ES2361606T3 - COMPOSITIONS WITH GROWTH HORMONE RELEASE PROPERTIES. - Google Patents

Abstract

Compound of the general formula I ** Formula ** where R1 is hydrogen or C1-5 alkyl; R2 is C1-6 alkyl; L is ** Formula ** where q, s, t and u are independently of each other 0 or 1; r is 0 or 1 the sum q + r + s + t + u is 2 or 3; R9, R10, R11 and R12 are independently from each other hydrogen or C1-6 alkyl; Q is> N-R13 or ** Formula ** where or is 0 or 1; T is -N (R15) (R16) or hydroxyl; R13, R15 and R16 are independently from each other hydrogen or C1-6 alkyl; R14 is hydrogen; G is ** Formula ** where R17, R18, R20 and R21 are independently from each other hydrogen, where R19 is independently hydrogen or aryl; J is ** Formula ** where R22, R23 R25 and R26 independently of each other are hydrogen; where R24 is hydrogen or halogen; a is 1; b is 1; c is 0; d is 0 and is 0 or 1; fis1; R5 is hydrogen or C1-6 alkyl optionally substituted with one or more hydroxyl, aryl or heteroaryl; R6 and R7 are independently from each other hydrogen or C1-6 alkyl; or R6 and R7 form - (CH2) i-U- (CH2) j-, where i and j independently from each other are 1 or 2, and U is a valence bond; R8 is hydrogen or C1-6 alkyl; and M is arylene or -CR27 = CR28-; where R27 and R28 are independently from each other hydrogen or C1-6 alkyl; or a pharmaceutically acceptable derivative salt.

Description

FIELD OF THE INVENTION

[0001] The present invention relates to new compounds, compositions containing them, and their use to treat medical disorders that result from a deficiency of growth hormone.

BACKGROUND OF THE INVENTION

[0002] Growth hormone is a hormone that stimulates the growth of all tissues capable of growing. In addition, it is known that growth hormone has several effects on metabolic processes, for example, it stimulates protein synthesis and mobilization of free fatty acids and causes a change in the energy metabolism of carbohydrate metabolism to fatty acids. Growth hormone deficiency can result in a number of severe medical disorders, for example, dwarfism.

[0003] The pituitary gland releases growth hormone. The release is subject to strict control of various hormones and neurotransmitters either directly or indirectly. Growth hormone release can be stimulated with growth hormone releasing hormone (GHRH) and inhibited by somatostatin. In both cases the hormones are released by the hypothalamus but their action is mediated mainly by specific receptors located in the pituitary gland. Other compounds that stimulate the release of growth hormone from the pituitary gland have also been described. For example, arginine, L-3,4-dihydroxyphenylalanine (L-Dopa), glucagon, vasopressin, PACAP (pituitary adenylyl cyclase activating peptide), muscarinic receptor agonists and a synthetic hexapeptide, GHRP (hormone releasing peptide growth) releases endogenous growth hormone either by direct effect on the pituitary gland or by affecting the release of GHRH and / or somatostatin from the hypothalamus.

[0004] In disorders or conditions where increased levels of growth hormone are required, the nature of the growth hormone protein does anything except non-viable parenteral administration. In addition, other natural secretagogues that act directly, for example, GHRH and PACAP, are longer polypeptides, which is why parenteral administration is preferred.

[0005] The use of determined compounds to increase growth hormone levels in mammals has been previously proposed, for example in EP 18 072, EP 83 864, WO 8302272, WO 8907110, WO 8907111, WO 8910933, WO 8809780, WO 9118016, WO 9201711, WO 9304081, WO 9413696, WO 9517423, WO 9514666, WO 9615148, WO 9622997, WO 9635713, WO 9700894, WO 9722620, WO 9723508, WO 9740023, and WO 9810653.

[0006] WO 98/58950 refers to growth hormone release compounds. The compounds are used to regulate and / or release growth hormone.

[0007] WO 97/23508 refers to growth hormone compounds with improved bioavailability. The invention specifically relates to compounds that act in pituitary cells to release growth hormone.

[0008] WO98 / 03473 refers to compounds, their pharmaceutical compositions to increase the rate and extent of growth of animals to increase their milk and wool production, or for the treatment of ailments and to use the compounds for the preparation of medicaments.

[0009] The composition of growth hormone release compounds is important for their growth hormone release potency as well as their bioavailability. It is therefore an object of the present invention to provide novel compounds with growth hormone release properties. On the other hand, it is an object to provide new growth hormone release compounds (growth hormone secretagogues) that are specific and / or selective and have substantially no side effects, such as for example release of LH, FSH, TSH, ACTH, vasopressin, oxytocin, cortisol and / or prolactin. It is also an object to provide compounds that have good oral bioavailability.

Summary of the Invention

[0010] According to the present invention, new compounds are provided that act directly on the pituitary cells under normal experimental conditions in vitro to release growth hormone thereof.

[0011] These growth hormone release compounds can be used in vitro as unique research tools to understand, among other things, how segregation of growth hormone is regulated at the level of the pituitary gland.

[0012] On the other hand, the growth hormone release compounds of the present invention can also be administered in vivo to increase the release of endogenous growth hormone.

[0013] An embodiment of the invention relates to a compound of the general formula I

image 1

where

one

R is hydrogen or C-- alkyl;

16

2

R is C-- alkyl;

16

5 L is

image 1

where q, s, t and u are independently of each other 0 or 1; R is 0 or 1

10 the sum q + r + s + t + u is 2 or 3; R9, R10, R11 and R12 are independently from each other hydrogen or C1-6 alkyl; Q is> N-R13 or

image 1

15 where or is 0 or 1;

T is -N (R15) (R16) or hydroxyl;

R13, R15 and R16 are independently from each other hydrogen or C alkyl; R14 is hydrogen;

1-6

G is where R17, R18, R20 and R21 independently of each other are hydrogen where R19 is independently hydrogen or aryl;

image 1

J is

image 1

5 where R22, R23 R25 and R26 are independently from each other hydrogen; where R24 is hydrogen or halogen; a is 1; b is 1; 10 c is 0; d is 0;

ees0o1; f is 1; R5

 it is hydrogen or C1-6 alkyl optionally substituted with one or more hydroxy, aryl or heteroaryl; R6 and R7 are independently from each other hydrogen or C1-6 alkyl; or R6 and R7 form - (CH2) i-U- (CH2) j-, where i and j independently from each other are 1 or 2, and U is a valence bond; R8

 is hydrogen or C1-6 alkyl; Y M is arylene or -CR27 = CR28-; where R27 and R28 are independently from each other hydrogen or C1-6 alkyl

20 or a pharmaceutically acceptable derivative salt. [0014] In one embodiment the invention relates to a pharmaceutical composition comprising, as

active substance, a compound or pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier or diluent. [0015] In one embodiment of the invention it refers to a use of a compound or pharmaceutically salt.

It is acceptable for the preparation of a medicament for stimulating the release of growth hormone from the pituitary gland of a mammal. DESCRIPTION OF THE INVENTION [0016] Accordingly, the present invention relates to a compound of the general formula I

image 1

30 where R1

  is hydrogen or C1-8 alkyl; R2

  it is C1-6 alkyl;

image 1

L is where in q, s, t, u are independently of each other 0 or 1 r is 0 or 1; the sum q + r + s + t + u is 2 or 3 R9, R10, R11, and R12 are independently from each other hydrogen or C1-6 alkyl;

image 1

Q is> N-R13 or

10 where or is 0 or 1, T is -N (R15) (R16) or hydroxyl; R13

, R15, and R16 are independently from each other hydrogen or C1-6 alkyl; R14

  it's hydrogen

image 1

15 G is where R17, R18, R20 and R21 independently of each other are hydrogen where R19 is hydrogen or aryl;

image 1

5

10

fifteen

twenty

25

30

35

J is where R22, R23, R25 and R28 independently of each other are hydrogen, where R24 is hydrogen or halogen a is 1 b is 1 c is 0 d is 0 e is 0 or 1 f is 1;

R5

 it is hydrogen or C1-6 alkyl optionally substituted with one or more hydroxyl, aryl or heteroaryl; R6 and R7 are independently from each other hydrogen or C1-6 alkyl, R8

  is hydrogen or C1-6 alkyl

R6 and R7 can optionally form - (CH2) i -U- (CH2) j-, where i and j independently from each other are 1 or 2 and U is a chemical bond;

M is arylene or -CR27 = CR28-; R27

 and R28 are independently from each other hydrogen or C1-6 alkyl,

or a pharmaceutically acceptable derivative salt.

[0017] On the other hand, the compounds of the formula I may comprise any optical isomer thereof, in the form of separate, pure optical isomers or partially purified optical isomers or racemic derived mixtures. Provided that one or more chiral carbon atoms are present, such chiral center or centers (s) may be in the R and / or S configuration, or a mixture of R and S.

[0018] In addition, the compounds of the formula I may have one or more carbon-carbon double bonds with the possibility of geometric isomers, and it is intended that the possible stereoisomers (E or Z isomers) be included within the scope of the invention, unless a special geometric isomer is specified.

[0019] In one embodiment of the compound of the formula R1 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl. In a second embodiment R1 is hydrogen.

[0020] In another embodiment of the compound of formula I R2 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl.

[0021] In another embodiment of the compound of formula I L is

image 1

where q, s, t, or independently of each other are 0 or 1; R is 0 or 1; The sum q + r + s + t + u is 2 or 3 R9, R10, R11, and R12 are independently from each other hydrogen or C1-6 alkyl; Q is> N-R13 or

where or is 0 or 1,

image 1

5

10

fifteen

twenty

25

30

35

T is -N (R15) (R16) or hydroxyl; R13R15

, and R16 are independently from each other hydrogen or C1-6 alkyl; R14

  is hydrogen in one embodiment q is 0. In a second embodiment q is 1. In a third embodiment s is 0. In another embodiment s is 1. In another embodiment t is 0. In another embodiment t is 1. In another embodiment more u is 0. In another embodiment u is 1. In another embodiment r is 0. In another embodiment r is 1. In another embodiment of embodiment R9 is hydrogen. In yet another embodiment, R9 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl. In another embodiment R10 is hydrogen. In yet another embodiment, R 10 is C 1-6 alkyl, such as C 1-4 alkyl, in particular methyl. In another embodiment R11 is hydrogen. In yet another embodiment, R 11 is C 1-6 alkyl, such as C 1-4 alkyl, in particular methyl. In another embodiment R12 is hydrogen. In yet another embodiment R12 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl. In another embodiment, Q is> N-R13. In another embodiment, more R13 is hydrogen. In yet another embodiment R13 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl. In another embodiment Q is

image 1

where R4 is hydrogen. In another embodiment or is 0. In another embodiment or is 1. In yet another embodiment, T is hydroxyl. In another embodiment, T is -N (R15) (R16). In yet another embodiment R15 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl. In another embodiment R15 is C1-6 alkyl, such as C1-4 alkyl, in particular methyl.

