BACKGROUND OF THE INVENTION Growth hormone is a hormone which stimulates growth of all tissues capable of growing. In addition, growth hormone is known to have a number of effects on metabolic processes, e.g., stimulation of protein synthesis and free fatty acid mobilization and to cause a switch in energy metabolism from carbohydrate to fatty acid metabolism. Deficiency in growth hormone can re- sult in a number of severe medical disorders, e.g., dwarfism.

Growth hormone is released from the pituitary. The release is under tight control of a number of hormones and neurotransmitters either directly or indirectly. Growth hormone release can be stimulated by growth hormone releasing hormone (GHRH) and inhibited by somatostatin. In both cases the hormones are released from the hypothalamus but their action is mediated pri- marily via specific receptors located in the pituitary. Other compounds which stimulate the re- lease of growth hormone from the pituitary have also been described. For example arginine, L-3,4-dihydroxyphenylalanine (L-Dopa), glucagon, vasopressin, PACAP (pituitary adenylyl cy- clase activating peptide), muscarinic receptor agonists and a synthethic hexapeptide, GHRP (growth hormone releasing peptide) release endogenous growth hormone either by a direct effect on the pituitary or by affecting the release of GHRH and/or somatostatin from the hypo- thalamus.

In disorders or conditions where increased levels of growth hormone is desired, the protein nature of growth hormone makes anything but parenteral administration non-viable. Further- more, other directly acting natural secretagogues, e.g., GHRH and PACAP, are longer polypeptides for which reason oral administration of them is not viable.

The use of certain compounds for increasing the levels of growth hormone in mammals has previously been proposed, e.g. in EP 18 072, EP 83 864, WO 89/07110, WO 89/01711, WO 89/10933, WO 88/9780, WO 83/02272, WO 91/18016, WO 92/01711, WO 93/04081, WO 95/17422, WO 95/17423 and WO 95/14666.

The composition of growth hormone releasing compounds is important for their growth hor- mone releasing potency as well as their bioavailability. It is therefore an object of the present invention to provide novel compounds with growth hormone releasing properties. Moreover it is an object to provide growth hormone releasing compounds with no or substantially no side- effects. It is also an object to provide compounds which have good oral bioavailability.

SUMMARY OF THE INVENTION In accordance with the present invention there is provided compounds which act directly on the pituitary cells under normal experimental conditions in vitro to release growth hormone there- from.

These growth hormone releasing compounds can be utilised in vitro as unique research tools for understanding, inter alia, how growth hormone secretion is regulated at the pituitary level.

Moreover, the growth hormone releasing compounds of the present invention can also be ad- ministered in vivo to increase growth hormone release.

Accordingly, the present invention relates to a compound of general formula I wherein R3, R4, R5, X, Y, M, T and Q are as defined below, or a pharmaceutically acceptable salt thereof, having growth hormone releasing properties.

DESCRIPTION OF THE INVENTION In a broad aspect the present invention relates to a compound of general formula I wherein R3, R4 and R5 independently of each other are hydrogen,or C,6 alkyl optionally substitued with C,-6 alkyl, X is aryl optionally substituted with halogen, C16 alkyl or phenyl, Y is aryl or hetaryl optionally substituted with halogen,C,6 alkyl, C1-6 alkoxy or phenyl, M is M', M2, M3, M4 or M6, wherein M1 is -C(=O)-CH=CH-(CH2)m-C(R16R17)-N(R1 R2), M2 is -C(=O)-CH((CH2)m-hetaryl)-NH-C(=O)-C(R'6R'7)-N(R'R2), wherein m is 1, 2 or 3, and R16 and R17 independently of each other are C,6 alkyl, optionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C1-6 alkoxy or phenyl; M3 is -C(=O)-D1-CH2-N(R1R2), M4 is -C(=O)-D1-C(R6R7)-N(R1R2), and M5 is -C(=O)-(CH2)s-O-(CH2)p-C(R6R7)q-A, wherein R6 and R7 independently of each other are hydrogen, C,4 alkyl, D1 is arylene, such as phenylene or naphthylene, preferably phenylene, p and s independently of each other are 1, 2 or 3, q is 0 or 1, and A is -N(R'R2) or a saturated heterocyclic ring containing 5 or 6 ring members, one ring member being a heteroatom such as N, S and/or 0, preferably N, wherein R' and R2 independently of each other are hydrogen, -C(=O)-C14 alkyl, C,4 alkyl op- tionally substituted with halogen, amino, hydroxyl, C1-6 alkyl, C1-6 alkoxy or phenyl; or benzyl, optionally substituted with halogen, amino, hydroxyl, C1.6 alkyl, C1-6 alkoxy or phenyl, T is hydrogen, T1, T2 or T3, wherein T1 is -(CH2)n-NH2, wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10,

T2 is -(CH2)n-N(R8R9), wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R8 and R9 independently of each other are C,6 alkyl optionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C,6 alkoxy or phenyl, or R8 and Re may be joined to form a saturated heterocyclic ring containing 5 or 6 ring members, one of the ring members being N and the other being carbon atoms; and T3 is -(CH2)n-NHZ, wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and Z is -(C=O)-R10, -C(=O)-O-R10, -SO2R'°, -C(=NH)-NR11R12 or -C(=O)-NR11R12, wherein R10 is hydrogen, C,6 alkyl optionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C1-6 alkoxy or phenyl; benzyl, optionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C1-6 alkoxy or phenyl; or 3a,7a,12a- trihydroxy-5b-cholanyl, and R" and R12 independently of each other are hydrogen, C,6 alkyl optionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C1-6 alkoxy or phenyl; or benzyl, optionally substituted with halogen, amino, hydroxyl, C1-6 alkyl, C,6 alkoxy or phenyl, Q is hydrogen, Q1, Q2 or wherein Q1 is -C(=O)-NHR13, Q2 is -C(=O)-NH2, and Q3 is -C(=O)-NR14R15, wherein R13, R14 and R15 independently of each other are C,6 alkyl op- tionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C1-6 alkoxy or phenyl; or a pharmaceutically acceptable salt thereof; with the proviso(s) that if M is M2 then T cannot be hydrogen, T' or T2, if T is hydrogen then Q cannot be hydrogen, if M is M' or M3 then R4 cannot be hydrogen, if M is M3 and X is 2-naphthyl and Y is phenyl and T is T' then Q cannot be Q2, if T is T2 and n is 2 then R5 cannot be hydrogen, if Q is Q3 and Y is phenyl and X is 2-naphthyl and R3, R4 and R5 are methyl, then M cannot be M1, or if T is T2 and n is 3 and Y is phenyl and X is 2-naphthyl and R3, R4 and R5 are methyl, then M cannot be M'.

In a more narrow aspect the present invention relates to a compound of general formula I wherein R3, R4 and R5 independently of each other are hydrogen,or C1-6 alkyl optionally substitued with C1-6 alkyl, X is aryl optionally substituted with halogen, C16 alkyl or phenyl, Y is aryl or hetaryl optionally substituted with halogen, C1-6 alkyl, C16 alkoxy or phenyl, M is M', M2, M3, M4 or M5, wherein M1 is -C(=O)-CH=CH-(CH,),-C(R1GR'7)~N(R' R2), M2 is -C(=O)-CH((CH2)m-hetaryl)-NH-C(=O)-C(R16R17)-N(R1R2), wherein m is 1, 2 or 3, and R16 and R'7 independently of each other are C,4 alkyl, optionally substituted with halogen, amino, hydroxyl, C16 alkyl, C1-6 alkoxy or phenyl; M3 is -C(=O)-D1-CH2-N(R1R2), M4 is -C(=O)-D1-C(R6R7)-N(R1 R2), and M5 is -C(=O)-(CH2)s-O-(CH2)p-C(RGR7),-A, wherein R6 and R7 independently of each other are hydrogen, C,4 alkyl, D' is arylene, such as phenylene or naphthylene, preferably phenylene, p and s independently of each other are 1, 2 or 3, q is 0 or 1, and A is -N(R'R2) or a saturated heterocyclic ring containing 5 or 6 ring members, one ring member being a heteroatom such as N, S and/or 0, preferably N, wherein R1 and R2 independently of each other are hydrogen, -C(=O)-C1-6 alkyl, C1-6 alkyl op- tionally substituted with halogen, amino, hydroxyl, C1-6 alkyl, C16 alkoxy or phenyl; or benzyl, optionally substituted with halogen, amino, hydroxyl, C1-6 alkyl, C16 alkoxy or phenyl, T is hydrogen, T1, T2 or T3, wherein T1 is -(CH2)n-NH2, wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, T2 is -(CH2)n-N(R8R9), wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R8 and R9 independently of each other are C1-6 alkyl optionally substituted with halogen, amino, hydroxyl, C16 alkyl, C1-6

alkoxy or phenyl, or R8 and R9 may be joined to form a saturated heterocyclic ring containing 5 or 6 ring members, one of the ring members being N and the other being carbon atoms; and T3 is -(CH2)n-NHZ, wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and Z is -C(=O)-R10, -C(=O)-O-R10, -SO2R'° or -C(=O)-NR11R12, wherein R10 is hydrogen, C16 alkyl optionally substituted with halo- gen, amino, hydroxyl, C16 alkyl, C,6 alkoxy or phenyl; benzyl, optionally substituted with halo- gen, amino, hydroxyl, C,6 alkyl, C,6 alkoxy or phenyl; or 3a,7a,12a-trihydroxy-5b-cholanyl, and R11 and R12 independently of each other are hydrogen, C16 alkyl optionally substituted with halogen, amino, hydroxyl, C,4 alkyl, C,6 alkoxy or phenyl; or benzyl, optionally substituted with halogen, amino, hydroxyl, C1-6 alkyl, C1-6 alkoxy or phenyl, Q is hydrogen, Q1, Q2 or Q3, wherein Q' is -C(=O)-NHR13, Q2 is -C(=O)-NH2, and Q3 is -C(=O)-NR14R15, wherein R'3, R14 and R15 independently of each other are C,6 alkyl op- tionally substituted with halogen, amino, hydroxyl, C,6 alkyl, C,6 alkoxy or phenyl; or a pharmaceutically acceptable salt thereof; with the proviso(s) that if M is M2 then T cannot be hydrogen, T' or T2, if T is hydrogen then Q cannot be hydrogen, if M is M' or M3 then R4 cannot be hydrogen, if M is M3 and X is 2-naphthyl and Y is phenyl and T is T' then Q cannot be Q2, if T is T2 and n is 2 then R5 cannot be hydrogen, if Q is Q3 and Y is phenyl and X is 2-naphthyl and R3, R4 and R5 are methyl, then M cannot be M1, or if T is T2 and n is 3 and Y is phenyl and X is 2-naphthyl and R3, R4 and R5 are methyl, then M cannot be M1.

The compounds of formula I comprise any optical isomers thereof, in the form of separated, pure or partially purified optical isomers or racemic mixtures thereof. For instance, whenever

one or more chiral carbonatoms are present such chiral center(s) may be in the R- or S- configuration, or a mixture of R and S.

In one embodiment of the compound of the above formula I there is the following further pro- viso(s) that if M is M3 and X is phenyl and Y is phenyl and T is T' then Q cannot be Q2, if T is H and X is napthyl and Y is phenyl and R6 is H then M cannot be M1, or if n is 2 and Q is H and T is T2 and X is napthyl and Y is phenyl then M cannot be M,.

In one further embodiment of the compound of the above formula I R' and R2 are independ- ently of each other hydrogen, C,6alkyl, or -C(=O)-C,6alkyl, preferably hydrogen, methyl or - C(=O)-CH3. In a particular embodiment R1 and R2 are both hydrogen or one is hydrogen and the other is methyl or -C(=O)-C,6 alkyl e.g. -C(=O)-CH3. Especially preferred R1 is hydrogen and R2 is hydrogen, methyl or -C(=O)-CH3.

In another embodiment of the compound of the above formula I M is hydrogen, -C(=O)- CH=CH-(CH2)-C(R16R17)-N(R1R2), -C(=O)-CH(CH2-hetary)-NH-C(=O)-C(R16R17)-N(R1R2), wherein R16 and R17 independently of each other are C,6 alkyl, optionally substituted with halo- gen, amino, hydroxyl, C,6 alkyl, C,6 alkoxy or phenyl; -C(=O)-D1-C(R6R7)-N(R1R2), -C(=O)- CH2-O-CH2-C(R6R7)-N(R1R2) or -C(=O)-CH2-O-CH2-A, wherein R6 and R7 independently of each other are hydrogen or C,4 alkyl, A is a saturated heterocyclic ring containing 5 or 6 ring members, one ring member being a nitrogen atom, and D', R' and R2 are as defined above.

When M is -C(=O)-CH=CH-(CH2)-C(R16R17)-N(R1R2), it is in either cis or trans configuration, trans being preferred. When M is -C(R16R17)-C(=O)-NH-CH(CH2-hetaryl)-C(=O)-N(R1R2), hetaryl is preferably imidazolyl, which may be attached to the methylene group in any of the five positions, the 5-position being preferred. Moreover hetaryl is connected through a meth- ylen group to a chiral carbonatom, the hetaryl-CH2-moiety being in either R- or S- configuration, preferably S-configuration. In one embodiment of M1 or M2, R16 and R'7 are in- dependently of each other C,6 alkyl, preferably methyl, and in a particular embodiment R16 and R'7 are both methyl. When M is -C(=O)-D1-C(R6R7)-N(R1R2), D1 is preferably phenylene, which may be attached to the carbonyl group through any of its five ring positions, the 3-

position being preferred. In one embodiment of M4, R6 and R7 are independently of each other hydrogen or C16 alkyl, preferably hydrogen or methyl. When R6 and R7 are both hydro- gen, then M4 is M3. In a particular embodiment R6 and R7 are both hydrogen or one is hydro- gen and the other is C16 alkyl, preferably methyl. When M is -C(=O)-CH2-O-CH2-C(R6R7)- N(R1R2), R6 and R7 are independently of each other hydrogen or C16 alkyl, preferably hydro- gen or ethyl. When M is -C(=O)-CH2-O-CH2-A, A is a saturated heterocyclic ring containing 5 ring members, one ring member being a nitrogen atom (N), and most preferred A is pyrrolidi- nyl, such as pyrrolidin-2-yl in the S-configuration. Especially preferred M is hydrogen, -C(=O)- CH=CH-CH2-C(CH3)2-NH2 in trans configuration, -C(=O)-CH(CH2-(1 H-imidazol-5-yl))-NH- C(=O)-C(CH3)2-NH2, -C(=O)-CH(CH2-(1 H-imidazol-5-yl))-NH-C(=O)-C(CH3)2-NH(C(=O)-CH3), - C(=O)-(m-phenylene)-CH2-NH2, -C(=O)-(m-phenylene)-CH2-NH-CH3, -C(=O)-(m-phenylene)- CH2-NH(C(=O)-CH3), -C(=O)-(m-phenylene)-CH(CH3)-NH2, -C(=O)-CH2-O-CH2-CH(CH2CH3)- NH2 or-C(=O)-CH2-O-CH2-((2S)-pyrrolidin-2-yl).

In a further embodiment of the compound of the above formula I R3, R4 and R5 independently of each other are hydrogen or C,6 alkyl, preferably hydrogen or methyl. In a particular em- bodiment R3 is methyl, R4 is methyl and R5 is methyl, or R3 is methyl, R4 is methyl and R5 is hy- drogen, or R3 is hydrogen, R4 is methyl and R5 is hydrogen, or R3 is hydrogen, R4 is hydrogen and R5 is hydrogen.

In a still further embodiment of the compound of the above formula I X is aryl optionally substi- tuted with C16 alkyl or phenyl. Preferably X is naphthyl, such as 1-naphthyl or 2-naphthyl, or phenyl substituted with C16 alkyl or phenyl, preferably naphthyl or phenyl substituted with phenyl, which C,4 alkyl or phenyl may be attached to phenyl through any one of the five ring positions, the 4-position being preferred. Especially preferred X is 2-naphthyl or biphenylA-yl.

Moreover, aryl, such as phenyl or naphthyl, is connected through a methylene group to a chiral carbonatom, the aryl-CH2-moiety being in either R- or S-configuration, preferably R- configuration.

In a further embodiment of the compound of the above formula I Y is hetaryl or phenyl option- ally substituted with halogen, such as chloro or fluoro. Preferably Y is phenyl optionally substi- tuted with halogen, such as fluoro; or thienyl. When Y is thienyl it may be attached to the meth- ylene group through any one of its four ring positions, the 2-position being preferred. Especially preferred Y is phenyl, 2-thienyl or 4-flouro-phenyl. Moreover, hetaryl or phenyl is connected

through a methylene group to a chiral carbonatom, the (hetaryl or phenyl)-CH2-moiety being in either R- or S-configuration, preferably R-configuration.

In a still further embodiment of the compound of the above formula I Q is hydrogen, -C(=O)- NHR'3, -C(=O)-NH2 or -C(=O)-NR14R15, wherein R13, R14 and R'5 independently of each other are C,6 alkyl. When Q is -C(=O)-NHR'3, R'3 is preferably C1.4alkyl, such as methyl. When Q is - C(=O)-NR14R16, R14 and R15 are independently of each other preferably C14alkyl, and in par- ticular methyl. Especially preferred Q is hydrogen, -C(=O)-NH-CH3, -C(=O)-NH2 or -C(=O)- N(CH3)2.

