AU2370801A - Non peptide tachykinin receptor antagonists - Google Patents

Description

WO 01/46176 1 PCT/EPOO/13349 NON PEPTIDE TACHYKININ RECEPTOR ANTAGONISTS 5 BACKGROUND OF THE INVENTION The mammalian tachykinins are a family of small peptides which share the common C-terminal sequence Phe-X-Gly-Leu-Met-NH 2 (Nakanishi S., Physiol 10 Rev., 67:117, 1987). It is now well established that substance P, neurokinin A (NKA) and neurokinin B (NKB) are widely distributed throughout the periphery and central nervous system (CNS), where they appear to interact with at least three receptor types referred to as NKI, NK 2 and NK3 (Guard S., et al., Neurosci. Int., 18:149, 1991). Substance P displays the highest affinity for the NKI 15 receptor, whereas NKA and NKB bind preferentially to the NK 2 and NK 3 receptors, respectively. All three receptors have been cloned and sequenced and shown to be members of the G-protein-linked 'super family' of receptors (Nakanishi S., Annu. Rev. Neurosci., 14:123, 1991). In the years since 1991, a number of high-affinity nonpeptide tachykinin receptor 20 antagonists have been reported (IDrugs, Vol.1, No.1, p. 73-91, 1998). A wealth of evidence supports the involvement of tachykinin neuropeptides in a variety of biological activities including pain transmission, neuronal excitation, secretion of saliva, angiogenesis,vasodilation, smooth muscle contraction, bronchoconstriction, activation of the immune system and neurogenic 25 inflammation (Pernow B, Pharmacol. Rev. 35:85, 1983). Accordingly, compounds capable of antagonising the effects of substance P at NKI receptors may be useful in treating or preventing a variety of CNS disorders including pain (inflammatory, surgical and neuropathic), anxiety, panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, 30 sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and psoriasis; WO 01/46176 2 PCT/EPOO/13349 gastrointestinal disorders including colitis, Crohn's disease, irritable bowel syndrome and satiety; allergic responses such as eczema and rhinitis; vascular disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis. 5 SUMMARY OF THE INVENTION The invention provides tachykinin receptor antagonists; the compounds have proved to be highly selective and functional tachykinin receptor antagonists. 10 These compounds are unique in the substitution at the C4 carbon. Compounds of the invention are those of Formula (I): 1 P4 F Ar R C H 2 ) mF7X - C I COZ R6 (CH 2 ) n R5 R2 (I) or a pharmaceutically acceptable salt thereof wherein R, m, X, RI, R2, n, Y, R3, 15 R4, R5 and R6 are as described below: * and A indicate all stereoisomers; R is phenyl, pyridyl, thienyl, 20 furyl, quinolyl isoquinolyl naphthyl, benzofuryl, WO 01/46176 3 PCT/EP00/13349 benzo[1,3]dioxole benzothienyl or, benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, 5 hydroxy, alkoxy, halogen,

CF

3 or

OCF

3 ; 10 m is an integer from 1 to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; R1 is (CH 2 )pY where p is 0 to 3 and Y is OH, OCH 3 , F, CF 3 , CO 2 H, N(CH 3

)

2 ,

NHCH

3 , NH 2 , COCF 3 , COCH 3 or NO 2 ; n is an integer from 1 to 2; 15 R2 is naphthyl or indolyl unsubstituted or N-substituted with hydroxy, alkyl,

CH

2

N(CH

3

)

2 , CH 2 N O or CH 2 N N formyl, CH 2 OH, Z is NR3 or 0, where R3 is H or CI-C 4 alkyl; R4 and R5 are each independently hydrogen, or (CH 2 )pR7 where: p is an integer of I to 3, and 20 R7 is H, CH 3 , CN, OH, OCH 3 , CO 2

CH

3 , NH 2 , NHCH 3 , or N(CH3)2; R6 is phenyl, pyridyl, thienyl, furyl, 25 pyrrolyl, pyrazolyl, imidazolyl, quinolyl, isoquinolyl, 30 naphthyl, WO 01/46176 4 PCT/EPOO/13349 indolyl, benzofuryl, benzothiophenyl, benzimidazolyl, or 5 benzoxazolyl, wherein each of the foregoing is unsubstituted, mono-, di or trisubstituted by alkyl, hydroxy, alkoxy, halogen, 10

CF

3 ,

NO

2 , N(CH3)2,

OCF

3 ,

SONH

2 , 15

NH

2 ,

CONH

2 ,

CO

2

CH

3 or CO 2 H, or R6 is: 20 straight alkyl of from 1 to 3 carbons, branched alkyl of from 3 to 8 carbons, cycloalkyl of from 5 to 8 carbons or heterocycloalkyl, each of which can be substituted with up to one or two substituents 25 selected from OH, CO 2 H, N(CH3)2,

NHCH

3 and 30 CH 3 ; or R5 and R6, when joined by a bond, can form a ring.

WO 01/46176 5 PCT/EPOO/13349 Preferred compounds of the invention are those of Formula I above wherein * is R or S, and A is R or S; -R is phenyl, 5 pyridyl, thienyl, furyl, quinolyl isoquinolyl 10 benzofuryl, benzo[1,3]dioxole benzothienyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, 15 hydroxy, alkoxy, halogen,

CF

3 or

OCF

3 ; 20 m is an integer from 1 to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; R1 is (CH 2 )pY where p is 0 to 3 and Y is OH, F, CF 3 , OCH 3 , CO 2 H, N(CH 3

)

2 ,

NHCH

3 , NH 2 , COCF 3 , COCH 3 or NO 2 ; n is an integer from 1 to 2; 25 R2 is naphthyl or indolyl unsubstituted or N-substituted with hydroxy, alkyl,

CH

2

N(CH

3

)

2 , CH 2 N O or CH 2 N N formyl, CH 2 OH, Z is NR3 or 0, where R3 is H or CH 3 ; R4 and R5 are each independently hydrogen, CH 3 or CH 2 OH; R6 is phenyl, 30 pyridyl, WO 01/46176 6 PCT/EPOO/13349 thienyl, furyl, pyrrolyl, cyclohexyl or 5 benzimidazolyl, wherein each of the foregoing is unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, 10 alkoxy, halogen,

CF

3 ,

NO

2 , or

N(CH

3

)

2 . 15 More preferred compounds of the invention are those of Formula I above wherein * is R or S, and A is R or S; R is phenyl, 20 pyridyl, thienyl, furyl, benzofuryl, benzo[1,3]dioxole 25 benzothienyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, 30 alkoxy, halogen, WO 01/46176 PCT/EPOO/13349

CF

3 or

OCF

3 ; m is an integer from I to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; 5 RI is (CH 2 )pY where p is 0 to 3 and Y is OH, OCH 3 , F, CF 3 , CO 2 H, N(CH 3

)

2 ,

NHCH

3 , NH2, COCF 3 , COCH 3 or NO 2 ; n is an integer from I to 2; R2 is indolyl unsubstituted or N-substituted with alkyl or formyl; Z is NR3 or 0, where R3 is H or CH 3 ; 10 R4 and R5 are each independently hydrogen, CH 3 , or CH 2 OH; R6 is phenyl, pyridyl, thienyl, furyl, 15 pyrrolyl, cyclohexyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, 20 hydroxy, alkoxy, halogen,

CF

3 ,

NO

2 or 25

N(CH

3

)

2 . Most preferred compounds of the invention are: 2-[(Benzofuran-2-ylmethyl)-amino]-3-(1H-indol-3-yl)-2-methoxymethyl-N-(1 phenyl-ethyl)-propionamide [S-(R*,R*)]; 30 2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H-indol-3-yl) N-(1-phenyl-ethyl)-propionamide (S,S); WO 01/46176 8 PCT/EPOO/13349 2-[(Benzofuran-2-ylmethyl)-amino]-2-hydroxymethyl-3-(1H-indol-3-yl)-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)]; 2-(3-Benzofuran-2-ylmethyl-ureido)-2-hydroxymethyl-3-(1H-indol-3-yl)-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)]; 5 2-(3-Benzofuran-2-ylmethyl-ureido)-3-(1H-indol-3-yl)-2-methoxymethyl-N-(1 phenyl-ethyl)-propionamide [S-(R*,R*)]; (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1-hydroxymethyl 1H-indol-3-ylmethyl)-N-((S)-1-phenyl-ethyl)-propionamide; (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-( 1 10 dimethylaminomethyl-1H-indol-3-ylmethyl)-N-((S)- 1 -phenyl-ethyl) propionamide. The invention additionally provides pharmaceutical formulations comprising a compound of Formula I admixed with a pharmaceutically acceptable carrier, 15 diluent or excipient therefor. The invention also provides a method for antagonizing NKI receptors in a mammal in need of treatment comprising administering to a mammal an effective amount of a compound of Formula I. The invention further provides a method for treating or preventing a variety of 20 CNS disorders including pain (inflammatory, surgical and neuropathic), anxiety, panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, obesity and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and 25 psoriasis; gastrointestinal disorders including colitis, Crohn's disease, irritable bowel syndrome and satiety; allergic responses such as eczema and rhinitis; vascular disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis comprising administering to a mammal in need of treatment an effective amount of a compound of Formula I. 30 The compounds of the invention, NKI receptor antagonists, being useful as anti angiogenic agents , the invention further provides a method for treating or WO 01/46176 9 PCT/EPOO/13349 preventing conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth, which comprises administering to a mammal in need of treatment an effective amount of a compound of Formula I. 5 The invention further provides agents for imaging NKI receptors in vivo in conditions such as ulcerative colitis and Crohn's disease. The invention further provides the use of a compound of Claim 1 for the preparation of a medicament intended for preventing or treating CNS disorders such as pain (inflammatory, surgical and neuropathic), anxiety, panic, 10 depression, major depression with anxiety, schizophrenia, neuralgia, stress, sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, obesity and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and psoriasis; gastrointestinal disorders including colitis, Crohn's disease, irritable bowel 15 syndrome and satiety; allergic responses such as eczema and rhinitis; vascular disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis; conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth. 20 This invention also concerns a process for the preparation of ((S)-2 benzylideneamino)-3-(lH-indol-3-yl)-propionic acid methyl ester which comprises reacting (S)-tryptophan methyl ester with benzaldehyde and recovering the desired product. The present invention also further concerns a process for the preparation of a 25 dimethylaminomethyltryptophan methyl ester, wherein ((S)-2-benzylidene amino)-3-(lH-indol-3-yl)-propionic acid methyl ester is reacted with 1 dimethylaminomethylbenzotriazole to give racemic a dimethylaminomethyltryptophan methyl ester. The present invention also discloses a process wherein the racemic methyl ester 30 obtained is separated into the (R)- and (S)-enantiomers.

WO 01/46176 10 PCT/EPOO/13349 Another embodiment of the present invention is the preparation of 2 [(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H-indol-3-yl)-N (1-phenyl-ethyl)-propionamide (S,S) wherein (S)-2-[(Benzofuran-2-ylmethyl) amino] -2-dimethylaminomethyl-3 -(1 H-indol-3 -yl)-propionic acid bis 5 hydrochloride is reacted with (S)-alpha-methylbenzylamine. A further embodiment of this invention is the preparation of (R)-C [(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1 -hydroxymethyl-1H indol-3 -ylmethyl)-N-((S)- 1 -phenyl-ethyl)-propionamide wherein 2 10 [(Benzofuran-2-ylmethyl)-amino] -2-dimethylaminomethyl-3 -(1H-indol-3 -yl)-N (1-phenyl-ethyl)-propionamide (S,S) is reacted with potassium hexamethyldisilazide and formaldehyde. The invention also concerns the preparation of (R)-C-[(Benzofuran-2-ylmethyl) 15 amino] -dimethylamino-C-(1 -dimethylaminomethyl- 1H-indol-3 -ylmethyl)-N ((S)-l-phenyl-ethyl)-propionamide wherein 2-[(Benzofuran-2-ylmethyl)-amino] 2-dimethylaminomethyl-3-(1H-indol-3-yl)-N-(1-phenyl-ethyl)-propionamide (S,S) is reacted with lithium hexamethyldisilazide and Eschenmoser's salt. 20 BRIEF DESCRIPTION OF FIGURES Figure 1 shows the evaluation of the compound of Example 2 in the carrageenan induced hypersensitivity model in the guinea-pig. 25 DETAILED DESCRIPTION OF THE INVENTION 30 The invention provides tachykinin receptor antagonists; the compounds have proved to be highly selective and functional tachykinin receptor antagonists. These compounds are unique in the substitution at the C carbon. Compounds of the invention are those of Formula (I): WO 01/46176 1 PCTIEPOO/13349 1 4 R -CH 2 ) M7X _C I COZ R6

(CH

2 )n R5 R2 (1) or a pharmaceutically acceptable salt thereof wherein R, m, X, R1, R2, n, Y, R3, R4, R5 and R6 are as described below: * and A indicate all stereoisomers; 5 R is phenyl, pyridyl, thienyl, furyl, quinolyl 10 isoquinolyl naphthyl, benzofuryl, benzo[1,3]dioxole benzothienyl or, 15 benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, alkoxy, halogen, 20 CF 3 or

OCF

3 ; m is an integer from I to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; WO 01/46176 12 PCT/EPOO/13349 RI is (CH 2 )pY where p is 0 to 3 and Y is OH, OCH 3 , F, CF 3 , CO 2 H, N(CH 3

)

2 ,

NHCH

3 , NH 2 , COCF 3 , COCH 3 or NO 2 ; n is an integer from 1 to 2; R2 is naphthyl or indolyl unsubstituted or N-substituted with hydroxy, alkyl,

CH

2

N(CH

3

)

2 , CH 2 N 0 or CHN N 5 formyl, CH 2 OH, Z is NR3 or 0, where R3 is H or C 1

-C

4 alkyl; R4 and R5 are each independently hydrogen, or (CH 2 )pR7 where: p is an integer of I to 3, and R7 is H, CH 3 , CN, OH, OCH 3 , CO 2