[0022] In the compound of the formula I above L is preferably 4-hydroxy-4- (2-thienyl) piperidino, (3-hydroxycyclohexyl) amino, 4- (N, N-dimethylamino) piperidino, N-methyl-N- ( 1-methylpiperidino-4-yl) amino), 4 - ((N, N-dimethylamino) methyl) piperidino, 4-methylpiperazino, (2,2,6,6-tetramethylpiperidine-4-yl) amino, 4-hydroxypiperidine, (3S ) -3 ((N, N-dimethylamino) methyl) -piperidino, (2S) -2 - ((N, N-dimethylamino) methyl) pyrrolidino,

[0023] In yet another embodiment of the compound of formula I G is

image 1

where R17, R16, R20 and R21 independently of each other are hydrogen and where R19 is hydrogen or aryl. In one embodiment R17 is hydrogen. In a second embodiment R18 is hydrogen. In a third embodiment R19 is hydrogen. In another embodiment R19 is aryl, in particular phenyl. In yet another embodiment R20 is hydrogen. In another embodiment R21 is hydrogen. In the compound of the formula I above G is preferably 2-naphthyl or biphenyl-4-yl. In another embodiment of the compound of formula I J is

image 1

where R22, R23, R25 and R26 independently of each other are hydrogen and where R24 is hydrogen or halogen. In one embodiment R22 is hydrogen. In a second embodiment R23 is hydrogen. In a third embodiment R24 is hydrogen. In another embodiment R24 is halogen, in particular fluorine. In yet another embodiment R25 is hydrogen. In another embodiment R26 is hydrogen. In the compound of the formula I above, J is preferably phenyl, 4-fluorophenyl or 2-thienyl.

[0024] In yet another embodiment of the compound of formula I a is 1. [0025] In another embodiment of the compound of formula I b is 1. [0026] In yet another embodiment of the compound of the compound of formula I c is 0. [0027] In another embodiment of the compound of formula I d is 0.

[0028] In yet another embodiment of the compound of the formula I M is arylene or -CR27 = CR28-, where R27 and R28 independently of one another are hydrogen or C1-8 alkyl, optionally substituted with aryl or hetaryl. In one embodiment M is arylene, in particular phenylene. In another embodiment, M is -CR27 = CR28-, where R27 and R28 are independently hydrogen or C1-6 alkyl. In another embodiment R27 is hydrogen. In another way more than

Embodiment R27 is C1-6 alkyl, in particular methyl. In another embodiment R28 is hydrogen. In another embodiment M is the E-isomer of -CR27 = CR28-. In the compound of the formula I above M is preferably ethenylene, 1,3-phenylene or 1,2-propenylene

[0029] In another embodiment of the compound of the formula I e is 0.

[0030] In yet another embodiment of the compound of the formula I e is 1.

[0031] In yet another embodiment of the compound of the formula I f is 1.

[0032] In another embodiment of the compound of the formula I R6 and R7 are independently from each other hydrogen or C1-6 alkyl. In one embodiment R6 is hydrogen. In a second embodiment R8 is C1-6 alkyl, in particular methyl. In a third embodiment R7 is hydrogen. In another embodiment R7 is C1-6 alkyl, in particular methyl.

[0033] In yet another embodiment of the compound of the formula I R6 and R7 form - (CH2) r U- (CH2) j-, where i and j independently of one another are 1 or 2 and U is a chemical bond. In one embodiment the sum i + j is 3. In a second embodiment U is a chemical bond. In a particular embodiment (CR6R7) it is cyclobutyl.

[0034] In another embodiment of the compound of formula I R8 is hydrogen. In a second embodiment R8 is C1-6 alkyl, in particular methyl.

[0035] In a special embodiment the present invention relates to a compound of the general formula I

image 1

where R1

 is hydrogen or C1-6 alkyl; R2

 it is C1-6 alkyl; 25 L is

image 1

q, s, t, u are independently from each other 0 or 1; r is 0 or 1; The sum q + r + s + t + u is 2 or 3;

R9, R10, R11, and R12 are independent of each other hydrogen or C1-6 alkyl; Q is> N-R13 or where or is 0 or 1;

image 1

T is -N (R15) (R16) or hydroxyl; R13

, R15, and R16 are independently from each other hydrogen or C1-6 alkyl; R14

 it's hydrogen G is

image 1

where R17, R18, R20 and R21 independently of each other are hydrogen, where R19 is independently hydrogen or aryl;

10 J is

image 1

where R22, R23, R25 and R26 independently of each other are hydrogen where R24 is hydrogen or halogen; a is 1 b is 1,

15 c is 0, d is 0 e is 0 or 1 f is 1;

R6 is hydrogen or C1-6 alkyl optionally substituted with one or more hydroxyl, aryl or hetaryl;

R6 and R7 are independently of each other hydrogen or C1-6 alkyl, R8

is hydrogen or C1-6 alkyl,

R6 and R7 may optionally form - (CH2) i-U- (CH2) j-, where i and j independently of each other are 1 or 2 and U is a chemical bond;

M is arylene or -CR21 = CR28-; R27

25 and R28 are independently from each other hydrogen or C1-6 alkyl,

or a pharmaceutically acceptable derivative salt. [0036] Preferred compounds of formula I of the invention are: (2E) -5-Amino-5-methylhexo-2- oenic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4 - ((dimethylamino) -methyl) piperidine -1-il) -2

oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3- (dimethylaminomethyl) piperidin -1-il) - 2

(2E) -4- (1-Aminocyclobutyl) but-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3- (dimethylaminomethyl) piperidin-1- and l) 2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image2

image 1

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethyl- amino) methyl) pyrrolidin-1- and l) 2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N- methylcarbamoyl} -2- (2-naphthyl) ethyl)

image3

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4- (dimethyl-amino) piperidin-1 -il) -2-oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide.

image 1

3-Aminomethyl-N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- ( 2-naphthyl) ethyl) -Nmethylbenzamide

image 1

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-methylpiperazin-1-yl) -2 -oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -4- (1-Aminocyclobutyl) but-2-enoic acid N - ((1R) 1- {N - [(1R) 1-benzyl-2- (4-methylpiperazin-1-yl) -2- oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-3,5-dimethylhexo-2-enoic acid N - ((1R) 1- {N - [(1R) 1-benzyl-2- (4-methylpiperazin-1-yl) -2 -oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -4- (1-Aminocyclobutyl) but-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxy-piperidin-1-yl ) -2-oxoethyl] -Nmethylcarbamoyl} -2- (biphenyl-4-yl) ethyl) -N-methylamide

image 1

(2E) -5-amino-5-methylexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2 -oxoethyl] -N

image4

(2E) -5-amino-3,5-dimethylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -Nmethylcarbamoyl} -2- (biphenyl-4-yl) ethyl) -N-methylamide

image 1

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2 -oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-3,5-dimethylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxy-piperidin-1- il) -2-oxoethyl] -N

image5

(2E) -4- (1-Aminocyclobutyl) but-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxy-piperidin-1-yl ) -2-oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1- (4-fluorobenzyl) -2- (4-hydroxy-piperidin- 1-yl) -2-oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-3,5-dimethylhexo-2-enoic acid N - ((1R) -1- {N - [(1R) -1- (4-fluorobenzyl) -2- (4-hydroxypiperidin- 1-il) -2

image6

(2E) 4- (1-Aminocyclobutyl) but-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4- (dimethyl-amino) piperidin-1 -il) -2- oxoethyl] N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

image 1

(2E) -5-amino-3,5-dimethylhexo-2-enoic acid N - ((1R) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo -1 - ((2-thienyl) methyl)

image7

(2E) -5-amino-5-methylhexo-2-enoic acid N - ((1R) -2- (biphenyl-4-yl) -1- {N - [(2R) -2- (4-hydroxy- piperidin-1-yl) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} ethyl) -N-methylamide

image 1

(2E) -5-amino-3,5-dimethylhexo-2-enoic acid N - ((1R) -2- (biphenyl-4-yl) -1- {N - [(1R) -2- (4- hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} ethyl) -N-methylamide

image 1

(2E) -5-methyl-5- (methylamino) hex-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl ) -2-oxoethyl] -N

image4

(2E) -4- (1-Aminocyclobutyl) but-2-enoic acid ((1R) -1- {N - [(1R) -1-benzyl) -2- (4-hydroxypiperidin-1-yl) -2 -oxoethyl] -Nmethylcarbamoyl} -2- (biphenyl-4-yl) ethyl) amide

image 1

10 and pharmaceutically acceptable salts thereof.

General Methods

[0037] The methods illustrated in Schemes I-III below are not limiting of the present invention in any aspect, they should only be seen as a guide to know how the present compounds can be prepared.

image 1

 Type amines [0038]

image 1

5 can be synthesized from a protected BOC amino acid (cf. scheme I). The acid is transformed into an ester by reaction with or without a reagent such as for example 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3 benzotriazol-4 (3H) -one and a reagent such as N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide or diisopropylcarbodiimide hydrochloride and a catalyst such as N, N-dimethylaminopyridine. The ester can be reduced with a suitable reagent, such as diisobutylaluminum hydride in an appropriate solvent such as, for example, toluene, dichloromethane,

10 ether, or tetrahydrofuran to give the aldehyde protected by BOC or alcohol. If alcohol is obtained, it can be oxidized to the corresponding aldehyde, by a suitable method, such as, for example, dimethylsulfoxide / oxalyl chloride / triethylamine or dimethylsufloxide / sulphotroxide / pyridine, pyridinium dichromate, or pyridinium chlorochromate. A reductive amination with an appropriate N (R15) (R16) amine and a suitable reagent, such as sodium cyanoborohydride or sodium triacetoxyborohydride in a suitable solvent such as for example alcohols can produce the protected BOC amine. If at least one of R15 or R16 is hydrogen, the amino group can be protected by a method known to those skilled in the art and described in the literature such as TW Greene, PGM Wuts Protective groups in organic synthesis, 2nd edition, Wiley , New York, before performing the following steps. The removal of the BOC protection group can be achieved by a method known to those skilled in the art as described in TW Greene, PGM Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York, such as for example hydrogen chloride in ethyl acetate, or trifluoroacetic acid in dichloromethane.

image8

[0039] Compounds of the type of formula I can be synthesized by coupling an amine of type

image 1

and a suitable acid protected with or without a binding reagent such as for example 1-hydroxybenzotriazole, 1-hydroxy-7azabenzotriazole, or 3-hydroxy-1,2,3-benzotriazol-4 (3H) -one and a reagent such as for example N- (3-dimethylaminopropyl) -N'ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent, such as N, N-dimethylformamide or dichloromethane (cf. scheme II). The product can be deprotected in the acid nitrogen by a method known to one skilled in the art and described in the literature for example in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York. The product is coupled with a suitable acid protected with or without a binding reagent such as for example 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3benzotriazola-4 (3H) -one and a reagent such as for example N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride or 10 diisopropylcarbodiimide in a suitable solvent, such as N, N-dimethylformamide or dichloromethane. The product can be deprotected in the acid nitrogen by a method known to one skilled in the art and described in the literature for example in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York. The product is coupled with suitable acid protected with or without a binding reagent such as for example 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3-benzotriazol-4 (3H) -one and a reagent as per

Example N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent, such as N, N-dimethylformamide or dichloromethane. All protection groups can be eliminated by a method known to those skilled in the art and described in the literature, for example in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York.

image 1

[0040] Compounds of the type of formula I can be synthesized by coupling an amine of type

image 1

and a suitable acid protected with or without a binding reagent such as for example 1-hydroxybenzotriazole, 1-hydroxy-7azabenzotriazole, or 3-hydroxy-1,2,3-benzotriazol-4 (3H) -one and a reagent as per example N- (3-dimethylaminopropyl) -N'ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent, such as N, N-dimethylformamide or dichloromethane (cf. scheme III). The product can be deprotected in the acid nitrogen by a method known to one skilled in the art and described in the literature for example in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York. The product is coupled with a suitable acid protected with or without a binding reagent such as 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3benzotriazol-4 (3H) -one and a reagent such as N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent, such as N, N-dimethylformamide or dichloromethane. The product can be deprotected in the acid nitrogen by a method known to one skilled in the art and described in the literature for example in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York. The product is coupled with suitable acid protected with or without a binding reagent such as for example 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxyl 1,2,3-benzotriazol-4 (3H) -one and a reagent such as N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent, such as N, N-dimethylformamide or dichloromethane. All protection groups can be removed with a method known to one skilled in the art and described in the literature for example in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis, 2nd edition, Wiley, New York.

