.

ESI-MS: 492.4 (M+H)

^-NMR (300MHz, CDC1₃) δ 1.27(6H,d,J=6Hz) , 1.37(9H,s),

2.45(2H,dd,J=llHz,J=13Hz), 3.37(3H,s), 3.39-3.53(4H,m) , 3.75-3.87(2H,m), 5.32-5.42 (lH,m) , 5.47-5.53 ( lH,m) , 6.92(lH,s), 6.93(2H,d,J=9Hz), 7.07-7.15(2H,m) , 7.18 ( 2H,d,J=9Hz ), 7.55(lH,dt,J=2Hz , J=8Hz ) , 8.54(lH,d,J=5Hz). Preparation 14

( IS )-l-[ l-Methyl-5- [ ( 2R^*, 6S*)-4- ( 2 , 6-dimethylmorpholin-4- yl )phenyl ] imidazol-2-yl] -2- ( 2-pyridyl)ethylamine

To a suspension of 1-(tert-butoxy)-N- [ (lS)-l-[l-methyl-5-

[ ( 2R*, 6S*) -4- ( 2 , 6-dimethylmorpholin-4-yl )phenyl] imidazol-2-yl ]-2- (2-pyridyl)ethyl] formamide (2.36 g) in methanol (5.0 ml) was added

4N hydrogen chloride in ethyl acetate (20 ml) at 0°C and the solution was stirred at 0°C for 10 minutes. The temperature of the reaction mixture was raised to ambient temperature and the suspension was stirred for 1.5 hours. The mixture was diluted with water (80 ml), and the organic layer and the aqueous layerwere separated. The aqueous layer was basified with sodium hydrogencarbonate (3.0 g), then extracted with chloroform ( 30 ml X 2 ) . The organic layer was dried over magnesium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent; 2-10% methanol in chloroform) to give (lS)-l-[l-methyl-5-[ (2R*,6S^*)-4-(2,6- dimethylmorpholin-4-yl )phenyl] imidazol-2-yl]-2- ( 2- pyridyl )ethylamine (1.70 g) as pale-yellow syrup. ESI-MS: 392.3 (M+H) ^-NMR (300MHz, CDC1₃) δ 1.28 ( 6H,d,J=6Hz) ,

2.46(2H,dd,J=llHz,J=13Hz) , 3.32(lH,dd,J=8Hz,J=14Hz) , 3.41(lH,dd,J=6Hz, J=14Hz), 3.47(3H,s), 3.49(2H,dd,J=2Hz,J=13Hz) , 3.75-3.88(2H,m) , 4.57(lH,dd,J=6Hz,J=8Hz), 6.94(2H,d,J=9Hz) , 6.96(1H,S), 7.10-7.18(2H,m), 7.21(2H,d,J=9Hz) , 7.59(lH,dt,J=2Hz,J=8Hz), 8.57 ( lH,d,J=4Hz) .

Example 2

1- [ ( 2E )-2-( 4-Chlorophenyl )ethenyl ]-N- [ ( IS)-1-[ l-methyl-5- [ ( 2R*, 6S*) -4-( 2 , 6-dimethylmorpholin-4-yl )phenyl ] imidazol-2-yl]-2- (2-pyridyl)ethyl]formamide

To a solution of (lS)-l-[l-methyl-5-[ (2R*,6S*)-4-(2,6- dimethylmorpholin-4-yl)phenyl] imidazol-2-yl]-2- ( 2- pyridyl)ethylamine (1.02 g), 4-chlorocinnamic acid (500 mg) and 1-hydroxybenzotriazole (387 mg) in DMF (20.0 ml) was added l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (549 mg) at 0°C, and then the mixture was stirred at ambient temperature for 16 hours. The resulting mixture was diluted with water (40 ml) and extracted with ethyl acetate (40 ml) . The organic layer was extracted with IN hydrochloric acid (25 ml) and the aqueous layer was basified with saturated aqueous sodium hydrogencarbonate solution, then extracted with ethyl acetate (40 ml). The organic layer was washed with brine, dried over magnesium sulfate and concentrated in vacuo. The residual syrup was purified by silica gel column chromatography (eluent; 5% methanol in chloroform). The syrup obtained was solidified with diisopropyl ether and the solid was collected by filtration to give l-[ (2E)-2-(4-chlorophenyl) ethenyl ]-N-[ (IS) -1-

[ l-methyl-5-[ ( 2R^*, 6S^*)-4-( 2 , 6-dimethylmorpholin-4- yl)phenyl ]imidazol-2-yl] -2- (2-pyridyl)ethyl] formamide (1.05 g) as an off-white powder.

ESI-MS: 556.2 (M+)

'H-NMR (300MHZ, CDC1₃) δ 1.27 (6H,d,J=6Hz) ,

2.46(2H,dd,J=llHz,J=13Hz), 3.47(3H,s), 3.48-3.69(4H,m) , 3.74-3.88(2H,m), 5.83 ( lH,q,J=8Hz) , 6.37 ( lH,d,J=16Hz ) , 6.92(1H,S), 6.94(2H,d,J=9Hz), 7.09-7.20(4H,m) , 7.25- 7.37(4H,m), 7.46( lH,d,J=16Hz) , 7.52-7.60(2H,m) , 8.54(lH,d,J=4Hz). Preparation 15

( 2S, 3R)-2- [ (tert-Butoxy)carbonylamino ]-N-[2- [4- (morpholin- 4-yl)phenyl ]-2-oxoethyl] -3-(phenylmethoxy)butanamide

To a stirred solution of ( 2S,3R)-2-[ (tert- butoxy)carbonylamino]-3-(phenylmethoxy)butanoic acid (3 g) in DMF (25 ml) were added diphenylphosphoryl azide (2.2 ml) and 2-amino-l- [ 4-(morpholin-4-yl)phenyl]ethan-l-one dihydrochloride (2.83 g) at 4°C. N,N-Diisopropylethylamine (5.6 ml) was added dropwise at 0°C (orange suspension) . After removing ice bath, the reaction mixture was stirred at room temperature for 8 hours (from orange suspension to yellow solution). After evaporation of the solvent, the residue was diluted with a mixture of ethyl acetate and tetrahydrofuran, washed with saturated aqueous sodium hydrogencarbonate solution, water and brine, dried over anhydrous magnesium sulfate and concentrated in vacuo. The residue was washed with ethyl acetate and n-hexane to give ( 2S, 3R)-2-[ (tert-butoxy)carbonylamino ]-N-[ 2- [4- (morpholin-4- yl)phenyl]-2-oxoethyl]-3-(phenylmethoxy)butanamide (4.60 g) as light yellow crystals.