In a further embodiment of the compound of the above formula I T is hydrogen, -(CH2)n-NH2, - (CH2)n-N(R8R9), wherein R8 and R9 independently of each other are C16 alkyl or R8 and R9 are joined to form a saturated heterocyclic ring containing 5 ring members, one of the ring mem- bers being nitrogen and the other four being carbon atoms; or -(CH2)n-NHZ, wherein n is 1, 2, 3, 4, 5 or 6, and Z is -C(=O)-R10, -C(=O)-O-R10, SO2R'°, -C(=NH)-NR11R12 or -C(=O)-NR"R'2, wherein R10 is hydrogen, C1-6 alkyl, benzyl or 3a,7a,12a-trihydroxy-5b-cholanyl, and R" and R12 independently of each other are hydrogen, C16 alkyl or benzyl. When T is -(CH2)n-NH2, n is preferably 1, 2, 3, 4 or 5. When T is -(CH2)n-N(R8R9), R8 and R9 are independently of each other C14 alkyl, preferably methyl, and n is 1, 2, 3, 4, 5 or 6. In particular when T is T2, n is 1, 2 or 3. When T is -(CH2)n-NHZ, wherein n is 1, 2, 3, 4, 5 or 6, Z is -C(=O)-R10, -C(=O)-O-R10, SO2R'°, wherein R10, is Cr4 alkyl, preferably C1-4 alkyl such as methyl or t-butyl, 3a,7a,12a- trihydroxy-5b-cholanyl or benzyl; or -C(=O)-NR"R'2, wherein R" and R12 independently of each other are hydrogen or C,4 alkyl, preferably hydrogen or ethyl; or -C(=NH)-NR11R12, wherein R" and R12 independently of each other are hydrogen or Cr4 alkyl, preferably hydro- gen. When T is -(CH2)n-NHZ, n is preferably 1, 2, 3, 4 or 5, most preferred 3 or 4. Especially preferred T is hydrogen, -CH2-NH2, -(CH2)2-NH2, -(CH2)3-NH2, -(CH2)4-NH2, -(CH2)5-NH2, -CH2- N(CH3)2,-(CH2)2-N(CH3)2, -(CH2)3-N(CH3)2,-(CH2)3-NH-C(=O)-NH2, -(CH2)4-NH-C(=O)-CH3, - (CH2)4-NHSO2CH3, -(CH2)4-NH-C(=NH)-NH2, -(CH2)4-NH-C(=O)-NH-CH2-CH3, -(CH2)4-NH- C(=O)-O-t-butyl or -(CH2)4-NH-C(=O)-O-CH2-phenyl. Moreover, T is connected to a carbon atom, which dependent on the remaining substituents may be in either R- or S-configuration, in which case the S-configuration is preferred.

Preferred compounds of the invention are: 3-Aminomethyl-N-((1 R)-1 -(((1 R)-2-(2-thienyl)-1 -((1 S)-1 -carbamoyl-5- aminopentylcarbamoyl)ethyl)-N-methylcarbamoyl)-2-(2-napthyl) ethyl)benzamide, 3-((1 R/S)-1 -Aminoethyl)-N-((1 R)-1 -(N-[(1 R)-1 -(N-(carbamoylmethyl)-N-methylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methy lbenzamide, 2-Amino-N-(((1 S)-1 -(((1 R)-1 -(((1 R)-1 -(((1 S)-1 -carbamoyl-5- (methylsulfonylamino)pentyl)carbamoyl)-2-phenylethyl)carbamo yl)-2-(2- naphthyl)ethyl)carbamoyl)-2-(4-imidazolyl)ethyl)carbamoyl)-2 -methylpropanamide, [(5S)-5-((2R)-2-(N-[(2R)-2-(3-(Aminomethyl)benzoylamino)-3-( 2-naphthyl)propionyl]-N- methylamino)-3-(2-thienyl)propionylamino)-5-carbamoylpentyl] carbamic acid benzyl ester, N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]- N-methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-3-aminomethylbenz amide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(5-aminopentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methy lamide, 3-((1 R/S)- 1 -Aminoethyl)-N-1(1 R)-2-(biphenyl-4-yl)- 1 -(N-((1 R)- 1 -[N-(3-dimethylaminopropyl)- N-methylcarbamoyl]-2-phenylethyl}-N-methylcarbamoyl)ethyl]-N -methylbenzamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-[( 1 R)-2-(biphenyl-4-yl)-1 -(N-((l R)-1-[N-(3- dimethylaminopropyl)-N-methylcarbamoyl]-2-phenylethyl)-N-met hylcarbamoyl)ethyl]-N- methylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-(( 1 R)-1 -(N-[( 1 R)-1 -((1 S)-5-amino-1 - carbamoylpentylcarbamoyl)-2-phenylethyl]-N-methylcarbamoyl}- 2-(2-naphthyl)ethyl)-N- methylamide, N-((1 R)-l-(N-[(l R)-1 -((1 S)-5-Amino-1 -carbamoylpentylcarbamoyl)-2-phenylethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-3-((1 R/S)-1 -aminoethyl)-N-methylbenzamide, (2E)-5-Amino-5-methylhex-2-enoic acid ((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-amino-1 - (dimethylcarbamoyl)pentyl)carbamoyl)-2-phenylethyl]-N-methyl carbamoyl}-2-(2- naphthyl)ethyl)amide, N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-Amino-1 -(dimethylcarbamoyl)pentyl)carbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-3-am inomethyl-N-methylbenzamide, (2E)-5-Amino-5-methyl-hex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -((1 S)-5-amino-1 - (dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyl-N-methylca rbamoyl}-2-(biphenyl-4- yl)ethyl)-N-methylamide, N-((1 R)-1 -([(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-3-aminomethylbenzamide ,

(2E)-5-Amino-5-methylhex-2-enoic acid ((1 R)-1 -(N-[( 1 R)-1 -(((5S)-5-amino-1 - carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]-N-methyl carbamoyl}-2-(2- naphthyl)ethyl)amide, N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]- N-methylcarbamoyl}-2-(2-naphtyl)ethyl)-3-aminomethyl-N-methy lbenzamide, (2S)-6-Amino-2-[(2R)-3-(4-fluorophenyl)-2-(N-methyl-N-{(2R)- 3-(2-naphthyl)-2-[2-(((2S)- pyrrolidin-2-yl)methoxy)acetylamino]-propionyl}amino)propion ylamino]hexanoic acid amide, (2S)-6-Amino-2-[(2R)-2-(N-{(2R)-2-[2-((2R/S)-2-aminobutoxy)a cetylamino]-3-(2- naphthyl)propionyl}-N-methylamino)-3-(4-fluorophenyl)propion ylamino]hexanoic acid amide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(((1 S)-5-amino-1 - <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl) -2-(2-naphthyl)ethyl)amide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-amino-1 - <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-phenyIethyl-N-methylcarbamoyl)- 2-(biphenylA-yl)ethyl)amide, N-((1 R)-l-(N-[(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-phenylethyl]-N- methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-3-aminomethylbenzam ide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(((1 S)-5-amino-1 - carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylcarba moyl}-2-(2- naphthyl)ethyl)amide, 3-Aminomethyl-N-((1 R)-1 -(((1 R)-2-(2-thienyl)-1 -((1 S)-1 -carbamoyl-5- aminopentylcarbamoyl)ethyl)-N-methylcarbamoyl)-2-(2-napthyl) ethyl)benzamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(((1 S)-5-amino-1 - carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylcarba moyl}-2-(biphenyl-4- yl)ethyl)amide, N-(( 1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N- methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-3-aminomethylbenzam ide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 {N-[(1 R)-1 -(((1 S)-5-acetylamino-1 - <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl) -2-(2-naphthyl)ethyl)amide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-acetylamino-1 - carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylcarba moyl)-2-(biphenyl-4- yl)ethyl)amide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1{N-[(1 R)-1 -(((1 S)-5-acetylamino-1 - carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl} -2-(biphenyl-4- yl)ethyl)amide,

(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-l-(N-[(1 R)-1 -((1 S)-5-acetylamino-1 - carbamoylpentylcarbamoyl)-2-(2-thienyl)ethyl]-N-methylcarbam oyl}-2-(2- naphthyl)ethyl}amide, (2)-5-Amino-5-methylhex-2-enoic acid N-((1R)-l-(N-[(l R)-1 -(6-aminohexylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(4-aminobutylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(3-aminopropylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(2-aminoethylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide, 3-(1 -Aminoethyl)-N-((l R)-1 -(N-((1 R)-1 -(N-(3-dimethylaminopropyl)-N-methylcarbamoyl)-2- phenylethyl)-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lbenzamide, 3-(1 -Aminoethyl)-N-((1 R)-1 -(N-((1 R)-1 -(N-((dimethylcarbamoyl)methyl)-N- methylcar- bamoyl)-2-phenylethyl)-N-methylcarbamoyl)-2-(2-naphthyl)ethy l)-N-methylbenzamide, <BR> <BR> <BR> <BR> (2S)-6-Acetylamino-2-((2R)-2-((2R)-2-(2S)-2-(2-amino-2-methy lprnpionylamino)-3-(imidazol- 4-yl)propionylamino]-3-(2-naphthyl)propionylamino}-3-phenylp ropionylamino)hexanoic acid amide, <BR> <BR> <BR> <BR> (2S)-5-Ureido-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2-methylpro pionylamino)-3-(3H-imidazol-4- yl)propionylamino]-3-(2-naphthyl)propionylamino}-3-phenylpro pionylamino)pentanoic acid amide, (2S)-6-tert Butyloxycarbonylamino-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2- methylpropionylamino)-3-(imidazol-4-yl)propionylamino]-3-(2- naphthyl)propionylamino}-3- phenylpropionylamino)hexanoic acid amide, (2S)-6-Acetylamino-2-((2R)-2{N-((2R)-2-[(2S)-2-(2-amino-2-me thylpropionylamino)-3- (imidazol-4-yl)propionylamino]-3-(2-naphthyl)propionyl)-N-me thylamino}-3- phenylpropionylamino)hexanoic acid amide, (2S)-6-(3a,7a,12a-trihydroxy-5b-cholanoylamino)-2-((2R)-2-{( 2R)-2-[(2S)-2-(2-amino-2- <BR> <BR> <BR> <BR> methylpropionylamino)-3-(3H-imidazol-4-yl)propionylamino]-3- (2-naphthyl)propionylamino)-3- phenylpropionylamino)hexanoic acid amide, (2E)-5-Amino-5-methylhex-2-enoic acid ((1 R)-1 -(N-[(1 R)-1 -(5-aminopentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)amide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(5-aminopentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methy lamide,

N-((1R)-1-{N-[(1R)-1-(4-Aminobutylcarbamoyl)-2-phenylethyl]- N-methylcarbamoyl}-2- (biphenyl-4-yl)ethyl)-3-aminomethyl-N-methylbenzamide, 3-Aminomethyl-N-({1R}-1-{N-[(1R)-1-(5-aminopentylcarbamoyl)- 2-phenylethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)benzamide, 3-Aminomethyl-N-((1 R)-1 -(N-[(1 R)-1 (5-aminopentylcarbamoyl)-2-phenylethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methylbenzamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1-((4- <BR> <BR> <BR> <BR> dimethylaminobutyl)carbamoyl)-2-phenylethyl]-N-methylcarbamo yl)-2-(2-naphthyl)ethyl)-N- methylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(5-guanidinopentylcarbamoyl)- 2-phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-met hylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-(( 1 R)- 1 -(N-((1 R)-1-[5-(3- ethylureido)pentylcarbamoyl]-2-phenylethyl}-N-methylcarbamoy l)-2-(2-naphthyl)ethyl]-N- methylamide, 3-Aminomethyl-N-[(1 R)- 1 -(N-{(1 R)- 1 -[N-(2-(dimethylamino)ethyl)-N-methylcarbamoyl]-2- phenylethyl}-N-methylcarbamoyl)-2-(2-naphthyl)ethyl]-N-methy lbenzamide, N-[(1R)-1-(N-{(1R)-1-[N-(2-(Dimethylamino)ethyl)-N-methylcar bamoyl]-2-phenylethyl}-N- methylcarbamoyl)-2-(2-naphthyl)ethyl]-N-methyl-3-(methylamin omethyl)benzamide, 3-((1 R/S)-1 -Aminoethyl)-N-[(1 R)-1 -(N-((1 R)-1 -[N-(2-(dimethylamino)ethyl)-N- methylcarbamoyl]-2-phenylethyl}-N-methylcarbamoyl)-2-(2-naph thyl)ethyl]-N- methylbenzamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-2-(biphenyl-4-yl)-1 -(N-[(1 R)-1-(4- <BR> <BR> <BR> <BR> (dimethylamino)butylcarbamoyl)-2-phenylethyl]-N-methylcarbam oyl)ethyl)-N-methylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-[(1 R)-1 -(N-((1 R)-1 -[N-(2-dimethylaminoethyl)-N- <BR> <BR> <BR> <BR> methylcarbamoyl]-2-phenylethyl)-N-methylcarbamoyl)-2-(2-naph thyl)ethyl]-N-methylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1-((4- <BR> <BR> <BR> <BR> dimethylaminobutyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylca rbamoyl)-2-(2-naphthyl)ethyl)- N-methylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-2-(4-biphenyl-4-yl)-1-{N-[(1R)-1-((4- <BR> <BR> <BR> <BR> dimethylaminobutyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylca rbamoyl)ethyl)-N-methylamide, (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -((1 S)-5-(acetylamino)-1 - (dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methylc arbamoyl)-2-(2- naphthyl)ethyl)-N-methylamide, and

(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -((1 S)-5-acetylamino-1 - <BR> <BR> <BR> <BR> (methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methylcar bamoyl)-2-(2-naphthyl)ethyl)- N-methylamide.

The compounds of formula I exhibit an improved resistance to proteolytic degradation by en- zymes compared to that of the peptides suggested in the prior literature, due to the lack of natural peptide bonds. The increased resistance to proteolytic degradation of the compounds of the invention in comparison with known growth hormone releasing peptides is expected to improve their bioavailability compared to that of the peptides suggested in the prior literature.

In the above structural formulas and throughout the present specification, the following terms have the indicated meanings: The C,6-alkyl, C16-alkylene, C,4-alkyl or C,-alkylene groups specified above are intended to include those alkyl or alkylene groups of the designated length in either a linear or branched or cyclic configuration. Examples of linear alkyl are methyl, ethyl, propyl, butyl, pentyl, and hexyl and their corresponding divalent moieties, such as ethylene. Examples of branched alkyl are isopropyl, sec-butyl, tert-butyl, isopentyl, and isohexyl and their corresponding divalent moie- ties, such as isopropylene. Examples of cyclic alkyl are C,-cycloalkyl such as cyclopropyl, cy- clobutyl, cyclopentyl and cyclohexyl and their corresponding divalent moieties, such as cyclo- propylene.

The C,6-alkoxy groups specified above are intended to include those alkoxy groups of the designated length in either a linear or branched or cyclic configuration. Examples of linear alk- oxy are methoxy, ethoxy, propoxy, butoxy, pentoxy, and hexoxy. Examples of branched alkoxy are isopropoxy, sec-butoxy, tert-butoxy, isopentoxy, and isohexoxy. Examples of cyclic alkoxy are cyclopropyloxy, cyciobutyloxy, cyclopentyloxy and cyclohexyloxy.

In the present context, the term "aryl" is intended to include monovalent carbocyclic aromatic ring moieties, being either monocyclic, bicyclic or polycyclic, e.g. selected from the group con- sisting of phenyl and naphthyl, optionally substituted with one or more C,.6-alkyl, C,4-alkoxy, halogen, amino or aryl.

In the present context, the term "arylene" is intended to include divalent carbocyclic aromatic ring moieties, being either monocyclic, bicyclic or polycyclic, e.g. selected from the group con-

sisting of phenylene and naphthylene, optionally substituted with one or more C,6-alkyl, C1.6- alkoxy, halogen, amino or aryl.

In the present context, the term "hetaryl" is intended to include monovalent heterocyclic aro- matic ring moieties, being either monocyclic, bicyclic or polycyclic, e.g. selected from the group consisting of pyridyl, 1-H-tetrazol-5-yl, thiazolyl, imidazolyl, indolyl, pyrimidinyl, thiadiazolyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thienyl, quinolinyl, pyrazinyl, or isothiazolyl, option- ally substituted by one or more C,6-alkyl, C,6-alkoxy, halogen, amino or aryl.

In the present context, the term "hetarylene" is intended to include divalent heterocyclic aro- matic ring moieties, being either monocyclic, bicyclic or polycyclic, e.g. selected from the group consisting of pyridinediyl, 1-H-tetrazolediyl, thiazoldiyl, imidazolediyl, indolediyl, pyrimidinediyl, thiadiazolediyl, pyrazolediyl, oxazolediyl, isoxazolediyl, oxadiazolediyl, thiophenediyl, quinolin- ediyl, pyrazinediyl, or isothiazolediyl, optionally substituted by one or more C,6-alkyl, C1.6- alkoxy, halogen, amino or aryl.