[0041] The compounds of the formula I show improved resistance to proteolytic degradation by enzymes because they are not natural, in particular because the natural amide bonds are replaced by unnatural amide bond mimetics. The increased resistance to proteolytic degradation of the compounds of the invention compared to known hormone releasing peptides is expected to improve their bioavailability compared to that of the peptides suggested in the preceding literature.

[0042] In the above structural formulas and throughout this description, the following terms have the indicated meanings:

The C1-6 alkyl, C1-6 alkylene, C1-4 alkyl or C1-4 alkylene groups specified above are intended to include those alkylene or alkyl groups of the length designated either in a cyclic or linear or branched configuration. Examples of linear alkyl are methyl, ethyl, propyl, butyl, pentyl, and hexyl and their corresponding bivalent fractions, such as ethylene. Examples of branched alkyl are isopropyl, sec-butyl, tert-butyl, isopentyl, and isohexyl and their corresponding bivalent fractions, such as isopropylene. Examples of cyclic alkyl are C3-6 cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl and their corresponding bivalent fractions, such as cyclopropylene., Such as cyclopropylene.

The C1-6 alkoxy groups specified above are intended to include those alkoxy groups of designated length in a cyclic or linear or branched configuration. Examples of linear alkoxy 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 C3-6 cycloalkoxy such as cyclopropyloxy, cyclobutyloxy, cyclopentyloxy and cyclohexyloxy.

[0043] In the present context, the term "aryl" includes carbocyclic monovalent aromatic ring fractions, being, polycyclic, monocyclic or bicyclic, for example selected from the group consisting of phenyl and naphthyl, optionally substituted with one or more C1- alkyl 6, C1-6 alkoxy, halogen, amino or aryl.

[0044] In the present context, the term "arylene" includes bivalent carbocyclic aromatic ring moieties, being, polycyclic, monocyclic or bicyclic, for example selected from the group consisting of phenylene and naphthylene, optionally substituted with one or more C1- alkyl 6, C1-6 alkoxy, halogen, amino or aryl.

[0045] In the present context, the term "heteroaryl" is intended to include heterocyclic monovalent aromatic ring fractions, being, for example, polycyclic, monocyclic or bicyclic, selected from the group consisting of pyridyl, 1H-tetrazol-5- yl, thiazolyl, imidazolyl, indolyl, pyrimidinyl, thiadiazolyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thienyl, quinoleinyl, pyrazinyl, or isothiazolyl optionally substituted with one or more C1-6 alkyl, C1-6 alkoxy, halogen, amino or aryl.

[0046] In the present context, the term "hetarylene" includes bivalent heterocyclic aromatic ring moieties, being, polycyclic, monocyclic or bicyclic, for example selected from the group consisting of pyridinediyl, 1-H

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tetrazolediyl, thiazoldiyl, imidazolediyl, indolediyl, pyrimidinediyl, thiadiazolediyl, pyrazolediyl, oxazolediyl, isoxazolediyl, oxadiazolediyl, thiophenediyl, quinolein-ediyl, pyrazinediyl, or isoxy6-alkoxy, C1-alkoxy-substituted alkoxy or aryl.

[0047] In the present context, the term "heterocyclic system" includes aromatic as well as non-aromatic ring fractions, which may be polycyclic, monocyclic or bicyclic, and contains in its ring structure at least one, such as one, two or three, nitrogen atom (s), and optionally one or more, such as one or two, other hetero atoms, for example sulfur or oxygen atoms. The heterocyclic system is preferably selected from pyrazole, pyridazine, triazine, indazol, phthalazine, cinnoline, pyrazolidine, pyrazoline, aziridine, dithiazine, pyrrole, imidazole, pyrazole, isoindole, indole, indazole, purine, pyrrolidine, pyrroline, imidazolidine, iminazolidine, iminazolidine, iminazolidine, pyrazoline , pyrazoline, piperidine, piperazine, indoline, isoindoline, or morpholine, optionally substituted with one or more C1-6 alkyl, C1-6 alkoxy, halogen, amino, oxy or aryl.

[0048] The term "halogen" includes chlorine (Cl), fluorine (F), bromine (Br) and iodine (I).

[0049] In the context of the present application, the term "growth hormone secretagogue" is intended to include any compound that has the ability, directly or indirectly, to induce (ie, stimulate or increase) the release of hormone from the pituitary gland growth The term "growth hormone secretagogue" includes growth hormone releasing peptides, growth hormone release peptidomimetics, and growth hormone release compounds of a non-peptidyl nature. The compounds of the present invention may optionally be in the form of a pharmaceutically acceptable salt such as pharmaceutically acceptable acid addition salts of compounds of formula I including those prepared by reacting the compound of formula I with an inorganic or organic acid such as hydrochloric acid , hydrobromic, sulfuric, acetic, phosphoric, lactic, malic, maleic, mandelic, phthalic, citric, glutaric, gluconic, methanesulfonic, salicylic, succinic, tartaric, fumaric or toluenesulfonic, trifluoroacetic, sulfamic and / or water.

[0050] The compounds of the formula I may be administered in the form of a pharmaceutically acceptable acid addition salt or, where appropriate, as an alkali metal or alkaline earth metal or lower alkylammonic salt. It is believed that such salt forms show approximately the same order of activity as the free base forms.

[0051] In another aspect, the present invention relates to a pharmaceutical composition comprising, as active substance, a compound of the general formula I or a pharmaceutically acceptable derivative salt together with a pharmaceutically acceptable carrier or diluent.

[0052] Pharmaceutical compositions containing a compound of the present invention can be prepared by conventional techniques, for example as described in Remington's Pharmaceutical Sciences, 1985 or in Remington: The Science and Practice of Pharmacy, 19th Edition (1995). The compositions may appear in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or topical applications.

[0053] The pharmaceutical support or diluent used can be a conventional solid or liquid carrier. Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pectin, 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.

[0054] 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.

[0055] If a solid carrier is used for oral administration, the preparation may be in tablets, placed in a hard gelatin capsule in powder or granule form or in the form of a tablet or dragee. The amount of solid support will vary greatly 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 syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.

[0056] A typical tablet that can be prepared with conventional tablet disposal techniques can

contain:

Nucleus:

Active compound (as free compound or derivative of

10mg

Salt)

Colloidal Silicon Dioxide (Aerosil)

1.5mg

Cellulose, microcrist. (Avicel)

70mg

Modified cellulose gum (Ac-Di-Sol)

7.5mg

Magnesium stearate

Covering:

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HPMC approx. 9mg

* Mywacett 9-40 T approx. 0.9mg

* Acylated monoglyceride used as plasticizer for film coating.

[0057] 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 carrier may contain additives such as solubilizing agents, for example propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives such as parabens.

[0058] It has been shown that the compounds of the general formula I possess the ability to release the endogenous growth hormone in vivo. The compounds can therefore be used in the treatment of conditions that require increased plasma growth hormone levels such as in human beings deficient in growth hormone or in older patients or livestock.

[0059] Thus, in a particular aspect, the present invention relates to a pharmaceutical composition for stimulating the release of growth hormone from the pituitary gland, the composition comprising, as active substance, a compound of the general formula I or a pharmaceutically acceptable derivative salt together with a pharmaceutically acceptable carrier or diluent.

[0060] In another aspect, the present invention relates to a method of stimulating the release of growth hormone from the pituitary gland, said method comprising the administration to a subject in need of an effective amount of a compound of the formula general I or a pharmaceutically acceptable derivative salt.

[0061] In a further aspect, the present invention relates to a method of treating growth retardation in relation to asthma, the method comprises administering to an individual in need thereof an effective amount of a growth hormone secretagogue or a pharmaceutically acceptable derivative salt. In a particular embodiment, the present invention relates to a method of treatment of growth retardation in relation to asthma, the method comprises administering to an individual in need thereof an effective amount of a compound of the general formula I or a pharmaceutically acceptable derivative salt.

[0062] In a further aspect, the present invention relates to a method of treatment of growth retardation in relation to rheumatic juvenile arthritis or systolic fibrosis, the method comprises administering to an individual in need thereof an effective amount of a hormone secretagogue. of growth or a pharmaceutically acceptable derivative salt. In one embodiment, the present invention relates to a method of treating growth retardation in relation to rheumatic juvenile arthritis, the method comprises administering to an individual in need thereof an effective amount of a growth hormone secretagogue or a salt. Pharmaceutically acceptable derivative. In a second embodiment, the present invention relates to a method of treating growth retardation in relation to systolic fibrosis, the method comprises administering to an individual in need thereof an effective amount of a growth hormone secretagogue or a pharmaceutically acceptable derivative salt. In a particular embodiment, the present invention relates to a method of treating growth retardation in relation to rheumatic juvenile arthritis, the method comprising administering to a subject in need thereof an effective amount of a compound of the general formula I or a pharmaceutically acceptable derivative salt. In another particular embodiment, the present invention relates to a method of treating growth retardation in relation to systolic fibrosis, the method comprises administering to an individual in need thereof an effective amount of a compound of the general formula I

or a pharmaceutically acceptable derivative salt.

[0063] In a 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 gland.

[0064] It is known to those skilled in the art that the current and potential uses of growth hormone in humans are varied and numerous. Thus, the compounds of the formula I can be administered to stimulate the release of growth hormone from the pituitary gland and then have effects or uses similar to the growth hormone itself. The compounds of formula I are useful for: stimulating the release of growth hormone itself, a greater prevention of catabolic side effects of glucocorticoids, prevention and treatment of osteoporosis, treatment of chronic fatigue syndrome (CFS), treatment of acute fatigue syndrome and muscle loss after elective surgery, immune system stimulation, acceleration of wound healing, accelerated bone fracture repair, acceleration of complicated fractures, for example distraction osteogenesis, degeneration treatment resulting from fractures , growth retardation treatment, growth retardation treatment resulting in renal failure, cardiomyopathy treatment, degeneration treatment in relation to chronic liver disease, thrombocytopenia treatment, growth retardation treatment in relation to Crohn's disease, treatment of short bowel syndrome, degenerative treatment related to chronic obstructive pulmonary disease (COPD), treatment of complications associated with transplantation, treatment of short physiological stature including children deficient in growth hormone and short stature associated with chronic disease, treatment of obesity and growth retardation associated with obesity, treatment of anorexia, treatment of growth retardation associated with Prader-Willi syndrome and Turner syndrome; increase in the growth rate of a patient with insensitive partial growth hormone syndrome, acceleration of recovery and reduction of hospitalization of burn patients; treatment of intrauterine growth retardation, skeletal dysplasia, hypercortisolism and Cushing's syndrome; induction of pulsatile growth hormone release; growth hormone replacement in stressed patients, treatment of osteo-chondrodysplasia, Noonan syndrome, schizophrenia, depressions, Alzheimer's disease, delayed wound healing and psychosocial deprivation, catabolism treatment in relation to pulmonary dysfunction and ventilator dependence; treatment of cardiac failure or related vascular dysfunction, treatment of impaired cardiac function, treatment or prevention of myocardial infarction, decrease in blood pressure, protection against ventricular dysfunction or prevention of reperfusion events; treatment of adults with chronic dialysis; attenuation of catabolic protein responses after major surgery, reduction of cachexia and loss of protein due to chronic disease such as cancer or AIDS; hyperinsulinemia treatment including nesidioblastosis, adjuvant treatment for ovulation induction; stimulation of thymic development and prevention of age-related decline in thymic function, treatment of immunosuppressed patients; sarcopenia treatments, degeneration treatment in relation to AIDS; improvement in muscular endurance, mobility, maintenance of skin thickness, treatment of metabolic homeostasis and renal homeostasis in frail elderly, osteoblast stimulation, bone remodeling and cartilage growth; regulation of food intake; stimulation of the immune system in companion animals and treatment of aging disorder in companion animals, promotion of cattle growth and stimulation of sheep wool growth, increase in milk production in cattle, treatment of metabolic syndrome ( syndrome X), insulin resistance treatment, including NIDDM, in mammals, for example humans, treatment of insulin resistance in the heart, improvement of sleep quality and correction of relative senescence hyposomatotropism due to a high increase in MOR sleep and a latent reduction of ROM, hypothermia treatment, frailty treatment associated with aging, congestive heart failure treatment, hip fracture treatment, immunological deficiency treatment in individuals with depressed T4 / T8 cell proportion, muscle atrophy treatment, degradation treatment Musculoskeletal in elderly people, increased activity of protein kinase B (PKB), improvement of overall pulmonary function, and treatment of sleep disorders.