MS(m/z) 511(M⁺+1), 131(bp)

'H-NMR (CDC1₃) δ 1.22(3H,d,J=7Hz), 1.48(9H,s),

3.34(4H,d,J=5Hz), 3.86(4H,d, J=5Hz ) , 4.22-4.35(2H,m) , 4.59 ( 2H,AB, J=10.5Hz,J=10Hz ) , 4.68 ( 2H,d,J=7Hz ) , 5.52(lH,d,J=7Hz), 6.89(2H,d, J=8Hz) , 7.24-7.33 ( 5H,m) , 7.55(lH,s), 7.90(2H,d,J=8Hz) . Preparation 16

1- (tert-Butoxy) -N-[ ( IR, 2R)-1- [ l-methyl-5-[ 4- (morpholin-4- yl )phenyl]-lH-imidazol-2-yl] -2-(phenylmethoxy)propyl ] formamide

In a flask equipped with a Dean-Stark trap, a mixture of (2S, 3R)-2- [ (tert-butoxy)carbonylamino]-N-[2- [4- (morpholin-4- yl)phenyl]-2-oxoethyl]-3-(phenylmethoxyJbutanamide (3.58 g) and 40% methylamine solution (12 ml) in acetic acid (12 ml) and xylene (72 ml ) was refluxed for 2 hours and allowed to cool to ambient temperature in a nitrogen atmosphere. Acetic acid (12 ml) and 40% methylamine solution (12 ml) were added to the mixture and the mixture was refluxed again for 4 hours. The mixture was concentrated, neutralized with aqueous sodium hydrogencarbonate solution, and extracted three times with ethyl acetate. The combined extracts were washed successively with saturated aqueous sodium hydrogencarbonate solution and brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel, chloroform/methanol=70/l) to give 1- (tert-butoxy)-N- [ (1R,2R)-1- [ l-methyl-5-[ 4- (morpholin-4-yl )phenyl ]-lH-imidazol-2-yl ]-2- (phenylmethoxy)propyl] formamide (830 mg) as a pale yellow amorphous solid.

MS (ESI) m/z 507(M+H)⁺

'H-NMR (300MHz,CDCl₃) δ 1.18(3H,d,J=6Hz), 1.45(9H,s), 3.15- 3.25(4H,m), 3.51(3H,s), 3.82-3.92 (4H,m) ,

4 . 01 ( lH ,qd, J=6Hz , J=5Hz ) , 4 .58 ( lH,A of AB, JAB=12Hz , OCH₂Ph ) , 4 . 65 ( lH, B of AB, JAB=12Hz , OCH₂Ph) , 5 .02 ( lH,dd, J=8Hz , J=5Hz ) , 5.70(lH,br d, J=8Hz), 6.94 (2H,d,J=9Hz) , 6.96(lH,s), 7.17-7.36(7H,m) . Preparation 17

1- (tert-Butoxy) -N-[ ( IR, 2R) -2-hydroxy-l-[ l-methyl-5-[4- (morpholin-4-yl )phenyl]-lH-imidazol-2-yl ]propyl ] formamide

A solution of l-(tert-butoxy)-N-[ (lR,2R)-l-[ l-methyl-5-[4- (morpholin-4-yl )phenyl ] -lH-imidazol-2-yl ]-2-

(phenylmethoxy)propyl] formamide (808 mg) in acetic acid (4 ml) was hydrogenated (3.5 atm) over 20% palladium hydroxide on carbon (173 mg) at 40°C for 10 hours. After the catalyst was filtered off, the filtrate was concentrated, neutralized with saturated aqueous sodium hydrogencarbonate solution, and extracted three times with ethyl acetate. The combined extracts were washed with brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel, chloroform/methanol=50/l) to give l-(tert-butoxy)-N-[ (lR,2R)-2- hydroxy-1-[l-methyl-5-[4-(morpholin-4-yl)phenyl]-lH-imidazol-2- y1 ]propyl] formamide (413 mg) as a white amorphous solid.

MS(ESI) m/z 417(M+H)⁺

'H-NMR (300MHz,CDCl₃) δ 1.28(3H,d,J=6Hz) , 1.45(9H,s), 3.19- 3.24(4H,m), 3.59(3H,s), 3.85-3.91 (4H,m) , 4.44 ( lH,qd, J=6Hz ,J=2Hz ) , 4.74 ( 1H,dd, J=10Hz ,J=2Hz ) , 5.35(lH,br d, J=10Hz), 6.88(lH,s), 6.96(2H,d,J=9Hz) , 7.27(2H,d,J=9Hz). Preparation 18

( IR, 2R)-2-Hydroxy-l- [ l-methyl-5-[ 4-(morpholin-4-yl )phenyl ]- lH-imidazol-2-yl]propylamine trihydrochloride