CH

3 , NH 2 , NHCH 3 , or N(CH3)2; 10 R6 is phenyl, pyridyl, thienyl, furyl, pyrrolyl, 15 pyrazolyl, imidazolyl, quinolyl, isoquinolyl, naphthyl, 20 indolyl, benzofuryl, benzothiophenyl, benzimidazolyl, or benzoxazolyl, wherein each of the foregoing is unsubstituted, mono-, di 25 or trisubstituted by alkyl, hydroxy, alkoxy, halogen,

CF

3 , 30

NO

2

,

WO 01/46176 13 PCT/EPOO/13349 N(CH3)2,

OCF

3 ,

SONH

2 ,

NH

2 , 5

CONH

2 ,

CO

2

CH

3 or

CO

2 H, or R6 is: straight alkyl of from 1 to 3 carbons, 10 branched alkyl of from 3 to 8 carbons, cycloalkyl of from 5 to 8 carbons or heterocycloalkyl, each of which can be substituted with up to one or two substituents selected from 15 OH,

CO

2 H,

N(CH

3

)

2 ,

NHCH

3 and

CH

3 ; or 20 R5 and R6, when joined by a bond, can form a ring. Preferred compounds of the invention are those of Formula I above wherein 0 is R or S, and A is R or S; 25 -R is phenyl, pyridyl, thienyl, furyl, quinolyl 30 isoquinolyl benzofuryl, WO 01/46176 14 PCT/EPOO/13349 benzo[1,3]dioxole benzothienyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, 5 hydroxy, alkoxy, halogen,

CF

3 or

OCF

3 ; 10 m is an integer from 1 to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; RI is (CH 2 )pY where p is 0 to 3 and Y is OH, F, CF 3 , OCH 3 , CO 2 H, N(CH 3

)

2 ,

NHCH

3 , NH2, COCF 3 , COCH 3 or NO 2 ; n is an integer from 1 to 2; 15 R2 is naphthyl or indolyl unsubstituted or N-substituted with hydroxy, alkyl,

CH

2

N(CH

3

)

2 , CH 2 N 0 or CH2 N N formyl, CH 2 OH, Z is NR3 or 0, where R3 is H or CH 3 ; R4 and R5 are each independently hydrogen, CH 3 or CH 2 OH; R6 is phenyl, 20 pyridyl, thienyl, furyl, pyrrolyl, cyclohexyl or 25 benzimidazolyl, wherein each of the foregoing is unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, 30 alkoxy, WO 01/46176 15 PCT/EP00/13349 halogen,

CF

3 ,

NO

2 , or

N(CH

3

)

2 . 5 More preferred compounds of the invention are those of Formula I above wherein 0 is R or S, and A is R or S; R is phenyl, 10 pyridyl, thienyl, furyl, benzofuryl, benzo[1,3]dioxole 15 benzothienyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, 20 alkoxy, halogen,

CF

3 or

OCF

3 ; m is an integer from 1 to 3; 25 X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; RI is (CH 2 )pY where p is 0 to 3 and Y is OH, OCH 3 , F, CF 3 , CO2H, N(CH 3

)

2 ,

NHCH

3 , NH1 2 , COCF 3 , COCH 3 or NO 2 ; n is an integer from 1 to 2; R2 is indolyl unsubstituted or N-substituted with alkyl or formyl; 30 Z is NR3 or 0, where R3 is H or CH 3 ; R4 and R5 are each independently hydrogen, CH 3 , or CH 2 OH; R6 is phenyl, WO 01/46176 16 PCT/EPOO/13349 pyridyl, thienyl, furyl, pyrrolyl, 5 cyclohexyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, 10 alkoxy, halogen,

CF

3 ,

NO

2 or

N(CH

3

)

2 15 Most preferred compounds of the invention are: 2-[(Benzofuran-2-ylmethyl)-amino]-3-(1H-indol-3-yl)-2-methoxymethyl-N-(1 phenyl-ethyl)-propionamide [S-(R*,R*)]; 2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H-indol-3-yl) 20 N-(1-phenyl-ethyl)-propionamide (S,S); 2-[(Benzofuran-2-ylmethyl)-amino]-2-hydroxymethyl-3 -(1H-indol-3-yl)-N-(1 phenyl-ethyl)-propionamide [S-(R*,R*)]; 2-(3-Benzofuran-2-ylmethyl-ureido)-2-hydroxymethyl-3-(1H-indol-3-yl)-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)]; 25 2-(3-Benzofuran-2-ylmethyl-ureido)-3-(1H-indol-3-yl)-2-methoxymethyl-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)]; (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1-hydroxymethyl 1H-indol-3-ylmethyl)-N-((S)-1-phenyl-ethyl)-propionamide; (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1 30 dimethylaminomethyl-1H-indol-3-ylmethyl)-N-((S)-1-phenyl-ethyl) propionamide.

WO 01/46176 17 PCT/EPOO/13349 The present invention also concerns prodrugs of a compound of Formula I such as would be contemplated by to one skilled in the art (see Bundgaard et al., Acta Pharma Suec., 1987; 24: 233-246.) for example a suitable moiety may be attached to a nitrogen of the linker X, to the nitrogen of the Z linker, or that of an 5 indolyl radical of R2 when R2 is an indolyl radical. The invention additionally provides pharmaceutical formulations comprising a compound of Formula I admixed with a pharmaceutically acceptable carrier, diluent or excipient therefor. 10 The invention also provides a method for antagonizing NK 1 receptors in a mammal in need of treatment comprising administering to a mammal an effective amount of a compound of Formula I. The invention further provides a method for treating or preventing a variety of CNS disorders including pain (inflammatory, surgical and neuropathic), anxiety, 15 panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, obesity and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and psoriasis; gastrointestinal disorders including colitis, Crohn's disease, irritable 20 bowel syndrome and satiety; allergic responses such as eczema and rhinitis; vascular disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis comprising administering to a mammal in need of treatment an effective amount of a compound of Formula I. The compounds of the invention, NK 1 receptor antagonists, being useful as anti 25 angiogenic agents, the invention further provides a method for treating or preventing conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth, which comprises administering to a mammal in need of treatment an effective amount of a compound of Formula I. 30 The invention further provides agents for imaging NKI receptors in vivo in conditions such as ulcerative colitis and Crohn's disease.

WO 01/46176 18 PCT/EPOO/13349 The invention further provides the use of a compound of formula 1 for the preparation of a medicament intended for preventing or treating CNS disorders such as pain (inflammatory, surgical and neuropathic), anxiety, panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, 5 sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, obesity and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and psoriasis; gastrointestinal disorders including colitis, Crohn's disease, irritable bowel syndrome and satiety; allergic responses such as eczema and rhinitis; vascular 10 disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis; conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth. Throughout this application, the following abbreviations have the meanings 15 listed below: CBZ carbobenzoxy CNS central nervous system COPD chronic obstructive pulmonary disease 20 DCC 1,3-dicyclohexyl carbodiimide DIPEA NN-diisopropylethylamine DMAP NN-dimethyl-4-amino pyridine DMF NN-dimethylformamide DMPU NN'-dimethyl-N,N'-propylene urea 25 HBTU O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate HRMS high resolution mass spectrometry LHMDS lithium hexamethyl disilazide Met methionine 30 PEI poly(ethylene imine) Sar sarcosine s.c. subcutaneous WO 01/46176 19 PCT/EPOO/13349 SEM-Cl 2-(trimethylsilyl)ethoxymethyl chloride RT room temperature TBAF tetrabutylammonium fluoride TFA trifluoroacetic acid 5 THF tetrahydrofuran TIPS triisopropylsilyl Tris tris(hydroxymethyl)aminomethane Trp tryptophan 10 The compounds of Formula I are further defined as follows. The term "alkyl" means a straight or branched hydrocarbon having from one to 12 carbon atoms and includes, for example, methyl, ethyl, n-propyl, isopropyl, n butyl, secbutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, undecyl, dodecyl, and the like unless stated specifically otherwise. 15 The term "cycloalkyl" means a saturated hydrocarbon ring which contains from 3 to 12 carbon atoms, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl except as otherwise stated. The term "alkoxy" means an alkyl as described above attached through an oxygen atom. 20 The term "halogen" is chlorine, bromine, fluorine or iodine. The ring formed by joining R5 and R6 is from 4 to 6 atoms total and is unsubstituted. The compounds of Formula I are capable of forming pharmaceutically acceptable acid addition salts. All of these forms are within the scope of the 25 present invention. Pharmaceutically acceptable acid addition salts of the compound of Formula I include salts derived from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, hydrofluoric, phosphorous, and the like as well as the salts derived from nontoxic organic 30 acids, such as the aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy-alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus include sulfate, WO 01/46176 20 PCT/EPOO/13349 pyrosulfate, bisulfate sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, fluoride, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, 5 maleate, mandalate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, tartrate, methanesulfonate, and the like. Also contemplated are salts of amino acids such as arginate and the like. For example, see Berge S.M., et al., Pharmaceutical Salts, J. Pharm. Sci., 66:1-19 (1977) incorporated herein by 10 reference. The acid addition salts of said basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner. Preferably, a compound of Formula I can be converted to an acidic salt by treating an aqueous solution of the desired acid, such that the 15 resulting pH is less than four. The solution can be passed through a C18 cartridge to absorb the compound, washed with copious amounts of water, the compound eluted with a polar organic solvent such as, for example methanol acetonitrile, aqueous mixtures thereof, and the like, and isolated by concentrating under reduced pressure followed by lyophilisation. The free base form may be 20 regenerated by contacting the salt form with a base and isolating the free base in the conventional manner. The free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free base for the purpose of the present invention. 25 Certain of the compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms, including hydrated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain of the compounds of the present invention possess one or more chiral 30 centres and each centre may exist in the R (D) or S (L) configuration. The present invention includes all enantiomeric and epimeric forms as well as the appropriate mixtures thereof.

WO 01/46176 21 PCT/EPOO/13349 The compounds of the present invention can be prepared and administered in a wide variety of oral and parenteral dosage forms. Thus, the compounds of the present invention can be administered by injection, that is intravenously, intramuscularly, intracutaneously, subcutaneously, intraduodenally, or 5 intraperitoneally. In addition, the compounds of the present invention can be administered by inhalation, for example intranasally. Additionally, the compounds of the present invention can be administered transdermally. It will be obvious to those skilled in the art that the following dosage forms may comprise as the active component, either a compound of Formula I or a corresponding 10 pharmaceutically acceptable salt of the compound of Formula I. For preparing pharmaceutical compositions from the compounds of the present invention, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, pills, tablets, capsules, cachets, suppositories and dispersible granules. A solid carrier can be one or more 15 substances which may also act as diluents, flavouring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is a finely divided solid which is a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary 20 binding properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain from 5% or 10% to about 70% of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, 25 methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets 30 and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

WO 01/46176 22 PCT/EPOO/13349 For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein, as by stirring. The molten homogeneous mixture is then poured into convenient sized moulds, allowed to cool, and the thereby to 5 solidify. Liquid form preparations include solutions, suspensions and emulsions, for example, water or water propylene glycol solutions. For parenteral injection liquid preparations can be formulated in solution in aqueous polyethylene glycol solution. 10 Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavours, stabilising and thickening agents as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or 15 synthetic gums, resins, methylcellulose sodium carboxymethylcellulose, and other well-known suspending agents. Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These preparations 20 may contain, in addition to the active component, colorants, flavours, stabilisers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilising agents and the like. The pharmaceutical preparation is preferably in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of 25 the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampules. Also, the unit dosage form can be a capsule, tablet, cachet or lozenge itself, or it can be the appropriate number of any of these in packaged form. 30 The quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 200 mg preferably 0.5 mg to 100 mg according to the WO 01/46176 23 PCT/EPOO/13349 particular application and the potency of the active component. The composition can, if desired also contain other compatible therapeutic agents. In therapeutic use, the highly selective and competitive antagonists of the NKI receptor and compounds of this invention are administered at the initial dosage 5 of about 0.01 mg to about 500 mg/kg daily. A daily dose range of about 0.01 mg to about 100 mg/kg is preferred. The dosages, however, may be varied depending upon the requirements of the patient, the severity of the condition being treated and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment 10 is initiated with smaller doses which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired. 15 The compounds of Formula I or Intermediates for their synthesis and particularly compounds for which R2 is an indolyl unsubstituted or N-substituted with hydroxy, alkyl, formyl, CH2OH, CH2N(CH3)2,

CH

2 N 0 CH 2 N N or , can be prepared by any several synthetic 20 processes well known to those skilled in the art of organic chemistry. The synthesis can be carried out on racemic reactants, to provide invention compounds in racemic form, which can then be resolved by conventional methods, if desired. Alternatively, the invention compounds can be prepared in optically active form by using enantiomeric reactants and asymmetric reactions. 25 A process for the obtention of an Intermediate for the synthesis of a compound of the present invention is the process of scheme 1.

WO 01/46176 24 PCT/EP00/13349 Scheme 1 describes the synthesis of the tricyclic Intermediate 1, which is one of the possible key Intermediates in the synthesis of compounds of the invention and particularly the compounds of Examples I to 7.

WO 01/46176 25 PCT/EPOO/13349 Scheme 1: OH ONHP H H O 0 N, N,, N P 1 N P1 H

P

1 Intermediate 1 where P1 is an N-protecting group such as benzyloxy carbonyl, alkyloxy carbonyl (eg, Methyl, Ethyl, halogenated alkyl), arylsufonyl (eg toluyl, phenyl), alkylsulfonyl (Methyl, Ethyl), RCO-. 5 i) Allyl alcohol, DCC, DMAP, CH 2 Cl 2 , 89 % ii) TFA, 52 % iii) P1-Cl, Na 2

CO

3 (aq), dioxan, 92 % 10 In this synthesis: - in step i) the allyl ester of P1-Trp is formed using DCC and DMAP; - in step ii) the compound obtained is then cyclised using TFA; and - in step iii) the indole nitrogen is then protected with a second protecting group. 15 In a preferred embodiment of the present invention P1 is a benzyloxy carbonyl group. Other Intermediates useful in the synthesis of compounds of the present invention can be obtained as shown in scheme 2. 20 WO 01/46176 26 PCT/EPOO/13349 Scheme 2: R1 N _ 0 N N P 1 N P 1 P

P

1 | 0 0 -R1 M RI \/OH O / N NHP 1 N NHP 1 IV 0 R4 N R5 R5 H N R6 ~ NHPNR NH H N 1N 2 H wherein R1, R4, R5, R6 and P1 are as defined previously. 5 In this synthesis: -In step i) the tricyclic Intermediate 1 is asymmetrically alkylated; -In step ii) the product obtained is then ring opened for instance using TFA; -In step iii) the allyl ester is removed; 10 -In step iv) the acid obtained is coupled with the appropriate amine; and -In step v) the N-terminal protecting groups are removed to yield the desired Intermediates.