[0065] In the context of the present application, the term "treatment" also includes prophylactic treatment.

[0066] In another aspect the present invention relates to the use of a growth hormone secretagogue or a pharmaceutically acceptable salt derived for the preparation of a medicament for the treatment of growth retardation in relation to asthma. In a particular embodiment the invention relates to the use of a compound of the general formula I or a pharmaceutically acceptable salt derived for the treatment of growth retardation in relation to asthma. In a second particular embodiment the invention relates to the use of growth hormone releasing peptides, growth hormone releasing peptidomimetics, or growth hormone releasing compounds of a non-peptide nature or a pharmaceutically acceptable salt derived for the treatment. of stunted growth in relation to asthma.

[0067] In a further aspect the present invention relates to the use of a growth hormone secretagogue or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of growth retardation in relation to rheumatic juvenile arthritis or systical fibrosis. . In one embodiment, the invention relates to the use of a growth hormone secretagogue or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of growth retardation in relation to rheumatic juvenile arthritis. In a second embodiment, the invention relates to the use of a growth hormone secretagogue or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of growth retardation in relation to systolic fibrosis. In a particular embodiment the invention relates to the use of a compound of the general formula I or a pharmaceutically acceptable salt derived for the treatment of growth retardation in relation to rheumatic juvenile arthritis. In another particular embodiment the invention relates to the use of a compound of the general formula I or a pharmaceutically acceptable salt derived for the treatment of growth retardation in relation to systolic fibrosis. In another particular embodiment, the invention relates to the use of growth hormone releasing peptides, growth hormone releasing peptidomimetics, or growth hormone releasing compounds of a non-peptide nature or a pharmaceutically acceptable salt derived for the treatment of retardation. of growth in relation to rheumatic juvenile arthritis. In yet another particular embodiment the invention relates to the use of growth hormone releasing peptides, growth hormone releasing peptidomimetics, or growth hormone releasing compounds of a non-peptide nature or a pharmaceutically acceptable salt derived for the treatment of growth retardation in relation to systolic fibrosis.

For the above mentioned indications the dosage will vary depending on the growth hormone secretagogue employed, for example of the compound of the formula I employed, the mode of administration and the desired therapy. However, generally the dosage levels between 0.0001 and 100 mg / kg of daily body mass are administered to patients and animals to obtain an effective release of endogenous growth hormone. In addition, the compounds of the formula I have substantially no side effects, when administered at the above dosage levels, such side effects are for example release of LH, FSH, TSH, ACTH, vasopressin, oxytocin, cortisol and / or prolactin. Normally, dosage forms suitable for transdermal, oral, nasal or pulmonary 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 the formula

mixed with a pharmaceutically acceptable carrier or diluent.

[0069] The dosage of the compounds according to this invention is suitably 0.01-500 mg / day, for example from about 5 to about 50 mg, such as about 10 mg per dose, when administered to patients, for example humans, as a medicine

[0070] Optionally, the pharmaceutical composition of the invention may comprise a compound of formula I combined with one or more compounds exhibiting a different activity, for example, an antibiotic or other pharmacologically active material.

[0071] The route of administration may be any route that effectively transports the active compound to the desired or appropriate site of action, such as the parenteral, oral, nasal, pulmonary, transdermal route, with the oral route being preferred.

[0072] In addition to the pharmaceutical use of the compounds of formula I, in vitro tools may be useful for investigating the regulation of growth hormone release.

[0073] Compounds of the formula I may also be useful in vivo tools for assessing the ability to release growth hormone from the pituitary gland. For example, serum samples taken before and after administration of these compounds to humans for growth hormone can be evaluated. The comparison of growth hormone in each serum sample would directly determine the ability of the pituitary gland of patients to release growth hormone.

[0074] The compounds of the formula I can be administered to commercially important animals to increase their rate and extent of growth, and to increase milk and wool production.

[0075] Another use of growth hormone secretagogue compounds of the formula I is in combination with other secretagogues such as GHRP (2 or 6), GHRH and its analogs, growth hormone and its analogues or inclusive somatomedins IGF-1 and IGF-2

Pharmacological methods

[0076] Compounds of formula I can be evaluated in vitro for their efficacy and strength to release growth hormone in primary cultures of rat pituitary gland, and such evaluation can be performed as described below.

[0077] Isolation of rat pituitary gland cells is a modification of O. Sartor et al., Endocrinology 116, 1985, pp. 952-957. Albino male Sprague-Dawley rats (250 +/- 25 grams) were purchased by Møllegaard, Lille Skensved, Denmark. The rats were housed in a group in cages (four animals / cage) and placed in spaces with a 12-hour light cycle. The ambient temperature varied from 19-24 ° C and the humidity from 30 - 60%.

[0078] The rats were decapitated and the pituitary sectioned. The neurointermediary lobes were removed and the remaining tissue was immediately placed in a frozen isolation buffer (Gey medium (Gibco 041-04030) supplemented with 0.25% D-glucose, 2% non-essential amino acids (Gibco 043-01140) and 1% bovine serum albumin (ABS) (Sigma A-4503)). The tissue was cut into small pieces and transferred to the isolation buffer supplemented with 3.8 mg / ml trypsin (Worthington # 3707 TRL-3) and 330 mg / ml DNase (Sigma D-4527). This mixture was incubated at 70 rotations / min for 35 min at 37 ° C in a 95/5% O2 / CO2 atmosphere. The tissue was then washed three times in the previous buffer. Using a standard pasteur pipette, the tissue was then aspirated into single cells. After dispersion, the cells were filtered through a nylon filter (160 mm) to remove non-assimilated tissue. The cell suspension was washed 3 times with an isolation buffer supplemented with a trypsin inhibitor (0.75 mg / ml, Wortington # 2829) and finally resuspended in a 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 % essential calcium serum (FCS, Gibco 01106290), 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, at a density of 2 x 105 cells / ml. The cells were seeded in microtiter plates (Nunc; Denmark), 200 ml / well, and cultured for 3 days at 37 ° C and 8% CO2.

Compound Test

[0079] After culturing, the cells were washed twice with a Hanks Balanced Salt Solution stimulation buffer (Gibco 041-04020) supplemented with 1% ABS (Sigma A-4503), 0.25% D-glucose (Sigma G -5250) and 25 mM HEPES (Sigma H-3375) pH 7.3) and pre-incubated for 1 hour at 37 ° C. The buffer was exchanged for 90 ml of stimulation buffer (37 ° C). Ten ml of test compound solution were added and the plates were incubated for 15 min at 37 ° C and 5% CO2, the medium was decanted and analyzed for GH content in a rGH SPA test system.

[0080] All compounds were evaluated in doses ranging from 10 pM to 100 mM. A dose response relationship was constructed using the Hill equation (Fig P, Biosoft). The efficacy (maximum GH released, Emax) was expressed in% of the Emax of GHRP-6. The force (EC50) was determined as the concentration induced a maximum mean stimulation of GH release.

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[0081] The compounds of the formula I can be evaluated for their metabolic stability using the procedure described below:

[0082] The compound is dissolved in a concentration of 1 mg / ml in water. 25 ml of this solution are 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 37 ° C overnight. 10 ml of the different degradation solutions are analyzed against a corresponding zero sample using mass spectrometry by flow injection electrospray (ESMS) with ion control selected from the molecular ion. If the signal has decreased more than 20% compared to the zero sample, the rest of the solution is analyzed by HPLC and mass spectrometry identifying the extent and degradation site (s) precisely.

[0083] Different standard peptides (ACTH 4-10, angiotensin 1-14 and glucagon) have been included in the stability tests to verify the ability of various solutions to degrade peptides.

[0084] Standard peptides (angiotensin 1-14, ACTH 4-10 and glucagon) were purchased from Sigma, MO, USA)

[0085] Enzymes (trypsin, chymotrypsin, elastase aminopeptidase M and carboxypeptidase Y and B) were all purchased from Boehringer Mannheim GmbH (Mannheim, Germany).

[0086] Pancreatic enzyme mixture: trypsin, chymotrypsin and elastase in 100 mM ammonia bicarbonate pH 8.0 (all concentrations 0.025 mg / ml).

[0087] Carboxypeptidase mixture: Carboxypeptidase Y and B in 50 mM ammonia acetate pH 4.5 (all concentrations 0.025 mg / ml).

[0088] Solution of aminopeptidase M: aminopeptidase M (0.025 mg / ml) in 100 mM ammonia bicarbonate pH 8.0

[0089] A mass spectrometric analysis was performed using two different mass spectrometers. An Sciex API III triple quadrupole LC-MS instrument (Sciex instruments, Thomhill, Ontario) equipped with an electrospray ion source and a time-of-flight plasma desorption instrument (Bio-Ion Nordic AB, Uppsala, Sweden ).

[0090] The quantification of the compounds (before and after degradation) was done in the API III instrument using single ion monitoring of the molecular ion in question with analyte flow injection. 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 established under standard operating conditions, and SIM monitoring was performed using the most intense molecular ion (in most cases this corresponded to the double charged molecular ion).

[0091] The identification of degradation products also involved the use of plasma desorption mass spectrometry (MSPD) with application of samples on nitrocellulose coated targets and standard instrumental settings. The accuracy of the masses determined here is generally better than 0.1%.

[0092] Separation and isolation of degradation products was done using a phase HPLC column Inverse HY-TACH C-18 4.6x105 mm (Hewlett-Packard Company, Palo Alto, CA) with a standard acetonitrile: TFA separation gradient. The HPLC system used was HP1090M (Hewlett-Packard Company, Palo Alto, CA).

Peptide derivative MW / SIM ion (amu) Mixture of carboxy-peptidase Mixture of pan enzyme. Standards ACTH 4-10 1124.5 / 562. 8 + - Glucagon 3483 / 871.8 - Insulin (B23-29) 859.1 / 430.6 Angiotensin 1-14 1760.1 / 881. 0 -GHRP-2 817.4 / 409.6 -GHRP-6 872.6 / 437.4 - +: Stable (less than a 20% decrease in the SIM signal after 24 hours in the degradation solution). -: Unstable (more than 20% decrease in the SIM signal after 24 hours in the degradation solution)

[0093] Any new feature or combination of features described here is considered essential for this invention. EXAMPLES: [0094] The process for preparing the compounds of the formula I and the preparations containing them is subsequently

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illustrated in the following examples, which, however, are not limiting.