To an ice-cooled mixture of 1-(tert-butoxy)-N-[ (lR,2R)-2- hydroxy-1-[l-methyl-5- [4-(morpholin-4-yl)phenyl ]-lH-imidazol-2- yl]propyl] formamide (403 mg) and methanol (0.8 ml) was added 4N hydrogen chloride in ethyl acetate ( 3.2 ml ) , and the mixture was stirred at room temperature for 3 hours . The mixture was concentrated and the residue was dried in vacuo to give ( 1R,2R) -2-hydroxy-l-[1-methyl- 5- [4-(morpholin-4-yl )phenyl] -lH-imidazol-2-yl ]propylamine trihydrochloride (411 mg) as a pale yellow amorphous solid. MS (ESI) m/z 317(M-3HC1+H)⁺ -NMR (300MHz,DMSO-d₆) δ 1.17 ( 3H,d, J=6Hz ) , 3.15-3 ,30(4H,m) , 3.70-3.82(4H,m), 3.83(3H,s), 4.41 ( lH,qd,J=6Hz, J=6Hz ) , 4.87(lH,brd, J=6Hz), 7.13 (2H,d,J=9Hz) , 7.41 (2H,d,J=9Hz ) , 7.85(lH,s), 9.20(3H,br s). Example 3 l-(5-Chlorobenzo[b]furan-2-yl)-N-[(lR,2R)-2-hydroxy-l-[l- methyl-5-[4- (morpholin-4-yl) henyl]-lH-imidazol-2- yl]propyl]formamide

To an ice-cooled mixture of (lR,2R)-2-hydroxy-l-[l-methyl- 5-[4-(morpholin-4-yl )phenyl ] -lH-imidazol-2-yl]propylamine trihydrochloride (213 mg), 5-chlorobenzo[b]furan-2-carboxylic acid (109 mg), 1-hydroxybenzotriazole (68 mg), and l-(3- di_methylaminopropyl)-3-ethylcarbodiimide hydrochloride (120 mg) in DMF (1.7 ml) was added dropwise N,N-diisopropylethylamine (0.28 ml). The mixture was stirred at room temperature for 24 hours. Saturated aqueous sodium hydrogencarbonate solution was added to the mixture, and the mixture was extracted three times with chloroform. The organic layer was washed successively with saturated aqueous sodium hydrogencarbonate solution and brine, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel, chloroform/methanol=50/l) . The eluate was evaporated and triturated with ethyl acetate-hexane to give 1- ( 5-chlorobenzo[b] furan-2-yl )-N- [ ( IR, 2R)-2-hydroxy-l-[ 1-methyl- 5-[4-(morpholin-4-yl )phenyl] -lH-imidazol-2-yl]propyl ] formamide (159 mg) as a white powder, mp 188-189°C MS (ESI) m/z 495(M+H)⁺

^-NMR (300MHz, CDC1₃) δ 1.33 (3H,d,J=6Hz) , 3.15-3.26(4H,m) , 3.66 (3H,d,J=lHz ) , 3.82-3.92 ( 4H,m) ,

4.55(lH,qd,J=6Hz,J=2Hz), 5.30(lH,dd,J=10Hz,J=2Hz) , 6.94(1H,S), 6.95(2H,d,J=9Hz), 7.26(2H,d,J=9Hz) , 7.34- 7.50(4H,m), 7.62-7.68(lH,m) . Exampl 4

1-[ (E) -2-(4-Chlorophenyl )ethenyl] -N-[ ( IR,2R) -2-hydroxy-l- [ l-methyl-5-[4- (morpholin-4-yl)phenyl ]-lH-imidazol-2- yl]propyl]formamide The title compound was obtained from the compound of Preparation

18 in substantially the same manner as in Example 3. white powder mp 218-219°C MS (ESI) m/z 481(M+H)⁺

^-NMR (300MHz, CDC1₃) δ 1.30(3H,d,J=6Hz) , 3.14-3.29(4H,m) , 3.64(3H,s), 3.80-3.94(4H,m), 4.49( lH,qd,J=6Hz,J=2Hz ) , 5.25(lH,dd,J=10Hz,J=2Hz), 6.43 ( lH,d, J=16Hz ) , 6.52(lH,brd, J=10Hz), 6.91(lH,s), 6.96(2H,d,J=9Hz ) , 7.26(2H,d,J=9Hz) , 7.35(2H,d,J=8Hz), 7.43(2H,d,J=8Hz ) , 7.62 ( lH,d, J=16Hz) . Preparation 19

(2S ) -2-Benzyloxycarbonylamino-N- [ 2-[ 4- (morpholin-4- yl )phenyl ]-2-oxoethyl]-3- (tert-butoxycarbonylamino )propanamide

The title compound was obtained from the compound of Preparation 4 and ( 2S)-2-benzyloxycarbonylamino-3-(tert- butoxycarbonylamino)propanoic acid in substantially the same manner as in Preparation 12.

ESI-MS: 541.0 (M+H)

'H-NMR (300MHz, DMSO-d₆) δ 1.36(9H,s), 3.14-3.25( lH,m) , 3.28-3.35(5H,m), 3.70-3.77(4H,m) , 4.14-4.24 (lH,m) , 4.47-4.54(2H,m), 5.02 (lH,d,J=14Hz ) , 5.06( lH,d, J=14Hz) , 6.73-6.79(lH,m), 7.01 (2H,d,J=9Hz) , 7.28-7.39(6H,m) , 7.85(2H,d,J=9Hz), 8.11-8.17 ( lH,m) . Preparation 20

1-Benzyloxy-N-[ ( IS )-1-[ l-methyl-5-[4- (morpholin-4- yl )phenyl] imidazol-2-yl ]-2-(tert-butoxycarbonylamino )ethyl] - formamide

The title compound was obtained from the compound of Preparation

19 in substantially the same manner as in Preparation 13.

ESI-MS: 536.5 (M+H)

'H-NMR (300MHZ, CDC1₃) δ 1.42(9H,s), 3.18-3.25(4H,m) , 3.54- 3.72(2H,m), 3.58(3H,s), 3.85-3.92 (4H,m) , 5.05-5.16( lH,m) , 5.13(2H,s), 5.43-5.52(lH,m), 5.66-5.74 ( lH,m) , 6.91(lH,s), 6.96(2H,d,J=9Hz), 7.24(2H,d, J=9Hz ) , 7.30-7.39(5H,m) .