The term "halogen" is intended to include chlorine (Cl), fluorine (F), bromine (Br) and iodine (I).

The compounds of the present invention may have one or more asymmetric centres and it is intended that stereoisomers, as separated, pure or partially purified stereoisomers or racemic mixtures thereof are included in the scope of the invention.

The compounds of the present invention may optionally be on a pharmaceutically acceptable salt form such as the pharmaceutically acceptable acid addition salts of compounds of formula I which include those prepared by reacting the compound of formula I with an inorganic or or- ganic acid such as hydrochloric, hydrobromic, sulfuric, acetic, phosphoric, lactic, maleic, man- delic phthalic, citric, glutaric, gluconic, methanesulfonic, salicylic, succinic, tartaric, toluenesul- fonic, trifluoracetic, sulfamic or fumaric acid.

The compounds of formula I may be administered in pharmaceutically acceptable acid addition salt form or, where appropriate, as a alkali metal or alkaline earth metal or lower alkylammo- nium salt. Such salt forms are believed to exhibit approximately the same order of activity as the free base forms.

In another aspect, the present invention relates to a pharmaceutical composition comprising, as an active ingredient, a compound of the general formula I or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier or diluent.

Pharmaceutical compositions containing a compound of the present invention may be pre- pared by conventional techniques, e.g. as described in Remington's Pharmaceutical Sciences, 1985 or in Remington: The Science and Practice of Pharmacy, 19th Edition (1995). The com- positions may appear in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or topical applications.

The pharmaceutical carrier or diluent employed may be a conventional solid or liquid carrier.

Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pec- tin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, polyoxyethylene or water.

Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.

If a solid carrier is used for oral administration, the preparation may be tabletted, placed in a hard gelatin capsule in powder or pellet form or it can be in the form of a troche or lozenge.

The amount of solid carrier will vary widely but will usually be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.

A typical tablet which may be prepared by conventional tabletting techniques may contain: Core: Active compound (as free compound or salt thereof) 100mg Colloidal silicon dioxide (Aerosil) 1.5mg Cellulose, microcryst. (Avicel) 70mg Modified cellulose gum (Ac-Di-Sol) 7.5mg Magnesium stearate

Coating: HPMC approx. 9mg *Mywacett 940 T approx. 0.9mg *Acylated monoglyceride used as plasticizer for film coating.

For nasal administration, the preparation may contain a compound of formula I dissolved or suspended in a liquid carrier, in particular an aqueous carrier, for aerosol application. The car- rier may contain additives such as solubilizing agents, e.g. propylene glycol, surfactants, ab- sorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives such as parabenes.

Generally, the compounds of the present invention are dispensed in unit dosage form com- prising 50-200 mg of active ingredient together with a pharmaceutically acceptable carrier per unit dosage.

It has been demonstrated that compounds of the general formula I possess the ability to re- lease endogenous growth hormone in vivo. The compounds may therefore be used in the treatment of conditions which require increased plasma growth hormone levels such as in growth hormone deficient humans or in elderly patients or livestock.

Thus, in a particular aspect, the present invention relates to a pharmaceutical composition for stimulating the release of growth hormone from the pituitary, the composition comprising, as an active ingredient, a compound of the general formula I or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier or diluent.

In a further aspect, the present invention relates to a method of stimulating the release of growth hormone from the pituitary, the method comprising administering to a subject in need thereof an effective amount of a compound of the general formula I or a pharmaceutically ac- ceptable salt thereof.

In a still further aspect, the present invention relates to the use of a compound of the general formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for stimulating the release of growth hormone from the pituitary.

To those skilled in the art, it is well known that the current and potential uses of growth hor- mone in humans are varied and multitudinous. Thus, compounds of formula I can be adminis- tered for purposes stimulating release of growth hormone from the pituitary and would then have similar effects or uses as growth hormone itself. The uses of growth hormone may be summarized as follows: stimulation of growth hormone release in the elderly, prevention of cataboiic side effects of glucocorticoids, prevention and treatment of osteoporosis, treatment of chronic fatigue syndrom (CFS), treatment of acute fatigue syndrom and muscle loss following election surgery, stimulation of the immune system, acceleration of wound healing, accelerat- ing bone fracture repair, accelerating complicated fractures, e.g. disctraction osteogenesis, treatment of wasting secondary to fractures, treatment of growth retardation, treating growth retardation resulting from renal failure or insufficiency, treatment of cardiomyopathy, treatment of chronic liver disease, treatment of thrombocytopenia, treatment of Crohn's disease, treat- ment of short bowel syndrome, treatment of chronic obstructive pulmonary disease (COPD), treatment of complications associated with transplantation, treatment of physiological short stature including growth hormone deficient children and short stature associated with chronic illness, treatment of obesity and growth retardation associated with obesity, treatment of ano- rexia, treating growth retardation associated with the Prader-Willi syndrome and Turner's syn- drome; accelerating the recovery and reducing hospitalization of burn patients; treatment of intrauterine growth retardation, skeletal dysplasia, hypercortisolism and Cushing's syndrome; induction of pulsatile growth hormone release; replacement of growth hormone in stressed pa- tients, treatment of osteochondrodysplasias, Noonan's syndrome, schizophrenia, depressions, Alzheimer's disease, delayed wound healing and psychosocial deprivation, treatment of pul- monary dysfunction and ventilator dependency, attenuation of protein cataboiic responses after major surgery, reducing cachexia and protein loss due to chronic illness such as cancer or AIDS; treatment of hyperinsulinemia including nesidioblastosis, adjuvant treatment for ovulation induction; to stimulate thymic development and prevent the age-related decline of thymic func- tion, treatment of immunosuppressed patients, improvement in muscle strength, mobility, maintenance of skin thickness, metabolic homeostasis, renal homeostasis in the frail elderly, stimulation of osteoblasts, bone remodelling and cartilage growth, stimulation of the immune system in companion animals and treatment of disorder of aging in companion animals, growth

promoter in livestock and stimulation of wool growth in sheep, and treatment of metabolic syn- drom (syndromex). Moreover the compounds of formula I may be used in the treatment of in- sulin resistance, including NIDDM, in mammals, e.g. humans. It is furthermore believed that the present compounds of formula I may improve sleep quality and correct the relative hy- posomatotropism of senescence due to high increase in REM sleep and a decrease in REM latency.

For the above indications the dosage will vary depending on the compound of formula I em- ployed, on the mode of administration and on the therapy desired. An effective amount of the compounds according to this invention will depend, in part, on the particular condition to be treated, age, weight, and general health of the patient, and other factors evident to those skilled in the art. However, generally dosage levels between 0.0001 and 100 mg/kg body weight daily are administered to patients and animals to obtain effective release of endogenous growth hormone. Morever the compounds of formula I have no or substantially no side-effects, when administered in the above dosage levels, such side-effects being e.g. release of cortisol, LH and/or prolactin. Usually, dosage forms suitable for oral, nasal, pulmonal or transdermal administration comprise from about 0.0001 mg to about 100 mg, preferably from about 0.001 mg to about 50 mg of the compounds of formula I admixed with a pharmaceutically acceptable carrier or diluent.

Optionally, the pharmaceutical composition of the invention may comprise a compound of for- mula I combined with one or more compounds exhibiting a different activity, e.g., an antibiotic or other pharmacologically active material.

The route of administration may be any route which effectively transports the active compound to the appropriate or desired site of action, such as oral, nasal, pulmonary, transdermal or par- enteral, the oral route being preferred.

Apart from the pharmaceutical use of the compounds of formula I, they may be useful in vitro tools for investigating the regulation of growth hormone release.

Compounds of formula I may also be useful in vivo tools for evaluating the growth hormone releasing capability of the pituitary. For example, serum samples taken before and after ad- ministration of these compounds to humans can be assayed for growth hormone. Comparison

of the growth hormone in each serum sample would directly determine the ability of the pa- tients pituitary to release growth hormone.

Compounds of formula I may be administered to commercially important animals to increase their rate and extent of growth, and to increase milk production.

A further use of growth hormone secretagogue compounds of formula I is in combination with other secretagogues such as GHRP (2 or 6), GHRH and its analogues, growth hormone and its analogues or somatomedins including IGF-1 and IGF-2.

Pharmacological Methods Compounds of formula I may be evaluated in vitro for their efficacy and potency to release growth hormone in rat pituitary primary cultures, and such evaluation may be performed as de- scribed below.

The isolation of rat pituitary cells is a modification of O. Sartor et al., Endocrinology 116, 1985, pp. 952-957. Male albino Sprague-Dawley rats (250 +/- 25 grams) were purchased from Mllegaard, Lille Skensved, Denmark. The rats were housed in group cages (four ani- mals/cage) and placed in rooms with 12 hour light cycle. The room temperature varied from 19-24"C and the humidity from 30 - 60%.

The rats were decapitated and the pituitaries dissected. The neurointermediate lobes were re- moved and the remaining tissue was immediately placed in icecold isolation buffer (Gey's me- dium (Gibco 041-04030) supplemented with 0.25% D-glucose, 2% non-essential amino acids (Gibco 043-01140) and 1% bovine serum albumine (BSA) (Sigma A-4503)). The tissue was cut into small pieces and transferred to isolation buffer supplemented with 3.8 mg/ml of trypsin (Worthington #3707 TRL-3) and 330 mg/ml of DNase (Sigma D-4527). This mixture was incu- bated at 70 rotations/min for 35 min at 37"C in a 95/5% atmosphere of O2/CO2. The tissue was then washed three times in the above buffer. Using a standard pasteur pipette, the tissue was then aspirated into single cells. After dispersion, cells were filtered through a nylon filter (160 mm) to remove undigested tissue. The cell suspension was washed 3 times with isolation buffer supplemented with trypsin inhibitor (0.75 mg/ml, Worthington #2829) and finally resu- spended in culture medium; DMEM (Gibco 041-01965) supplemented with 25 mM HEPES

(Sigma H-3375), 4 mM glutamine (Gibco 043-05030H), 0.075% sodium bicarbonate (Sigma S- 8875), 0.1% non-essential amino acid, 2.5% fetal calf serum (FCS, Gibco 011-06290), 3% horse serum (Gibco 034-06050), 10% fresh rat serum, 1 nM T3 (Sigma T-2752) and 40 mg/l dexamethasone (Sigma D-4902) pH 7.3, to a density of 2 x 105 cells/ml. The cells were seeded into microtiter plates (Nunc, Denmark), 200 mlAvell, and cultured for 3 days at 37"C and 8% CO2.

Compound testing After culturing, the cells were washed twice with stimulation buffer (Hanks Balanced Salt Solu- tion (Gibco 041-04020) supplemented with 1% BSA (Sigma A-4503), 0.25% D-glucose (Sigma G-5250) and 25 mM HEPES (Sigma H-3375) pH 7.3) and preincubated for 1 hour at 37"C. The buffer was exchanged with 90 ml stimulation buffer (37"C). Ten ml test compound solution was added and the plates were incubated for 15 min at 37"C and 5% CO2. The medium was de- canted and analyzed for GH content in an rGH SPA test system.

All compounds were tested in doses ranging from 10 pM to 100 mM. A dose-response relation was constructed using the Hill equation (Fig P, Biosoft). The efficacy (maximal GH released, Emax) was expressed in % of the Ernax of GHRP-6. The potency (EC60) was determined as the concentration inducing half maximal stimulation of the GH release.

Compounds of formula I may be evaluated for their metabolic stability using the procedure de- scribed below: Compounds is dissolved at a concentration of 1 mg/ml in water. 25 ml of this solution is added to 175 ml of the respective enzyme-solution (resulting in an enzyme:substrate ratio (w/w) of approximately 1:5). The solution is left at 370C overnight. 10 ml of the various degradation so- lutions is analyzed against a corresponding zero-sample using flow injection electrospray mass spectrometry (ESMS) with selected ion monitoring of the molecular ion. If the signal has de- creased more than 20% compared to the zero-sample, the remainder of the solution is ana- lyzed by HPLC and mass spectrometry in order to identify the extent and site(s) of degradation precisely.

Several standard peptides (ACTH 4-10, Angiotensin 1-14 and Glucagon) have been included in the stability tests in order to verify the ability of the various solutions to degrade peptides.

Standard peptides (angiotensin 1-14, ACTH 4-10 and glucagon) were purchased from Sigma, MO, USA) Enzymes (trypsin, chymotrypsin, elastase aminopeptidase M and carboxypeptidase Y and B) were all purchased from Boehringer Mannheim GmbH (Mannheim, Germany) Pancreatic enzyme mix: trypsin, chymotrypsin and elastase in 100 mM ammoniumbicarbonate pH 8.0 (all concentrations 0.025 mg/ml).

Carboxypeptidase mix: carboxypeptidase Y and B in 50 mM ammoniumacetate pH 4.5 (all concentrations 0.025 mg/ml).

Aminopeptidase M solution: aminopeptidase M (0.025 mg/ml) in 100 mM ammoniumbicarbon- ate pH 8.0 Mass spectrometric analysis was performed using two different mass spectrometers. A Sciex API III triple quadrupole LC-MS instrument (Sciex instruments, Thornhill, Ontario) equipped with an electrospray ion-source and a Bio-lon 20 time-of-flight Plasma Desorption instrument (Bio-lon Nordic AB, Uppsala, Sweden).

Quantification of the compounds (before and after degradation) was done on the API Ill in- strument using single ion monitoring of the molecular ion in question with flow injection of the analyte. The liquid flow (MeOH:water 1:1) of 100 ml/min was controlled by an ABI 140B HPLC unit (Perkin-Elmer Applied Biosystems Divisions, Foster City, CA). The instrument parameters were set to standard operation conditions, and SIM monitoring was performed using the most intense molecular ion (in most cases this corresponded to the doubly charged molecular ion).

Identification of degradation products furthermore involved the use of plasma desorption mass spectrometry (PDMS) with sample application on nitrocellulose coated targets and standard instrumental settings. The accuracy of the hereby determined masses is generally better than 0.1%.

Separation and isolation of degradation products was done using a HY-TACH C-18 reverse phase 4.6x105 mm HPLC column (Hewlett-Packard Company, Palo Alto, CA) with a standard acetonitril: TFA separation gradient. The HPLC system used was HP1090M (Hewlett-Packard Company, Palo Alto, CA). Peptide de- MW/SIM Carboxy Pan. rivative ion (amu) - enzyme peptidas mix e mix Standards ACTH 4-10 1124.5/562. + - 8 Glucagon 3483/871.8 - - Insulin (B23- 859.1/430.6 29) Angiotensin 1- 1760.1/881. - - 14 0 GHRP-2 817.4/409.6 - - GHRP-6 872.6/437.4 - - +: Stable (less than 20% decrease in SIM signal after 24 h in degradation solution) -: Unstable (more than 20% decrease in SIM signal after 24 h in degradation solution) Any novel feature or combination of features described herein is considered essential to this invention.

EXAMPLES: The process for preparing compounds of formula I and preparations containing them is fur- ther illustrated in the following examples, which however, are not to be construed as limiting.

The structures of the compounds are confirmed by either elemental analysis (MA) nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR shifts (d) are given in parts per million (ppm) and only selected peaks are given. mp is melting point and is given in OC.

Column chromatography was carried out using the technique described by W.C. Still et al, J. Org. Chem. 1978, 43, 2923-2925 on Merck silica gel 60 (Art 9385). Compounds used as starting materials are either known compounds or compounds which can readily be prepared by methods known per se.

HPLC-Analysis: Method A.

The RP-HPLC analysis was performed using UV detection at 214nm and a Hibar LiChrosorb RP-18 (5uM) 250-4 (Merck) column, which was eluted at 1 ml/minute. Two solvent systems were used: Solvent system 1: 0.1% Trifluoroacetic acid in acetonitrile. Solvent system II: 0.1% Trifluoroacetic acid in water.

The column was equilibrated with a mixture composed of 20% of solvent system I and 95% of solvent system II. After injection of the sample a gradient of 20% to 80% of solvent system I in solvent system li was run over 30 minutes. The gradient was then extended to 100% of solvent system I over 5 minutes followed by isocratic elution with 100% of this system for 5 minutes.

Method Al.

The RP-analysis was performed using UV detections at 214, 254, 276, and 301 nm on a 218TP54 4.6 mm x 250 mm 5m C-18 silica column (The Seperations Group, Hesperia), which was eluted at 1 mL/min at 420C. The column was equilibrated with 5% acetonitrile in a buffer consisting of 0.1 M ammonium sulfate, which was adjusted to pH 2.5 with 4M sulfuric

acid. after injection the sample was eluted by a gradient of 5% to 60% acetonitriie in the same buffer during 50 min.

Method B1.

The RP-analysis was performed using UV detections at 214, 254, 276, and 301 nm on a 218to54 4.6 mm x 250 mm 5m C-18 silica column (The Seperations Group, Hesperia), which was eluted at 1 mL/min at 420C. The column was equilibrated with 5% (acetonitrile + 0.1 % TFA) in an aqueous solution of TFA in water (0.1%). After injection the sample was eluted by a gradient of 5% to 60% (acetonitrile + 0.1% TFA) in the same aqueous buffer during 50 min.