[0095] The structures of the compounds are confirmed either by elementary analysis (EA) or by nuclear magnetic resonance (NMR) or by mass spectrometry (MS). NMR changes (d) are given in parts per million (ppm) and only selected maximum values are given. MP is melting point and is given in ° C. Column chromatography was performed using the technique described by W.C. Still et al, J. Org. Chem. 1978, 43, 2923-2925 On silica gel 60. The compounds used as raw materials are either known compounds or compounds that can be easily prepared by methods known per se.

HPLC analysis:

Method A1

[0096] RP analysis was performed using UV detections at 214, 254, 276, and 301 nm on a C-18 218TP54 4.6 mm x 250 mm 5m silica column (The Seperations Group, Hesperia), which was eluted in 1 mL / min at 42 ° C. 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% acetonitrile in the same buffer for 50 min.

Method B1

[0097] RP analysis was performed using UV detections at 214, 254, 276, and 301 nm on a C-18 218TP54 4.6 mm x 250 mm 5m silica column (The Seperations Group, Hesperia), which was eluted in 1 mL / min at 42 ° C. The column was equilibrated with 5% (acetonitrile + 0.1% TLC) 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% TLC) in the same aqueous buffer

for 50 min.

Abbreviations:

[0098]

FTA:

Thin layer chromatography

DMSO:

 dimethylsulfoxide

Min:

 minutes

h:

 hours

Boc:

tert-butyloxycarbonyl

DMF:

 dimethylformamide

THF:

 tetrahydrofuran

EDAC:

N-ethyl-N '- (3-dimethylaminopropyl) carbodiimide hydrochloride

HOAt:

 1-hydroxy-7-azabenzotriazole

DIEA:

 diisopropylethylamine

TFA:

 trifluoroacetic acid

Functional Blocks:

[0099] The N-methylated amino acids used in the following examples were prepared as in Can. J. Chem. 1977,55,906. 3-hydroxy-1,1-dimethylpropylcarbamic acid tert-butyl ester: [0100]

image 1

[0101] At 0 ° C, ethyl chloroformate (1.10 mL, 11. 5 mmol) was poured dropwise into 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 precipitate formed was filtered and washed with tetrahydrofuran (20 mL). The liquid was immediately cooled to 0 ° C. A 2M solution of lithium borohydride in tetrahydrofuran (14.4 mL, 28. 8 mmol) was added dropwise. The solution was stirred at 0 ° C for 2 h, and then heated to temperature.

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room for 4 h and was cooled to 0 ° C. Methanol (5 mL) was added carefully. 1N 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 with magnesium sulfate. The solvent was removed in vacuo. The crude product was chromatographed on silica (110 g) with 1: 2 ethyl acetate / heptane to give 1.84 g of 3-hydroxy-1,1-dimethylpropylcarbamic acid tert-butyl ester.

1 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:

[0102]

image 1

[0103] 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-dimethylpropylcarbamic acid tert-butyl ester (1.75 g, 8.6 mmol) in dichloromethane (10 mL) was added dropwise over 15 min. The solution was stirred at -78 ° C for another 15 min. Triethylamine (6.0 mL, 43 mmol) was added. The solution was stirred at -78 ° C for 5 min and then heated to room temperature. The solution was diluted with dichloromethane (100 mL) and extracted with 1 N 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 with 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. MHz-1H-NMR (CDCl3): d 1.39 (s, 6 H); 1.45 (s, 9 H); 2.85 (d, 2 H); 4.73 (br. 1 H);

9.80 (t, 1 H).

(2E) -5- (ethyl tert-Butoxycarbonylamino) -5-methylhexo-2-enoate:

[0104]

image 1

[0105] Triethylphosphonoacetate (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) was added to tetrahydrofuran (6 ml). The solution was stirred at room temperature for 75 min. It was diluted in 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 a saturated sodium hydrogen carbonate solution (60 ml) and dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by silica column chromatography (90 g) with ethyl acetate / heptane (1: 4) to give 1.27 g of (2E) -5- (tert-butoxycarbonylamino) -5-methylhex-2-enoate of ethyl.

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).

(2E) -5- (tert-butoxycarbonylamino) -5-methylhex-2-enoic acid:

[0106]

image 1

[0107] (2E) -5- (ethyl tert-butoxycarbonylamino) -5-methylhex-2-enoate (1233 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

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ml) and was extracted with tert-butyl methyl ether (2 x 100 ml). The aqueous phase was acidified with a 1 N sodium hydrogen sulfate solution (pH = 1) and extracted with tert-butyl methyl ether (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 more synthesis.

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).

1-Aza-spiro [3.3] heptan-2-one:

[0108]

image 1

[0109] Methylenecyclobutane (40.0g, 0.587 mol) was dissolved in diethyl ether (250 ml). At - 40 ° C chlorosulfonyl isocyanate (26 ml,

0.294 mol) was added dropwise. The reaction mixture was heated to 10 ° C, an exothermic reaction was observed, and a precipitation formed. The reaction mixture was cooled to -20 ° C. It was stirred for 16 h, while warming to room temperature. A saturated aqueous solution of sodium sulphite (100 ml) was added dropwise. The reaction mixture was vigorously stirred for 1 h. Another saturated aqueous solution of sodium sulphite (100 ml) was added dropwise. Solid sodium hydrogen carbonate was added, until pH 7. Dichloromethane (500 ml) was added. The phases were separated. The organic layer was dried with magnesium sulfate. The solvent was removed in vacuo to give 23.59 g of 1-aza-spiro [3.3] heptan-2-one.

1H-NMR (CDCl3): d 1.75 (m, 2 H); 2.26 (m, 2 H); 2.39 (m, 2 H); 2.96 (s, 2 H); 6.55 (br, 1 H).

2-oxo-1-azaspiro [3.3] heptane-1-carboxylic acid tert-butyl ester

[0110]

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[0111] A solution of di-tert-butyl dicarbonate (55.7 g, 0.211 mol) in dichloromethane (100 ml) was added dropwise to a solution of 1-aza-spiro [3.3] heptan-2-one, triethylamine (36 ml, 0.255 mol), and 4-dimethylaminopyridiene (2.6 g,

0.021 mol) in dichloromethane (100 ml). The reaction mixture was stirred for 16 h at room temperature. It was washed with 10% aqueous solution of ammonium chloride (100 ml), water (100 ml) and a saturated aqueous solution of sodium hydrogen carbonate (100 ml). The organic layer was dried with magnesium sulfate. The solvent was removed in vacuo to give 48.24 g of crude 2-oxo-1-azaspiro [3.3] heptane-1-carboxylic acid tert-butyl ester, which was used for the next step without purification.

1 H-NMR (CDCl3): d 1.55 (s, 9 H); 1.78 (m, 1 H); 1.92 (m, 1 H); 2.18 (m, 2 H); 2.90 (m, 2 H); 3.04 (s, 1 H).

(1- (tert-Butoxycarbonylamino) cyclobutyl) acetic acid:

[0112]

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[0113] 1 N aqueous lithium hydroxide solution (227 ml, 227 mmol) was added to a 2-oxo-1- azaspiro [3.3] heptane-1- carboxylic acid tert-butyl solution (48 g, 0.227 mmol) in tetrahydrofuran (200 ml). The reactive mixture

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It was stirred for 2 h. Diethyl ether (200 ml) and water (200 ml) were added. The mixture was stirred for 16 h. The organic layer was isolated. The aqueous phase was extracted with diethyl ether (200 ml). The aqueous phase was acidified with 10% aqueous sodium hydrogen sulfate solution to pH 3. The precipitation formed was filtered, washed with water, and dried in vacuo, to give 38.84 g of acid (1- (tert-butoxycarbonylamino) cyclobutyl) acetic.

1H-NMR (CDCl3): d 1.45 (s, 9 H); 1.85 (m, 1 H); 1.95 (m, 1 H); 2.25 (m, 4 H); 2.87 (m, 2 H); 5.15 and 6.20 (both br, together 1 H).

(2E) -4- (1- (tert-butoxycarbonylamino) cyclobutyl) but-2-enoic acid:

[0114]

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[0115] (2E) -4- (1- (tert-butoxycarbonylamino) cyclobutyl) but-2-enoic acid was synthesized starting with (1- (tert-butoxycarbonylamino) cyclobutyl) acetic acid analogous to the synthesis of (2E) -5 acid - (tert-butoxycarbonylamino) -5-methylhex2-enoic starting with 3-tert-butoxycarbonylamino-3-methylbutanoic acid.

1H-NMR (CDCl3): d 1.43 (s, 9 H); 1.84 (m, 1 H); 1.95 (m, 1 H); 2.10 (m, 2 H); 2.20 (m, 2 H); 2.70 (m, 2 H); 4.75 (br, 0.5 H); 5.90 (m, 1 H); 6.35 (br, 0.5 H); 6.95 (m, 1 H).

Ethyl ester of (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid:

[0116]

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[0117] Hydrogen oxalate diacetonamine (30.0 g; 146 mmol) was suspended in tetrahydrofuran (400 ml). An aqueous solution of sodium hydroxide (1 N; 146 ml) was added. Di-tert-butyl dicarbonate (38.3 g; 175 mmol) was dissolved in tetrahydrofuran (100 ml) and added dropwise to the reaction mixture. The reaction mixture was stirred for 2 h at room temperature. Sodium hydroxide (1 N; 146 ml) was added and the reaction mixture was stirred for 12 h at room temperature. Water (200 ml) and ethyl acetate (200 ml) were added. The aqueous phase was extracted with ethyl acetate (4 x 200 ml). The combined organic phases were dried with magnesium sulfate, and the solvent was removed in vacuo. The residue was purified by flash column chromatography on silica (200 g), using ethyl acetate / heptane (1: 3) as eluent, to provide 28.4 g of acid tert-butyl ester (1,1-dimethyl- 3oxobutyl) carbamic.

[0118] Triethyl phosphonoacetate (4.7 g; 20.9 mmol) was dissolved in tetrahydrofuran (36 ml). Tert-postasium butoxide (2.3 g; 20.9 mmol) was added and the reaction mixture was stirred for 40 min at room temperature. (1,1-dimethyl-3-oxobutyl) carbamic acid tert-butyl ester (2.5 g; 11.6 mmol) was dissolved in tetrahydrofuran (15 ml) and added dropwise to the reaction mixture that was heated with reflux for 12 h. Ethyl acetate (100 ml) and hydrochloric acid (1 N; 100 ml) were added and the phases were separated. The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic phases were washed with an aqueous solution of sodium hydrogen carbonate (saturated; 100 ml), dried (magnesium sulfate) and evaporated in vacuo. The residue was purified by flash column chromatography on silica (120 g) using ethyl acetate / heptane (1: 2) as eluent to provide 2.0 g of ethyl ester of acid (2E) -5-tert butoxycarbonylamino-3, 5-dimethylhex-2-enoic.

1 H-NMR (CDCl3) d 1.25 (t, 3H); 1.30 (s, 6H); 1.44 (s, 9H); 2.21 (s, 3H); 2.58 (s, 2H); 4.14 (q, 2H); 4.48 (s, 1 H); 5.65 (s, 1 H).

(2E) -5-tert-Butoxycarbonylamino-3,5-dimethylhex-2- oenic acid:

[0119] [0120] Ethyl ester of (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid (1.95 g; 6.83 mmol) was dissolved in 1,4-dioxane (25 ml) and water (15 ml). Lithium hydroxide (0.18 g; 7.52 mmol) was added and the reaction mixture was stirred for 12 h at room temperature. Water (150 ml) and tert-butyl methyl ether (150 ml) were added. The aqueous phase was diluted with 10% aqueous sodium hydrogen sulfate solution to pH 2.5 and extracted with tert-butyl methyl ether (3 x

image9

5 100 ml). The combined organic phases were dried with magnesium sulfate and evaporated in vacuo. The residue was recrystallized from heptane (20 ml) to provide 0.6 g of (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid.