Preparation 21

( IS ) -1-[ l-Methyl-5-[ 4-(morpholin-4-yl )phenyl] imidazol-2- yl]-2-(tert-butoxycarbonylamino)ethylamine

A stirred solution of l-benzyloxy-N-[ (IS) -l-[ l-methyl-5- [4- (morpholin-4-yl )phenyl] imidazol-2-yl ]-2-(tert- butoxycarbonylamino) ethyl] formamide (3.37 g) in methanol (30 ml) was hydrogenated over palladium hydroxide (1.2 g) under 4 atm at 35°C for 4.5 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to give ( IS) -l-[ l-methyl-5- [4-(morpholin-4- yl)phenyl]imidazol-2-yl]-2- (tert-butoxycarbonylamino )ethylamine (2.52 g) as a brown amorphous solid.

ESI-MS: 402.2 (M+H)

"H-NMR (300MHz, DMSO-d₆) δ 1.34(9H,s), 3.14-3.21 (4H,m) , 3.28-3.53(2H,m), 3.60(3H,s), 3.72-3.79(4H,m) , 4.64- 4.71(lH,m), 7.02-7.10(3H,m), 7.27 (2H,d,J=8Hz) , 8.55- 8.68(2H,br) . Preparation 22

1- ( 5-Chlorobenzo[b] furan-2-yl )-N- [ ( IS )-1-[ l-methyl-5- [4- (morpholin-4-yl )phenyl] imidazol-2-yl ]-2-(tert- butoxycarbonylamino) ethyl] formamide

The title compound was obtained from the compound of Preparation

21 in substantially the same manner as in Example 2. mp 214-217°C

ESI-MS: 580.2 (M+)

'H-NMR (300MHz, CDC1₃) δ 1.40(9H,s), 3.18-3.25 (4H,m) , 3.59- 3.68(lH,m), 3.65(3H,s), 3.75-3.83 ( lH,m) , 3.85-3.92(4H,m) , 5.53-5.62(2H,m), 6.96(2H,d,J=9Hz) , 6.97(lH,s), 7.25(2H,d,J=9Hz), 7.34-7.45(3H,m) , 7.61( lH,d,J=8Hz) , 7.64(lH,d,J=2Hz). Preparation 23

1- (5-Chlorobenzo[b]furan-2-yl)-N- [ ( IS)-1-[ l-methyl-5-[4- (morpholin-4-yl )phenyl ] imidazol-2-yl ]-2-aminoethyl] formamide

The title compound was obtained from the compound of Preparation

22 in substantially the same manner as in Preparation 14. mp 184-188°C ESI-MS: 480.1(M+)

^-NMR (300MHz, DMSO-d₆) δ 3.04 ( lH,dd,J=6Hz,J=14Hz) , 3.11- 3.20(5H,m), 3.57(3H,s), 3.70-3.78(4H,m) , 5.18-5.26( lH,m) , 6.90(1H,S), 7.01(2H,d,J=9Hz), 7.29(2H,d,J=9Hz) , 7.49(lH,dd,J=2Hz,J=8Hz), 7.65(lH,s), 7 ,71( lH,d,J=8Hz) , 7.88(lH,d,J=2Hz). Example 5

1- ( 5-Chlorobenzo[b] furan-2-yl )-N- [ ( IS )-1-[ l-methyl-5- [4- (morpholin-4-yl )phenyl] imidazol-2-yl ]-2- methylsulfonylaminoethyl ] formamide

To a solution of l-(5-chlorobenzo[b]furan-2-yl)-N-[ (1S)-1- [ l-methyl-5-[4- (morpholin-4-yl )phenyl] imidazol-2-yl]-2- aminoethyl] formamide (120mg) and N,N-diisopropylethylamine (0.48ml) in dichloromethane (3 ml) was added methanesulfonyl chloride (30 mg) at 0°C . The reaction mixture was stirred at room temperature for 1.5 hours and diluted with chloroform. The organic layer was washed with saturated aqueous sodium hydrogencarbonate solution, dried over anhydrous magnesium sulfate and concentrated in vacuo. The residual solid was washed with ethyl acetate to give l-(5- chlorobenzo[b] furan-2-yl )-N- [ ( IS) -1- [ l-methyl-5- [4-(morpholin-4- yl )phenyl]imidazol-2-yl]-2-methylsulfonylaminoethyl ] formamide (87 mg) as colorless crystals. mp 231-234°C

ESI-MS: 558.2 (M+)

^-NMR (300MHz, DMSO-d₆) δ 2.91(3H,s), 3.12-3.20(4H,m) , 3.53-3.70(2H,m), 3.57(3H,s), 3.71-3.78(4H,m) , 5.40- 5.50(lH,m), 6.92(1H,S), 7.01 (2H,d,J=9Hz) , 7.23-7.34(3H,m) , 7.50(lH,dt,J=2Hz,J=8Hz), 7.65(lH,s), 7.71( lH,d,J=8Hz) , 7.90(lH,d,J=2Hz), 9.19(lH,d,J=8Hz ) . Preparation 24

( IR)-1-[ l-Methyl-5-[4-(morpholin-4-yl)phenyl]imidazol-2- yl ]-2- (phenylmethoxy)ethylamine trihydrochloride

The title compound was obtained from the compound of Preparation 4 in substantially the same manner as in Preparations 15, 16 and 18. Preparation 25

(2R)-2-Amino-2- [l-methyl-5-[4-(morpholin-4- yl )phenyl]imidazol-2-yl]ethan-1-ol triacetate