Abbrevations: TLC: thin layer chromatography DMSO: dimethylsulfoxide min: minutes h: hours Troc: 2,2,2-trichloroethoxycarbonyl Boc: tert butyloxycarbonyl Z: benzyloxycarbonyl OSu: N-oxysuccinimide Lys: lysine DMF: dimethylformamide THF: tetrahydrofuran EDAC: N-ethyl-N'-dimethylaminopropylcarbodiimide hydrochloride HOBt: 1 -hydroxybenzotriazole hydrate HOAt: 1 -hydroxy-7-azabenzotriazole DIEA: diisopropylethylamine TFA: trifluoroacetic acid Buildingblocks: Throughout the following examples some common buildingblocks were used for which ab- breviations, references and representative experimental details are given below.

N-methylated aminoacids used in the following examples were prepared as in Can. J. Chem.

1977, 55, 906.

Starting material of type buildingblock 1: Fmoc-Lys-OH Starting material of type buildingblock 2: Fmoc-NMe-D-Phe-OH Fmoc-NMe-D-(2-thienyl)-Ala-OH Starting material of type buildingblock 3: Fmoc-D-(4-biphenyl)-Ala-OH Fmoc-D-2Nal-OH Starting material of type buildingblock 4:

Boc-AEH-OH Boc-3-AMH-OH Synthesis of intermediates (buildingblocks): Buildingblock Boc-AEH-OH: (2E)-5-(tert-Butoxycarbonylamino)-5-methylhex-2-enoic acid: 3-Hydroxy- 1,1 -dimethylpropylcarbamic acid tert-butyl ester: Step A: At 0 OC, ethyl chloroformate (1.10 mL, 11.5 mmol) was given dropwise to a solution of 3-tert-butoxycarbonylamino-3-methylbutanoic acid (2.50 g, 11.5 mmol) and triethylamine (1.92 mL, 13.8 mmol) in tetrahydrofuran (10 mL). The solution was stirred for 40 min at 0 "C. The formed precipitate was filtered off and washed with tetrahydrofuran (20 mL). The liquid was immediately cooled to 0 OC. A 2M solution of lithium boronhydride in tetrahydrofuran (14.4 mL, 28.8 mmol) was added dropwise. The solution was stirred at 0 OC for 2 h, and then warmed to room temperature. over a period of 4 h. it was cooled to 0 OC. Methanol (5 mL) was added carefully. 1 N Hydrochloric acid (100 mL) was added. The solution was extracted with ethyl acetate (2 x 100 mL, 3 x 50 mL). The combined organic layers were washed with saturated

sodium hydrogen carbonate solution (100 mL) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was chromatographed on silica (110 g) with ethyl acetate/heptane 1:2 to give 1.84 g of 3-hydroxy-1,1-dimethylpropylcarbamic acid tert-butyl ester.

'H-NMR ((CDCl3): d 1.33 (s, 6 H); 1.44 (s, 9 H); 1.88 (t, 2 H); 1.94 (br, 1 H); 3.75 (q, 2 H); 4.98 (br, 1 H).

3-(tert-Butoxycarbonylamino)-3-methylbutanal: Step B: DMSO (1.22 mL, 17.2 mmol) was added to a solution of oxalyl chloride (1.1 mL, 12.9 mmol) at -78 "C in dichloromethane (15 mL). The mixture was stirred for 15 min at -78 "C. A solution of 3-hydroxy-1,1-dimethylpropylcarbamic acid tert-butyl ester (1.75 g, 8.6 mmol) in dichloromethane (10 mL) was added dropwise over a period of 15 min. The solution was stirred at -78 OC for another 15 min. Triethylamine (6.0 mL, 43 mmol) was added. The solution was stirred at -78 "C for 5 min and then warmed to room temperature. The solution was diluted with dichloromethane (100 mL) and extracted with 1N hydrochloric acid (100 mL). The aqueous phase was extracted with dichloromethane (50 mL). The combined organic layers were washed with saturated sodium hydrogen carbonate solution (100 mL) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by column chromatography on silica (140 g) with ethyl acetate/heptane (1:3) to give 1.10 g of 3- (tert-butoxycarbonylamino)-3-methylbutanal.

'H-NMR (CDCI3): d 1.39 (s, 6 H); 1.45 (s, 9 H); 2.85 (d, 2 H); 4.73 (br. 1 H); 9.80 (t, I H).

Ethyl (2E)-5-(tert-Butoxycarbonylamino)-5-methylhex-2-enoate:

Step C: Triethylphoshonoacetate (1.96 mL, 9.8 mmol) was dissolved in tetrahydrofuran (30 mL). Potassium tert-butoxide (1.10 g, 9.8 mmol) was added. The solution was stirred for 40 min at room temperature. A solution of 3-(tert-butoxycarbonylamino)-3-methylbutanal (1.10 g, 5.5 mmol) in tetrahydrofuran (6 mL) was added. The solution was stirred at room temperature for 75 min. It was diluted with ethyl acetate (100 mL) and 1 N hydrochloric acid (100 mL). The phases were separated. The aqueous phase was extracted with ethyl acetate (2 x 50 mL). The combined organic phases were washed with saturated sodium hydrogen carbonate solution (60 mL) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by column chromatography on silica (90 g) with ethyl acetate/hepatane (1:4) to give 1.27 g of ethyl (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-enoate.

1H-NMR (CDCl3): d 1.30 (s, 6 H); 1.30 (t, 3 H); 1.46 (s, 9 H); 2.62 (d, 2 H); 4.27 (q, 2 H); 4.42 (br, 1 H); 5.88 (d, 1 H); 6.94 (td, 1 H).

Step: Ethyl (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-enoate (1.233 g, 4.54 mmol) was dissolved in dioxane (20 mL). Lithium hydroxide (0.120 g, 5.00 mmol) was added as a solid. Water (10 mL) was added, until a clear solution was reached. The solution was stirred 16 h at room temperature. The solution was diluted with water (70 mL) and was extracted with tert-butyl methyl ether (2 x 100 mL). The aqueous phase was acidified with 1N sodium hydrogensulfate solution (pH = 1) and was extracted with tert-butylmethylether (3 x 70 mL).

The organic phases were combined and dried over magnesium sulfate. The solvent was removed in vacuo to give 1.05 g of (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-enoic acid. The crude product was used for further syntheses.

1H-NMR (DMSO d6): d 1.15 (s, 6 H); 1.35 (s, 9 H); 2.53 (d, 2 H); 5.75 (d, 1 H); 6.57 (br, 1 H); 6.75 (td, 1 H); 12.15 (s, 1 H).

Synthesis of buildingblock S: (2S)-2-(((Carboxy)methoxy)methyl)pyrrolidin-I -carboxylic acid tert-butylester: To a solution of N-t-butyloxycarbonyl-(S)-prolinol (5.0 g, 25 mmol) in 1,2- dichloroethane (500 ml) rhodium(ll)acetate (180 mg) was added and the mixture was heated to 80 "C. Ethyldiazoacetate (3.9 ml, 37 mmol) in 1,2-dichloroethane (180 ml) was added over a period of 90 min and the mixture was heated at 80 "C for 3 hours.

Then another portion of ethyl diazoacetate (1.3 ml, 12 mmol) in 1,2-dichloroethane (40 ml) was added and the mixture was refluxed for 6 hours. The mixture was cooled to room temperature and washed with saturated sodium bicarbonate (2 x 100 ml) and brine (100 ml), dried (magnesium sulfate) and concentrated in vacuo. The crude prod- uct was chromatographed on silica (300 g) with petrol ether/ethyl acetate 4:1 as eluent to give 4.7 g of (2S)-2-(((ethoxyCarbonyl)methoxy)methyl)pyrrolidin-1-carboxy lic acid tert-butylester. The obtained product was dissolved in 1 M lithium hydroxide in wa- ter/methanol 1:3 (50 ml) and stirred at room temperature overnight. The mixture was concentrated in vacuo, water (20 mL) was added and washed with ether (20 mL). The aqueous phase was acidified to pH 4 with 1 M aqueous hydrogen chloride, extracted with ethyl acetate (2 x 100 ml) and the combined organic layers were dried (magnesium sulfate) and concentrated in vacuo to give 3.6 g of (2S)-2- (((carboxy)methoxy)methyl)-pyrrolidin-I -carboxylic acid tert-butyl ester.

1H-NMR (CDCl3): d 1.45(2, 9H) 1.90 (m, 4H) 3.55 (t, 2H) 3.60 (m, 3H) 4.10 (s, 2H) 10.6 (s, 1H).

Synthesis of building block 6 (2-(tert-Butoxycarbonylamino)butoxy)acetic acid: To a solution of (1-(hydroxymethyl)propyl)carbamic acid tert-butylester (7.2 g, 39 mmol) in 1,2- dichloroethane (500 ml) rhodium(ll)acetate (180 mg) was added and the mixture was heated to 80 "C. Ethyldiazoacetate (6.0 ml, 57 mmol) in 1,2-dichloroethane (180 ml) was added over a period of 60 min and the mixture was heated at 80 "C for 6 hours. Then another portion of ethyldiazoacetate (2.0 ml, 19 mmol) in 1,2-dichloroethane (40 ml) was added and the mixture was refluxed for 7 hours. The mixture was cooled to room temperature and washed with sodium bicarbonate (2 x 100 ml) and brine (100 ml), dried over magnesium sulfate and concentrated in vacuo. The crude product was chromatographed on silica (300 g) with pentane/ethyl acetate 7:3 as eluent to give 4.3 g of (2-(tert-butoxycarbonylamino)butoxy)acetic acid ethylester. The product was dissolved in of 1 M lithium hydroxide in water/methanol 1:3 (40 ml) and stirred at room temperature for 4 hours. The mixture was concentrated in vacuo and water (100 mL) was added and the solution was washed with ether (20 mL). The aqueous phase was acidified to pH 4 with 1 M aqueous hydrogen chloride and extracted with ethyl acetate (200 ml), dried over magnesium sulfate and concentrated in vacuo to give 2.46 g of (2- (tert butoxycarbonylamino)butoxy)acetic acid.

1H-NMR (CDCl3): d 0.95 (t, 3H) 1.45 (s, 9H) 1.60 (m, 3H) 3.55 (m, 2H) 4.10 (s, 2H) Synthesis of buildingblock 7 3-(1-(N-tertbutoxycarbonylamino)ethyl)benzoic acid: Ammonium acetate (10.6 g, 138 mmol) was evaporated from dry ethanol (100 mL), and redissolved in dry methanol (100 mL) over molecular sieves (3A, 3 g). 3-

Acetylbenzonitrile (2.0 g, 13.8 mmol) was added. After 30 minutes at room tempera- ture sodium cyanoborohydride (0.87 g, 138 mmol) was added and the reaction mixture was stirred for 18 hours. The reaction mixture was concentrated in vacuo and redis- solved in water (100 mL). Concentrated hydrochloric acid was added until pH 2, and the aqueous solution was extracted with ethyl acetate (2 x 100 mL). The aqueous phase was adjusted to pH 11 with solid potassium hydroxide, and extracted with di- chloromethane (2 x 100 mL). The combined organic phases were dried (magnesium sulfate) and concentrated in vacuo. A concentrated solution of hydrogen chloride in ethyl acetate added (100 mL) was, and the solution was concentrated in vacuo. The residue was dissolved in ethanol (25 mL) and sulphuric acid (9N, 25 mL) was added.

After 16 hours at room temperature and 2 hours at reflux temperature the ethanol was removed by evaporation in vacuo and the residual aqueous mixture was adjusted to pH > 8 using solid potassium hydroxide. Ditertbutyldicarbonate (2.0 g) dissolved in tet- rahydrofuran (100 mL) was added at OOC. After 18 hours at room temperature the re- action mixture was concentrated in vacuo and redissolved in water (100 mL). Solid cit- ric acid was added until pH 5. The reaction mixture was extracted with di- chloromethane (2 x 100 mL), and the combined organic phases was dried (magnesium sulfate) and concentrated in vacuo. The residue was purified by column chromatogra- phy on silica gel (3 x 40 cm) using ethanol and dichloromethane (1 :9) as eluent to give 1.1 g of 3-(1-(N-tertbutoxycarbonyl)aminoethyl)benzoic acid.

General procedure for the conversion of N-Boc-protected buildinablocks to N-Troc-protected analogs: (Also valid for N-FMOC buildingsblocks using TrocOSu instead of FMOC-OSu) To a solution of hydrogen chloride in ethyl acetate (3M, 25 ml) was added the Boc-protected buildingblock (0.01 mol) and the mixture was stirred for 30 min. Diethyl ether (100 ml) was added and the precipitate was filtered and dissolved in a mixture of aqueous sodium hydrogencarbonate (5%) and dioxane (1:1,100 ml). Succinimidyl 2,2,2-trichloroethyl carbonate (0.01 mole) was added and the mixture was stirred overnight. The solvent was removed in vacuo and the residue was dissolved in water (100 ml) and washed once with diethyl ether (50 ml) and acidified to pH 3 with a solution of aqueous hydrogen sulphate. The aqueous phase was extracted with methylene chloride (3 x 100 ml) and the combined organic phases were

dried (magnesium sulphate) and the solvent was removed in vacuo to afford the Troc protected buildingblock.

Other buildingblocks: ½ CH3 0 H3C/I,AW H,C/g"" H3C ° H ) ooh3 ; Boc-D-Phe-OHO Boc-N-Me-D-Phe-OH OH3 0 OH 0 H H3O3 - H$OY0CN#%0H 3 H/ C NH¼$OH CH, O 0 Boc-D-2-Nai-OH Boc-N-Me-D-2-Nal-OH CH3 O OH3 H HO H OH HO 0 NHYF0H 3 N o OH3 O Boc-D-Bip-OH Boc-N-Me-D-Bip-OH OH 0 4½F OH3 0 H, H3O OH H OH 30 O t( < cn, D 0 OH3 0 Boc-D-4-F-Phe-OH Boc-N-Me-D-4-F-Phe-OH OH3 0 #4 H OH3 0 #4 '0/C H30O½0% OH HO 0 Na H3C 3 Nj CH3 O CH3 O Boc-D-2-thial-OH Boc-N-Me-D-2-thial-OH Example 1 3-Aminomethyl-N-((l R)-1 -(N-((1 R)-2-(2-thienyl)-l -((1S)-l -carbamoyl-5- aminopentylcarbamoyl)ethyl)-N-methylcarbamoyl)-2-(2-naphthyl )ethyl)benzamide. [(1S)-1-Carbamoyl-5-(2,2,2-trichloroethoxycarbonylamino)pent yl]carbamic acid tert butyl ester BocLys(Z)OSu (50 g; 0.10 mol) and ammonium hydrogencarbonate (25 g; 0.32 mole) were dissolved in DMF (300 ml) and stirred overnight. Water (1000 ml) was added and the pre- cipitate was isolated by filtration and washed with water (3 x 200 ml). The precipitate was dissolved in methanol (500 ml) and palladium on carbon (15 g; wet, 10%) was added. The mixture was hydrogenated for 4 h at ambient pressure, filtered and the solvent was removed

in vacuo. The residue was dissolved in THF (500 ml) and aqueous sodium hydroxide (4M, 50 ml) and succinimidyl 2,2,2-trichloroethyl carbonate was added (30.4 g; 0.10 mole) and the mixture was stirred for 3 h. The mixture was evaporated and methylene chloride (400 ml) was added. The organic phase was washed with water (300 ml), an aqueous solution of so- dium hydrogensulphate (300 ml), an aqueous solution of sodium hydrogencarbonate (300 ml) and water (300 ml). The organic phase was dried (magnesium sulphate) and the solvent was removed in vacuo to afford 39.5 g of [(1S)-l -carbamoyl-5-(2,2, 2-trichloro- ethoxycarbonylamino)pentyl]carbamic acid tert butyl ester.

1H-NMR: (CDCl3) d 1.43 (s, 9H); 1.53-1.89 (m, 6H); 3.23 (q, 2H); 4.15 (m, 3H); 4.72 (s, 2H).

((SS)-5-Amino-5-carbamoylpentyl)carbamic acid 2,2,2-trichloroethyl ester hydrochloride [(1S)-1-carbamoyl-5-(2,2,2-trichloroethoxycarbonylamino)pent yl]carbamic acid tert butyl es- ter (39.5 g; 94 mmole) was dissolved in a solution of hydrogen chloride in ethyl acetate (1.5 M, 300 ml) and stirred for 30 min. Diethyl ether (300 ml) was added and the precipitate was filtered and dried to afford 30.5 g of ((SS)-5-amino-5-carbamoylpentyl)carbamic acid 2,2,2- trichloroethyl ester hydrochloride.