1H-NMR (CDCl3) d 1.29 (s, 6H); 1.44 (s, 9H); 2.23 (s, 3H); 2.62 (s, 2H); 4.45 (s, 1 H); 5.66 (s, 1 H).

(2E) -5- (N- (tert-butoxycarbonyl) -N-methylamino) -5-methylhex-2-enoic acid.

[0121]

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[0122] (2E) -5- (tert-Butyloxycarbonylamino) -5-methylhex-2-enoic acid (5.00 g; 20.6 mmol) was dissolved in tetrahydrofuran

(70 ml). Metilyodide (10.3 ml; 164 mmol) was added and the solution was cooled to 0 ° C. Sodium hydride (60%) was added

in oil) (2.07 g; 61.6 mmol) in parts and the solution was stirred at room temperature for four days. Acetate

of ethyl (70 ml) and water (60 ml) was added dropwise and the solvent was removed in vacuo. The crude product was dissolved in water (40 ml) and ether (40 ml). The organic phase was washed with a saturated aqueous solution of hydrogen carbonate of

sodium (30 ml). The aqueous phases were mixed and 5% aqueous citric acid was added at pH 3. The aqueous phase

It was extracted with ethyl acetate (4 x 50 ml). The organic phase was washed with water (2 x 40 ml), an aqueous solution of

sodium thiosulfate (5%; 40 ml), water (40 ml), dried with MgSO4 and the solvent were removed in vacuo. The residue was

dissolved in ethyl acetate (45 ml) and washed with an aqueous solution of sodium hydrogen sulfate (10%; 3 x 30 ml), dried with MgSO4 and concentrated in vacuo to give 4.00 g of acid (2E) -5- (N- (tert butoxycarbonyl) -N-methylamino) -5-methylhex-2

Oenic

1 H-NMR (CDCl3) � 1.38 (s, 6H), 1.45 (s, 9H); 2.80 (d, 2H); 2.85 (s, 3 H); 5.88 (d, 1 H); 7.01 (q, 1H).

Example 1

N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2 - (2-Naphthyl) ethyl) -N25 (2E) -5-amino-5-methylhex-2-enoic acid methylamide

[0123]

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4- (dimethylcarbamoyl) piperidine-1-carboxylic acid tert-butyl ester [0124]

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[0125] 1- (tert-Butoxycarbonyl) piperidine-4-carboxylic acid (8.0 g, 35 mmol) was dissolved in dichloromethane (70 ml) and N, N

dimethylformamide (35 ml). 1-Hydroxy-7-azabenzotriazole (4.75 g, 35 mmol) was added. The solution was cooled to 0 ° C. Be

N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (6.69 g, 35 mmol) was added. The reaction mixture was stirred.

5 for 20 min at 0 ° C. A 5.6 M solution of dimethylamine in ethanol (37 ml, 209 mmol) was added. The reactive mixture

It was stirred for 3 days while it was warming to room temperature. It was diluted with ethyl acetate

(400 ml) and washed with 10% aqueous sodium hydrogen sulfate solution (400 ml). The aqueous phase was extracted

with ethyl acetate (2 x 200 ml). The combined organic layers were washed with a saturated aqueous solution of

sodium hydrogen carbonate (300 ml) and dried with magnesium sulfate. The solvent was removed in vacuo. The crude product 10 was purified by flash column chromatography on silica (300 g), using dichloromethane / methanol 20: 1 as

eluent, to give 4.56 g of 4- (dimethylcarbamoyl) piperidine-1-carboxylic acid tert-butyl ester.

1 H-NMR (CDCl3): 1.47 (s, 9 H); 1.70 (m, 4 H); 2.60-2.90 (m, 3 H); 2.96 (s, 3 H); 3.08 (s, 3 H); 4.17 (m, 2 H).

4 - ((dimethiamino) methyl) piperidine-1-carboxylic acid tert-butyl ester

[0126]

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[0127] At 0 ° C a solution of 4 - ((dimethylamino) methyl) piperidine-1-carboxylic acid tert-butyl ester (4.56 g, 18 mmol) in tetrahydroofuran (80 ml) was added dropwise to a suspension of sodium borohydride (1.61 g, 43 mmol) in tetrahydrofuran (80 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of iodine 4.51 g, 18 mmol) in tetrahydrofuran (80 ml) was added dropwise at 0 ° C. The reaction mixture was heated at reflux for 16 h. It was cooled to 4 ° C. Methanol (200 ml) was added dropwise. The solvent was removed in vacuo. The residue was dissolved in a 20% aqueous solution of sodium hydroxide (200 ml) and tert-butyl ether of carboxylic acid (150 ml). The phases were separated. The aqueous phase was extracted with carboxylic acid tert-butyl ether (3 x 100 ml). The combined organic layers were dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (100 g), using dichloromethane / methanol / 25% aqueous ammonia.

25 (100: 10: 1) as eluent, to give 4.07 g of 4 - ((dimethylamino) methyl) piperidine-1-carboxylic acid tert-butyl ester.

1 H-NMR (CDCl3): 1.22 (m, 2 H); 1.44 (s, 9 H); 1.85 (d, 2 H); 2.09 (m, 1 H); 2.61 (s, 6 H); 2.65 (m, 2 H); 2.78 (t, 2 H);

4.05 (d, 2 H).

N, N-Dimethyl-N - ((piperidin-4-yl) methyl) amine

[0128]

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[0129] A 3M solution of hydrogen chloride in ethyl acetate (120 ml, 360 mmol) was added to a solution of

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4 - ((dimethylamino) methyl) piperidine-1-carboxylic acid tert-butyl ester (2.0 g, 14 mmol) in ethyl acetate (50 ml). The reaction mixture was stirred for 30 min at room temperature. The solvent was removed in vacuo to give 2.3 g of crude N, N-dimethyl-N - ((piperidin-4-yl) methyl) amine dihydrochloride salt, which was used without purification for the next step.

1H-NMR (CDCl3, selected values): � 1.48 (m, 2 H); 1.92 (s, 6 H); 3.22 (d, 2 H).

tert-butyl ester of N - [(1R) -1-benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamic acid.

[0130]

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[0131] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (1,158 g, 6.04 mmol) was added to a solution of (2R) -2- (N- (tert-butoxycarbonyl) acid -N-Methylamino) -3-phenylpropionic (1-69 g, 6.04 mmol) and 1-hydroxy-7azabenzotriazole (0.822 g, 6.04 mmol) in dichloromethane (25 ml) and N, N-dimethylformamide (12 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of crude dihydrochloride salt of N, N-dimethyl-N - ((piperidin-4-yl) methyl) amine

(1.3 g, 6.04 mmol) in N, N-dimethylformamide (10 ml) and dichloromethane (5 ml) and ethyldiisopropylamine (6.2 ml, 36. 25 mmol) were added successively. The reaction mixture was stirred for 16 h, while it was heated to an ambient temperature. It was diluted with ethyl acetate (100 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (100 ml). The organic layer was dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash chromatography on silica (100 g), using dichloromethane / methanol / 25% aqueous ammonia (200: 10: 1) as eluent, to give 1.22 g of tert-butyl ester of N - [( 1R) -1-Benzyl-2- (4 ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamic.

1H-NMR (CDCl3, selected values): � 1.28, 1.11, 1.37, and 1.38 (all s, together 9 H); 4.00 (m, 1 H); 4.57 (m, 1 H);

4.97 and 5.28 (both t, together 1 H); 7.10 - 7.40 (m, 5 H). MS: 404 [M + 1] +

(2R) -1- (4 - ((Dimethylamino) methyl) piperidin-1-yl) -2- (methylamino) -3-phenylpropan-1-one

[0132]

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[0133] At 0 ° C, trifluoroacetic acid (20 ml) was added to a solution of tert-butyl ester of N - [(1R) -1-benzyl-2- (4 ((dimethylamino) methyl) piperidine-1 -yl) -2-oxoethyl] -N-methylcarbamic (1.22 g, 3.02 mmol) in dichloromethane (20 ml). The reaction mixture was stirred for 1 h at 0 ° C. The solvents were removed in vacuo. The residue was dissolved in dichloromethane (70 ml) and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (100 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1), to give 659 mg of (2R) -1- (4- ( (dimethylamino) methyl) piperidin-1-yl) -2- (methylamino) -3-phenylpropan-1-one.

1 H-NMR (CDCl3, selected values): � 0.91 and 1.47 (m and d, together 1 H); 1.27 (m, 1 H); 4.62 (t, 1 H).

tert-butyl acid ester N - ((1R) -1- {N - [(1R) -1-Benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] - Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamic

[0134]

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[0135] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (416 mg, 2.17 mmol) was added to a solution of (2R) -2- (N- (tert-butoxycarbonyl) acid -N-methylamino) -3- (2-naphthyl) -propionic (715 mg, 2.17 mmol) and 1 hydroxy-7azabenzotriazole (296 mg, 2.17 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 (2R) -1- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2- (methylamino) -3-phenylpropan-1-one (659 mg, 2.17 mmol) in dichloromethane (10 ml) and N, N-dimethylformamide (5 ml) and ethyldiisopropylamine

(0.56 ml, 3.26 mmol) were added successively. The reaction mixture was stirred for 16 h, while it was warming up to room temperature. It was diluted with ethyl acetate (100 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (100 ml). The aqueous solution was extracted with acetate

10 ethyl (3 x 50 ml). The combined organic layers were dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (80 g), using ethyl acetate / heptane / triethylamine (1: 1: 0.08) as eluent, to give 1.05 g of tert-butyl acid ester N - (( 1R) -1- {N - [(1R) -1benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N- methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamic

1 H-NMR (CDCl3, selected values): � 1.24 and 1.42 (both s, together 9 H); 5.04, 5.28, 5.44, 5.54, 5.73 (m, dd, dd, dd, 15 and m, together 3 H);

(2R) -N - [(1R) -1-Benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino) -3- (2-naphthyl) propionamide

[0136]

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[0137] At 0 ° C, trifluoroacetic acid (18 ml) was added to a solution of tert-butyl ester of N - ((1R) -1- {N - [(1R) -1

Benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) -ethyl) -N-methylcarbamic (1.05 g, 1.71 mmol) in dichloromethane (18 ml). The reaction mixture was stirred for 50 min at 0 ° C. The solvents were removed in vacuo. The residue was dissolved in dichloromethane (50 ml) and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (80 g), using dichloromethane / methanol / 25% aqueous ammonia as eluent, to give 846 mg of (2R) -N - [(1R) -1-benzyl-2 -(4

25 ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino) -3- (2-naphthyl) propionamide.

1 H-NMR (CDCl3, selected values): � 0.60 (m, 1 H); 4.38 (t, 1 H); 5.72 and 5.79 (both t, together 1 H).

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acid tert-butyl ester {(3E) -4- [N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1- il) -2-oxoethyl] -Nmethylcarbamoyl} -2- (2- naphthyl) ethyl) -N-methylcarbamoyl] -1,1-dimethylbut-3-enyl} carbamic.

[0138]

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[0139] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (112 mg, 0.58 mmol) was added to a solution of (2E) -5- (tert-butoxycarbonylamino) -5- methylhex-2-enoic (142 mg, 0.58 mmol) and 1-hydroxy-7-azabenzotriazole (79 mg, 0.58 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 (2R) -N - [(1R) -1-benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino) -3 (2-Naphthyl) propionamide (300 mg, 0.58 mmol) in dichloromethane (5 ml) and N, N-dimethylformamide (5 ml) and ethyldiisopropylamine

0.10 ml, 0.58 mmol) were added successively. The reaction mixture was stirred for 3 days, while heating to room temperature. It was diluted with ethyl acetate (70 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (70 g), using dichloromethane / methanol / 25% aqueous ammonia (200: 10: 1) as eluent, to give 313 g of acid tert-butyl ester {(3E ) -4- [N - ((1R) -1- {N - [(1R) -1benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl } 2- (2-Naphthyl) ethyl) -N-methylcarbamoyl] -1,1-dimethylbut3-enyl} carbamic.