WO 01/46176 27 PCT/EPOO/13349 Scheme 2.1 below examplifies the synthesis of Intermediates 2 and 3. In this synthesis: - the tricyclic Intermediate 1 is asymmetrically alkylated; 5 - the product obtained is then ring opened using TFA; - the allyl ester is then removed and - the acid obtained is coupled with alpha-methylbenzylamine using HBTU activation; - Intermediates 2 and 3 are then formed by removal of the benzyloxy carbonyl N 10 terminal protecting group with palladium hydroxide on carbon.

WO 01/46176 28 PCT/EPOO/13349 Scheme 2.1: o'^w z /7 z aN N z z Intermediate 1 a, R = CH 2 OMe b, R = CH 2 NMe 2 R0 iii 0 0 OH NHZ NHZ N N I I z z a, R = CH 2 0Me a, R = CH 2 OMe b, R = CH 2 NMe 2 b, R = CH 2 NMe 2 ivR 0 R N Ph V N Ph NHZ H NH H N N 2 H z a, R = CH 2 0Me Intermediate 2, R = CH 2 OMe b, R = CH 2 NMe 2 Intermediate 3, R = CH 2 NMe 2 5 wherein Z is an N-protecting group such as benzyloxy carbonyl, alkyloxy carbonyl (eg, Methyl, Ethyl, halogenated alkyl), arylsufonyl (eg toluyl, phenyl), alkylsulfonyl (Methyl, Ethyl), RCO-. 10 i) LHMDS, RX, DMPU, THF ii) TFA, CH2Cl- or H-S04, MeOH, H20 iii) Pd(PPh 3

)

4 , morpholine, THF iv) amine, HBTU, DIPEA, DMF v) Pd(OH) 2 /C, EtOH 15 In a preferred embodiment of this invention Z is a benzyloxy carbonyl group.

WO 01/46176 29 PCT/EPOO/13349 Further Intermediates useful in the synthesis of compounds of the invention can be obtained as shown in scheme 3. Scheme 3:

P

2 N 1 0 N P 1 N P 1

P

1

P

1 II

P

3 0 HO 0

NHP

1 N NHP 1 NN

P

1

P

1 iv

P

3 O P 3 0 R4 V _ R5 OH N R6 H N NHPI 1H e N NHP

P

3 0 0 R4 R N R6 H N N 2 H wherein P1, R4, R5, R6 are as defined previously and wherein P2 and P3 are 5 protecting groups, P2 being SEM, P3 being TIPS, TBS, TBDMS, or DPS or an ether group such as MOM, TT{P.

WO 01/46176 30 PCT/EPOO/13349 In this synthesis: -In step i) a protected hydroxyl function is introduced; -In step ii) the ring is opened and the protecting group on the hydroxy function is removed; 5 -In step iii) the hydroxy moiety is protected with an appropriate protecting group; -In step iv) the allyl ester is removed; -In step v) the acid is coupled with the appropriate amine; -In step vi) the desired Intermediate is obtained by removing the N-terminal 10 protecting groups. In a prefered embodiment of the present invention P2 is SEM and P3 is TIPS. Scheme 3.1 belows examplifies the synthesis of Intermediate 4. 15 In this process: - the protected hydroxyl function is introduced by reaction of Intermediate 1 with LHMDS followed by SEM-Cl; - the ring is opened and the protecting group is removed using TFA in 20 dichloromethane; - the hydroxyl moiety is then protected with a TIPS group by TIPS-Cl in DMF; - the allyl ester is then removed and - the acid is coupled with alpha-methylbenzylamine using HBTU activation. - Intermediate 4 is then formed by removal of the benzyloxycarbonyl N-terminal 25 protecting group with palladium hydroxide on carbon.

WO 01/46176 31 PCT/EPOO/13349 Scheme 3.1: 0 SEMO O HO O N \ N N H I z I zI z z z Intermediate 1. TIPSO 0 TIPSO 0 v OH v NHZ N NHZ N N z z TIPSO N H TIPS 0 ZNH Ph Ph N ON N 2 H z H Intermediate 4. 5 wherein Z is an N-protecting group such as benzyloxy carbonyl, alkyloxy carbonyl (eg, Methyl, Ethyl, halogenated alkyl), arylsufonyl (eg toluyl, phenyl), alkylsulfonyl (Methyl, Ethyl), RCO-. i) LHMDS, SEM-Cl, THF 10 ii) TFA, CH 2 Cl 2 iii) Imidazole, TIPS-Cl, CH 2 Cl 2 iv)Pd(PPh 3

)

4 , morpholine, THF v) amine, HBTU, DIPEA, DMF vi) Pd(OH) 2 /C, EtOH 15 In a prefered embodiment of the present invention Z is a benzyloxy carbonyl group.

WO 01/46176 32 PCT/EPOO/13349 A process for the synthesis of compounds of the present invention is as shown in scheme 4. Scheme 4: 0 R4 0 R4 R9 R1 R5 R5 N - N H R6 H R6 N NH A O N 2 iN X R' R' (H2)m R a 0 R4 (CH2)p R5 ".N H R6 NX R' (H2)m R 5 wherein R9 is RI as defined above or (CH2)p P3 wherein p is an integer from 0 to 3 and P3 is as defined above; wherein R, m, R4, R5 and R6 are as defined above, and wherein R' is hydroxy, alkyl, formyl, CH2OH,

CH

2 N 0 CH 2 N N CH2N(CH3)2, or 10 -Step i) of this process requires reduction of the amino group into a secondary amine.

WO 01/46176 PCT/EPOO/13349 -Step ii) is required only when R9 is (CH2)p P3. In this second step the protecting group is removed by conventional methods known to the skilled person. Very most preferred compounds are compounds wherein R is benzofuryl. Scheme 4.1 outlines below the synthesis of the compounds of Examples 1. 2 and 3. 10 Examples I and 2 are prepared from a reductive amination of benzofuran-2 carboxaldehyde with Intermediates 2 and 3 respectively and sodium triacetoxy borohydride in 1.2-dichloroethane. Example 3 is prepared in an analogous manner with an additional step to remove the TIPS protection using TBAF in tetrahydrofuran. 15 Scheme 4.1: HN HN || HN R R OH

H

2 N H N\ HN~ H N +,-N\ ] N 0 4 /0 0 Intermediate 4, R = TIPSO a), R =TI PSO Example 3 Intermediate 2, R = OMe Example 1, R = OMe Intermediate 3, R = NMe 2 Example 2, R = W~e i) Benzofuran-'?-carboxaldehvde. NaBH(Oac)-;. (CH-,CI )-' 20 11) TBAF, THF WO 01/46176 34 PCT/EPOO/13349 Scheme 4.2 below examplifies the synthesis of the compounds of Examples 4 and 5. Example 4 is prepared by reaction of Intermediate 2 with (2 5 benzofuranyl)methylisocyanate in tetrahydrofuran. Example 5 is prepared in an analogous manner with an additional step to remove the TIPS protection using TBAF in tetrahydrofuran. Scheme 4.2: 10 HN HN R R

H

2 N-~ N tH H 2. N H N N N -N Intermediate 4, R = TIPSO a), R = TIPSO Intermediate 2, R = OMe Example 4, R = OMe .. HN OH HP N>H N N H 0 0 Example 5. i) (2-benzofuran)methyl isocyanate, THF ii) TBAF, THF WO 01/46176 35 PCTIEPOO/13349 Another process of the invention can be used to introduce a substituent on the N atom of the indolyl group of R2, scheme 5. Scheme 5: 0 R4 5 RO N 6 5 X H N/ 6 N X )H H N X (CH2)m I R (CH2)m R R 5 wherein RI, R4, R5, R6, R and m are as defined above, R' being hydroxy, alkyl,

CH

2 N 0 CH 2 N N formyl, CH2OH, CH2N(CH3)2, or . In this synthesis: 10 -in step i) reaction with potassium hexamethyldisilazide takes place. -in step ii) reaction with formaldehyde or Eschenmoser's salt takes place. Those two reactions could also be combined as in Scheme 2.1 (above) into one 'step' e.g. LHMDS, RX, THF. 15 Scheme 5.1 below examplifies the synthesis of the compounds of Examples 6 and 7. 20 Example 6 is prepared by reaction of Example 2 with potassium hexamethyldisilazide and formaldehyde in THF at -78 "C.

WO 01/46176 36 PCT/EPOO/13349 Example 7 is prepared in an analogous manner but with Eschenmoser's salt in place of the formaldehyde.

WO 01/46176 37 PCT/EPOO/13349 Scheme 5.1: Q ,Me 2 N o Q ,Me 2 N o H \ H 0 N Ph i 0 N Ph NH

N

R Example 6, R = OH Example 7, R = N(CH 3

)

2 5 i) Potassium hexamethyldisilazide, Example 2, THF (-78 'C) ii) formaldehyde or Eschenmoser's salt Alternative Intermediates useful in the synthesis of compounds of the invention 10 can be obtained as shown in scheme 6.

WO 01/46176 38 PCT/EPOO/13349 Scheme 6:

NH

2 O R' O H N 0 N H20 R1 NH O R' iv RR Ri 0 R:0 0 NHH N N2

N

WO 01/46176 39 PCT/EPOO/13349 wherein RI is as described previously and R' is hydroxy, alkyl, formyl, CH2OH,

CH

2 N 0 CH 2 N N CH2N(CH3)2, or In this synthesis: 5 -In step i) reaction with benzaldehyde takes place. This is an imine formation where the water by-product is removed with a dehydrating reagent (eg MgSO 4 ), molecular sieves or azeotropic removal (Dean-Stark trap). -In step ii) an alpha aminoalkylation takes place; -In step iii) the hydrolysis of the benzylimine takes place: 10 -In step iv) the racemate is separated in the two corresponding diastereoisomers chiral HPLC phase. The racemate could also be separated in the two corresponding diastereoisomers by crystallisation after reaction with a chiral acid. 15 Scheme 6.1 below examplifies the synthesis of Intermediate 6.

WO 01/46176 40 PCT/EPOO/13349 Scheme 6.1: H2< H Hcj KN R,S H 0' 0 H H Intermediate 6 H Intermediate 5 WO 01/46176 41 PCT/EPOO/13349 Ways were sought to prepare a-dimethylaminomethyltryptophan methyl ester without the use of protective group chemistry contrary to literature references, the desired ax-aminoalkylation with 1-dimethylaminomethylbenzotriazole, starting from the Schiffs base ((S)-2-benzylideneamino)-3-(1H-indol-3-yl) 5 propionic acid methyl ester), could be achieved. It was very surprising that a-dimethylaminomethyltryptophan can only be esterified in conventional manner with extreme difficulty. Surprisingly, we have now found that ((S)-2-benzylidene-amino)-3-(1H-indol-3 10 yl)-propionic acid methyl ester (Schiffs base), which can be prepared in one synthesis step from the cheap (S)-tryptophan methyl ester ((S)-Trp-OMe), can, by reaction with 1-dimethylaminomethylbenzotriazole, be converted into racemic a-dimethylaminomethyltryptophan methyl ester without using protection group chemistry. 15 It is known that N-(a-aminoalkyl)-benzotriazole derivatives, which can be prepared very easily from benzotriazole, an aldehyde and a primary or secondary amine, can be used as aminoalkylation reagents (see A. Katritzky et al., Tetrahedron, 46, No. 24, 8153-8160/1990). The preparation of 1-dimethylaminomethylbenzotriazole has been described by 20 J.H. Bruckhalter et al. (J.A.C.S., 74, 3868-3369/1952). B.E. Love and B.T. Nguyeri (Synlett, 1123, October, 1998) have described the reaction of 1-methylaminomethylbenzotriazole and indole. As main reaction, there hereby takes place an aminoalkylation on the indole nitrogen atom, an aminoalkylation on the 3-position only taking place as a secondary reaction. 25 Surprisingly, in the case of the tryptophan derivative (Schiffs base) used in the case of the present invention, this aminoalkylation reaction with 1 dimethylaminomethyl-benzotriazole only takes place on the a-C atom. In contradistinction to the above-mentioned literature references, an alkylation on the indole nitrogen atom was not observed. 30 Furthermore, we have found that the racemic ax-dimethylaminomethyltryptophan methyl ester cannot be separated into the enantiomers either enzymatically or WO 01/46176 42 PCTIEPOO/13349 after formation of diastereomeric salts. It was even more surprising that the two enantiomers, i.e. the (S)-enantiomer and the (R)-enantiomer, can be separated on a preparative scale on a chiral HPLC phase. The racemic ax-dimethylaminomethyltryptophan methyl ester can then be further 5 reacted in the usual way with a second chiral component to give a diastereomeric mixture which can be separated by crystallisation into the two diastereomenic compounds. Further compounds of the invention can be obtained by an alternative process as 10 follows, scheme 7.

WO 01/46176 43 PCT/EPOO/13349 Scheme 7: R1 0 0 N H 2 R'Ri R1 0 0 x NI R' (CH2)m R R1 0 0 NJ x aN (CH2)m R R1 0 R4 N R6 NI FR' (CH2)m

R

WO 01/46176 PCT/EPOO/13349 CH 2 N wherein R' is hydroxy, alkyl, formyl, CH2OH, CH2N(CH3)2, or

CH

2 N N and wherein R1, m, R, R4, R5, R6 are as described previously. In this synthesis: 5 -In step i) the amino group is reduced into a secondary amine. -In step ii) the methyl ester is hydrolysed with a base (LiOH, NaOH, KOH) in an appropriate solvent system -In step iii) the acid is coupled with the appropriate amine. 10 Scheme 7.1 below examplifies the alternative synthesis of the compound of Example 2. Scheme 7.1: WO 01/46176 45 PCT/EPOO/13349 0 NH 2 H N 0 0 N H N H O N 00 H NH N H O 0 N 0 N N NH H -0

N.