'H-NMR: (DMSO) d 1.31 (m, 2H); 1.45 (m, 2H); 1.75 (m, 2H); 3.02 (dd, 2H); 3.67 (t,lH); 4.77 (s, 2H); 7.51 (s, 1H); 7.70 (t, 1H); 7.96 (s, 1H); [(5S)-5-Carbamoyl-5-((2R)-2-methylamino-3-(2-thienyl)-propio nylamino)pentyl]carbamic acid 2,2,2-trichloroethyl ester

2-(tert-Butoxycarbonylmethylamino)-3-(2-thienyl)propionic acid (10.07 g; 35 mmole), HOBt (5.72 g; 42 mmole) and EDAC (7.09 g; 37 mmole) were added to methylene chloride (200 ml) and stirred for 15 min. ((SS)-5-amino-5-carbamoylpentyl)carbamic acid 2,2,2- trichloroethyl ester hydrochloride (12.61 g; 35 mmole) was added and the mixture was stirred overnight. Methylene chloride (200 ml) was added and the organic phase was washed with water (200 ml), an aqueous solution of sodium hydrogensulphate (200 ml), an aqueous solution of sodium hydrogencarbonate (200 ml) and water (200 ml). The organic phase was dried (magnesium sulphate) and the solvent was removed in vacuo. The residue was dissolved in a solution of hydrogen chloride in ethyl acetate (3 M; 100 ml) and stirred for 30 min. Diethyl ether and water (500 ml) was added, the phases were separated and the acidic aqueous phase was treated with a solution of sodium carbonate to pH 10. The aque- ous phase was extracted with methylene chloride (4 X 700 ml) and the phases were sepa- rated. The organic phase was dried (magnesium sulphate) and the solvent was removed in vacuo to afford 11.0 g of [(5S)-5-carbamoyl-5-((2R)-2-methylamino-3-(2-thienyl)- propionylamino)-pentyl]-carbamic acid 2,2,2-trichloroethyl ester.

1H-NMR: (DMSO) d 1.11-1.68 (m, 6H); 2.20 (s, 3H); 2.86-3.08 (m, 4H); 3.18 (t, 1H); 4.18 (dd, 1 H); 4.78 (s, 1 H); 6.88 (d, 1 H); 6.92 (t, 1 H); 7.30 (d, 1 H) ((5R)-5-((2R)-2-[((2R)-2-Amino-3-(2-naphthyl)propionyl)methy lamino]-3-(2- thienyl)propionylamino)-5-carbamoylpentyl)carbamic acid 2,2,2-trichloroethyl ester

(2R)-2-(tert-Butoxycarbonylmethylamino)-3-(2-napthyl)propion ic acid (5.00 g; 15.8 mmole), HOAt (2.35 g; 17 mmole) and EDAC (3.03 g; 15.8 mmole) were added to methylene chloride (200 ml) and stirred for 15 min. [(5S)-5-carbamoyl-5-((2R)-2-methylamino-3-(2- thienyl)propionylamino)pentyl]carbamic acid 2,2,2-trichloroethyl ester (7.01 g; 14.5 mmole) was added and the mixture was stirred overnight. Methylene chloride (200 ml) was added and the organic phase was washed with water (200 ml), an aqueous solution of sodium hy- drogensulphate (200 ml), an aqueous solution of sodium hydrogencarbonate (200 ml) and water (200 ml). The organic phase was dried (magnesium sulphate) and the solvent was removed in vacuo. The residue was dissolved in a mixture of trifluoroacetic acid (30 ml) and methylene chloride (70 ml) and stirred for 15 min. Water (150 ml) and sodium hydrogencar- bonate were added to neutral pH, the phases were separated and the aqueous phase was extracted with methylene chloride (2 X 100 ml) and the phases were separated. The com- bined organic phase were dried (magnesium sulphate) and the solvent was removed in vacuo and the residue was chromatographed on silica using a mixture of aqueous ammonia in water (25%), ethanol and methylene chloride (0.7:9.3:90) as eluent to afford 6.3 g of ((SS)-S-((2R)-2-((2R)-2-amino-3-(2-naphthyl)prnpionyl)methyl amino]-3-(2- thienyl)propionylamino)-5-carbamoylpentyl)carbamic acid 2,2,2-trichloroethyl ester.

'H-NMR: (CDC13; selected peaks for major rotamer): d 1.36-1.61 (m, 6H); 2.88 (s, 3H); 4.00 (dd; 1 H); 4.65 (s, 2H).

MS(ES): m/z: 684 (M+H)+ 3-[(2,2,2-Trichloroethoxycarbonylamino)methyl]benzoic acid (8.52 g; 26.1 mmole) and EDAC (2.49 g; 13.0 mmole) were dissolved in methylene chloride (150 ml) and stirred for 15 min.

((SS)-S-((2R)-2-[((2R)-2-Amino-3-(2-naphthyl)propionyl)methy lamino]-3-(2-

thienyl)propionylamino}-5-carbamoylpentyl)carbamic acid 2,2,2-trichloroethyl ester (8.92 g; 13.0 mmole) was added and the mixture was stirred for 30 min. DIEA (3.6 ml) was added and the mixture was stirred overnight. The mixture was washed with an aqueous solution of sodium hydrogencarbonate (10%, 2 x 100 ml) and with an aqueous solution of sodium hy- drogensulphate (10%, 1 x 100 ml). The organic phase was dried (magnesium sulphate) and the solvent removed in vacuo. The residue was chromatograped on silica using ethyl acetate as eluent and the protected intermediate was dissolved in a mixture of THF (100 ml) and an aqueous solution of potassium dihydrogenphosphate (1 M, 40 ml). Zinc powder (50 g) was added and the mixture was stirred overnight. The mixture was filtered and the solvent was removed in vacuo. The residue was chromatograped on reverse phase silica (Waters, RP- 18,125 A) using gradient elution with acetonitrile and water (10% to 30%) with 0.1 % TFA added to afford 7.01 g of the title compound.

1H-NMR: (DMSO;): d 2.88; 3.08; (twos, 3H); 4.05 (s, 3H); 4.15; 4.20 (two dd, 1H); 5.02; 5.12; 5.15; 5.25 (four dd; 2H).

HPLC: (method Al): Rt = 26.90 min.

MS(ES): m/z 643.4 (M+H)+.

Example 2 3-((1 R/S)-1-Aminoethyl)-N-((1 R)-1 -(N-t(1 R)-1 -(N-(carbamoylmethyl)-N-methylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lbenzamide

The title compound was prepared as in example 1 using sarcosine amide instead of ((5S)-5- Amino-5-carbamoylpentyl)carbamic acid 2,2,2-trichloroethyl ester hydrochloride. Boc-N-Me- D-Phe-OH, Boc-N-Me-D-2-Nal-OH and building block 7 were used.

'H-NMR: ((CDCl3; selected peaks for major rotamer): d 1.25 (d, 3H); 2.51 (s, 3H); 2.65 (s, 3H); 3.03 (s, 3H); HPLC: (method Al): Rt = 27.30 min.

MS(ES): m/z 608.4 (M+H)+.

Example 3 2-Amino-N-(((1 S)-1 -(((1 R)-1 -(((1 R)-1 -(((1 S)-1 -carbamoyl-5- (methylsulfonylamino)pentyl)carbamoyl)-2-phenylethyl)carbamo yl)-2-(2- naphthyl)ethyl)carbamoyl)-2-(4-imidazolyl)ethyl)carbamoyl)-2 -methylpropanamide 2-Amino-N-(((1 S)-I -(((1 R)-1 -(((1 R)-1 -(((1 S)-5-amino-1 -carbamoylpentyl)carbamoyl)-2- phenylethyl)carbamoyl)-2-(2-naphthyl)ethyl)carbamoyl)-2-(4-i midazolyl)ethyl)carbamoyl)-2- methylpropanamide (217 mg, 0.30 mmol prepared as in WO9517423) was dissolved in 1,4- dioxane (2 ml) and water(2 ml). Potassium carbonate (207 mg, 1.50 mmol) was added. A solution of methanesulfonyl chloride (0.023 ml, 0.29 mmol) in 1,4-dioxane (0.5 ml) was added. The reaction mixture was stirred for 24 h at room temperature. The solvent was re- moved in vacuo. The residue was purified by three HPLC-chromatographic runs on a 25 mm x

250 mm 1 Om C18 silica column at 40 °C with a gradient of 20 to 40% acetonitrile in a 0.1M ammonium sulfate buffer, which was adjusted to pH 2.5 with 4M sulfuric acid. The peptide containing fractions were collected, diluted with 3 volumes of water and applied to a Sep-Pake C18 cartridge (Waters part. #: 51910) which was equilibrated with 0.1% trifluoroacetic acid.

The peptide was eluted from the Sep-Paks cartridge with 70% acetonitrile in a 0.1% trifluoroacetic acid solution in water. The product was lyophilized to afford 61 mg of the title compound as a trifluoroacetate salt HPLC: 23.68 min ( method Al).

25.55 min (B1).

MS: 790.4 ([M+H]+) Example 4 [(5S)-5-((2R)-2-{N-[(2R)-2-(3-(Aminomethyl)benzoylamino)-3-( 2-naphthyl)propionyl]-N- methylamino}-3-(2-thienyl)propionylamino)-5-carbamoylpentyl] carbamic acid benzyl ester The title compound was prepared as in example 1, except that the Z-protecting group was not transferred into the Troc-protecting group in the first step.

HPLC (method Al): rt 39.38 min.

Example 5 N-((1 R)-1-(N-[(1 R)-1-(((1 S)-5-Amino-l -carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]- N-methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-3-(aminomethyl)be nzamide The title compound was prepared as in example 1. Boc-N-Me-D-(4-F-Phe)-OH and Boc-D- Bip-OH were employed.

HPLC: (method Al): Rt = 30.78 min.

MS(ES): m/z 681 (M+H)+.

Example 6 (2E)-5-Amino-5-methylhex-2-enoic acid N-((I R)- I -(N-[(I R)- 1 -(5-aminopentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lamide The title compound was prepared as in example 1 substituting ((5S)-5-Amino-5-carbamoyl- pentyl)-carbamic acid 2,2,2-trichloro-ethyl ester hydrochloride with N-Troc-1,5-

diaminopentane (prepared as in Atwell, G. J. ; Denny, A. D.; Synthesis, 1032 (1984)) and Boc-AEH-OH.

1H-NMR: ((CDCl3; selected peaks for major rotamer): d 0.92 (s, 6H); 2.91 (s, 3H); 2.95 (s, 3H); 4.22 (dd, 1H); 6.22 (d, 1H) HPLC: (method Al): Rt = 27.30 min.

MS(ES): m/z 600.2 (M+H)+.

Example 7 3-((1 R/S)-1 -Aminoethyl)-N-[(1 R)-2-(biphenyl-4-yl)-1 -(N-((l R)-l -[N-(3-dimethylaminopropyl)- N-methylcarbamoyl]-2-phenylethyl}-N-methylcarbamoyl)ethyl]-N -methylbenzamide The title compound was prepared as in example 1, omitting acidic washes and using N- methyl-N'-dimethyldiaminopropane, buildingblock 7, Boc-N-Me.D-Phe-OH and Boc-N-Me-D- Bip-OH as the starting materials.

1H-NMR: ((CDCl3, selected peaks): d 1.36 (d, 3H); 2.26 (s, 3H); 2.32 (s, 6H); 2.52 (s, 3H); 3.03 (s, 3H) HPLC: (method Al): Rt = 28.88 min.

MS(ES): m/z 662.2 (M+H)+.

Example 8 (2E)-5-Amino-5-methylhex-2-enoic acid N-[(1 R)-2-(biphenyl-4-yl)-1 -(N-((1 R)-1 -[N-(3- <BR> <BR> <BR> dimethylaminopropyl)-N-methylcarbamoyl]-2-phenylethyl)-N-met hylcarbamoyl)ethyl]-N- methylamide The title compound was prepared as in example 1, omitting acidic washes and using N- methyl-N'-dimethyldiaminopropane, Boc-N-Me-D-Phe-OH and Boc-N-Me-D-Bip-OH and Boc-AEH-OH as the starting materials 'H-NMR: ((CDCl3, selected peaks): d 1.17 (s, 6H); 2.10 (s, 6H); 2.19 (s, 3H); 2.29 (s, 3H) 2.48 (s, 3H); 5.82 (m, 1 H); 6.18 (d, 1 H).

HPLC: (method A1): Rt = 28.93 min.

MS(ES): m/z 640.2 (M+H)+.

Example 9 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-((1S)-5-amino-1- <BR> <BR> <BR> carbamoylpentylcarbamoyl)-2-phenylethyl]-N-methylcarbamoyl)- 2-(2-naphthyl)ethyl)-N- methylamide

The title compound was prepared as in example 1. Boc-AEH-OH and Boc-N-Me-D-2-Nal-OH were used as starting materials.

HPLC: (method Al): Rt = 23.48 min.

MS(ES): m/z 643.2 (M+H)+.

Example 10 N-((1 R)-1-{N-[(1 R)-1 -((1 S)-5-Amino-1 -carbamoylpentylcarbamoyl)-2-phenylethyl]-N- methylcarbamoyl}-2-(2-naphthyl)ethyl)-3-((1R/S)-1-aminoethyl )-N-methylbenzamide The title compound was prepared as in example 1. Boc-N-Me-D-Phe-OH, Boc-N-Me-D-2- Nal-OH and buildingblock 7 were used as starting materials.

1H-NMR: ((CDCl3; selected peaks for major rotamer): d 1.54 (d, 3H); 2.77 (s, 3H); 3.01 (s, 3H); 4.42 (q, 1 H); 5.42 -5.72 (m, 2H) HPLC: (method Al): Rt = 26.61 min.

MS(ES): m/z 665.2 (M+H)+.

Example 11 (2E)-5-Amino-5-methylhex-2-enoic acid ((1R)-1-{N-[(1R)-1-(((1S)-5-amino-1- (dimethylcarbamoyl)pentyl)carbamoyl)-2-phenylethyl]-N-methyl carbamoyl)-2-(2- naphthyl)ethyl)amide The title compound was prepared as in example 1. Dimethylamine was used instead of am- monium hydrogencarbonate. Boc-N-Me-D-Phe-OH, Boc-N-Me-D-2-Nal-OH and Boc-AEH- OH were used as starting materials.

1H-NMR: ((CDCl3; selected peaks): d 1.32 (s, 6H); 1.81 (dd, 2H); 2.81; (s, 3H); 2.92 (s, 3H); 3.04 (s, 3H); 3.11 (s, 3H); 4.96 (dd, 1 H); 5.45 (dd, 1H); 5.60 (dd, 1 H); 6.11 (d, 1 H).

HPLC: (method Al): Rt = 26.82 min.

MS(ES): m/z 671.2 (M+H)+.

Example 12 N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-Amino-1 -(dimethylcarbamoyl)pentyl)carbamoyl)-2- <BR> <BR> phenylethyl]-N-methylcarbamoyl}-2-(biphenyl4-yl)ethyl)-3-ami nomethyl-N-methylbenzamide

The title compound was prepared as in example 1. Dimethylamine was used instead of am- monium hydrogencarbonate. Boc-N-Me-D-Phe-OH, Boc-N-Me-D-Bip-OH and Boc-AMB-OH were used as starting materials.

1H-NMR: ((CDCl3; selected peaks): d 2.85 (s, 3H); 2.94 (s, 3H); 2.96 (s, 3H); 3.05 (s, 3H); 3.96 (dd, 2H); 4.89 (dd, 1H); 5.48 (dd, 1H).

HPLC: (method Al): Rt = 30.30 min.

MS(ES): m/z 705.4 (M+H)+.

Example 13 (2E)-5-Amino-5-methyl-hex-2-enoic acid N-((1R)-1-{N-[(1R)-1-((1S)-5-amino-1- (dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methylc arbamoyl)-2-(biphenyl-4- yl)ethyl)-N-methylamide

The title compound was prepared as in example 1. Dimethylamine was used instead of am- monium hydrogencarbonate. Boc-N-Me-D-Phe-OH, Boc-N-Me-D-Bip-OH and Boc-AEH-OH were used as starting materials.

1H-NMR: ((CDCl3; selected peaks for major rotamer): d 1.12 (s, 6H); 2.45 (s, 3H); 2.90 (s, 3H); 3.03 (s, 3H); 3.05 (s, 3H); 4.82 (dd, 1H); 5.38 (m, 1H); 6.18 (d, 1H).

HPLC: (method Al): Rt = 30.22 min.

MS(ES): m/z 697.6 (M+H)+.

Example 14 N-((1 R)-1 -([(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-3-aminomethylbenzamide The title compound was prepared as in example 1. Boc-N-Me-D-4-F-Phe-OH, Boc-D-2-Nal- OH and Boc-AMB-OH were used as starting materials.

1H-NMR: (MeOH; selected peaks for major rotamer): d 1.10-1.90 (m, 6H); 3.00 (s, 3H); 4.12 (s, 2H); 4.55 (dd, 1 H); 5.26 (dd, 1 H) HPLC: (method Al): Rt = 28.23 min.

MS(ES): m/z 655.2 (M+H)+.

Example 15 (2E)-5-Amino-5-methylhex-2-enoic acid ((1R)-1-{N-[(1R)-1-(((1S)-5-amino-1- carbamoylpentyl)carbamoyl)-2-(4-fluorophenyl)ethyl]-N-methyl carbamoyl}-2-(2- naphthyl)ethyl)amide The title compound was prepared as in example 1. Boc-N-Me-D-4-F-Phe-OH, Boc-D-2-Nal- OH and Boc-AEH-OH were used as starting materials 1H-NMR: (MeOH, selected peaks for major rotamer): d 1.32 (s, 6H); d, 2H); 2.95 (s, 3H); 4.31 (dd, 1H); 4.50 (dd, 1H); 5.12 (t, 1H); 6.12 (d, 1H); 6.60 m, 1H) .