1 H-NMR (CDCl3, selected values): � 1.28 and 1.30 (both s, together 6 H); 1.42 (s, 9H); 2.23, 2.27, 2.38, 2.43, 2.51, 2.52, 2.81, and 2.82 (all s, together 12 H); 5.56, 5.76, and 5.90 (m, m, and dd, together 2 H); 6.17 and 6.19 (both dd, together 1 H);

6.94 (m, 1 H).

[0140] At 0 ° C, trifluoroacetic acid (6 ml) was added to a solution of acid tert-butyl ester {(3E) -4- [N - ((1R) -1- {N [(1R) - 1-Benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] -1,1-dimethylbut -3-enyl} carbamic (212 mg, 0.29 mmol) in dichloromethane (6 ml). The reaction mixture was stirred for 20 min at 0 ° C. It was diluted with dichloromethane (30 ml). A saturated aqueous solution of sodium hydrogen carbonate (30 ml) was added dropwise. Solid sodium hydrogen carbonate was added, obtained until pH 7. The phases were separated. The aqueous phase was extracted with dichloromethane (3 x 50 ml). The combined organic layers were dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (20 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 5 mg of the title compound.

1 H-NMR (CDCl3, selected values): � 1.20 (s, 6 H); 2.28, 2.32, 2.41, 2.49, 2.56, 2.57, 2.82, and 2.83 (all s, together 12 H); 5.58, 5.78, and, 5.92 (m, m, and dd, together 2 H); 6.16 and 6.19 (both d, together 1 H); 7.00 (m, 1 H).

HPLC: 39. 23 min (A1).

41. 55 min (B1).

MS: 640.4 [M + 1] +.

[0141] For biological tests, the title compound was transferred into its acetate salt by lyophilization with 0.5 M acetic acid (40 ml).

Example 2

N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3- (dimethylaminomethyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} - (2E) -5-Amino-5-methylhex-2- enoic acid 2- (2-naphthyl) ethyl) -N-methylamide

[0142] 1-tert-butyl ester 3-ethyl ester of (3R) -piperidine-1,3-dicarboxylic acid [0143]

image 1

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[0144] (R) -ethyl nipetcotate tartrate (10.0 g, 32.5 mmol) were suspended in tetrahydrofuran (90 ml). A 1 N solution of sodium hydroxide in water (98 ml, 98 mmol) was added. A solution of di-tert-butyl dicarbonate (7.10 g, 32.5 mmol) was added in tetrahydrofuran (90 ml). The reaction mixture was stirred for 16 h at room temperature. Ethyl acetate (400 ml) was added. The reaction mixture was washed with a 10% aqueous solution of sodium hydrogen sulfate (400 ml). The aqueous solution was extracted with ethyl acetate (2 x 200 ml). The combined organic layers

10 were washed with a saturated aqueous solution of sodium hydrogen carbonate (200 ml) and dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (90 g), using 1: 4 ethyl acetate / heptane as eluent, to give 4.13 g of 1-tert-butyl ester 3-ethyl acid ester (3R ) -piperidine-1,3-dicarboxylic.

1 H-NMR (CDCl3): 1.27 (t, 3 H); 1.48 (s, 9 H); 1.54 (m, 1 H); 1.62 (m, 1 H); 1.73 (m, 2H); 2.05 (m, 1 H); 2.45 (m, 1 H);

15 2.81 (m, 1 H); 2.98 (br, 1 H); 3.93 (m, 1 H); 4.14 (q, 1 H).

(3R) -3-Formylpiperidine-1-carboxylic acid tert-butyl ester

[0145] [0146] A 1.2 M solution of diisobutylaluminum hydride in toluene (30.8 ml, 36.9 mmol) was added at -78 ° C to a solution of 1-tert-butyl ester 3-ethyl ester of (3R) -piperidine-1,3-dicarboxylic acid (4.13 g, 16. 1 mmol) in diethyl ether (30 ml). The reaction mixture was stirred for 2.5 h at - 78 ° C. Water (9.6 ml) was added dropwise. The reactive mixture

image 1

5 was heated to room temperature. Precipitation was removed by filtration through a plug of celite. The celite was washed with tert-butyl methyl ether (3 x 100 ml). The liquids were combined and dried with magnesium sulfate. The solvent was removed in vacuo, to give 1.94 g of crude (3R) -3-formylpiperidine-1-carboxylic acid tert-butyl ester, which was used for the next step without further purification.

1 H-NMR (CDCl3): 1.45 (s, 9H); 1.67 (m, 2 H); 1.95 (m, 1 H); 2.43 (m, 1 H); 3.10 (m, 1 H); 3.32 (dd, 1 H); 3.52 (d, 1 H);

10 3.66 (m, 1 H); 3.95 (m, 1 H); 9.69 (s, 1 H).

(3S) -3- (Dimethylaminomethyl) piperidine-1-carboxylic acid tert-butyl ester

[0147]

image 1

[0148] A solution of crude tert-butyl ester of (3R) -3-formylpiperidine-1-carboxylic acid (1.94 g, 9.1 mmol) was

15 prepared in dichloromethane (80 ml). A 5.6 M solution of dimethylamine in ethanol (3.2 ml, 18. 2 mmol) and molecular sieves were added successively. Sodium triacetoxyborohydride (5.78 g, 27.3 mmol) was added to this mixture. Acetic acid (1.04 ml, 18. 2 mmol) was added. The reaction mixture was stirred for 16 h at room temperature. A 1 N aqueous solution of sodium hydroxide (70 ml) and tert-butyl methyl ether (70 ml) were added. The phases were separated. The aqueous solution was extracted with tert-butyl methyl ether (3 x 70 ml). Organic layers

20 combined were dried with magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 866 mg of acid tert-butyl ester (3S) -3- (dimethylaminomethyl) piperidine-1-carboxylic.

1H-NMR (CDCl): 1.10 (m, 1 H); 1.45 (s, 9 H), 1.45 (m, 1 H); 1.64 (m, 2 H); 1.85 (m, 1 H); 2.10 (m, 2 H); 2.20 (s, 6 H);

3

2.50 (br, 1 H); 2.79 (m, 1 H); 3.95 (m, 2 H).

25 N, N-Dimethyl-N - (((3R) -piperidin-3-yl) methyl) amine

[0149]

image 1

[0150] tert-butyl ester of (3S) -3- (dimethylaminomethyl) piperidine-1-carboxylic acid (1.25 g, 5.15 mmol) was dissolved in ethyl acetate (30 ml). A 2.7 M solution of hydrogen chloride in ethyl acetate (75 ml, 203 mmol) was added. The reaction mixture was stirred for 45 min at room temperature. The solvent was removed in vacuo to give 976 mg.

5

10

fifteen

twenty

25

30

35

of N, N-dimethyl-N - (((3R) -piperidin-3-yl) methyl) amine dihydrochloride salt, which was used for the next step without further purification.

1 H-NMR (CD3OD): 1.42 (m, 1 H); 1.86 (m, 1 H); 2.00 (m, 2 H); 2.38 (m, 1 H); 2.85 (t, 1 H); 2.95 (s, 6 H); 2.98 (m, 1 H);

3.16 (m, 2 H); 3.42 (m, 1 H); 3.53 (m, 1 H). N- [(1R) -1-Benzyl-2 - ((3S) -3- (dimethylaminomethyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamic acid tert-butyl ester [0151]

image 1

[0152] At 0 ° C N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (870 mg, 4.54 mmol) was added to a solution of (2R) -2- (N- (tert-butoxycarbonyl) -N acid -methylamino) -3-phenylpropionic (1.27 g, 4.54 mmol) and 1-hydroxy-7-azabenzotriazole (617 mg, 4.54 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 dihydrochloride salt of N, N-dimethyl-N - (((3R) -piperidin-3-yl) methyl) amine (976 mg,

4.54 mmol) in dichloromethane (20 ml) and N, N-dimethylformamide (10 ml) and ethyldiisopropylamine (3.9 ml, 22. 7 mmol) were added successively. The reaction mixture was stirred for 3 days, while heating until it reached an ambient temperature. Ethyl acetate (300 ml) was added. The solution was washed with a saturated aqueous solution of sodium hydrogen carbonate (300 ml). The aqueous phase was extracted with ethyl acetate (2 x 200 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 1.69 g deterc-butyl ester of N - [( 1R) -1-Benzyl-2 - ((3S) -3 (dimethylaminomethyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamic.

1H-NMR (CDCl3, selected values): � 1.20, 1.24, 1.31, and 1.32 (all s, together 9 H); 2.12, 2.13, and 2.18 (all s, together 6 H); 2.81 (m, 3 H); 4.97 and 5.30 (both m, together 1 H); 7.05 - 7.35 (m, 5 H).

(2R) -1 - ((3S) -3 - ((Dimethylamino) methyl) piperidin-1-yl) -2-methylamino-3-phenylpropan-1-one

[0153]

image 1

[0154] At 0 ° C, trifluoroacetic acid (25 ml) was added to a solution of tert-butyl ester of N - [(1R) -1-benzyl-2 acid ((3S) -3- (dimethylaminomethyl) piperidine- 1-yl) -2-oxoethyl] -N-methylcarbamic (1.69 g, 4.2 mmol) in dichloromethane (25 ml). The reaction mixture was stirred for 30 min at 0 ° C. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (100 ml) and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 1.15 g of (2R) -1 - ((3S ) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-methylamino-3-phenylpropan-1-one.

1 H-NMR (CDCl3, selected values): � 0.38, 1.11, 1.37, and 1.65 (all m, together 4 H); 2.11, 2.19, 2.25, and 2.31 (all s, together 9 H); 4.37 and 4.53 (both m, together 1 H); 7.10 - 7.35 (m, 5 H).

acid tert-butyl ester N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2 -oxoethyl] -Nmethylcarbamoyl} -2- (2- naphthyl) ethyl) -N-methylcarbamic

[0155]

image 1

[0156] At 0 ° C N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (379 mg, 1.98 mmol) was added to a solution of (2R) -2- (N- (tert-butoxycarbonyl) - N-methylamino) -3- (2-naphthyl) -propionic (651 mg, 1.98 mmol) and 1-hydroxy-7azabenzotriazole (269 mg, 1.98 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 (2R) -1 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-methylamino-3-phenylpropan-1-one (600 mg, 1.98 mmol) in dichloromethane (10 ml ) and ethyldiisopropylamine (0.51 ml, 2.97 mmol) were added successively. The reaction mixture was stirred for 3 days, while heating to room temperature. Ethyl acetate (100 ml) was added. The solution was washed with a saturated aqueous solution of sodium hydrogen carbonate (100 ml). The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methane / 25% aqueous ammonia.

(100: 10: 1) as eluent, to give 1.18 g of tert-butyl acid ester N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((35) -3 ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamic.

1 H-NMR (CDCl3, selected values): � 0.45 and 0.71 (both m, together 1 H); 1.03, 1.05, 1.15, 1.20, 1.28, 1.36, and 1.42 15 (all s, together 9 H); 2.12, 2.15, 2.21, 2.26, 2.29, 2.85 (all s, together 6 H); 5.05, 5.44, 5.58, 5.71, 5.85, and 6.00 (all s, together 2 H); 7.10 - 7.80 (m, 12 H).