WO 01/46176 46 PCT/EPOO/13349 i) benzofuran-2-carbaldehyde, sodium triacetoxyborohydride, (CH 2 Cl) 2 ii) NaOH, 1,4-dioxan/water. iii) HBTU, amine, DIPEA, DMF. 5 The compounds of Examples 6 and 7 can be obtained from the compound of example 2 synthesised said process, through the process outlined in scheme 5.1. The present invention is further illustrated, but in no case limited, by the figures and the examples below.

WO 01/46176 47 PCT/EPOO/13349 EXAMPLES EXAMPLE 1 2-[(Benzofuran-2-ylmethyl)-amino]-3-(1H-indol-3-yl)-2-methoxymethyl-N-(1 5 phenyl-ethyl)-propionamide [S-(R*,R*)] HN 0 H / NH N 0 0 Step 1 A solution of (R)-N-CBZ-tryptophan (1.00 g, 29.6 mmol), N, N' dicyclohexylcarbodiimide (640 mg, 31.1 mmol), N, N-dimethyl-4-aminopyridine 10 (36 mg, 2.96 mmol) and dichloromethane (10 ml) was stirred for 10 min, then allyl alcohol (0.22 ml, 32.5 mmol) was added. After 30 min the mixture was filtered and the solvent removed in vacuo. The products was purified by chromatography (33 % EtOAc in heptane) to yield a clear oil which solidified on standing. Recrystallisation (EtOAc/heptane) gave an amorphous solid (1.00 g, 15 89 %); mp 83-85 "C; 8H 3.32 (2H, d, IndCH 2 , J5.2 Hz), 4.56 (2H, bs, CH 2 O), 4.74 (1H, dt, a-H, J7.8, 7.8 Hz), 5.11 (2H, m, CH 2 0), 5.20-5.32 (3H, m, NH, =CH 2 ), 5.82 (1H, m, CH), 6.96 (1H, d, arom, J2.0 Hz), 7.08 (1H, t, arom, J7.4 Hz), 7.18 (1H, d, arom, J 20 7.2 Hz), 7.34 (6H, m, arom), 7.52 (1H, d, arom, J 8.0 Hz), 8.04 (1H, bs, NH); Vmax 3410, 3361, 3060, 1713, 1512, 1205, 743 cm- 1 ; m/z 378 (MH+, 18 %), 130 (100 %); Anal. calc. for C 2 2

H

22

N

2 0 4 C 69.82, H 5.86, N 7.40 % found C 69.88, H 5.86, N 7.44 %; [Ia]D 2 0 (c = 0.75, MeOH): + 14.50. 25 WO 01/46176 48 PCT/EP00/13349 Step 2. The ester (17.25 g, 45.6 mmol) was dissolved in trifluoroacetic acid (100 ml) and stirred at RT for 3 h. The mixture was concentrated (~ 50 ml) in vacuo, then added dropwise to a well stirred mixture of NaHCO 3 (15 %, 1 1) and 5 dichloromethane (500 ml). After the addition the organics were washed with saturated NaHCO 3 , brine and dried (MgSO 4 ). The product was purified by chromatography (20-50 % Et 2 O in heptane) to yield a clear oil (8.90 g, 52 %); 6 H 2.64 (2H, m, CH 2 ), 3.83-4.25 (3H, m, CH, CH 2 0), 4.58 (0.5H, dd, a-H, J 1.9, 8.3 Hz), 4.68 (0.5H, m, a-H), 4.75 (0.5H, bs, 0.5 NH), 5.05-5.26 (4.5H, m, 10 CH 2 0, =CH 2 , 0.5 NH), 5.50 (1H, m, CH=), 5.60 (1H, t, CH, J6.4 Hz), 6.58 (1H, m, arom), 6.67 (1H, m, arom), 7.01 (2H, m, arom), 7.28-7.40 (4H, m, arom), 7.41 (1H, m, arom) ; Vmax 3400, 1702, 1416, 747 cm- 1 ; m/z 378 (MH*, 90 %), 130 (100 %); 15 Anal. calc. for C 2 2

H

22

N

2 0 4 C 69.82, H 5.86, N 7.40 % found C 69.54, H 5.85, N 7.79 %;

[C]D

20 (c = 1.62, MeOH): -144.9 Step 3. Intermediate 1. 20 Benzyl chloroformate (8.01 g, 47.0 mmol, 6.7 ml) was added to a stirred mixture of the amine (8.90 g, 23.5 mmol), Na 2

CO

3 .10 H 2 0 (13.43 g, 47.0 mmol), 1,4 dioxan (100 ml) and water (10 ml) at 0 "C. The resulting mixture was allowed to warm to RT and stirred for 16 h. The solvent was removed in vacuo and the product extracted into EtOAc, the organics were washed with water, 10 % HCl, 25 brine and dried (MgSO 4 ). The product was purified by chromatography (25 % EtOAc in heptane) to give a clear oil (10.39 g, 86 %); 6 H 2.55 (1H, m, CH 2 ), 2.65 (1H, d, CHH, J13.2 Hz), 3.85 (1H, dd, OCHH, J5.2, 13.2 Hz), 4.01 (1H, t, CH, J 7.0 Hz), 4.12 (1H, m, OCHH), 4.69 (1H, t, a-H, J 8.0 Hz), 4.80-5.24 (6H, m, 2x CH 2 0, =CH 2 ), 5.50 (1H, m, CH=), 6.51 (1H, d, 30 CH, J 6.0 Hz), 6.98 (1H, t, arom, J 7.4 Hz), 7.10 (1H, m, arom), 7.19 (1H, t, arom, J7.6 Hz), 7.27-7.38 (10H, m, arom), 7.63 (1H, bs, arom); WO 01/46176 49 PCT/EPOO/13349 Vmax 3065, 3033, 1716, 1483, 1416, 1266, 1173, 753 cm- 1 ; m/z 513 (MH*, 100); [c]D 2 0 (c = 0.11, MeOH): + 2.60. 5 Step 4. LHMDS (7.81 ml, 7.81 mmol, IM in THF) was added to a solution of Intermediate 1 (2.00 g, 3.91 mmol), DMPU (0.47 ml, 3.91 mmol) in THF (30 ml) at -78 "C under dry N 2 . After 2 h at this temperature, iodomethyl methyl ether (1.34 g, 7.81 mmol) was added and the mixture allowed to warm to RT 10 overnight. The solvent was removed in vacuo and the product extracted into EtOAc, washed with 10 % HCl, brine and dried (MgSO 4 ). Purification was achieved by chromatography (15-20 EtOAc in heptane) to leave a clear oil (1.66 g, 76 %); 6 H 2.52 (1H, dd, CHH, J 1.2, 13.2 Hz), 2.82 (1H, dd, CHH, J 8.0, 13.6 Hz), 3.26 15 (3H, s, OCH 3 ), 3.58 (1H, d, CHHO, J 7.6 Hz), 3.78 (1H, dddd, OCHH, J 1.6, 1.6, 5.6, 13.2 Hz), 3.90 (1H, t, CH, J 7.0 Hz), 4.06 (1H, bs, CHHO), 4.17 (1H, dd, OCHH, J 5.6, 13.2 Hz), 4.96-5.16 (6H, m, 2x CH 2 0, =CH 2 ), 5.38 (1H, m, =CH), 6.44 (1H, d, CH, J 6.0 Hz), 6.99 (1H, m, arom), 7.08 (1H, d, arom, J 7.2 Hz), 7.19 (1H, t, arom, J7.6 Hz), 7.25-7.34 (10H, m, arom), 7.59 (1H, d, arom, J 20 8.0 Hz); vmax 1717, 1483, 1412, 1335, 1274, 751 cm- 1 ; m/z 557 (MH*, 100 %); Anal. calc. for C 3 2

H

32

N

2 0 7 C 69.05, H 5.80, N 5.03 % found C 68.82, H 5.52, N 4.88 %; 25 [CID 2 0 (c = 0.75, MeOH): + 9.6 Step 5. TFA (2 ml) was added to a solution of the above oil (1.66 g, 3.07 mmol) in dichloromethane (10 ml) and the resulting solution stirred at RT for 24 h. The 30 solvent was removed and the residue diluted with EtOAc, the organics were washed with saturated NaHCO 3 , brine and dried (MgSO 4 ). The product was WO 01/46176 50 PCT/EPOO/13349 purified by chromatography (15 % EtOAc in heptane) to yield a clear oil (1.19 g, 72 %); 6 H 3.32 (3H, s, OCH 3 ), 3.24 (1H, d, IndCHH, J 14.4 Hz), 3.60 (1H, d, IndCHH, J 14.4 Hz), 3.77 (1H, d, CHHO, J 9.2 Hz), 4.13 (1H, d, CHHO, J 9.2 Hz), 4.49 5 (1H, dd, OCHH, J4.8, 12.8 Hz), 4.61 (1H, dd, OCHH, J5.2, 12.8 Hz), 5.09 (2H, s, CH 2 0), 5.11 (1H, d, =CHH, J 10.8 Hz), 5.28 (1H, d, =CHH, J 17.2 Hz), 5.41 (2H, dd, CH 2 0, J 12.0, 14.8 Hz), 5.75 (2H, m, =CH, NH), 7.14 (1H, t, arom, J 8.0 Hz), 7.28-7.47 (13H, m, arom), 8.15 (1H, bd, arom, J6.4 Hz); vmax 3418, 3352, 1736, 1501, 1456, 1399, 1250, 1087, 749 cm- 1 ; 10 m/z 557 (MH*, 100 %); [MD20 (c = 0.67, MeOH): + 13.0'. Step 6. Tetrakis(triphenylphosphine)palladium (0) (50 mg, 43 trnol) was added to a solution of the alpha substituted amino ester (1.14 g, 2.11 mmol) in THF (10 ml), 15 after 5 min morpholine (1.84 g, 21.1 mmol) was added and the mixture stirred at RT for 30 min. EtOAc was added and the organics washed with 10 % HCl, brine and dried (MgSO 4 ). After removal of the solvent in vacuo a clear glass was obtained (1.11 g, 100 %); 6 H 3.33 (1H, d, IndCHH, J 14.7 Hz), 3.37 (3H, s, OCH 3 ), 3.60 (1H, d, IndCHH, J 20 14.4 Hz), 3.84 (1H, d, CHHO, J 9.3 Hz), 3.99 (1H, d, CHHO, J 8.8 Hz), 5.09 (2H, s, CH 2 O), 5.40 (2H, s, CH 2 0), 5.71 (1H, s, NH), 7.14 (1H, t, arom, J 7.6 Hz), 7.27-7.52 (13H, m, arom), 8.18 (1H, dd, arom, J6.8, 6.8 Hz); vmax 3411, 1732, 1456, 1399, 1250, 1086, 748 cm- 1 ; m/z (MH+, 100 %); 25 HRMS for C 2 9

H

2 9

N

2 0 7 requires 517.1975 found 499.187 (MH-H 2 O*). Step 7. A mixture of the acid (1.01 g, 2.02 mmol), HBTU (766 mg, 2.02 mmol), DIPEA (0.70 ml, 2.02 mmol) in DMF (10 ml) was stirred at RT for 10 min then (S) 30 methylbenzylamine (244 mg, 2.02 mmol) and DIPEA (0.70 ml, 2.02 mmol) added and the resulting solution stirred for 8 h. The solvent was removed and WO 01/46176 51 PCT/EPOO/13349 the product extracted into EtOAc, washed with 10 % HCl, 10 % K 2 C0 3 , brine and dried (MgS04). Purification by chromatography gave a clear glass which was recrystallised (EtOAc/heptane) to give a clear glass (1.04 g, 78 %); 6 H 1.37 (3H, d, CHCH 3 , J 6.8 Hz), 3.35 (3H, s, OCH 3 ), 3.39 (1H, d, IndCHH, J 5 15.2 Hz), 3.47 (1H, d, CHHO, J 9.2 Hz), 3.66 (1H, d, IndCHH, J 14.4 Hz), 4.16 (1H, d, CHHO, J8.0 Hz), 4.98 (1H, m, CHCH 3 ), 5.03 (2H, bs, CH 2 O), 5.40 (2H, bs, CH 2 O), 6.02 (1H, bs, NH), 7.14-7.53 (20 H, m, arom, NH), 8.14 (1H, bs, arom); vmax 3350, 1732, 1653, 1488, 1455, 1398, 1249, 1084, 748 cm- 1 ; 10 m/z 620 (MH*, 100 %); Anal. calc. for C 37

H

37

N

3 0 6 : C 71.71, H 6.02, N 6.78 Found: C 71.85, H 6.04, N 6.59 %; [a]D 20 (c = 0.53, MeOH): -21.7 Step 8. Intermediate 2. 15 A mixture of the amide (980 mg, 1.63 mmol), 10 % palladium hydroxide on carbon and methanol (20 ml) were hydrogenated at 50 psi (345 kPa) at 30 *C. After 90 min the mixture was filtered through Kieselguhr and upon removal of the solvent in vacuo to give a pink coloured foam (630 mg, quant.); 6 H (DMSO-d 6 ) 1.36 (3H, d, CHCH 3 , J6.8 Hz), 3.26 (3H, s, OCH 3 ), 3.37 (2H, s, 20 IndCH 2 ), 3.66 (1H, d, CHHO, J 10.0 Hz), 4.17 (1H, d, CHHO, J 10.0 Hz), 4.90 (1H, dq, CHCH 3 , J 6.8, 6.8 Hz), 7.00-7.38 (8H, m, arom), 7.70 (1H, d, arom, J 7.6 Hz), 8.17 (3H, bs, NH, NH 2 ), 8.94 (1H, d, arom, J7.6 Hz), 11.17 (1H, d, NH, J1.2 Hz); Vmax 3419, 3213, 3057, 1667, 1494, 1458, 1106, 746 cm- 1 ; 25 HRMS for C 2 1

H

26

N

3 0 2 requires 352.2025 found 352.2025 (MH); Anal. calc. for C 2 1

H

2 5

N

3 0 2 .0.4 H 2 0: C 70.33, H 7.25, N 11.72 % Found: C 70.32, H 6.94, N 11.66 %; [a]D 1 9 (c = 0.66, MeOH): -9.2 .