HPLC: (method Al): Rt = 28.05 min.

MS(ES): m/z 647.4 (M+H)+.

Example 16 N-((1R)-1-{N-[(1R)-1-(((1S)-5-Amino-1-carbamoylpentyl)carbam oyl)-2-(4-fluorophenyl)ethyl]- N-methylcarbamoyl)-2-(2-naphtyl)ethyl)-3-aminomethyl-N-methy lbenzamide

The title compound was prepared as in example 1. Boc-N-Me-D-4-F-Phe-OH, Boc-N-Me-D- 2-NaI-OH and Boc-AMB-OH were used as starting materials 1H-NMR: ((CDCl3;): d 2.61 (s, 3H); 2.87 (s, 3H); 4.30 (m, 1H); 4.70 (m, 1H); HPLC: (method Al): Rt = 27.03 min.

MS(ES): m/z 669.4 (M+H)+.

Example 17 (2S)-6-Amino-2-!(2R)-3-(44luorophenyl)-2-(N-methyl-N-((2R)-3 -(2-naphthyl)-2-t2-(((2S)- pyrrolidin-2-yl)methoxy)acetylamino]-propionyl}amino)proiony lamino]hexanoic acid amide The title compound was prepared as in example 1. Boc-N-Me-D-4-F-Phe-OH, Boc-D-2-Nal- OH and buildingblock 6 were used as starting materials

1H-NMR: (MeOH, selected peaks): d 1.22-1.90 (m, 10H); 3.02 (s, 3H); 3.49 (dd, 1H); 3.95 (ab-syst., 2H); 4.32 (dd, 1H); 4.94 (dd, 1H); 5.03, (dd, 1H); 5.15 (dd, 1H).

HPLC: (method Al): Rt = 26.52 min.

MS(ES): m/z 663.4 (M+H)+.

Example 18 (2S)-6-Amino-2-[(2R)-2-(N-{(2R)-2-[2-((2R/S)-2-aminobutoxy)a cetylamino]-3-(2- naphthyl)propionyl}-N-methylamino)-3-(4-fluorophenyl)propion ylamino]hexanoic acid amide The title compound was prepared as in example 1. Boc-N-Me-D-4-F-Phe-OH, Boc-D-2-Nal- OH and buildingblock 5 were used as starting materials 1H-NMR: (MeOH, selected peaks;): d 0.95 (t, 3H); 1.25-1.86 (m, 8H); 2.98 (s, 3H); 3.95 (m, 2H); 4.32 (dd, 1H); 4.90 (t, 1H); 5.17 (t, 1H): HPLC: (method Al): Rt = 26.90 min and 27.05 min (diastereomeric mixture) MS(ES): m/z 651.2 (M+H)+.

The following examples (example 19 to example 25) where prepared on solid support using the general procedure described below:

o Xj Fmoc deprotechon g HO =}NHFmoc Resin Resin H coupling reagents 0 deprotection ° HO w NRFmoc Sb~lik3- Rink Resin building reagents coupling reagents deprotection ° o Rink (+ -Luffct-#H ~ -NH NHR lOx HOL-n- NHFmoc coupling reagents o 0 0 1 buildingblock2-i~~b~block-NHFmoc 0 HO buitdingblcck4 NHBoc deprotection . . ~, blode . (t<}H<}NHR ~ H 9t M=K4 NHBoc 0 0 O 0 0 cleavage buitdingbtocktN building block H H H2N g}H = NHR< }Fi sDNH2 HN - NHR N 2 Fmoc deprotection procedure: The resin is swelled and shaken in 20% piperidine in DMF (5 ml) for 3 min. The resin is drained and the deprotection process is repeated, with 15 min reaction time.

Standard washing procedure: The drained resin is washed using the following standard washing procedure: The resin is repeatedly swelled, shaken and drained three times with 5mL of DMF, methyl- ene chloride, DMF, 2-propanol, methanol and finally with diethyl ether.

Standard protocol for solid phase synthesis of example 19-25: Deprotection:

Rink amide resin (Novabiochem 01-64-0013, lg , 0,46 mmol) is deprotected using the method described above followed by the standard washing procedure.

Attachment of building block 1: Building block 1 (0.9mmol), HOBt (138mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 1Smin, and then transferred to the drained resin, followed by addition of DMF (3mL). DIEA (116mg, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and the washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Attachment of building block 2: Building block 1 (0.9mmol), HOBt (138mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 15min, and then transferred to the drained resin, followed by addition of DMF (3mL). DIEA (116mug, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and the washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Attachment of building block 3: Building block 2 (0.9mmol), HOAt (177mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 15min, and then transferred to the drained resin followed by addition of DMF (3mL). DIEA (116mg, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and the washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Attachment of building block 4: Building block 1 (0.9mmol), HOBt (138mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 1Smin, and then transferred to the drained resin, followed by addition of

DMF (3mL). DIPEA (116mg, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and the washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Detachment of compound from the resin: The resin which has been treated as described above is swelled and shaken in a mixture of 5% water in trifluoro acetic acid (IOmL) for Ih. The resin is then drained and repeatedly washed with triflouro acetic acid (2x5mL) and DCM (4x5mL). All drained solutions are com- bined and concentrated under nitrogen to a 4mL volume. Et2O (50mL) is the heterogeneous mixture is centrifuged. The centrifuge pellet is washed with Et2O (50mL) and centrifuged.

This treatment is repeated, and the final compound is dried in exicator for 18h.

Using this procedure and building blocks 1-4 the following compounds (19-25 were synthe- sised: Example 19 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-(((1S)-5-amino-1- <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl) -2-(2-naphthyl)ethyl)amide: Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-Phe-OH Buildingblock 3: Fmoc-D-2Nal-OH Buildingblock 4: Boc-AEH-OH HPLC: Rt = 17.1 min (method A) Rt = 27.8 min (method Al) ESMS: M+1 = 629.2 Example 20 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-amino-1- <BR> <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl) -2-(biphenyl-4-yl)ethyl)amide: Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-Phe-OH Buildingblock 3: Fmoc-D-(4-biphenyl)-Ala-OH Buildingblock 4: Boc-AEH-OH HPLC: Rt = 18.4 min (method A) Rt = 30.3 min (method Al) ESMS: M+1 = 655.2 Example 21 N-((1 R)-l -(N-[(1 R)-1 -(((1 S)-5-Amino-1 -carbamoylpentyl)carbamoyl)-2-phenylethyl]-N- methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-3-aminomethylbenzam ide:

Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-Phe-OH Buildingblock 3: Fmoc-D-(4-biphenyl)-Ala-OH Buildingblock 4: Boc-3-AMH-OH HPLC: Rt = 18.9 min (method A) Rt = 30.4 min (method Al) ESMS: M+1 = 663.4 Example 22 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(((1 S)-5-amino-1- carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylcarba moyl}-2-(2- naphthyl)ethyl)amide: Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-(2-thienyl)-Ala-OH Buildingblock 3: Fmoc-D-2Nal-OH Buildingblock 4: Boc-AEH-OH HPLC: Rt = 16.7 min (method A) Rt = 26.7 min (method Al) ESMS: M+1 = 635.4 Example 23 3-Aminomethyl-N-((1 R)- 1 -(N-((1 R)-2-(2-thienyl)- 1 -((1S)-l -carbamoyl-S- aminopentylcarbamoyl)ethyl)-N-methylcarbamoyl)-2-(2-naphthyl )ethyl)benzamide: Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-(2-thienyl)-Ala-OH Buildingblock 3: Fmoc-D-2Nal-OH Buildingblock 4: Boc-3-AMH-OH HPLC: Rt = 17.1 min (method A) Rt = 26.8 min (method Al) ESMS: M+1 = 643.4

Example 24 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-(((1S)-5-amino-1- <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylcarba moyl}-2-(biphenyl-4- yl)ethyl)amide: Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-(2-thienyl)-Ala-OH Buildingblock 3: Fmoc-D-(4-biphenyl)-Ala-OH Buildingblock 4: Boc-AEH-OH HPLC: Rt = 18.7 min (method A) Rt = 29.6 min (method Al) ESMS: M+1 = 661.4 Example 25 N-((1R)-1-{N-[(1R)-1-(((1S)-5-Amino-1-carbamoylpentyl)carbam oyl)-2-(2-thienyl)ethyl]-N- methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-3-aminomethylbenzam ide:

Starting material used: Buildingblock 1: Fmoc-Lys-OH Buildingblock 2: Fmoc-NMe-D-(2-thienyl)-Ala-OH Buildingblock 3: Fmoc-D-(4-biphenyl)-Ala-OH Buildingblock 4: Boc-3-AMH-OH HPLC: Rt = 18.8 min (method A) Rt = 29.7 min (method Al) ESMS: M+1 = 669.4 Acetylation of the Lysine epsilon amino group of example 26-29 was performed using the methodology described below.

The compound to be acetylated at the Lysine epsilon amino group (0.16mmol) is disolved or suspended in 2% Na2CO3 aqueous solution (40mL) and acetic acid anhydride (0.8mmol, 75mL) is added. This mixture is heavily stirred at 200C for 18h. Additional acetic acid anhy- dride (0.8mmol, 75mL) and THF (2.5mL) are added. After 2h the reaction mixture is con- centrated to 30mL, and extracted with DCM (2x20mL). The combined organic phases are concentrated in vacuo, and the crude product is purified on silica (35g) using 10% ammonia in ethanol and DCM (7:3) as eluent to give the monoactylated compound.

Example 26 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1 -(N-[(1 R)-1 -(((1 S)-5-acetylamino-1- <BR> <BR> <BR> <BR> carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl) -2-(2-naphthyl)ethyl)amide:

This compound was prepared by acetylation of the epsilon amino group in the lysine frag- ment using the general method described above and using the compound prepared in ex- ample 19 as starting material.

HPLC: Rt = 19.4 min (method A) Rt = 31.5 min (method Al) ESMS: M+1 = 671.4 Example 27 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-(((1S)-5-acetylamino-1- carbamoylpentyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylcarba moyl)-2-(biphenyl-4- yl)ethyl)amide:

This compound was prepared by acetylation of the epsilon amino group in the lysine frag- ment using the general method described above and using the compound prepared in ex- ample 24 as starting material.

HPLC: Rt = 21.0 min (method A) Rt = 33.6 min (method Al) ESMS: M+1 = 703.4 Example 28 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-(((1S)-5-acetylamino-1- carbamoylpentyl)carbamoyl)-2-phenylethyl]-N-methylcarbamoyl} -2-(biphenyl-4- yl)ethyl)amide: This compound was prepared by acetylation of the epsilon amino group in the lysine frag- ment using the general method described above and using the compound prepared in ex- ample 20 as starting material.

HPLC: Rt = 21.2 min (method A) Rt = 33.9 min (method Al) ESMS: M+1 = 697.4 Example 29 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -((1 S)-5-acetylamino-1 - carbamoylpentylcarbamoyl)-2-(2-thienyl)ethyl]-N-methylcarbam oyl}-2-(2- naphthyl)ethyl)amide: This compound was prepared by acetylation of the epsilon amino group in the lysine frag- ment using the general method described above and using the compound prepared in ex- ample 22 as starting material.

HPLC: Rt = 19.5 min (method A) Rt = 30.4 min (method Al) ESMS: M+1 = 677.2 The following examples (30-33) were prepared on solid support using the general procedure described below:

o + Fmoc deprotection Resin H HO + NHFmoc coupling reagents 0 deprotection ° HO ç > NRFMOC 0 NH2 --NH, coupling reagents deprotection 0 0 Rate buildingbtocki ---NHR H H ~ NHR ingo 0 HO building block coupling reagents 0 0 0 H 1? building blode NHR M v NHFMOC 1.deprotection 0 0 o 2.cleavage t 9b42 f N > 2 e NHR - bd3 NH2 Fmoc deprotection procedure: The resin is swelled and shaken in 20% piperidine in DMF ( 5 ml) for 3 min. The resin is drained and the deprotection process is repeated, with 15 min reaction time.

Standard washing procedure: The drained resin is washed using the following standard washing procedure: The resin is repeatedly swelled, shaken and drained three times with 5mL of DMF, methyl- ene chloride, DMF, 2-propanol, methanol and finally with diethyl ether

Standard protocol for solid phase synthesis of example 30-33.

Deprotection: Commercial diaminoalkyltrityl resins (Novabiochem 01-64-0082, 01-64-0083, 01-64-0081 and 01-34-0132) were employed using the method described above and below.

Attachment of building block 1 (FMOC-N-Me-D-Phe-OH): Building block 1 (0.9mmol), HOBt (138mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 15min, and then transferred to the drained resin, followed by addition of DMF (3mL). DIPEA (116mg, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and the washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Attachment of building block 2 (Fmoc-N-Me-D-Bip-OH): Building block 2 (0.9mmol), HOAt (137mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 15min, and then transferred to the drained resin followed by addition of DMF (3mL). DIPEA (116mg, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and the washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Attachment of buildingblock 3 (FMOC-AEH-OH) Building block 3 (0.9mmol), HOAt (138mg, 0.9mmol) and EDAC (173mg, 0.9mmol) is mixed in DMF (2mL) for 1Smin, and then transferred to the drained resin, followed by addition of DMF (3mL). DlEA (116mg, 0.9mmol) is added after 30min, and the reaction mixture is shaken for 20h.

The resin is drained and washed using the standard procedure described above.

The resin is deprotected using the method described above followed by the standard wash- ing procedure.

Detachment of compound from the resin: The resin which has been treated as described above is swelled and shaken in a mixture of 5% triflouroacetic acid in DCM for 30 min. The resin is then drained and the procedure is re- peated. All drained solutions are combined and concentrated in vacuo.

Using this procedure and FMOC-N-Me-D-Phe-OH and FMOC-N-Me-D-Bip-OH and FMOC- AMH as building blocks, the following four compounds were synthesised: Example 30 (2)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-l -(6-aminohexylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide Diaminohexyl trityl resin was employed.

MS(ES): m/z 640.2 (M+H)+.

Example 31 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-l-(N-[(l R)-l -(4-aminobutylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide

Diaminobutyl trityl resin was employed.

MS(ES): m/z 612.2 (M+H)+.

Example 32 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-(3-aminopropylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-N-me thylamide Diaminopropyl trityl resin was employed.

MS(ES): m/z 598.2 (M+H)+.

Example 33 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(2-aminoethylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-me thylamide Diaminoethyl trityl resin was employed.

MS(ES): m/z 584.2 (M+H)+.

Example 34 3-(1 -Aminoethyl)-N-((l R)- 1 -(N-((1 R)- 1 -(N-(3-dimethylaminopropyl)-N-methylcarbamoyl)-2- phenylethyl)-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lbenzamide

The title compound was prepared as in example 1, omitting acidic washes and using N- methyl-N'-dimethyldiaminopropane as the starting material. Buildingblock 7 and Boc-N-Me- D-2-Nal-OH was employed. The two stereoisomers could not be separated by HPLC. How- ever buildingblock 7 was readily separated into two enantiomers by chiral HPLC and by re- peating the procedure above the two stereoisomers were obtained.

EA: (+ 1H2O) Calc. for C39H51N504: C: 71.55%; H: 7.80%; N: 10.70% Found: C: 71.32%; H: 7.86%; N: 10.70% 'H-NMR: ((CDCl3; selected peaks for major rotamer): d 1.31 (d, 3H); 2.15 (s, 6H); 2.30 (s, 3H); 2.32 (s, 3H); 3.00 (s, 3H) HPLC: (method Al): R, = 29.16 min.

MS(ES): m/z (M+H)+.

Example 35 3-(1 -Aminoethyl)-N-((1 R)- 1 -(N-(( 1 R)-l -(N-((dimethylcarbamoyl)methyl)-N- methylcar- bamoyl)-2-phenylethyl)-N-methylcarbamoyl)-2-(2-naphthyl )ethyl)-N-methylbenzamide.

The title compound was prepared as in example 1 using sarcosine dimethyl amide instead of ((SS)-5-Amino-5-carbamoylpentyl)carbamic acid 2,2,2-trichloroethyl ester hydrochloride.

Boc-N-Me-D-Phe-OH, Boc-N-Me-D-2-Nal-OH and buildingblock 7 were used as starting materials.

1H-NMR: ((CDC13; selected peaks for major rotamer): d 1.40 (d, 3H); 2.90 (1, 6H); 3.07 (s, 3H); 5.71 (dd, 1H); 5.89 (dd, 1H); 5.95 (dd, 2H) HPLC: (method Al): Rt = 31.98 min.

MS(ES): m/z (M+H)+.