To a solution of ( lR)-l-[ l-methyl-5- [4- (morpholin-4- yl )phenyl] imidazol-2-yl] -2-(phenylmethoxy)ethylamine trihydrochloride (500 mg) in acetic acid (5 ml) was added palladium hydroxide (200 mg) , and the mixture was stirred at 45°C under a hydrogen atmosphere (4 atm) for 4 hours. After removal of the catalyst, the solvent was removed in vacuo to give (2R)-2-amino-2-[ l-methyl-5- [4- (morpholin-4-yl)phenyl] imidazol-2-yl]ethan-l-ol triacetate (370 mg) as a brown amorphous solid. Further purification was not attempted. Example 6

1- ( 5-Chlorobenzo[b] furan-2-yl ) -N- [2-hydroxy- ( IR)-1-[ 1- methyl-5- [4-(morpholin-4-yl)phenyl] imidazol-2-yl ]ethyl] formamide

The title compound was obtained from the compound of Preparation 25 in substantially the same manner as in Example 3. mp 188-190°C MS 481 (ES+)

^-NMR (CDC1₃) δ 3.18-3.24(4H,m), 3.65(3H,s), 3.85-3.90(5H,m) , 4.01(lH,dd,J=13.5Hz, J=3.0Hz),

4.34(lH,dd,J=13.5Hz,J=1.5Hz), 5.45-5.51 (lH,m) , 6.94- 6.97(2H,m), 7.23-7.27(2H,m), 7.35-7.46(3H,m) , 7.62- 7.65(2H,m).

[α_]D25° = +247.2° (c 1.04, CHCl₃:MeOH = 10:1) Example 7

( 2R)-2-[ ( 5-Chlorobenzo[b] furan-2-yl )carbonylamino]-2- [ 1- methyl-5-[4-(morpholin-4-yl )phenyl] imidazol-2-yl ]ethyl acetate

To a solution of l-(5-chlorobenzo[b]furan-2-yl)-N-[2- hydroxy- ( IR)-1- [ l-methyl-5-[ 4-(morpholin-4-yl )phenyl] imidazol-2- yl ]ethyl] formamide (31 g) in pyridine (120 ml) was added acetic anhydride (20 ml) at room temperature. After 1 hour, pyridine was removed in vacuo. Water (200 ml) was added to the residual oil, and the resulting precipitate was collected by filtration and washed with water (100 ml). The wet solid was recrystallized from ethanol (150 ml) and washed with ethanol (50 ml) to give (2R)-2-[(5- chlorobenzo [b] furan-2-yl )carbonylamino]-2- [ l-methyl-5-[4- (morpholin-4-yl)phenyl]imidazol-2-yl]ethyl acetate (27 g) as white crystals . mp 163-164°C

^:H-NMR (CDC1₃) δ 1.65(3H,s), 3.20-3.24 (4H,m) , 3.65(3H,s), 3 . 85-3 .90 ( 4H ,m) , 4 .53-4 . 57 ( lH ,m) , 5. 70-5 . 77 ( lH ,m) , 6 . 93-7 .00 ( 3H ,m) , 7 .24-7 .28 ( 2H ,m) , 7 .35-7 .65 ( 5H ,m) .

[ α ]D24 ° = +162 .72° ( c 1 . 14 , CHC1₃ ) Preparation 26

6-Chloro-N-methoxy-N-methylnicotinamide

To a mixture of 6-chloronicotinic acid (29 g), N,0- dimethylhydroxylamine hydrochloride (26.9 g) and 1- hydroxybenzotriazole (36.6 g) in DMF (300 ml) were added l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (42.3 g) and N,N-diisopropylethylamine (48 ml) at 5°C. The reaction mixture was stirred at 5°C for 1 hour. To the reaction mixture were added saturated aqueous sodium hydrogencarbonate solution (300 ml) and ethyl acetate (300 ml). The organic layer was separated, washed with saturated aqueous sodium hydrogencarbonate solution and dried over magnesium sulfate. After removal of the solvent in vacuo, 6-chloro-N- methoxy-N-methylnicotinamide (33.3 g) was obtained as a pale brown oil.

MS 200.99(ES+)

'H-NMR (CDC1₃) δ 3.40(3H,s), 3.56(3H,s), 7.40(lH,d,J=7.5Hz) , 8.03(lH,dd,J=7.5Hz,J=lHz), 8.78(lH,d,J=lHz ) . Preparation 27

1- ( 6-Chloro-3-pyridyl)ethan-l-one

6-Chloro-N-methoxy-N-methylnicotinamide (33 g) was placed in a three neck flask. Tetrahydrofuran (300 ml) was added thereto, and the mixture was cooled to -30°C. To the mixture was added a solution (165 ml) of 1.1M methyllithium in diethyl ether dropwise over a 10-minute period. After 5 minutes, the reaction mixture was poured into saturated brine. The organic layer was separated, washed with saturated aqueous ammonium chloride solution and dried over magnesium sulfate. After removal of the solvent in vacuo, the residual solid was triturated with a mixture of diisopropyl ether and n-hexane to give l-(6-chloro-3-pyridyl)ethan-l-one (16 g) as a tan color powder.

MS 155.94(ES+)

^XH-NMR (CDC1₃) δ 2.65(3H,s), 7.45( lH,d,J=7.5Hz) , 8.20(lH,dd,J=7.5Hz,J=lHz), 8.93 ( lH,d,J=lHz ) . 1_ [ 6-( 4-Morpholinyl) -3-pyridyl ]ethan-1-one A mixture of l-(6-chloro-3-pyridyl)ethan-l-one (5 g), morpholine (8.4 g) and potassium carbonate (4.4 g) in dimethyl sulfoxide (100 ml) was heated at 130°C (bath temperature) for 2 hours. Thin-layer chromatography showed a perfect consumption of the starting material (l-(6-chloro-3-pyridyl)ethan-l-one) . The reaction mixture was poured into saturated aqueous sodium hydrogencarbonate solution (200 ml), and the mixture was extracted with ethyl acetate (200 ml). The organic layer was concentrated. The residue was dissolved in diethyl ether (200 ml) and washed with saturated aqueous ammonium chloride solution ( 100 ml) . The organic layer was dried overmagnesium sulfate and filtered. The filtrate was concentrated in vacuo, and the residue was triturated with diisopropyl ether (50 ml) to give 1- [ 6- (4-morpholinyl)-3-pyridyl ]ethan-l-one (5.99 g) as a pale yellow powder.