WO 01/46176 52 PCT/EPOO/13349 Step 9. Benzofuran-2-carboxaldehyde (83 mg, 568 ptmol), Intermediate 2 (200 mg, 406 pmol) and sodium triacetoxyborohydride (172 mg, 811 ptmol) were stirred in 5 1,2-dichloroethane (2 ml) at RT for 16 h. The mixture was diluted with CH 2 Cl 2 , washed with 0.5 M NaOH, brine and dried (MgSO 4 ). The product was purified by chromatography (5-15 % EtOAc in heptane) to yield a clear glass (60 mg, 44 mp 50-53 "C; 10 6 H 1.45 (3H, d, CHCH 3 , J 6.8 Hz), 2.15 (1H, bs, NH), 3.15 (2H, dd, IndCH 2 , J 14.6, 51.6 Hz), 3.39 (3H, s, OCH 3 ), 3.69 (2H, dd, CH 2 O), J 9.6, 44.0 Hz), 3.92 (2H, dd, CH 2 N, J 13.8, 67.6 Hz), 5.02 (1H, dq, CHCH 3 , J7.6, 7.6 Hz), 6.49 (1H, s, arom), 6.59 (1H, s, NH, J 2.0 Hz), 7.03-7.27 (10H, m, arom), 7.39 (1H, m, arom), 7.50 (1H, m, arom), 7.57 (1H, d, arom, J 8.6 Hz), 7.77 (1H, d, arom, J 8.4 15 Hz), 7.85 (1H, s, NH); vmax 3338, 2925, 1659, 1512, 1455, 1106, 742 cm- 1 ; m/z 482 (MH*, 100 %); Anal. calc. for C 30

H

3 1

N

3 0 3 C 74.82, H 6.49, N 8.73 % found C 74.57, H 6.36, N 8.74 %; [a]D 1 9 (c = 0.31, MeOH): -37.7 *.

WO 01/46176 53 PCT/EPOO/13349 EXAMPLE 2. 2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H-indol-3-yl) N-(1-phenyl-ethyl)-propionamide (S,S) 5 HN N~ H / N H N 0 0 Step 1. 10 Method as for Example 1, step 4 to give a clear oil (1.90g, 76 %); 8H 2.23 (6H, s, 2x CH 3 ), 2.40 (1H, d, IndCHH, J 13.2 Hz), 2.66 (11H, d, CHHN, J 14.4 Hz); 3.00 (1H, dd, IndCHH, J 8.2, 13.4 Hz), 3.29 (1H, bs, CHHN), 3.69 (1H, dddd, OCHH, J 1.6, 2.9, 5.8, 13.3 Hz), 3.94 (1H, t, CH, J7.0 Hz), 4.11 (1H, bs, CHHN), 4.93-5.33 (7H, m, 2x CH 2 0, =CH, =CH 2 ), 6.40 (1H, d, CH, J 6.4 15 Hz), 6.99 (1H, t, arom, J7.4 Hz), 7.06 (1H, d, arom, J7.6 Hz), 7.18 (1H, t, arom, J7.6 Hz), 7.26-7.37 (10H, m, arom), 7.58 (1H, bs, arom); Vmax 2947, 1717, 1483, 1412, 1331, 1267, 1043, 1020, 750 cm-1; HRMS for C 33

H

36

N

3 0 6 requires 570.2604 found 570.2604 (MH*, 100 %); [aID 19 (c = 0.49, MeOH): -0.4'. 20 Step 2. Method as for Example 1, step 5 to give a straw coloured gum (3.46g, 59 %); 6 H 2.26 (6H, s, 2x CH 3 ), 2.83 (1H, d, CHHN, J 13.6 Hz), 3.23 (1H, d, IndCHH, J 14.4 Hz), 3.32 (1H, d, CHHN, J 13.6 Hz), 3.64 (1H, dd, IndCHH, J 14.4 Hz), 25 4.49 (1H, d, CHHO, J 13.2 Hz), 4.59 (1H, d, CHHO, J 6.0 Hz), 5.08 (2H, dd, WO 01/46176 54 PCT/EPOO/13349

CH

2 0, J 12.4, 27.6 Hz), 5.23 (1H, d, =CHH, J 10.4 Hz), 5.34 (1H, d, =CIH, J 14.4 Hz), 5.41 (2H, s, CH 2 0), 5.85 (1H, m, =CH), 6.00 (1H, s, NH), 7.14 (2H, m, arom), 7.25-7.48 (12H, m, arom), 8.15 (1H, bd, arom, J6.4 Hz); Vmax 3418, 1736, 1456, 1248, 1084, 1037, 748 cm- 1 ; 5 m/z 570 (MH*, 100 %); [ca]D1 9 (c = 0.27, MeOH): -12.6 . Step 3. Method as for Example 1, step 6 to yield a straw coloured foam (690 mg, 10 quant.); 6 H of little use due to impurities and zwitter-ion; Vmax 3373, 1731. 1633, 1485, 1456, 1401, 1388, 1248 cm- 1 ; HRMS for C 3 0

H

32

N

3 0 6 requires 530.2291 found 530.229 (MH*). 15 Step 4. Method as for Example 1, step 7 to afford white crystals (EtOAc/heptane) (150 mg, 34 %); mp 102-107 'C; 6 H 1.38 (3H, d, CHCH 3 , J 6.8 Hz), 2.14 (6H, s, 2x CH 3 ), 2.43 (1H, d, CHHN, J 20 14.4 Hz), 3.35 (1H, d, CHHN, J 14.4 Hz), 3.38 (1H, d, IndCHH, J 15.2 Hz), 3.63 (1H, d, IndCHH, J 15.2 Hz), 4.98 (1H, dq, CHCH 3 , J7.2, 7.2 Hz), 5.02 (2H, dd,

CH

2 0, J 12.4, 28.8 Hz), 5.40 (2H, s, CH 2 0), 6.40 (1H, s, NH), 7.15-7.55 (19H, m, arom, NH), 8.16 (H, s, arom), 8.28 (1H, s, arom); Vmax 3373, 1732, 1666, 1486, 1250, 1077, 747 cm- 1 ; 25 m/z 633 (MH, 100 %), 486 (37 %); Anal. calc. for C 3 8

H

4 0

N

4 0 5 C 72.13, H 6.36, N 8.86 % found C 71.77, H 6.16, N 8.66 %; [cX]D 20 (c = 0.36, MeOH): 34.6 0. Step 5. Intermediate 3 30 Method as for Example 1, step 8 to give a clear glass (342 mg, quant.); WO 01/46176 55 PCT/EP00/13349 6 H 1.43 (3H, d, CHCH 3 , J7.6 Hz), 2.32 (6H, s, 2xNCH 3 ) 2.46 (1H, d, CHHN, J 12.4 Hz), 2.83 (1H, d, IndCHH, J 14.4 Hz), 3.13 (1H, d, CHHN, J 12.4 Hz), 3.20 (1H, d, IndCHH, J 14.4 Hz), 5.00 (11H, dq, CHCH 3 , J 7.6, 7.6 Hz), 6.74 (1H, s, arom), 7.04-7.26 (7H, m, arom), 7.33 (1H, d, arom, J 7.6 Hz), 7.61 (1H, d, arom, 5 J7.8 Hz), 7.89 (1H, bs, NHInd), 8.14 (1H, d, NH); [a]D 1 9 (c = 0.56, MeOH): 4.5 Step 6 Method as for Example 1, step 9 to yield a yellow glass (30 mg, 19 %); 10 6 H 1.44 (3H, d, CHCH 3 , J 7.0 Hz), 1.59 (1H, bs, NH), 2.34 (6H, s, N(CH 3

)

2 ), 2.67 (1H, d, CHHN, J 13.4 Hz), 2.96 (1H, d, CHH, J 13.4 Hz), 3.06 (1H, d, IndCHH, J 15.2 Hz), 3.29 (1H, d, IndCHH, J 15.2 Hz), 3.99 (2H, dd, CH 2 N, J 14.0, 24.4 Hz), 5.00 (1H, m, CHCH 3 ), 6.45 (1H, s, arom), 6.90 (1H, d, arom, J 7.4 Hz), 7.02 (2H, m, arom), 7.08-7.26 (7H, m, arom), 7.31 (1H, m, arom), 7.42 15 (1H, d, arom, J 8.0 Hz), 7.49 (1H, m, arom), 7.64 (1H, d, arom, J 8.0 Hz), 7.80 (1H, bs, NHind), 7.99 (1H, d, NH, J8.0 Hz); Vmax 3312, 1655, 1454, 741 cm- 1 ; m/z 495.3 (MH*, 100 %); 20 EXAMPLE 3. 2-[(Benzofuran-2-ylmethyl)-amino]-2-hydroxymethyl-3-(1H-indol-3-yl)-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)] H N OH H / N H N 0205 0 25 WO 01/46176 56 PCT/EPOO/13349 Step 1. LHMDS (7.81 ml, 7.81 mmol, IM in THF) was added to a solution of Intermediate 1 (2.00 g, 3.91 mmol), DMPU (0.47 ml, 3.91 mmol) in THF (30 ml) at -78 'C under dry N 2 . After 2 h at this temperature, iodoethane (1.22 g, 5 7.81 mmol) was added and the mixture allowed to warm to RT overnight. The solvent was removed in vacuo and the product extracted into EtOAc, washed with 10 % HCl, brine and dried (MgSO 4 ). Purification was achieved by chromatography (15-20 EtOAc in heptane) to leave a clear oil (1.53 g, 61 %); 6 H 0.04 (9H, s, Si(CH 3

)

3 ),0.85 (2H, t, CH 2 Si, J 8.0 Hz), 2.52 (1H, d, IndCHH, J 10 13.2 Hz), 2.84 (1H, dd, IndCHH, J 7.6, 13.2 Hz), 3.47 (2H, t, OCH 2 , J 8.0 Hz), 3.58 (1H, d, CHH, J 9.6 Hz), 3.78 (1H, dd, OCHH, J 5.6?, 13.2 Hz), 3.90 (1H, t, CH, J7.0 Hz), 4.15 (2H, m, OCHH, CHH), 4.96-5.15 (6H, m, 2x CH 2 0, CH 2 =), 5.38 (iH, m, CH=), 6.44 (1H, d, CH, J6.0 Hz), 7.00 (1H-, t, arom, J6.4 Hz), 7.07 (IH, d, arom, J 7.6 Hz), 7.19 (1H, t, arom, J7.6 Hz), 7.26-7.34 (10H, m, arom), 15 7.60 (1H, d, arom, J7.6 Hz) ; Vmax 2952, 1720, 1483, 1412, 1275, 838, 751 cm- 1 ; HRMS for C 36

H

4 3

NO

7 Si requires 643.2840 found 643.2840; [a]D 2 1 (c = 0.45, MeOH): + 13.9 20 Step 2. TFA (2 ml) was added to a solution of the above compound (1.66 g, 3.07 mmol) in dichloromethane (10 ml) and the resulting solution stirred at RT for 24 h. The solvent was removed and the residue diluted with EtOAc, the organics were washed with saturated NaHCO 3 , brine and dried (MgSO 4 ). The product was 25 purified by chromatography (15 % EtOAc in heptane) to yield a straw coloured oil (910 mg, 71 %); 6 H 3.01 (1H, bs, OH), 3.24 (1H, d, IndCHH, J 14.7 Hz), 3.54 (1H, d, IndCHH, J 14.7 Hz), 4.01 (1H, m, CHHO), 4.35 (1H, m, CHHO), 4.56 (2H, m, CH 2 0), 5.08 (iH, s, CH 2 O), 5.22 (2H, m, CH 2 =), 5.42 (1H, s, CH 2 0), 5.73 (1H1, s, NH), 5.79 30 (iH, m, CH=), 7.16 (1H, t, arom, J7.2 Hz), 8.14 (1H, bs, arom); vmax 3413, 1732, 1456, 1399, 1251, 1085, 749 cm- 1

;

WO 01/46176 57 PCT/EPOO/13349 HRMS for C 3 1

H

3 1

N

2 0 7 requires 543.2130 found 543.2130; [a]D 19 (c = 0.2, MeOH): + 2.4 o Step 3. 5 Imidazole (351 mg, 5.24 mmol) was added to a solution of triisopropylsilyl chloride (606 mg, 3.14 mmol) in DMF (5 ml), followed by the amino acid (1.42 g, 2.62 mmol). The resulting mixture warmed to 80 "C for 36 h, then the solvent was removed in vacuo and the residue diluted with EtOAc, washed with 10 % HCl, brine and dried (MgSO 4 ). The crude material was purified by 10 chromatography (10 % EtOAc in heptane) to yield a clear oil which solidified on standing. (1.36 g, 74%); mp 72-74 'C (pentane/Et 2 0); 6 H 1.02 (18H, s, 6xCH 3 ), 1.26 (3H, m, 3xCH), 3.17 (1H, d, IndCHH, J 14.4 Hz), 3.67 (1H, s, IndCHH, J 14.4 Hz), 4.09 (1H, d, CHHO, J 9.2 Hz), 4.49 (3H, m, 15 CH 2 0, CHHO), 5.05 (2H, s, CH 2 0), 5.20 (2H, m, CH 2 =), 5.41 (2H, s CH 2 O), 5.83 (2H, m, CH=, NH), 7.13 (lH, t, J 7.6 Hz), 7.26 -7.48 (13, m, arom), 8.17 (1H, bs, arom); Vmax 3422, 2944, 2866, 1741, 1248, 1086 cm- 1 ; Anal. calc. for C 4 0