Example 36 <BR> <BR> <BR> <BR> (2S)-6-Acetylamino-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2-meth ylpropionylamino)-3-(imidazol- 4-yl)propionylamino]-3-(2-naphthyl)propionylamino)-3-phenylp ropionylamino)hexanoic acid amide (2S)-6-Amino-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2-methylprop ionylamino)-3-(3H-imidazol-4- yl)propionylamino]-3-(2-naphthyl)propionylamino)-3-phenylpro pionylamino)hexanoic acid amide (200 mg, 0.28 mmol) (prepared as in W09517423) was suspended in DMF (10 ml) and DIEA (0.24 ml 1.4 mmole) and acetic anhydride (0.032 ml; 0.33 mmole) was added. The mixture was stirred for 20 h and was diluted with TFA in water (0.2 %, 290 ml) and purified by preparative HPLC in three consecutive runs on a 25 X 250 mm 10u RP-18 column at 40"C with a gradient of 20-40% 0.1% TFA/acetonitrile in 0.1% aqueous TFA. The peptide containing fractions were pooled and lyophilised to afford 114 mg of the title compound.

HPLC (method Al) Rt = 22.13 min (method B1) Rt = 24.60 min ESMS: m/z 754.2 (M+H)+

Example 37 <BR> <BR> <BR> <BR> (2S)-5-Ureido-2-((2R)-2-{(2R)-2-[(2S)-2-(2-amino-2-methylpro pionylamino)-3-(3H-imidazol-4- yl)propionylamino]-3-(2-naphthyl)propionylamino)-3-phenylpro pionylamino)pentanoic acid amide The title compound was prepared as in WO9517423 starting from FMOC-citruline-OH (Bachem B2090).

HPLC (method Al) Rt = 20.83 min (method B1) Rt = 22.97 min ESMS: m/z 742.0 (M+H)+ Example 38 (2S)-6-tert Butyloxycarbonylamino-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2 methylpropionylamino)-3-(imidazol-4-yl)propionylamino]-3-(2- naphthyl)propionylamino}-3- phenylpropionylamino)hexanoic acid amide

(2S)-6-Amino-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2-methylprop ionylamino)-3-(3H-imidazol-4- yl)propionylamino]-3-(2-naphthyl)propionylamino)-3-phenylpro pionylamino)hexanoic acid amide (712 mg, 1.0 mmol) (prepared as in WO9517423) was suspended in water (10 ml) and di-tert-butyldicarbonate (240 mg, 1.0 mmol) was added. The mixture was stirred for 20 h and was diluted with TFA in water (0.2 %, 185 ml) and purified on a Sep-Pac (Waters #36925) RP-18 column washed with 28 % acetonitrile in 0.1% aqueous TFA and eluted with 35% acetonitrile in 0.1% aqueous TFA. The peptide containing fractions were pooled, diluted and lyophilised to afford 408 mg of the title compound.

HPLC (method Al) Rt = 30.53 min (method B1) Rt = 32.75 min ES-MS: m/z 812.8 (M+H)+ Example 39 <BR> <BR> <BR> <BR> <BR> <BR> <BR> (2S)-6-Acetylamino-2-((2R)-2-(N-((2R)-2-(2S)-2-(2-amino-2-me thylprnpionylamino)-3- <BR> <BR> <BR> <BR> <BR> <BR> (imidazol-4-yl)propionylamino]-3-(2-naphthyl)propionyl)-N-me thylamino}-3- phenylpropionylamino)hexanoic acid amide

The title compound was prepared as in example 36.

HPLC (method Al) Rt = 26.38 min (method B1) Rt = 28.70 min ESMS: m/z 769.1 (M+H)+ Example 40 (2S)-6-(3a,7a,12a-trihydroxy-5b-cholanoylamino)-2-((2R)-2-{( 2R)-2-[(2S)-2-(2-amino-2- <BR> <BR> <BR> methylpropionylamino)-3-(3H-imidazol-4-yl)propionylamino]-3- (2-naphthyl)propionylamino)-3- phenyipropionylamino)hexanoic acid amide 3a,7a,12a-trihydroxy-5b-cholanic acid (409 mg; 1 mmole) and HOBt (153 mg; Immole were dissolved in DMF (7 ml) and EDAC (192 mg, 1 mmole) were added and the mixture was stirred for 15 min. (2S)-6-Amino-2-((2R)-2-((2R)-2-[(2S)-2-(2-amino-2- <BR> <BR> <BR> <BR> methylpropionylamino)-3-(3H-imidazol-4-yl)propionylamino]-3- (2-naphthyl)propionylamino)-3- phenylpropionylamino)hexanoic acid amide (712 mg, 1.0 mmol) (prepared as described in

WO9517423) was added and the mixture was stirred for 20 h and was diluted with TFA in water (0.2 %, 100 ml) and purified on a Sep-Pac (Waters #43345) RP-18 column washed with 35 % acetonitrile in 0.1% aqueous TFA and eluted with 55% acetonitrile in 0.1% aque- ous TFA. The peptide containing fractions were pooled, diluted and lyophilised to afford 803 mg of the title compound.

HPLC (method Al) Rt = 35.43 min (method B1) Rt = 37.97 min ES-MS: m/z 1103.0 (M+H)+ Example 41 (2E)-5-Amino-5-methylhex-2-enoic acid ((1 R)-l -(N-[(l R)-l -(5-am i nope ntylcarbam oyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)amide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 28.08min.

(method B1): R, = 29.05min.

Example 42 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-(5-aminopentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methy lamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): R, = 27.30min.

MS(ES): m/z 600.2 (M+H)+.

Example 43 N-((1R)-1-{N-[(1R)-1-(4-Aminobutylcarbamoyl)-2-phenylethyl]- N-methylcarbamoyl}-2- (biphenyl-4-yl)ethyl)-3-aminomethyl-N-methylbenzamide

The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method A1): Rt = 30.40 min.

MS(ES): m/z 620.4 (M+H)+.

Example 44 3-Aminomethyl-N-({1R}-1-{N-[(1R)-1-(5-aminopentylcarbamoyl)- 2-phenylethyl]-N- methylcarbamoyl}-2-(2-naphthyl)ethyl)benzamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): R, = 28.47min.

(method B1): R, = 29.95min.

MS(ES): m/z 594.4 (M+H)+.

Example 45 3-Aminomethyl-N-((1R)-1-{N-[(1R)-1-(5-aminopentylcarbamoyl)- 2-phenylethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methylbenzamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 27.53 min.

MS(ES): m/z 608.4 (M+H)+.

Example 46 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -((4- dimethylaminobutyl)carbamoyl)-2-phenylethyl]-N-methylcarbamo yl}-2-(2-naphthyl)ethyl)-N- methylamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 28.02 min.

MS(ES): m/z 614.7 (M+H)+.

Example 47 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -(5-guanidinopentylcarbamoyl)- 2-phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-met hylamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 25.27 min.

MS(ES): m/z 642.6 (M+H)+.

Example 48 (2E)-5-Amino-5-methylhex-2-enoic acid N-[(1 R)-1 -(N-((I R)-1 -[5-(3- ethylureido)pentylcarbamoyl]-2-phenylethyl}-N-methylcarbamoy l)-2-(2-naphthyl)ethyl]-N- methylamide

The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 29.62 min.

(method B1): Rt = 31.65 min.

MS(ES): m/z 672.4 (M+H)+.

Example 49 3-Aminomethyl-N-[(1R)-1-(N-{(1R)-1-[N-(2-dimethylamino)ethyl )-N-methylcarbamoyl]-2- phenylethyl}-N-methylcarbamoyl)-2-(2-naphthyl)ethyl]-N-methy lbenzamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (methodAl): Rt = 26.33 min.

(method B1): Rt = 29.08 min.

MS(ES):m/z 608.4 (M+H)+.

Example 50 N-[(1R)-1-(N-{(1R)-1-[N-(2-(Dimethylamino)ethyl)-N-methylcar bamoyl]-2-phenylethyl}-N- methylcarbamoyl)-2-(2-naphthyl)ethyl]-N-methyl-3-(methylamin omethyl)benzamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method A1): Rt = 26.68 min.

(method B1): Rt = 29.25 min.

MS(ES): m/z 622.4 (M+H)+.

Example 51 3-((1R/S)-1-Aminoethyl)-N-[(1R)-1-(N-{(1R)-1-[N-(2-(dimethyl amino)ethyl)-N- methylcarbamoyl]-2-phenylethyl}-N-methylcarbamoyl)-2-(2-naph thyl)ethyl]-N- methylbenzamide

The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 27.30 min.

(method B1): Rt = 29.87 min.

MS(ES): m/z 622.0 (M+H)+ Example 52 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-2-(biphenyl-4-yl)-1 -(N-[(l R)-1-(4- (dimethylamino)butylcarbamoyl)-2-phenylethyl]-N-methylcarbam oyl}ethyl)-N-methylamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 30.92 min.

MS(ES): m/z 640.6 (M+H)+.

Example 53 (2E)-5-Amino-5-methylhex-2-enoic acid N-[(I R)-1 -(N-((1 R)-l -[N-(2-dimethylaminoethyl)-N- methylcarbamoyl]-2-phenylethyl}-N-methylcarbamoyl)-2-(2-naph thyl)ethyl]-N-methylamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method Al): Rt = 26.43 min.

(method B1): Rt = 29.02 min.

MS(ES): m/z 600.2 (M+H)+.

Example 54 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1-((4- <BR> <BR> <BR> <BR> dimethylaminobutyl)carbamoyl)-2-(2-thienyl)ethyl]-N-methylca rbamoyl)-2-(2-naphthyl)ethyl)- N-methylamide The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method A1): Rt = 27.30 min.

(method B1): Rt = 29.39 min.

MS(ES): m/z 620.4 (M+H)+.

Example 55 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-2-(4-biphenyl-4-yI)-1 -(N-[(l R)-1-((4- <BR> <BR> <BR> dimethylaminobutyl)carbamoyl)-2-(2-thienyl)ethyll-N-methylca rbamoyl)ethyl)-N-methylamide

The title compound was prepared as in example 1 using the appropriate amine and the buildingblocks listed previously.

HPLC: (method A1): Rt = 30.23 min.

(method B1): Rt = 32.27 min.

MS(ES): m/z 646.4 (M+H)+.

Example 56 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1 R)-1 -((1 S)-5-(acetylamino)-l - (dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methylc arbamoyl}-2-(2- naphthyl)ethyl)-N-methylamide

((1 S)-5-Acetylamino-1 -(dimethylcarbamoyl)pentyl)carbamic acid tert-butyl ester (2S)-6-Acetylamino-2-(tert-butoxycarbonylamino)hexanoic acid (Purchased at Bachem, 5.0 g, 17.3 mmol) was dissolved in dichloromethane (100 ml) and N,N-dimethylformamide (50 ml). 1-Hydroxybenzotriazole hydrate (2.65 g, 17.3 mmol) was added. The solution was cooled to 0 "C. N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.32 g, 17.3 mmol) was added. The reaction mixture was stirred for 20 min at 0 OC. A 33% solution or di- methylamine in methanol (16.4 ml, 121 mmol) was added. The reaction mixture was stirred for 16 h, while it was warming up to room temperature. It was diluted with ethyl acetate (250 ml) and washed with a 10% aqueous solution of sodium hydrogensulfate (200 ml). The aqueous phase was extracted with ethyl acetate (2 x 100 ml). The combined organic layers were washed with a saturated aqueous solution of sodium hydrogen carbonate (200 ml) and dried over magnesium sulfate. The solvent was removed. The crude product was purified by flash chromatography on silica (120 g), using dichloromethane/methanol (10:1) as eluent to give 3.00 g of ((1 S)-5-acetylamino-1-(dimethylcarbamoyl)pentyl)carbamic acid tert-butyl es- ter.

'H-NMR (CDCl3): d 1.49 (s, 9H); 1.34 - 1.75 (m, 6 H); 1.97 (m, 3 H); 2.96 (s, 3 H); 3.07 (s, 3 H); 3.23 (m, 2 H); 4.60 (m, 1 H); 4.99 (br d, 1 H); 5.83 (br, 1 H).

(2S)-6-Acetylamino-2-aminohexanoic acid dimethylamide ((1 S)-5-Acetylamino-1 -(dimethylcarbamoyl)pentyl)carbamic acid tert-butyl ester was dis- solved in a solution of 3.1 M hydrogen chloride in ethyl acetate (15 ml, 46 mmol). The mix- ture was stirred for 1 h at room temperature. The solid was isolated by filtration and dried in vacuo to give 1.86 g of the hydrochloride salt of (2S)-6-acetylamino-2-aminohexanoic acid dimethylamide as crude product, which was used without further purification.

1H-NMR (DMSO-d6, selected values): d 1.39 (m, 4 H); 2.92 (s, 3 H); 3.04 (s, 3 H). <BR> <BR> <BR> <BR> <BR> <BR> <P>N-[(l R)-1 -((1 S)-5-(Acetylamino)-1 -(dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N- methylcarbamic acid tert-butyl ester N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (824 mg, 4.30 mmol) was added at 0 "C to a solution of (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3- phenylpropionic acid (1.20 g, 4.30 mmol) and 1-hydroxybenzotriazole hydrate (659 mg, 4.30

mmol) in dichloromethane (15 ml) and N,N-dimethylformamide (15 ml). The reaction mixture was stirred for 20 min at 0 OC. A solution of the hydrochloride salt of (2S)-6-acetylamino-2- aminohexanoic acid dimethylamide (1.86 g, 6.44 mmol) in N,N-dimethylformamide (10 ml) was added. Ethyldiisopropylamine (2.98 ml, 17.2 mmol) was added. The reaction mixture was stirred for 16 h, while it was warming up to room temperature. The reaction mixture was diluted with ethyl acetate (150 ml) and washed with a 10% aqueous solution of sodium hy- drogen sulfate (200 ml). The aqueous solution was extracted with ethyl acetate (2 x 100 ml).

The combined organic layers were washed with a saturated aqueous solution of sodium hy- drogen carbonate (200 ml) and dried over magnesium sulfate. The solvent was removed.

The crude product was purified by flash chromatography on silica (60 g), using di- chloromethane/methanol (10:1) as eluent to give 1.81 g of N-[(1R)-1-((1S)-5-(acetylamino)- 1 -(dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methyl carbamic acid tert-butyl es- ter.

'H-NMR (CDCl3, selected values): d 1.31 (br, 9 H); 1.55 (m, 4 H); 1.81 and 1.97 (both s, to- gether 3 H); 4.76, 4.91, and 6.02 (all m, together 2 H).

(2S)-6-Acetylamino-2-(((2R)-2-(methylamino)-3-phenylpropi onyl)amino)hexanoic acid di- methylamide N-[(1 R)-1 -((1 S)-5-(Acetylamino)-1 -(dimethylcarbamoyl)pentylcarbamoyl)-2-phenylethyli-N- methylcarbamic acid tert-butyl ester (1.44 g, 3.02 mmol) were dissolved in ethyl acetate (5

ml). A 3.1 M solution of hydrogen chloride in ethyl acetate (40 ml, 124 mmol) was added.

The reaction mixture was stirred for 1.5 h. The liquid was removed by decantation. The solid was washed with ethyl acetate (50 ml) and dried in vacuo to give 1.34 g of the hydrochloride salt of crude (2S)-6-acetylamino-2-(((2R)-2-(methylamino)-3-phenylpropiony l)amino)- hexanoic acid dimethylamide, which was used for the next step without purification.

'H-NMR (DMSO-d6, selected values): d 0.90 (m, 2 H); 1.21 (m, 4 H); 1.80 (s, 3 H); 3.27 (dd, 1 H); 4.10 (m, I H); 4.48 (m, 1 H).

N-((l R)-I -(N-[(1 R)-1 -((1 S)-5-(Acetylamino)-1 -(dimethylcarbamoyl)pentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lcarbamic acid tert-butyl ester N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (594 mg, 3.1 mmol) was added at 0 OC to a solution of (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-(2 naphthyl)propionic acid (1.02 g, 3.1 mmol) and 1-hydroxy-7-azabenzotriazole (422 mg, 3.1 mmol) in dichloromethane (10 ml) and N,N-dimethylformamide (5 ml). The reaction mixture was stirred for 20 min at 0 °C. A solution of the hydrochloride salt of crude (2S)-6- acetylamino-2-(((2R)-2-(methylamino)-3-phenylpropionyl)amino )hexanoic acid dimethyla- mide (1.28 g, 3.1 mmol) in dichloromethane (20 ml)and N,N-dimethylformamide (10 ml) and ethyldiisopropylamine (1.59 ml, 9.3 mmol) were added subsequently. The reaction mixture was stirred for 16 h, while it was warming up to room temperature. It was diluted with ethyl acetate (200 ml) and washed with a 10% aqueous sodium hydrogen sulfate solution (150

ml). The aqueous solution was extracted wtih ethyl acetate (3 x 100 ml). The combined or- ganic layers were washed with a saturated aqueous solution of sodium hydrogen carbonate (200 ml) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash chromatography on silica (120 g), usin di- chloromethane/methanol (10:1) as eluent, to give 1.57 g of N-((1R)-l-(N-[(l R)-1-((1S)-5- (acetylamino)-1-(dimethylcarbamoyl)pentylcarbamoyl)-2-phenyl ethyl]-N-methylcarbamoyl}-2- (2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester.