MS 207.05(ES+)

'H-NMR (CDC1₃) δ 3.64-3.89(4H,m), 3.80-3.84 (4H,m) , 6.62 ( 1H,d,J=7.5Hz ) , 8.04 ( 1H,dd,J=7.5Hz ,J=lHz ) , 8.77(lH,d,J=lHz). Preparation 29

2-Bromo-l-[ 6- (4-morpholinyl ) -3-pyridyl ]ethan-l-one To a solution of l-[6-(4-morpholinyl)-3-pyridyl]ethan-l-one (3 g) in 48% HBr (5 ml) was added a solution of bromine (0.75 ml) in 48% HBr (2 ml) dropwise at 65°C. The mixture was heated at 65°C for 0.5 hour. After cooling on the ice bath, the resulting precipitate was collected by filtration and washed with a small amount of HBr. The obtained pale brown solid was basified with saturated aqueous sodium hydrogencarbonate solution. The resulting precipitate was collected by filtration and washed with water. The wet solid was dried under vacuum to give 2-bromo-l-[6-(4-morpholinyl)-3-pyridyl]ethan-l-one (3.6 g) as a brown solid.

MS 285.10 & 287.10(ES+)

'H-NMR (CDC1₃) δ 3.70-3,74(4H,m), 3.80-3.84(4H,m) , 6.63(lH,d,J=7.5Hz), 8.07 (lH,dd,J=7.5Hz,J=lHz) , 8.80(lH,d,J=lHz). Preparation 30 2- [ 2-{6- (4-Morpholinyl ) -3-pyridyl}-2-oxoethyl ] -1H- isoindole-1 , 3 ( 2H) -dione

A mixture of 2-bromo-l-[6-(4-morpholinyl)-3-pyridyl]ethan- 1-one (3.6 g) and potassium lH-isoindole-l,3(2H)-dione (2.5 g) in DMF (40 ml) was heated at 130°C for 15 minutes. After cooling, the resulting precipitate was collected by filtration, washed with DMF (4 ml) and water (10 ml), successively to give 2-[2-{6-(4- morpholinyl )-3-pyridyl}-2-oxoethyl]-lH-isoindole-1 , 3 (2H)-dione (2.7 g) as a pale yellow powder.

MS 352.12(ES+)

'H-NMR (CDC1₃) δ 3.70-3.74(4H,m), 3.80-3.84(4H,m) , 5.03 (2H,s), 6.63(lH,d,J=7.5Hz), 7.72-7.77(2H,m) , 7.87-7.92 (2H,m) , 8.04(lH,dd,J=7.5Hz,J=lHz), 8.83( lH,d,J=lHz ) . Preparation 31

2-Amino-l- [ 6- (4-morpholinyl ) -3-pyridyl ]ethan-1-one trihydrochloride

A solution of 2-[2-{6-(4-morpholinyl)-3-pyridyl}-2- oxoethyl]-lH-isoindole-l,3(2H)-dione (2.7 g) in concentrated hydrochloric acid (30 ml) was heated under reflux for 8 hours. After removal of the solvent in vacuo, the residual oily solid was triturated with methanol ( 5 ml ) , collected by filtration and washed with methanol to give 2-amino-l-[ 6-(4-morpholinyl) -3-pyridyl ]ethan-l-one trihydrochloride (0.34 g) as a brown powder.

MS 222.1(ES+) (free mw 221)

^XH-NMR (DMSO-d₆) δ 3.68 ( 8H,br-s ) , 4.42-4.48(2H,m) , 6.98 ( 1H,d,J=7.5Hz ) , 8.06 ( 1H,dd,J=7.5Hz ,J=lHz ) , 8.75(lH,d,J=lHz).

Preparation 32 tert-Butyl 2-[ [2-{6-(4-morpholinyl)-3-pyridyl}-2- oxoethyl ]amino] -2-oxo- ( IS ) -1-( 2-pyridylmethyl )ethylcarbamate

To a mixture of 2-amino-l-[6-(4-morpholinyl ) -3- pyridyl]ethan-1-one trihydrochloride (1.5 g) and (2S)-2-[ (tert- butoxycarbonyl )amino]-3- ( 2-pyridyl)propanoic acid ( 1.2 g) in DMF were added diphenylphosphoryl azide (1.3 g) and N,N-diisopropylethylamine

(3.5 ml) at 5°C. The reaction mixture was stirred at 5°C for 0.5 hour and at room temperature for 1.5 hours. To the reaction mixture was added saturated aqueous sodium hydrogencarbonate solution (50 ml), and the mixture was extracted with ethyl acetate. The organic layer was separated and extracted with IN hydrochloric acid. The aqueous layer was washed with ethyl acetate, basified with aqueous sodium hydrogencarbonate solution and extracted with diethyl ether-ethyl acetate (10:1). The organic layer was dried over magnesium sulfate and filtered. The filtrate was concentrated in vacuo, and the residual solid was triturated with diisopropyl ether to give tert-butyl 2- [ [ 2-{6-(4-morpholinyl )-3-pyridyl}-2-oxoethyl ]amino]-2-oxo- ( IS )-1- (2-pyridylmethyl)ethylcarbamate (1.42 g) as a white powder. MS 470.31(ES+)

^XH-NMR (CDC1₃) δ 1.45(9H,s), 3.19-3.43 (2H,m) , 3.66-3.70 (4H,m) , 3.77-3.81(4H,m), 4.54-4.58(2H,m) , 4.67(lH,m), 6.40(lH,br-s), 6.58( lH,d,J=7.5Hz) , 7.10-7.13 (2H,m) , 7.55-7.63(lH,m), 7.87 ( lH,br-s) , 7.97 ( lH,dd,J=7.5Hz, J=lHz ) , 8.52-8.56(lH,m), 8.73( lH,d,J=lHz) .