H

5

ON

2 0 7 Si C 68.74, H 7.21, N 4.01 % found C 68.79, H 7.06, 20 N 4.08 %; m/z 699.2 (MH*). Step 4. Tetrakis(triphenylphosphine)palladium (0) (50 mg, 43 pmol) was added to a 25 solution of the allyl ester (1.14 g, 2.11 mmol) in THF (10 ml); after 5 min morpholine (1.84 g, 21.1 mmol) was added and the mixture stirred at RT for 30 min. EtOAc was added and the organics washed with 10 % HCl, brine and dried (MgSO 4 ). After removal of the solvent in vacuo a clear gum is obtained (845 mg, 94 %); 30 8H 1.03 (21H, m, Si(CHMe 2

)

3 ), 3.31 (1H, d, IndCHH, J 14.8 Hz), 3.63 (1H, d, IndCHH, J 14.8 Hz), 4.30 (2H, s, CH 2 O), 5.05 (2H, s, CH 2 0), 5.39 (2H, d, WO 01/46176 58 PCT/EPOO/13349

CH

2 0, J2.8 Hz), 5.76 (1H, s, NH), 7.12 (1H, t, arom, J7.6 Hz), 7.26-7.47 (13H, m, arom), 8.17 (1H, bs, arom); Vmax 3 4 11, 2 9 4 4 , 2 8 6 6 , 1733, 1456, 1400, 1249, 1086, 746 cm- 1 ; m/z 659.2 (MH+, 100 %); 5 Anal. calc. for C 37

H

4 6

N

3 0 7 Si C 67.45, H 7.04, N 4.25 % found C, 67.83, H 6.94, N 4.24 %; [a]D 2 1 (c = 0.40, MeOH): -1.0 0 Step 5. A mixture of the acid (1.01 g, 2.02 mmol), HBTU (766 mg, 2.02 mmol), DIPEA 10 (0.70 ml, 2.02 mmol) in DMF (10 ml) was stirred at RT for 10 min then (S) methylbenzylamine (244 mg, 2.02 mmol) and DIPEA (0.70 ml, 2.02 mmol) added and the resulting solution stirred for 8 h. The solvent was removed and the product extracted into EtOAc, washed with 10 % HCl, 10 % K 2 C0 3 , brine and dried (MgSO 4 ). Purification by chromatography gave a clear glass (533 mg, 15 92 %); 6 H 1.05 (21H, bs, Si(CHMe 2 )), 1.35 (3H, d, CHCH 3 , J 7.2 Hz), 3.40 (1H, d, IndCHH, J 13.6 Hz), 3.78 (1H, d. IndCHH, J 13.6 Hz), 3.80 (1H, m, CHHO), 4.65 (1H, m, CHHO), 4.93 (1H, m, CHCH 3 ), 5.03 (2H, s, CH 2 0), 5.40 (2H, d,

CH

2 0, J 5.2 Hz), 6.20 (1H, bs, NH), 7.17-7.46 (18H, m, arom), 8.18 (1H, bs, 20 arom); vmax 3372, 2944, 1733, 1674, 1486, 1456, 1398, 1249, 1077 cm- 1 ; m/z 620 (MH*, 100 %); Anal. calc. for C 4 5

H

5 5

N

3 0 6 Si.0.5 H 2 0 C 70.10, H 7.32, N 5.45 % found C 70.27, H 7.12, N 5.25 %; 25 [a]D 19 (c = 0.62, MeOH): -24.6 0 Step 6. Intermediate 4. A mixture of the amide (980 mg, 1.63 mmol), 10 % palladium hydroxide on carbon and methanol (20 ml) were hydrogenated at 50 psi (345 kPa) at 30 *C. 30 After 90 min the mixture was filtered through Kieselguhr and upon removal of the solvent in vacuo a white gum (325 mg, quant.) was obtained; WO 01/46176 59 PCT/EPOO/13349 51 1.10 (21H, m, Si(CHMe 2

)

3 ), 1.40 (3H, s, CHCH 3 , J 6.8 Hz), 1.61 (2H, bs, NH1 2 ), 3.02 (1H, d, IndCHJH, J 14.4 Hz), 3.32 (1H, d, IndCHH, J 14.4 Hz), 3.76 (1H, d, CHHO, J 8.8 Hz), 4.10 (11H, d, CHH, J 8.8 Hz), 5.01 (1H, m CHCH 3 ), 6.68 (1H, s, NH), 7.03 (3H, m, arom), 7.17-7.24 (4H, m, arom), 7.32 (1H, d, 5 arom, J7.6 Hz), 7.58 (1H, d, arom, J8.0 Hz), 7.83 (2H, bs, arom, NH); Vmax 3304, 2942, 2866, 1651, 1512, 1104, 741 cm- 1 ; m/z 494 (MH*); Anal. calc. for C 2 9

H

4 3

N

3 0 2 Si.0.3 H 2 0 C 69.78, H 8.80, N 8.42 % found C 69.84, H 8.49, N 8.11 %; 10 [at]D1 9 (c = 0.49, MeOH): -37.7 . Step 7. Method as for Example 2 step 6 to yield white fluffy crystals (103 mg, 41 %); 15 mp 110-111 C; 6 H 1.17 (21H, m, Si(CHMe 2

)

3 ), 1.36, (3H, d, CHCH 3 , J 6.8 Hz), 2.23 (1H, bs, NH), 3.17 (2H, dd, IndCH 2 , J 14.8, 29.2 Hz), 3.92 (1H, d, CHHN, J 14.0 Hz), 4.07 (1H, d, CHHN, J 14.0 Hz), 4.09 (1H, d, CHHO, J 10.0 Hz), 4.23 (1H, d, CHHO,J1O.0 Hz), 4.92 (1H, dt, CHCH 3 , J7.2, 7.2 Hz), 6.72 (1H, s, arom), 6.73 20 (1H, d, NH, J 2.4 Hz), 6.94 (2H, m, arom), 7.05 (1H, m, arom), 7.18 (7H, m, arom), 7.37 (1H, m, arom), 7.49 (1H, m, arom), 7.58 (2H, 2xd, arom, J 8.0 Hz), 7.69 (1H, s, NH); Vmax 3306, 2942, 2866, 1657, 1512, 1455, 1100, 741 cm- 1 ; m/z 624.1 (MH*); Anal. calc. for C 3 8

H

3 9

N

3 0 3 Si C 73.15, H 7.92, N 6.73 % 25 found C 73.48, H 7.81, N 6.73 %; [aID 19 (c = 0.49, MeOH): -22.0 .

WO 01/46176 60 PCT/EPOO/13349 Step 8. A solution of the TIPS-protected Intermediate (103 mg, 1.65 Imol) in THF (1 ml) was treated with TBAF (IM in THF, 331 tmol, 0.33 ml) and the resulting 5 solution left to stir for 1 h at RT. The solution was diluted with EtOAc, washed with brine and dried (MgSO 4 ). The product was purified by chromatography (20-40 % EtOAc in heptane) to yield a white foam (29 mg, 38 %); mp 53-56 'C; 6 H 1.52 (3H, d, CHCH 3 , J 7.2 Hz), 2.00 (1H, bs, NH), 2.98 (1H, d, IndCHH, J 10 14.6 Hz), 3.45 (1H, d, IndCHH, J 14.6 Hz), 3.75 (1H, d, CHHO, J 10.4 Hz), 3.81 (1H, bs, OH), 3.85 (2H, dd, CH 2 N, J 14.2, 24.3 Hz), 4.07 (1H, d, CHHO, J 10.4 Hz), 5.12 (1H, dq, CHCH 3 , J7.2, 7.2 Hz), 6.45 (1H, s, arom), 7.01 (1H, d, arom, J2.0 Hz), 7.09-7.40 (9H, m, arom, NH), 7.51 (1H, m, arom), 7.77 (1H, d, arom, J7.6 Hz), 8.00 (1H, d, arom, J8.0 Hz), 8.13 (1H, s, NH); 15 Vmax 3312, 1646, 1514, 1455, 740 cm- 1 ; m/z 468.1 (MH*, 100 %); Anal. calc. for C 29

H

2 9

N

3 0 3 .0.5 H 2 0 C: 73.09, H 6.34, N 8.82 % found C 73.02, H 6.19, N 8.92 %. 20 EXAMPLE 4. 2-(3-Benzofuran-2-ylmethyl-ureido)-3-(1H-indol-3-yl)-2-methoxymethyl-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)] HN 0 HH O H N N\ H 0_

_

WO 01/46176 61 PCT/EPOO/13349 A solution of Intermediate 2 (142 mg, 288 tmol) and (2 benzofuranyl)methylisocyanate (50 mg, 288 tmol) were stirred in THF (2 ml) under nitrogen for 2h. The solvent was removed in vacuo and the product purified by chromatography (20-25 % EtOAc in heptane) to give a white solid 5 (109 mg, 72 %); mp 78-82 'C; 6 H (DMSO) 1.38 (3H, d, CH 3 CH, J6.8 Hz), 3.37 (311, s, CH 3 0), 3.42, 3.68 (2H, dd, IndCH 2 , J 14.8 Hz), 3.52, 4.27 (2H, dd, CH 2 0, J 13.4 Hz), 4.38 (2H, 2, dd,

CH

2 N, J 6.0, 16.0 Hz), 4.93 (1H, dq, CHCH 3 , J 7.2 Hz), 5.26 (111, t, NH, J5.8 10 Hz), 5.68 (1H, s, NH), 6.67 (1H, d, NH, J2.4), 7.06 (1H, dt, arom, J 1.2, 8.0 Hz), 7.15 (1H, dt, arom, J 1.0, 68. Hz), 6.52 (1H, s, arom), 7.20-7.32 (811, m, arom), 7.41 (2H, m, arom), 7.51 (1H, m, arom), 7.60 (1H, d, arom, J7.6 Hz), 7.65 (1H, d, arom, J7.6 Hz), 10.80 (1H, bs, NH); Vmax 3327, 1644, 1557, 1455, 1253, 1106, 741 cm- 1 ; 15 Anal. calc. for C 3 1

H

3 2

N

4 0 4 .0.2 C 7

H

16 : C 71.45, H 6.51, N 10.29 % found C 71.34, H 6.38, N 10.02 %;

[X]D

1 9 (c 0.34, MeOH): -19.4 . EXAMPLE 5 20 2-(3-Benzofuran-2-ylmethyl-ureido)-2-hydroxymethyl-3-(1H-indol-3-yl)-N-( 1 phenyl-ethyl)-propionamide [S-(R*,R*)] HN OH O- N H 0

-

WO 01/46176 62 PCT/EPOO/13349 Step 1. Method as for Example 4 to give a clear glass (110 mg, 57 %); 6 H 1.07 (21H, m, 18xCH 3 ), 1.21 (3H, d, CH 3 CH, J 7.2 Hz), 3.42 (1H, d, IndCHH, J 14.8 Hz), 3.78 (1H, d, CHHO, J 10.0 Hz), 4.03 (1H, d, IndCHH, J 5 14.8 Hz), 4.12 (1H, m, CHHN, 4.30 (1H, dd, CHHN, J6.0, 15.6 Hz), 4.69 (1H, dq, CHCH 3 , J 7.2 Hz), 4.89 (1H, d, CHHO, J 9.6 Hz), 5.73 (1H, t, NH, J 5.6 Hz), 6.10 (1H, d, NH, J2.4 Hz), 7.05-7.25 (9H, m, arom), 7.33 (1H, m, arom), 7.43 (1H, m, arom), 7.62 (1H, d, arom, J 8.0 Hz), 7.91 (1H, d, arom, J 8.0 Hz) IndNH ?; 10 Vmax 3337, 1634, 1548, 1495, 1455, 1060, 741 cm- 1 ; HRMS for C 3 9

H

5 1

N

4 0 4 Si 667.368 found 667.3680 (MH+); [c]D 1 9 (c 0.70, MeOH): -18.9 . Step 2. 15 A solution of the TIPS-protected alcohol (110 mg, 165 tmol) in THF (1 ml) was treated with TBAF (0.33 ml, 330 pmol, 1M/THF) and the solution stirred at RT for 10 min. The mixture was diluted with EtOAc, washed with 10 % HCl, brine and dried (MgSO 4 ). The product was purified by chromatography (20-70% EtOAc in heptane) to yield a clear glass (20 mg, 24 %); 20 mp 82-82 *C; 6 H (DMSO) 1.27 (3H, d, CH 3 CH, J6.8 Hz), 3.27 (2H, dd, IndCH 2 , J 14.8, 38.0 Hz)3.79 (1H, dd, CHHO, J5.8, 12.0 Hz), 4.07 (1H, dd, CHHO, J7.0, 11.6 Hz), 4.29 (1H, dd, CHHN, J5.6, 16.0 Hz), 4.33 (lH, dd, CHHN, J5.6, 16.4 Hz), 4.91 (1H, dq, CHCH 3 , J 6.8 , 6.8 Hz), 4.93 (1H, bs, OH), 5.95 (1H, t, NHCH 2 , J 5.4 25 Hz), 6.02 (1H, s, NH), 6.43 (1H, s, arom), 6.68 (1H, d, NH, J 6.0 Hz), 7.04 (1H, m, arom), 7.09-7.27 (8H, m, arom), 7.35 (1H, d, arom, J 8.0 Hz), 7.47 (2H, m, arom), 7.54 (1H, s, arom), 8.01 (1H, d, NH, 17.6 Hz) IndH ?; Vmax 3346, 1644, 1557, 1455, 1253, 742 cm- 1 ; m/z 448 (M*-61, 100 %), 101 (16 %); 30 Anal. calc. for C 3 0

H

30

N

4 0 4 .0.5 EtOAc : C 69.30, H 6.18, N 10.10 % found C 69.18, H 6.03, N 10.29 %; WO 01/46176 63 PCT/EPOO/13349 [aX]D 19 (c 0.46, MeOH): -15.6 0. EXAMPLE 6 (R)-C- [(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1 -hydroxymethyl 5 1H-indol-3-ylmethyl)-N-((S)- 1 -phenyl-ethyl)-propionamide H nj",N O N Ph

N-

OH Potassium hexamethyldisilazide (4.4 ml, 2.02 mmol, 0.5 M in toluene) was 10 added to a solution of Example 2 (1.0 g, 2.02 mmol) in THF (15 ml) at --78 *C under N 2 . Stirring was continued at this temperature for 10 min then a freshly prepared solution of formaldehyde (approximatively 250 mg in 15 ml) was added and stirring continued at -78 *C for 3 h. The mixture was diluted with EtOAc, washed with saturated NaHCO 3 , brine and dried (MgSO 4 ). The product 15 was purified by chromatography (0-1 % MeOH in CH 2 Cl 2 ) to yield a yellow foam (182 mg, 17 %). MS m/z 525.6 (MH+, BP); IR v 3353, 1646, 1454, 1040, 741 cm-1; IH NMR 8 1.42 (3H, d, CHCH 3 , J 7.2 Hz), 2.33 (6H, s, N(CH 3

)

2 ), 2.55 (1H, bs, NH), 2.65 (1H, d, CHHN, J 13.6 Hz), 2.97 (2H, 2xd, CHHN, CHHInd, J 15.6 Hz), 3.25 20 (111, d, CHHInd, J 15.6 Hz), 3.94 (2H, dd, CH 2 N, J 13.6, 19.6 Hz), 4.98 (1H, dq,

CHCH

3 , J 7.6 Hz), 5.26 (2H, bs, CH 2 0), 6.44 (1H, s, ar), 6.84 (1H, s, ar), 7.05 (2H, m, ar), 7.12-7.26 (7H, m, ar), 7.41 (2H, m, ar), 7.51 (1H, d, ar, J 5.6 Hz), 7.60 (1H, d, ar, J8.0 Hz), 8.00 (1H, d, NH, J7.2).