'H-NMR (CDCl3, selected values): d 0.86, 0.95, 1.20, 1.24, and 1.36 (all s, together 9 H); 1.91, 1.99, 2.21, and 2.25 (all s, together 3 H). <BR> <BR> <BR> <BR> <BR> <BR> <P>(2S)-6-Acetylamino-2-((2R)-2-[N-methyl-N-(2-(methyl amino)-3-(2-naphthyl)propionyl)amino]- 3-phenylpropionylamino)hexanoic acid N, N-dimethylamide N-((1 R)-l-(N-[(l R)-1 -((1 S)-5-(Acetylamino)-1 -(dimethylcarbamoyl)pentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methy lcarbamic acid tert-butyl ester (1.56 g, 2.27 mmol) was dissolved in ethyl acetate (10 ml). A 3.1 M solution of hydrogen chloride (30 ml, 93 mmol) was added. The reaction mixture was stirred for45 min. The sol- vent was removed by decanatation. The residue was washed with ethyl acetate (50 ml) and dried in vacuo to give 1.08 g of the crude hydrochloride salt of (2S)-6-acetylamino-2-((2R)-2- [N-methyl-N-(2-(methylamino)-3-(2-naphthyl)propionyl)amino]- 3-

phenylpropionylamino)hexanoic acid N,N-dimethylamide, which was used for the next step without purification.

'H-NMR (DMSO-d6, selected values): d 1.93 and 2.00 (both s, together 3 H); 5.55 (dd, 1 H). <BR> <BR> <BR> <BR> <BR> <BR> <P>((3E)-4-[N-((1 R)-l-(N-[(l R)-1 -((1 S)-5Acetylamino-l -(dimethylcarbamoyl)pentylcarbamoyl)- 2-phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-met hylcarbamoyl]-1 , I - dimethylbut-3-enyl)carbamic acid tert-butyl ester N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (279 mg, 1.46 mmol) was added at 0 °C to a solution of (2E)-5-(tert-butyoxycarbonylamino)-5-methylhex-2-enoic acid (355 mg, 1.46 mmol) and 1-hydroxy-7-azabenzotriazole (199 mg, 1.46 mmol). The reaction mixture was stirred for 20 min at 0 OC. A solution of the crude hydrochloride salt of (2S)-6- acetylamino-2-((2R)-2-[N-methyl-N-(2-(methylamino)-3-(2-naph thyl)propionyl)amino]-3- phenylpropionylamino)hexanoic acid N,N-dimethylamide (857 mg, 1.46 mmol) in N,N- dimethylformamide (10 ml) and dichloromethane (20 ml) and ethyldiisopropylamine (0.75 ml, 4.38 mmol) were added subsequently. The reaction mixture was stirred for 16 h, while it was warming up to room temperature. It was diluted with ethyl acetate (200 ml) and washed wtih a 10% aqueous solution of sodium hydrogen sulfate (200 ml). The aqueous phase was ex- tracted with ethyl acetate (2 x 100 ml). The combined organic layers were wshed with a satu- rated aqueous solution of sodium hydrogen carbonate (200 ml) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash chro-

matography on silica (120 g), using dichloromethane/methanol (10:1) as eluent, to give 719 mg of ((3E)-4-[N-((1 R)-1-(N-[(1 R)-1 -((1 S)-5-acetylamino-1 -(dimethylcarbamoyl)pentyl- carbamoyl)-2-phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)e thyl)-N-methylcarbamoyl]-1,1- dimethylbut-3-enyl)carbamic acid tert-butyl ester.

1H-NMR (CDCl3, selected values): d 1.32 and 1.39 (both s, together 9 H); 5.38 (m, 1 H); 5.48 (m, 1 H); 6.02 (d, 1 H).

((3E)-4-[N-((1 R)-l-(N-[(1 R)-1 -((1 S)-5-Acetylamino-l -(dimethylcarbamoyl)pentylcarbamoyl)- 2-phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-met hylcarbamoyl]-1,1- dimethylbut-3-enyl)carbamic acid tert-butyl ester (897 mg, 1.10 mmol) was dissolved in di- chloromethane (4 ml). The solution was cooled to 0 "C. Trifluoroacetic acid (4 ml) was added. The reaction mixture was stirred for 35 min at 0 "C. A saturated aqueous solution of sodium hydrogen carbonate (10 ml) was added. Solid sodium hydrogen carbonate was added until pH 7. Water (50 ml) and dichloromethane (30 ml) were added. The phases were separated. The aqueous phase was extracted with dichloromethane (5 x 40 ml). The com- bined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo.

The crude product was purified by flash chromatography on silica (120 g), using dichloro- methane/methanol/25% aqueous ammonia (first: 100:10:1, then 50:10:1), as eluent to give 408 mg of the title compound.

'H-NMR (CDCl3, selected values): d 1.08 and 1.09 (both s, together 6 H); 2.90, 2.92, 3.04, and 3.07 (all s, together 12 H); 6.06 (d, 1 H).

HPLC: 32.07 min (Al).

34.0 min (B1).

LC-MS: 713.4 [M+1]+ at 9.54 min.

For biological testing, the title compound was transferred into its acetate salt by lyophilization with 0.5 M acetic acid (40 ml).

Example 57 (2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-((1S)-5-acetylamino-1- <BR> <BR> (methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methylcar bamoyl)-2-(2-naphthyl)ethyl)- N-methylamide ((1 S)-5-(Acetylamino)-I -(methylcarbamoyl)pentyl)carbamic acid tert-butyl ester

(2S)-5-(Acetylamino)-2-(tert-butoxycarbonylamino)hexanoic acid (purchased at Bachem, 5.0 g, 17.34 mmol) and 1-hydroxybenzotriazole hydrate (2.65 g, 17.34 mmol) were subsequently dissolved in dichloromethane (40 ml) and N,N-dimethylformamide (20 ml). The solution was cooled to 0 "C. N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.32 g, 17.34 mmol) was added. The reaction mixture was stirred at 0 OC for 20 min. A 8.0 M solution of methylamine in ethanol (13 ml, 104 mmol) was added. The reaction mixture was stirred for 16 h, while it was warming up to room temperature. It was diluted with ethyl acetate (300 ml) and washed with a 10% aqueous solution of sodium hydrogensulfate (300 ml). The aquoeus phase was extracted with ethyl acetate (2 x 150 ml). The combined organic layers were washed with a saturated aqueous solution of sodium hydrogen carbonate (300 ml) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash chromatography on silica (65 g), using dichloromethane/methanol (10:1) as eluent to give 792 mg of ((1S)-5-(acetylamino)-1-(methylcarbamoyl)pentyl)carbamic acid tert-butyl ester.

'H-NMR (CDCl3): d 1.30 - 1.90 (m, 6 H); 1.44 (s, 9H); 1.99 (s, 3 H); 2.82 (d, 3 H); 3.25 (q, 2 H); 4.07 (m, 1 H); 5.24 (br, d, 1 H);5.90(br,l H); 6.48 (br, 1H).

(2S)-6-Acetylamino-2-aminohexanoic acid methylamide ((1 S)-5-(Acetylamino)-1 -(methylcarbamoyl)pentyl)carbamic acid tert-butyl ester (792 mg, 2.63 mmol) were dissolved in ethyl acetate (50 ml). A 3 M solution of hydrogen chloride in ethyl acetate (50 ml, 150 mmol) was added. The reaction mixture was stirred for 20 min at room temperature. The solvent was removed by decantation. The residue was washed with ethyl acetate (2 x 20 ml) and dried in vacuo to give 566 mg of the crude hydrochloride salt of (2S)-6-acetylamino-2-aminohexanoic acid methylamide, which was used for the next step without purification.

1H-NMR (DMSO-d6): d 1.30 (m, 2 H); 1.40 (m, 2 H); 1.71 (m, 2 H); 1.81 (s, 3 H); 2.64 (d, 3H); 3.01 (m, 2 H); 3.72 (m, 1 H); 8.06 (br, 1 H); 8.47 (br, 3 H); 3.65 (q, 1 H).

N-[(1 R)-l -((1 S)-5-(Acetylamino)-1 -(methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N- methylcarbamic acid tert-butyl ester N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (411 mg, 2.30 mmol) was added at 0 OC to a solution of(2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3- phenylpropionic acid (643 mg, 2.30 mmol) and 1-hydroxybenzotriazole hydrate (311 mg, 2.30 mmol) in dichloromethane (10 ml) and N,N-dimethylformamide (5 ml). The reaction mixture was stirred for 20 min at O"C. A solution of the crude hydrochloride salt of (2S)-6- acetylamino-2-aminohexanoic acid methylamide (547 mg, 2.30 mmol) in N,N-dimethyl- formamide (5 ml) and ethyldiisopropylamine (1.18 ml, 6.90 mmol) were added subsequently.

The reaction mixture was stirred for 3 days, while it was warming up to room temperature. It was diluted with ethyl acetate (70 ml) and washed with a 10% aqueous solution of sodium hydrogen sulfate (70 ml). The aqueous phase was extracted with ethyl acetate (2 x 40 ml).

The combined organic layers were washed with a saturated aqueous solution of sodium hy- drogen carbonate (70 ml) and dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash chromatography on silica (80 g), using di- chloromethane/methanol (20:1) as eluent to give 1.04 g of N-[(I R)-I -((I S)-5-(acetylamino)- 1-(methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N-methylc arbamic acid tert-butyl ester.

'H-NMR (CDCl3, selected values): d 1.41 (br, 9 H); 1.99 (s, 3 H); 3.20 (q, 2 H); 3.37 (dd, 1 H); 4.39 (m, 1 H); 4.52 (m, 2 H).

(2S)-6-Acetylamino-2-((2R)-2-(methylamino)-3-phenylpropio nylamino)hexanoic acid methy- lamide N-[(1 R)-1 -((1 S)-5-(acetylamino)-1 -(methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N- methylcarbamic acid tert-butyl ester (1.04 g, 2.25 mmol) was dissolved in ethyl acetate (20 ml). A 3 M solution of hydrogen chloride in ethyl acetate (60 ml, 180 mmol) was added. The reaction mixture was stirred at room temperature for 45 min. Ethyl acetate (50 ml) was added. The solvent was removed by decantation. The residue was washed with ethyl ace- tate (2 x 20 ml) and dried in vacuo to give 741 mg of the crude hydrochloride salt of (2S)-6- acetylamino-2-((2R)-2-(methylamino)-3-phenylpropionylamino)h exanoic acid methylamide, which was used for the next step without purification.

1H-NMR (DMSO-d6, selected values): d 0.81 (m, 2 H); 1.10 - 1.55 (m, 4 H); 1.79 (s, 3 H); 3.23 (m, 1 H); 4.03 (m, 2 H).

N-((1 R)-l-(N-[(l R)- 1 -((1 S)-5-(Acetylamino)-1 -(methylcarbamoyl)pentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lcarbamic acid tert-butyl ester N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (313 mg, 1.63 mmol) was added at 0 OC to a solution of (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-(2- naphthyl)propionic acid (537 mg, 1.63 mmol) and 1-hydroxy-7-azabenzotriazole (222 mg, 1.63 mmol) in dichloromethane (20 ml) and N,N-dimethylformamide (10 ml). The reaction mixture was stirred for 20 min at 0 "C. A solution of the crude hydrochloride salt of (2S)-6- acetylamino-2-((2R)-2-(methylamino)-3-phenylpropionylamino)h exanoic acid methylamide (710 mg, 1.63 mmol) and ethyldiisopropylamine (0.84 ml, 4.89 mmol) were added subse- quently. The reaction mixture was stirred for 16 h, while it was warming up to room tem- perature. It was diluted with ethyl acetate (70 ml) and washed with a 10% aqueous solution of sodium hydrogen sulfate (70 ml). The aqueous solution was extracted with ethyl acetate (2 x 60 ml). The combined organic layers were washed with a saturated aqueous solution of sodium hydrogen carbonate (70 ml) and dried over magnesium sulfate. The solvent was re- moved in vacuo. The crude product was purified by flash chromatography on silica (80 g), using dichloromethane/methanol (10:1) as eluent to give 748 mg of N-((1R)-1-{N-[(1R)-1-

((1 S)-S-(acetylamino)-1 -(methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl]-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester.

'H-NMR (CDCl3, selected values): d 1.92 and 1.99 (both s, together 3 H); 4.32, 4.66, 4.82, 5.05, and 5.25 (all m, together 3 H).

(2S)-6-Acetylamino-2-((2R)-2-[N-methyl-N-((2R)-2-(methyla mino)-3-(2- naphthyl)propionyl)amino]-3-phenylpropionylamino)hexanoic acid methylamide N-((1 R)-l-(N-[(l R)-1 -((1 S)-5-(acetylamino)-l -(methylcarbamoyl)pentylcarbamoyl)-2- phenylethyl]-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lcarbamic acid tert-butyl ester (748 mg, 1.11 mmol) was dissolved inethyl acetate (10 ml). A 3 M solution on hydrogen chlo- ride in ethyl acetate (40 ml) was added. The reaction mixture was stirred for 30 min at room temperature. The solvent was removed by decantation. The residue was washed with ehyl acetate (2 x 40 ml). The residue was dried in vacuo to give 451 mg of the crude hydrochlo- ride salt of (2S)-6-acetylamino-2-((2R)-2-[N-methyl-N-((2R)-2-(methylamin o)-3-(2- naphthyl)propionyl)amino]-3-phenylpropionylamino)hexanoic acid methylamide, which was used without further purification.

'H-NMR (DMSO-d6, selected values): d 0.70 - 1.70 (m, 6 H); 4.38 (m); 5.50 (dd).

(3E)-3-(N-((1 R)-1 -(N-((1 R)-1 -((1 S)-5-(Acetylamino)-l -(methylcarbamoyl)pentylcarbamoyl)-2- <BR> <BR> <BR> phenylethyl)-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lcarbamoyl)-1 , I -dimethylbut- 3-enylcarbamic acid tert-butyl ester N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (142 mg, 0.74 mmol) was at 0 "C added to a solution of (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-enoic acid (180 mg, 0.74 mmol) and 1-hydroxy-7-azabenzotriazole (100 mg, 0.74 mmol) in dichloromethane (10 ml) and N,N-dimethylformamide (5 ml). The reaction mixture was stirred at 0 "C for 20 min. A solution of the crude hydrochloride salt of (2S)-6-acetylamino-2-((2R)-2-[N-methyl-N ((2R)-2-(methylamino)-3-(2-naphthyl)propionyl)amino]-3-pheny lpropionylamino}hexanoic acid methylamide (451 mg, 0.74 mmol) in dichloromethane (10 ml) and N,N-dimethyl- formamide (5 ml) and ethyldiisopropylamine (0.38 ml, 2.22 mmol) were added subsequently.

The reaction mixture was stirred for 16 h, while it was warming up to room temperature. It was diluted with ethyl acetate and washed with a 10% aqueous solution of sodium hydrogen sulfate (60 ml). The aqueous phase was extracted with ethyl acetate (2 x 50 ml). The com- bined organic layers were washed with a saturated aqueosu solution of sodium hydrogen carbonate (60 ml) and dried over magnesium sulfate. The solvent was removed in vacuo.

The crude product was purified by flash chromatography on silica (40g), using di- chloromethane/methanol (10:1) as eluent, to give 405 mg of (3E)-3-(N-((1R)-1 -(N-((l R)-l- ((1 S)-5-(acetylamino)-1 -(methylcarbamoyl)pentylcarbamoyl)-2-phenylethyl)-N- methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methylcarbamoyl)-1 , I -dimethylbut-3-enylcarbamic acid tert-butyl ester.

'H-NMR (CDCI3, selected values): d 1.10 - 2.05 (m, 21 H); 1.89, 1.90, 1,96, and 1.97 (all s, together 3 H); 6.05 and 6.10 (both d, toegether 1 H); 6.75 (m, 1 H).

(3E)-3-(N-((1 R)-1 -(N-((1 R)-1 -((1 S)-5-(Acetylamino)-1 -(methylcarbamoyl)pentylcarbamoyl)-2- phenylethyl)-N-methylcarbamoyl)-2-(2-naphthyl)ethyl)-N-methy lcarbamoyl)- 1,1 -dimethylbut- 3-enylcarbamic acid tert-butyl ester (391 mg, 0.49 mmol) was dissolved in dichloromethane (8 ml). The solution was cooled to 0 OC. Trifluoroacetic acid (8 ml) was added. The reaction mixture was stirred for 60 min at 0 "C. It was diluted with dichloromethane (30 ml). A satu- rated aqueous solution of sodium hydrogen carbonate (30 ml) was added carefully. Solid so- dium hydrogen carbonate was added until pH 7. The phases were separated. The aqueous phase was extracted with dichloromethane (3 x25 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash chromatography on silica (40 g), using dichloromethane/methanol/25% aqueous ammonia (first: 100:10:1, then 50:10:1) as eluent to give 124 mg of the title com- pound.

'H-NMR (CDCl3, selected values): d 1.04, 1.10 and 1.11 (all s, together 6 H); 1.90 and 1.96 (both s, together 3 H); 2.69 and 2.73 (both d, together 3 H); 2.78, 2.87, 2.92, and 3.05 (all s, together 6 H); 6.06 and 6.11 (both d, together 1 H); 6.85 (m, 1 H).

MS: 699.6 [M+1]+ HPLC 31.57 min (Al).

33.47 min (B1).

For biological testing, the title compound was transferred into its acetate salt, by lyophiliza- tion with 0.5 M acetic acid (40 ml).