Preparation 33

( IS)-l-[l-Methyl-5-{6- (4-morpholinyl )-3-pyridyl}-lH- imidazol-2-yl]-2-(2-pyridyl ) ethylamine tert-Butyl 2-[ [2-{6-(4-morpholinyl)-3-pyridyl}-2- oxoethyl]amino] -2-oxo-( IS ) -1- ( 2-pyridylmethyl )ethylcarbamate (0.2 g) was dissolved in hot methanol (1 ml), and 4N hydrogen chloride in ethyl acetate (5 ml) was added to the solution. After 0.5 hour, the solvent was removed in vacuo. The residual solid was treated with saturated aqueous sodium hydrogencarbonate solution, and the mixture was extracted with chloroform. The organic layer was separated, dried over magnesium sulfate and filtered. The filtrate was concentrated in vacuo to give ( IS )-l-[l-methyl-5-{6-(4-morpholinyl)-3- pyridyl}-lH-imidazol-2-yl]-2-(2-pyridyl)ethylamine (0.15 g) as a brown oil.

MS 365.21(ES+)

'H-NMR (CDC1₃) δ 3.38-3.43(2H,m), 3.48(3H,s), 3.53-3.57 (4H,m) , 3.79-3.84(4H,m), 4.63-4.68( lH,m) , 6.67 (lH,d, =7.5Hz ) , 6.97(1H,S), 7.1-7.19(2H,m), 7.44 ( lH,dd,J=7.5Hz,J=lHz) , 7.56-7.63(lH,m), 8.14( lH,d,J=lHz) , 8.55-8.57(lH,m) . Example 8 5-Chloro-N- [(IS)-l-[l-methyl-5-{6-(4-morpholinyl )-3- pyridyl}-lH-imidazol-2-yl ] -2- (2-pyridyl )ethyl ]benzo [b] furan-2- carboxamide

To a mixture of (lS)-l-[ l-methyl-5-{6- (4-morpholinyl )-3- pyridyl}-lH-imidazol-2-yl]-2- (2-pyridyl) ethylamine (140 mg), 5- chlorobenzo[b]furan-2-carboxylic acid (76 mg) and 1- hydroxybenzotriazole (62 mg) in DMF (3 ml) were added l-(3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (81 mg) and N,N-diisopropylethylamine (0.08 ml), successively at 5°C. The reaction mixture was stirred at room temperature for 1 hour. Water ( 10 ml ) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 ml). The organic layer was extracted with IN HCl (5 ml) . The aqueous layer was washed with ethyl acetate, basified with saturated aqueous sodium hydrogencarbonate solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and filtered. The filtrate was concentrated, and the residual oily solid was purified with thin-layer chromatography (chloroform:methanol = 20:1) . The resulting solid was trituratedwith diethyl ether to give 5-chloro-N-[ (lS)-l-[l-methyl-5-{6-(4- morpholinyl ) -3-pyridyl}-lH-imidazol-2-yl ]-2- ( 2- pyridyl)ethyl]benzo[b]furan-2-carboxamide (45 mg) as a pale yellow powder. mp 128-130°C

MS 543.25(ES+)

'H-NMR (CDC1₃) δ 3.45(3H,s), 3.51-3.57 (4H,m) , 3.58-3.62(2H,m) , 3.80-3.84(4H,m), 5.88-5.96( lH,m) , 6.67 (lH,d, =7.5Hz ) , 6.98(lH,s), 7.10-7.15(2H,m), 7.34-7.46(3H,m) , 7.52- 7.58(lH,m), 7.63(lH,d,J=lHz) , 7.73( lH,dd,J=7.5Hz,J=lHz) , 8.12(lH,d,J=lHz), 8.52-8.55( lH,m) .

Example 9

( 2E) -3-( 4-Chlorophenyl )-N-[ ( IS )-1-[ l-methyl-5-{6- ( 4- morpholinyl)-3-pyridyl}-lH-imidazol-2-yl]-2- (2-pyridyl) ethyl]-2- propenamide

To a mixture of ( IS) -l-[l-methyl-5-{6- (4-morpholinyl ) -3- pyridyl}-lH-imidazol-2-yl]-2- (2-pyridyl) ethylamine (100 mg), (2E)-3-(4-chlorophenyl)-2-propenoic acid (50 mg) and 1- hydroxybenzotriazole (45 mg) in DMF (3 ml) were added l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (58 mg) and N,N-diisopropylethylamine (0.08 ml), successively at 5°C. The reaction mixture was stirred at room temperature for 1 hour. Water ( 10 ml ) was added to the reactionmixture, and the mixture was extracted with ethyl acetate (10 ml). The organic layer was extracted with IN HCl (5 ml) . The aqueous layer was washed with ethyl acetate, basified with saturated aqueous sodium hydrogencarbonate solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and filtered. The filtrate was concentrated, and the residual oily solid was purified with thin-layer chromatography (chloroform:methanol = 20:1). The resulting solid was triturated with a mixture of diethyl ether and ethyl acetate to give (2E)-3-(4- chlorophenyl )-N- [ ( IS )-1-[ l-methyl-5-{6- ( 4-morpholinyl) -3- pyridyl}-lH-imidazol-2-yl]-2-( 2-pyridyl )ethyl ]-2-propenamide ( 27 mg) as a white powder. mp 172-173°C

MS 529.25(ES+)

'H-NMR (CDC1₃) δ 3.44 (3H,s), 3.47-3.52 (2H,m) , 3.52-3.56(4H,m) , 3.80-3.84(4H,m), 5.78-5.85(lH,m) , 6.35( lH,d,J=15Hz ) , 6.76(lH,d,J=7.5Hz),6.93(lH,s), 7.08-7.15(2H,m) , 7.26- 7.58(7H,m), 8.11(lH,d,J=lHz), 8.52-8.54( lH,m) .