WO 01/46176 64 PCT/EPOO/13349 EXAMPLE 7 (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1 dimethylaminomethyl- 1H-indol-3-ylmethyl)-N-((S)- 1 -phenyl-ethyl) propionamide 5 -N H N O N Ph

N-

zz--N- X // Lithium hexamethyldisilazide (1.1 ml, 1.01 mmol, 1 M in THF) was added to a solution of Example 2 (500 mg, 1.01 mmol) in THF (5 ml) at -78 *C under N 2 . 10 Stirring was continued at this temperature for 15 min, then Eschenmoser's salt (37 mg, 2.02 mmol) was added and stirring continued at -78 'C for 1 h and the mixture allowed to warm to RT overnight. The mixture was diluted with EtOAc, washed with saturated NaHC0 3 , brine and dried (MgSO 4 ). The product was purified by chromatography (0-2 % MeOH in CH 2 Cl 2 ) to yield a yellow gum 15 (68 mg, 12 %). MS m/z 552 (MH, BP); IR v 3366, 1661, 1494, 1454, 740 cm 1 ; 'H NMR 6 1.42 (3H, d, CHCH 3 , J7.2 Hz), 2.15 33 (6H, s, N(CH 3

)

2 ), 2.33 (6H, s, N(CH 3

)

2 ), 2.66 (1H, d, CHHN, J 13.6 Hz), 2.95 (1H, d, CHHN, J 13.6 Hz), 3.03 (1H, d, CHHInd, J 15.6 Hz), 3.30 (1H, d, CHHInd, J 15.2 Hz), 3.95 (2H, dd, CH 2 N, J 20 14.0, 25.2 Hz), 4.50 (2H, s, NCH 2 N), 4.98 (1H, dq, CHCH 3 , J7.2 Hz), 6.43 (1H, s, ar), 6.94 (1H, s, ar), 7.01 (2H, m, ar), 7.08(1H, t, ar, J7.6 Hz), 7.16-7.26 (6H, m, ar), 7.35 (1H, d, ar, J 8.4 Hz), 7.38 (1H, d, ar, J 8.0 Hz), 7.48 (1H, d, ar, 8.0 Hz), 7.61 (1H, d, ar, J7.6 Hz), 8.00 (1H, d, NH, J7.6 Hz).

WO 01/46176 65 PCT/EPOO/13349 EXAMPLE 8 Alternative synthesis of 2-[(Benzofuran-2-ylmethyl)-amino]-2 dimethylaminomethyl-3-(1H-indol-3-yl)-N-(1-phenyl-ethyl)-propionamide (S,S) 5 Example 8.A: synthesis of Intermediate 5 1) Preparation of (S)-2-(benzylidene-amino)-3-(1H-indol-3-yl)-propionic acid methyl ester (Schiffs base). 10 245 g (S)-Tryptophan methyl ester and 118 g benzaldehyde are dissolved in 1837 ml dichloromethane and mixed with 245 g dry magnesium sulphate. The reaction mixture is stirred for 4 hours at ambient temperature. After filtering off the magnesium sulphate, the solvent is distilled off on a rotary evaporator. The very viscous mass remaining behind (364 g) can be used directly in the next step. 15 2) Preparation of racemic a-dimethylaminomethyltryptophan methyl ester 117.9 g Diisopropylamine are dissolved in 1170 ml anhydrous tetrahydrofuran in a reaction flask gassed with nitrogen with the exclusion of moisture. 728 ml 20 20 of a 1.6 M butyl lithium/hexane solution are added dropwise at -30'C within the course of about 1 hour. After stirring for about 15 minutes, 340 g of the Schiffs base prepared in Example 1, dissolved in 1020 ml anhydrous tetrahydrofuran, are added dropwise at -30'C over the course of 1 hour. After a further 15 minutes, 205 g 1-dimethylaminomethyl-benzotriazole, dissolved in 1025 ml anhydrous 25 tetrahydrofuran are added dropwise at -30'C over the course of 1 hour. Subsequently, the reaction mixture is allowed to warm up slowly to ambient temperature. During the following approximately 16 hours stirring at ambient temperature, a thick slurry results. With ice cooling, a solution of 386 ml 37% hydrochloric acid in 1544 ml ice 30 water is allowed to run in in such a manner that the temperature in the reaction vessel does not exceed 25'C. The separating tetrahydrofuran phase is separated WO 01/46176 66 PCT/EPOO/13349 off and the aqueous phase extracted five times with 500 ml amounts of ethyl acetate. The aqueous phase is covered with 500 ml ethyl acetate and mixed portionwise, while stirring, with 247 g sodium carbonate. The organic phase is separated off 5 and the aqueous phase again extracted twice with, in each case, 500 ml ethyl acetate. The combined organic phases are washed twice with, in each case, 300 ml of a saturated aqueous solution of sodium chloride. After drying over anhydrous sodium sulphate, the solution was filtered clear and evaporated in a vacuum. 10 There remain 242 g (79.1% of theory) of a brown, viscous residue. According to HPLC analysis, this crude product contains about 62% of the desired product. For further purification, the crude product is recrystallised from diethyl ether to give 94 g (30.7% of theory) racemic x-dimethylaminomethyltryptophan methyl ester; m.p. 105*'C; HPLC purity 96.2 rel.%. 15 3) Separation of racemic c-dimethylaminomethyltryptophan methyl ester into the two enantiomeric compounds. 15 g of the racemic a-dimethylaminomethyltryptophan methyl ester obtained in 20 Example 2 are separated on a preparative HPLC apparatus on the chiral phase Chiracel OJ 20 ptm into 6.2 g of the (R)-enantiomer and 6.45 g of the (S) enantiomer. Example 8.B: synthesis of 2-[(Benzofuran-2-ylmethyl)-amino]-2 25 dimethylaminomethyl-3-(1H-indol-3-yl)-N-(1-phenyl-ethyl)-propionamide (S,S) Step 1: (S)-2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H indol-3-yl)-propionic acid methy 1 ester (S)-2-Amino-2-dimethylaminomethyl-3-(1H-indol-3-yl)-propionic acid methyl 30 ester (5.69 g, 20.7 mmol), benzofuran-2-carbaldehyde (4.77 g, 32.7 mmol) and sodium triacetoxyborohydride (9.24 g, 43. 6 mmol) were stirred in 1,2- WO 01/46176 67 PCTIEPOO/13349 dichloroethane (100 ml) for 8 h at RT. The mixture was diluted with CH 2

CI

2 , washed with NaOH (0.5 M), brine and dried (MgSO 4 ). After removal of the solvent the material was washed with heptane to leave fawn coloured solid (6.45 g, 77 %). 5 6 H 2.27 (6H, s, N(CH 3

)

2 ), 2.74 (2H, dd, CH 2 N, J 13.6, 20.4 Hz), 3.27 (2H, s,

CH

2 Ind), 3.63 (3H, s, OMe), 4.08 (2H, dd, CH 2 N, J 14.0, 38.8 Hz), 6.57 (lH, s, arom), 7.09-7.34 (6H, m, arom), 7.42 (111, m, arom), 7.51 (1H, m, arom), 7.63 (1H, d, arom, J 8.0 Hz), 8.03 (11H, s, NH). Vmax 1718, 142, 1174, 742 cm- 1 ; 10 m/z 406 (MH*, 100); Step 2: (S)-2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H indol-3-yl)-propionic acid bis-hydrochloride (S)-2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H-indol-3 15 yl)-propionic acid methy 1 ester (6.35 g, 15.7 mmol) and sodium hydroxide (608 mg, 15.7 mmol) in 1,4-dioxan/water were heated under reflux conitions for 4 days. The solvent was removed and the mixture acidified with 10 % HCl, the solvent was again removed to leave a brown foam (-10 g, > quant.) 6 H (DMSO-d 6 ) 2.18 (6H, s, N(CH 3

)

2 ), 2.48 (2H, s, CH 2 N), J 13.6, 20.4 Hz), 3.01 20 (2H, s, CH 2 Ind), 3.88 (2H, dd, CH 2 N), 6.57 (1H, d, arom, JO.4 Hz)), 6.89 (1H, m, arom), 6.96 (1H, m, arom), 7.14-7.28 (4H, m, arom), 7.45 (1H, m, arom), 7.52 (1H, m, arom), 7.57 (1H, d, arom, J 8.0 Hz), 10.69 (1H, s, NH). Step 3: (S)-2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H 25 indol-3-yl)-N-((S)-1-phenyl-ethyl)-propionamide (S)-2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H-indol-3 yl)-propionic acid bis-hydrochloride (6.13 g, 15.7 mml assumed), HBTU (5.95 g, 15.7 mmol), (S)-alpha-methylbenzylamine (2.85 g, 23.6 mmol) and DIPEA (10.13 g, 78.5 mmol) were stirred at RT in DMF (75 ml) for 6 h. The solvent 30 was removed and the material taken up in EtOAc, washed with 15 % K 2

CO

3 , brine and dried (MgS04). The crude material was purified by chromatography (20-50 % EtOAc in C 7 ) to leave a yellow solid which was washed with heptane WO 01/46176 68 PCT/EPOO/13349 to leave a yellow powder. Further material was obtained by re-purifying the impure fractions by RP-HPLC (0-100 % MeOH in H 2 0) to leave a yellow-white solid. Total yield (1.91 g , 25 %). 5 Data as example 2. EXAMPLE 9 Binding to NKI receptors 10 The compounds of the present invention are highly selective and competitive antagonists of the NKI 1 receptor. They have been evaluated in an NKI receptor binding assay which is described below. Methods 15 Human lymphoma IM9 cells are grown in RPMI 1640 culture medium supplemented with 10% foetal calf serum and 2mM glutamine and maintained under an atmosphere of 5% C02. Cells are passaged every 3-4 days by reseeding to a concentration of 4-8 million/40 ml per 175 cm 2 flask. Cells are harvested for experiments by centrifugation at 1000 g for 3 min. Pelleted cells are washed once 20 by resuspension into assay buffer (50 mM Tris HCl pH 7.4, 3 mM MnCl 2 , 0.02 % BSA, 40 mg/mL bacitracin, 2 mg/mL chymostatin, 2 mM phosphoramidon, 4 mg/mL leupeptin) and repeating the centrifugation step before resuspending at a concentration of 2.5 x 106 cells/mL assay buffer. Cells (200 ml) are incubated with

[

12 5 1]Bolton-Hunter Substance P (0.05-0.1 nM) in the presence and absence of 25 varying concentrations of test compounds for 50 min at 21 0 C. Non-specific binding (10% of the total binding observed under these conditions) is defined by 1 mM [Sar 9 ,Met(0 2 )11]substance P. Reactions are terminated by rapid filtration under vacuum onto GF\C filters presoaked in 0.2 % PEI for 1-2 h, using a Brandel cell harvester. Filters are washed with 6 x 1 ml ice-cold Tris HCl (50 mM, pH 7.4) and 30 radioactivity bound determined using a gamma counter. Results are analysed using iterative curve fitting procedures in RS 1 or Graphpad Inplot.

WO 01/46176 69 PCTIEPOO/13349 Results Table I: In Vitro Human NKI Receptor Binding Assay Example No. NKI binding

IC

5 0 (nM) 1 6.56 2 0.89 3 1.41 4 4.93 5 1.07 6 28.8 7 6.96 5 Because the compounds are potent ligands to the NK 1 receptor, they are effective at displacing substance P at that position, and therefore are useful for treating biological conditions otherwise mediated by substance P. Accordingly, compounds capable of antagonising the effects of substance P at NK 1 receptors will be useful in treating or preventing a variety of CNS disorders including pain 10 (inflammatory, surgical and neuropathic), anxiety, panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, COPD and psoriasis; gastrointestinal disorders including colitis, Crohn's disease, irritable 15 bowel syndrome and satiety; allergic responses such as eczema and rhinitis; vascular disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis. The compounds of the invention, NK 1 receptor antagonists, are also useful as anti-angiogenic agents, for the treatment of conditions associated with aberrant neovascularization such as rheumatoid 20 arthritis, atherosclerosis and tumour cell growth. They will also be useful as agents for imaging NK 1 receptors in vivo in conditions such as ulcerative colitis and Crohn's disease.