This application is based on application No. PQ 1425 filed in Australia on July 5 , 1999 , the content of which is incorporated hereinto by reference.

Claims

CLAIMS 1. A compound of the formula ( I ) :

wherein

X is CH or N, provided that ( i) R¹ is styryl substituted by halogen when R² is hydrogen, R³ is pyridyl, R⁵ and R⁶ are each hydrogen and X is CH, and (ii) X is N when R¹ is benzofuranyl substituted by halogen, R³ is pyridyl and R⁵ and R⁶ are each hydrogen, or a pharmaceutically acceptable salt thereof.

2. The compound of Claim 1 wherein R¹ is benzofuranyl substituted by halogen, and R³ is optionally protected hydroxy or lower alkylsulfonylamino, or a pharmaceutically acceptable salt thereof.

3. The compound of Claim 1 wherein R¹ is styryl substituted by halogen, or a pharmaceutically acceptable salt thereof.

4. The compound of Claim 3 wherein R³ is pyridyl or optionally protected hydroxy, or a pharmaceutically acceptable salt thereof.

5. The compound of Claim 1 wherein R¹ is benzofuranyl substituted by halogen, R³ is pyridyl, and X is N, or a pharmaceutically acceptable salt thereof.

6. A process for preparing a compound of the formula

wherein

X is CH or N, provided that ( i) R¹ is styryl substituted by halogen when R² is hydrogen, R³ is pyridyl, R⁵ and R⁶ are each hydrogen and X is CH, and (ii) X is N when R¹ is benzofuranyl substituted by halogen, R³ is pyridyl and R⁵ and R⁶ are each hydrogen, or a salt thereof, which comprises ( 1 ) reacting a compound of the formula

wherein R², R³, R⁴, R⁵, R⁶ and X are each as defined above, or its reactive derivative at the amino group, or a salt thereof, with a compound of the formula

R^COOH (III)

wherein R¹ is as defined above, or its reactive derivative at the carboxy group, or a salt thereof to give a compound of the formula

wherein R¹, R², R³, R⁴, R⁵, R⁶ and X are each as defined above, or a salt thereof, or

( 2 ) reacting a compound of the formula

wherein R¹, R², R⁴, R⁵, R⁶ and X are each as defined above, or a salt thereof, with a compound of the formula

R⁷-S0₂C1 (V)

wherein R⁷ is lower alkyl to give a compound of the formula

wherein R¹, R², R⁴, R⁵, R⁶, R⁷ and X are each as defined above, or a salt thereof, or

(3) subjecting a compound of the formula

wherein R¹, R², R⁴, R⁵, R⁶ and X are each as defined above, or a salt thereof to protection of hydroxy group to give a compound of the formula

wherein R¹, R², R⁴, R⁵, R⁶ and X are each as defined above, and R⁸ is hydroxy protective group, or a salt thereof.

6. A pharmaceutical composition comprising the compound of Claim 1 or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable carrier.

7. Use of the compound of Claim 1 or a pharmaceutically acceptable salt thereof as a medicament.

8. Use of the compound of Claim 1 or a pharmaceutically acceptable salt thereof as a medicament for prophylactic or therapeutic treatment of NO-mediated diseases.

9. Use of the compound of Claim 1 or a pharmaceutically acceptable salt thereof as a medicament for prophylactic or therapeutic treatment of NO-mediated disease selected from the group consisting of adult respiratory distress syndrome (ARDS); asthma; cardiovascular ischemia; myocarditis; heart failure; hypotension; atherosclerosis; diabetes including insulin-dependent diabetes mellitus; complications of diabetes mellitus including diabetic nephropathy, diabetic retinopathy and diabetic neuropathy; gout; glomerulonephritis; renal failure; peptic ulcer; inflammatory bowel diseases including ulcerative colitis and chronic colitis; pancreatitis; hepatitis; liver cirrhosis; synovitis; arthritis; osteoarthritis ; osteoporosis; autoimmune diseases including rheumatoid arthritis, systemic lupus erythematosus and multiple sclerosis; dermatitis; eczema; cancer; solid tumors; metastasis; rejection by organ transplantation; shock including septic shock; sepsis-induced systemic inflammatory response syndrome; CNS disorders; cerebral infarction; cerebral ischemia; cerebral hemorrhage; migraine; Alzheimer's disease; neuritis; postherpetic neuralgia; reflex sympathetic dystrophy (RSD); causalgia; deafferentation pain syndrome; neuropathic pain; allodynia; hyperalgesia; neurological disorders; neuroprotection; Parkinson's disease; amyotrophic lateral sclerosis; male sexual dysfunction including erectile dysfunction; female sexual dysfunction including orgasmic dysfunction related to clitoral disturbances; conjunctivitis including allergic conjunctivitis, vernal conjunctivitis, keratoconjunctivitis sicca, viral conjunctivitis and bacterial conjunctivitis; uveitis including Behcet disease, Harada disease, sympathetic ophthalmia, sarcoidosis and diabetic iritis; scleritis; corneal neovascularization; keratitis; corneal edema; corneal opacity; corneal dystrophy; keratoconus; neuroparalytic keratitis; diabetic retinopathy; retinal artery occlusion; retinal vein occlusion; central serous chorioretinopathy; central hemorrhagic chorioretinitis ; macular degeneration including age-related macular degeneration; retinal detachment; retinal pigmentary degeneration; macular neovascularization; macular hole; proliterative vitreoretinopathy; vitreous hemorrhage; vitreous opacity; cataract including senile cataract, traumatic cataract, diabetic cataract and atopic cataract; glaucoma including primary open-angle glaucome, primary angle-closure glaucoma, normal tension glaucoma and neovascular glaucoma; ocular hypertension; vision disorders including amblyopia, color vision defect and night blindness; refractive errors including astigmatism, hyperopia, myopia and presbyopia; dry eye syndromes; lacrimal duct obstruction and dacryocystitis .