WO 01/46176 70 PCTIEPOO/13349 EXAMPLE 10. Carrageenan-induced hypersensitivity model in the guinea-pig 5 Methods Male Dunkin Hartley guinea-pigs (200-250 g) are housed in groups of 4 under a 12 hour light/dark cycle (lights on at 7:00) with food and water ad libitum. Carrageenan-induced hypersensitivity: Guinea-pigs are administered carrageenan (100 pl of 20 mg/ml) by intraplantar 10 injection into the right hind paw. They are tested for hypersensitivity in the weight-bearing test, using an "Incapacitance tester" (Linton Instruments, U.K.): the animal is placed in the apparatus and the weight load exerted by the hind paws is noted. The measurements are taken three times at one-minute intervals and the average is calculated. The duration of the measurement is adjusted to 4 s 15 for the guinea-pig. The animals are tested before (baseline) and at different intervals after the injection of carrageenan. The compound of Example 2 was administered subcutaneously 1 h before carrageenan in a dosing volume of 1 ml/kg, in PEG 200 vehicle. Hypersensitivity was assessed using the weight bearing test. 20 The difference in weight-bearing between the ipsilateral and contralateral paws is calculated and is then subjected to a one-way-ANOVA followed by Dunnett's t test, for each time-point studied (*P<0.05, **P<0.01, significantly different from vehicle treated animals). Results are expressed as mean difference in weight load between ipsilateral and 25 contralateral paws ± SEM (g) (n per group = 6-19). Results The intraplantar injection of carrageenan (100 pl of 20 mg/ml) into the hindpaw induces hypersensitivity in the guinea-pig, as assessed by the weight bearing test. 30 The subcutaneous injection of the compound of Example 2 (0.1 and 1 mg/kg, in PEG 200 vehicle) 1 h before carrageenan dose-dependently prevents the development of hypersensitivity 3 h after carrageenan (Figure. 1).

WO 01/46176 71 PCT/EPOO/13349 As noted above, the compounds of Formula I will be best utilized in the form of pharmaceutical formulations. The following examples further illustrate specific formulations that are provided by the invention. 5 EXAMPLE 11 Tablet Formulation Ingredient Amount 2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H indol-3-yl)-N-(1 -phenyl-ethyl)-propionamide (S,S) 50 mg potato starch 100 mg talc 50 mg magnesium carbonate 20 mg dextrose 20 mg 240 mg 10 The above ingredients are blended to uniformity and pressed into a tablet. Such tablets are administered to human subjects from one to four times a day for the treatment of conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth.

WO 01/46176 72 PCT/EPOO/13349 EXAMPLE 12 Parenteral injection Ingredient Amount 2-(3-Benzofuran-2-ylmethyl-ureido)-3-(1H-indol-3-yl)-2 methoxymethyl-N-(1-phenyl-ethyl)-propionamide [S-(R*,R*)] 20 mg Citric acid monohydrate 0.75 mg Sodium phosphate 4.5 mg Sodium chloride 9 mg Water for injection to 10 ml 5 The sodium phosphate, citric acid monohydrate and sodium chloride are dissolved in a portion of the water. The active ingredient is dissolved in the solution and made up to volume. 10 EXAMPLE 13 Parenteral injection Ingredient Amount 2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3 (1H-indol-3-yl)-N-(1-phenyl-ethyl)-propionamide (S,S) 20 mg Citric acid monohydrate 0.75 mg Sodium phosphate 4.5 mg Sodium chloride 9 mg Water for injection to 10 ml WO 01/46176 73 PCT/EPOO/13349 The sodium phosphate, citric acid monohydrate and sodium chloride are dissolved in a portion of the water. The active ingredient is dissolved in the solution and made up to volume.

Claims (18)

1. A compound of Formula I R1 R4 R -(C H 2 ) m X C COZ -C :R6 (CH 2 ) n R5 5 R2 (I) or a pharmaceutically acceptable salt thereof wherein: * and A indicate all stereoisomers; R is phenyl, pyridyl, 10 thienyl, furyl, quinolyl isoquinolyl naphthyl, 15 benzofuryl, benzo[1,3]dioxole benzothienyl or, benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, 20 hydroxy, alkoxy, halogen, CF 3 or OCF 3 ; WO 01/46176 75 PCT/EPOO/13349 m is an integer from I to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; RI is (CH 2 )pY where p is 0 to 3 and Y is OH, OCH 3 , F, CF 3 , CO 2 H, N(CH 3 ) 2 , NHCH 3 , NH2, COCF 3 , COCH 3 or NO 2 ; 5 n is an integer from 1 to 2; R2 is naphthyl or indolyl unsubstituted or N-substituted with hydroxy, alkyl, CH 2 N O or CH 2 N N formyl, CH 2 OH, CH 2 N(CH 3 ) 2 , Z is NR3 or 0, where R3 is H or C 1 -C 4 alkyl; 10 R4 and R5 are each independently hydrogen, or (CH2)pR7 where: p is an integer of I to 3, and R7 is H, CH 3 , CN, OH, OCH 3 , CO 2 CH 3 , NH 2 , NHCH 3 , or N(CH 3 ) 2 ; R6 is phenyl, pyridyl, 15 thienyl, furyl, pyrrolyl, pyrazolyl, imidazolyl, 20 quinolyl, isoquinolyl, naphthyl, indolyl, benzofuryl, 25 benzothiophenyl, benzimidazolyl, or benzoxazolyl, wherein each of the foregoing is unsubstituted, mono-, di or trisubstituted by alkyl, hydroxy, 30 alkoxy, WO 01/46176 76 PCT/EPOO/13349 halogen, CF 3 , NO 2 , N(CH3)2, 5 OCF 3 , SONH 2 , NHl 2 , CONH 2 , CO 2 CH 3 or 10 CO 2 H, or R6 is: straight alkyl of from 1 to 3 carbons, branched alkyl of from 3 to 8 carbons, cycloalkyl of from 5 to 8 carbons or 15 heterocycloalkyl, each of which can be substituted with up to one or two substituents selected from OH, CO 2 H, 20 N(CH3)2, NHCH 3 and CH 3 ; or R5 and R6, when joined by a bond, form a ring. WO 01/46176 77 PCT/EPOO/13349

2. A compound of Formula I wherein R1 R4 R -CH 2 ) m X C COZ - C -R6 (CH 2 ) n R5 R2 e is R or S, and A is R or S; 5 -R is phenyl, pyridyl, thienyl, furyl, quinolyl 10 isoquinolyl benzofuryl, benzo[1,3]dioxole benzothienyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by 15 alkyl, hydroxy, alkoxy, halogen, CF 3 or 20 OCF 3 ; m is an integer from I to 3; X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; RI is (CH 2 )pY where p is 0 to 3 and Y is OH, F, CF 3 , OCH 3 , CO 2 H, N(CH 3 ) 2 , NHCH 3 , NH 2 , COCF 3 , COCH 3 or NO 2 ; WO 01/46176 78 PCT/EPOO/13349 n is an integer from 1 to 2; R2 is naphthyl or indolyl unsubstituted or N-substituted with hydroxy, alkyl, CH 2 N(CH 3 ) 2 , CH 2 N 0 or CH 2N N formyl, CH 2 OH, 5 Z is NR3 or 0, where R3 is H or CH 3 ; R4 and R5 are each independently hydrogen, CH 3 or CH 2 OH; R6 is phenyl, pyridyl, thienyl, 10 furyl, pyrrolyl, cyclohexyl or benzimidazolyl, wherein each of the foregoing is unsubstituted, mono-, di- or 15 trisubstituted by alkyl, hydroxy, alkoxy, halogen, 20 CF 3 , NO 2 , or N(CH 3 ) 2 . WO 01/46176 79 PCT/EPOO/13349

3. A compound of Formula I wherein R1 R4 R -CH 2 )m X __C -- COZ -- C :R6 (CH 2 )n R5 R2 (I) * is R or S, and A is R or S; R is phenyl, 5 pyridyl, thienyl, furyl, benzofuryl, benzo[1,3]dioxole 10 benzothienyl or benzimidazolyl, each unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, 15 alkoxy, halogen, CF 3 or OCF 3 ; m is an integer from I to 3; 20 X is NR8 or NHCONH where R8 is H or alkyl of 1 to 3 carbon atoms; Ri is (CH 2 )pY where p is 0 to 3 and Y is OH, OCH 3 , F, CF 3 , CO 2 H, N(CH 3 ) 2 , NHCH 3 , NH 2 , COCF 3 , COCH 3 or NO 2 ; n is an integer from 1 to 2; R2 is indolyl unsubstituted or N-substituted with alkyl or formyl; WO 01/46176 80 PCT/EPOO/13349 Z is NR3 or 0, where R3 is H or CH 3 ; R4 and R5 are each independently hydrogen, CH 3 , or CH 2 OH; R6 is phenyl, pyridyl, 5 thienyl, furyl, pyrrolyl, cyclohexyl or benzimidazolyl, 10 each unsubstituted, mono-, di- or trisubstituted by alkyl, hydroxy, alkoxy, halogen, 15 CF 3 , NO 2 or N(CH 3 ) 2

4. A compound according to Claim 1 selected from: 20 2-[(Benzofuran-2-ylmethyl)-amino]-3-(1H-indol-3-yl)-2-methoxymethyl N-(1-phenyl-ethyl)-propionamide [S-(R*,R*)], 2-[(Benzofuran-2-ylmethyl)-amino]-2-dimethylaminomethyl-3-(1H indol-3-yl)-N-(1-phenyl-ethyl)-propionamide (S,S), 2-[(Benzofuran-2-ylmethyl)-amino]-2-hydroxymethyl-3-(1H-indol-3-yl) 25 N-(1-phenyl-ethyl)-propionamide [S-(R*,R*)], 2-(3-Benzofuran-2-ylmethyl-ureido)-2-hydroxymethyl-3-(1H-indol-3-yl) N-(1-phenyl-ethyl)-propionamide [S-(R*,R*)], and 2-(3-Benzofuran-2-ylmethyl-ureido)-3-(1H-indol-3-yl)-2 methoxymethyl-N-(1-phenyl-ethyl)-propionamide [S-(R*,R*)]. 30 (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-(1 hydroxymethyl-1H-indol-3-ylmethyl)-N-((S)-1-phenyl-ethyl) propionamide WO 01/46176 81 PCTIEPOO/13349 (R)-C-[(Benzofuran-2-ylmethyl)-amino]-dimethylamino-C-( 1 dimethylaminomethyl-1H-indol-3-ylmethyl)-N-((S)-1 -phenyl-ethyl) propionamide 5

5. A method for antagonizing NK 1 receptors in a mammal in need of treatment comprising administering to a mammal an effective amount of a compound of Claim 1.

6. A method for treating or preventing: 10 - CNS disorders such as pain (inflammatory, surgical and neuropathic), anxiety, panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, obesity and addiction disorders; 15 - inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and psoriasis; - gastrointestinal disorders including colitis, Crohn's disease, irritable bowel syndrome and satiety; - allergic responses such as eczema and rhinitis; 20 - vascular disorders such as angina and migraine; - neuropathological disorders including scleroderma and emesis; and - conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth, 25 comprising administering to a mammal in need of treatment an effective amount of a compound of Claim 1.

7. A compound according to Claim 1 to be used as a medicament. 30

8. A pharmaceutical formulation comprising a compound of Claim 1 admixed with at least one pharmaceutically acceptable diluent, carrier or excipient. WO 01/46176 82 PCTIEPOO/13349

9. A pharmaceutical formulation comprising a compound of Claim 3 admixed with at least one pharmaceutically acceptable diluent, carrier or excipient. 5

10. A method for treating conditions associated with aberrant neovascularization such as rheumatoid arthritis, atherosclerosis and tumour cell growth. 10

11. A method for imaging NKI receptors in vivo in conditions such as ulcerative colitis and Crohn's disease.

12. Use of a compound of Claim 1 for the preparation of a medicament intended for preventing or treating CNS disorders such as pain 15 (inflammatory, surgical and neuropathic), anxiety, panic, depression, major depression with anxiety, schizophrenia, neuralgia, stress, sexual dysfunction, bipolar disorders, movement disorders, cognitive disorders, obesity and addiction disorders; inflammatory diseases such as arthritis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD) and 20 psoriasis; gastrointestinal disorders including colitis, Crohn's disease, irritable bowel syndrome and satiety; allergic responses such as eczema and rhinitis; vascular disorders such as angina and migraine; neuropathological disorders including scleroderma and emesis; conditions associated with aberrant neovascularization such as rheumatoid arthritis, 25 atherosclerosis and tumour cell growth.

13. Process for the preparation of ((S)-2-benzylideneamino)-3-(1H-indol-3 yl)-propionic acid methyl ester which comprises reacting (S)-tryptophan 30 methyl ester with benzaldehyde and recovering the desired product. WO 01/46176 83 PCT/EPOO/13349

14. Process for the preparation of a-dimethylaminomethyltryptophan methyl ester, wherein (S)-2-benzylidene-amino)-3-(1H-indol-3-yl)-propionic acid methyl ester is reacted with 1-dimethylaminomethylbenzotriazole to yield racemic c-dimethylaminomethyltryptophan methyl ester. 5

15. Process according to claim 14 wherein the racemic methyl ester obtained is separated into the (R)- and (S)-enantiomers.

16. Process for the preparation of 2-[(Benzofuran-2-ylmethyl)-amino]-2 10 dimethylaminomethyl-3-(1H-indol-3-yl)-N-(1-phenyl-ethyl) propionamide (S,S) wherein (S)-2-[(Benzofuran-2-ylmethyl)-amino]-2 dimethylaminomethyl-3-(1H-indol-3-yl)-propionic acid bis-hydrochloride is reacted with (S)-alpha-methylbenzylamine. 15

17. Process for the preparation of (R)-C-[(Benzofuran-2-ylmethyl)-amino] dimethylamino-C-(1-hydroxymethyl-1H-indol-3-ylmethyl)-N-((S)-1 phenyl-ethyl)-propionamide wherein 2-[(Benzofuran-2-ylmethyl) amino] -2-dimethylaminomethyl-3 -(1H-indol-3 -yl)-N-(1 -phenyl-ethyl) propionamide (S,S) is reacted with potassium hexamethyldisilazide and 20 formaldehyde.

18. Process for the preparation of (R)-C-[(Benzofuran-2-ylmethyl)-amino] dimethylamino-C-(1-dimethylaminomethyl-1H-indol-3-ylmethyl)-N-((S) 1-phenyl-ethyl)-propionamide wherein 2-[(Benzofuran-2-ylmethyl) 25 amino] -2-dimethylaminomethyl-3 -(1H-indol-3 -yl)-N-(1 -phenyl-ethyl) propionamide (S,S) is reacted with lithium hexamethyldisilazide and Eschenmoser's salt.