(2R) -N - [(1R) -1-Benzyl-2 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino ) -3- (2naphthyl) propionamide

[0157]

image 1

twenty

[0158] At 0 ° C, trifluoroacetic acid (20 ml) was added to a solution of tert-butyl ester of N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S ) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamic (1.18g,

1.92 mmol) in dichloromethane (20 ml). The reaction mixture was stirred for 50 min at 0 ° C. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (80 ml) and the solvent was removed in vacuo. This procedure was

25 repeated twice. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 788 mg of (2R) -N - [(1R ) -1-benzyl-2 ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino) -3- (2-naphthyl) - propionamide

1 H-NMR (CDCl3, selected values): � 2.01 and 2.25 (both s, together 9 H); 3.72 (m, 2 H); 3.95 and 4.27 (both m, together 1 H); 5.77, 5.86, and 6.03 (t, m, and dd, together 1 H); 7.10 and 7.85 (m, 12 H).

30 tert-butyl acid ester {(3E) -4- [N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3 - ((dimethylamino) methyl ) piperidin-1-yl) -2-oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] -1,1-dimethylbut-3-enyl} carbamic

5

10

fifteen

twenty

25

30

[0159]

image 1

[0160] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (105 mg, 0.55 mol) was added to a solution of (2E) -5- (tert-butoxycarbonylamino) -5-methylhex -2-enoic (136 mg, 0.55 mmol) and 1-hydroxy-7- azabenzotriazole (74 mg,

0.55 mmol) in dichloromethane (5 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of (2R) -N [(1R) -1-benzyl-2 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino) -3- (2-naphthyl) propionamide (281 mg, 0.55 mmol) in dichloromethane (5 ml) and N, N-dimethylformamide (5 ml) and ethyldiisopropylamine (0.094 ml, 0.55 mmol) were added successively. The reaction mixture was stirred for 16 h, while heating to room temperature. It was diluted with ethyl acetate (70 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia.

(100: 10: 1) as eluent, to give 398 mg of acid tert-butyl ester {(3E) -4- [N - ((1R) -1- {N - [(1R) -1-benzyl- 2 - ((3S) -3 ((dimethylamino) methyl) -piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] -1,1 -dimethylbut-3enyl} carbamic.

1 H-NMR (CDCl3, selected values): � 1.44 (s, 9 H); 5.58, 5.75, and 5.86 (all m, 2 H); 6.09 and 6.17 (both d, together 1 H); 6.84 (m, 1 H); 7.10 - 7.80 (m, 12 H).

[0161] At 0 ° C, trifluoroacetic acid (7 ml) was added to a solution of acid tert-butyl ester {(3E) -4- [N - ((1R) -1- {N [(1R) - 1-Benzyl-2 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] 1,1-dimethylbut-3-enyl} carbamic (398 mg, 0.54 mmol) in dichloromethane (7 ml). The reaction mixture was stirred for 40 min at 0 ° C. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (20 ml) and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 150 mg of the title compound.

1 H-NMR (CDCl3, selected values): � 1.08, 1.12, 1.14, and 1.15 (all s, together 6 H), 5.46, 5.59, 5.75, and 5.94 (all m, together 2 H); 6.15 (m, 1 H); 6.93 (m, 1 H).

HPLC

27. 5 min (A1).

30. 23min (B1).

LC-MS:

640.4 [M + 1] + at 8.54 min.

[0162] To carry out biological tests, the title compound was transferred in its acetate salt, by lyophilization of 0.5 M acetic acid (40 ml).

Example 3 N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3- (dimethylaminomethyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl } -2- (2-Naphthyl) ethyl) -N-Methylamide (2E) -4- (1-aminocyclobutyl) but-2-enoic acid

[0163] tert-butyl acid ester (1 - {(2E) -3- [N - ((1R) -1- {N - [(1R) -1-Benzyl-2 - ((3S) -3- ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] allyl} cyclobutyl) carbamic

image 1

image 1

[0164] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (85 mg, 0.44 mmol) was added to a solution of (2E) -5- (tert-butoxycarbonylamino) -5 acid -methylhex-2-enoic (113 mg, 0.44 mmol) and 1-hydroxy-7-azabenzotriazole (60 mg, 0.44 mmol) in dichloromethane (5 ml) and N, N-dimethylformamide (5 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of (2R) -N - [(1R) -1-benzyl-2 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methyl-2 (methylamino) -3- (2-naphthyl) propionamide (228 mg, 0.44 mmol) in dichloromethane (10 ml) and ethyldiisopropylamine (0.07 ml, 0.44

10 mmol) were added successively. The reaction mixture was stirred for 16 h, while heating to room temperature. It was diluted with ethyl acetate (70 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia.

15 (100: 10: 1) as eluent, to give 314 mg of acid tert-butyl ester (1 - {(2E) -3- [N - ((1R) -1- {N - [(1R) - 1-Benzyl-2 - ((3S) -3 ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] allyl } cyclobutyl) carbamic.

1 H-NMR (CDCl3, selected values): � 1.40 (m, 9 H); 4.22 and 4.67 (both m, together 2 H); 5.60, 5.75, 5.85, and 5.90 (dd, dd, m, and m, together 2 H); 6.10 and 6.19 (both d, together 1 H); 6.73 and 6.87 (both m, together 1 H); 7.22, 7.42, and 7.76

20 (all m, together 12 H).

[0165] At 0 ° C, trifluoroacetic acid (7 ml) was added to a solution of acid tert-butyl ester (1 - {(2E) -3- [N - ((1R) -1- {N [( 1R) -1-Benzyl-2 - ((3S) -3 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N -methylcarbamoyl] allyl} cyclobutyl) carbamic (314 mg, 0.42 mmol) in dichlormethane (7 ml). The reaction mixture was stirred for 20 min at 0 ° C. The solvent was removed in vacuo without heating. The residue was dissolved in dichloromethane (20 ml) and the solvent

25 was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / ammonia (100: 10: 1) as eluent, to give 180 mg of the title compound.

1 H-NMR (CDCl3, selected values): � 0.40 and 0.74 (both m, together 2 H); 3.73 and 4.22 (both m, together 2 H); 5.57, 5.77, and 5.91 (all m, together 2 H); 6.15 and 6.24 (both d, together 1 H); 6.85 and 6.96 (both m, together 1 H); 7.22, 7.92, and

30 7.74 (all m, together 12 H).

HPLC: 28. 03 min (A1).

29. 92min (B1). MS: 652.4 [M + 1] + [0166] To carry out biological tests, the title compound was transferred in its diacetate salt, by

lyophilization of 0.5 M acetic acid (40 ml).  Example 4

N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N- methylcarbamoyl} 2- (2-naphthyl) ethyl) -N-methylamide (2E) -5-amino-5-methylhex-2-enoic acid [0167]

image 1

1-tert-butyl ester 2-ethyl ester of (2S) -pyrrolidine-1,2-dicarboxylic acid [0168]

image 1

[0169] N-tert-Butoxycarbonylproline (24.38 g, 113 mmol) was dissolved in dichloromethane (60 ml). Ethanol (7.9 ml, 135 mmol) and 4-dimethylaminopyridine (1.52 g, 12.5 mmol) were added. The solution was cooled to 0 ° C. N- (3dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (23.88 g, 125 mmol) was added. The reaction mixture was stirred for 16 h, 15 while heating to room temperature. Ethyl acetate (400 ml) was added. It was washed with a 10% aqueous solution of sodium hydrogen sulfate (300 ml). The aqueous phase was extracted with ethyl acetate (3 x 200 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 column chromatography on silica (400 g), using ethyl acetate (1: 4) as eluent, to give 17.11 g of

1-tert-Butyl ester 2-ethyl ester of (2S) -pyrrolidin-1,2-dicarboxylic acid.

1 H-NMR (CDCl3): 1.28 (m, 3 H); 1.43 and 1.46 (both s, together 9 H); 2.95 (m, 3 H); 2.22 (m, 1 H); 3.50 (m, 2 H); 4.18 and

4.30 (m and dd, together 3 H).

N-t-butyloxycarbonyl- (S) -prolinal

[0170] [0171] At -78 ° C, a 1.2 M solution of diisobutylaluminum hydride (31.7 ml, 38 mmol) in toluene was added dropwise to a solution of 1-tert-butyl ester 2- (2S) -pyrrolidine-1,2-dicarboxylic acid ethyl ester (4.02 g, 16.5 mmol) in diethyl ether (15 ml). The reaction mixture was stirred for 3 h at -78 ° C. Water (9.9 ml) was added dropwise. The

image 1

5 reaction mixture was heated to room temperature. The mixture was filtered through a plug of celite. The celite was washed with tert-butyl methyl ether (3 x 100 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo to give 2.34 g of crude N-t-butyloxycarbonyl- (S) -prolinal, which was used for the next step without further purification.

1 H-NMR (CDCl3): 1.42 and 1.47 (both s, together 9 H); 1.70-2.20 (m, 4 H); 3.20-4.30 (m, 3 H); 9.45 and 9.55 (both s, 10 together 1 H).

(25) -2 - ((dimethylamino) methyl) pyrrolidine-1-carboxylic acid tert-butyl ester

[0172]

image 1

[0173] N-t-butyloxycarbonyl- (S) -prolinal crude (2.34 g, 11.7 mmol) was dissolved in dichloromethane (90 ml). A solution 5.6

15 M dimethylamine in ethanol (4.19 ml, 23.5 mmol) was added. 0.4 nm molecular sieves (10.0g) were added. Sodium triacetoxyborohydride 7.47 g, 35.2 mmol) and glacial acetic acid (1.34 ml, 23.5 mmol) were added successively. The reaction mixture was stirred for 3 days. It was filtered through a plug of celite. The celite was washed with methanol (150 ml). A 1 N aqueous solution of sodium hydroxide (150 ml) and tert-butyl methyl ether (150 ml) was added. The phases were separated. The aqueous phase was extracted with tert-butyl methyl ether (3 x 100 ml). The

20 combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 1.29 g of acid tert-butyl ester (2S) -2 - ((dimethylamino) methyl) pyrrolidine-1 carboxylic acid.

one

H-NMR (CDCl3): 1.48 (s, 9 H); 1.90 (m, 4 H); 2.15 and 2.23 (AB, 2 H); 2.26 (s, 6 H); 3.31 (br, 2 H); 3.85 (br, 1 H).

N-Dimethyl-N - (((2S) -pyrrolidin-2-yl) methyl) amine

[0174]

image 1

[0175] A 2.7 M solution of hydrogen chloride in ethyl acetate (75 ml, 202 mmol) was given to a solution of tert-butyl ester of (2S) -2 - ((dimethylamino) methyl) pyrrolidine-1- acid carboxylic (1.29 g, 5.65 mmol) in ethyl acetate (30 30 ml). The reaction mixture was stirred for 30 min at room temperature. The solvent was removed in vacuo to give

1.36 g of the crude N-dimethyl-N - (((2S) -pyrrolidin-2-yl) methyl) amine dihydrochloride salt, which was used for the next step without further purification.

H-NMR (CDCl3): 1.90 (m, 2 H); 2.17 (m, 1 H); 2.40 (m, 1 H); 2.90 (m, 2 H); 3.14 (s, 6 H); 3.55 (m, 2 H); 4.35 (m, 1 H). tert-butyl ester of N - [(1R) -1-benzyl-2 - ((28) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-carbamic [0176]

image 1

[0177] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (1.30 g, 6.76 mmol) was added to a solution of (2R) -2- (N- (tert-butoxycarbonyl) acid ) -N-Methylamino) -3-phenylpropionic (1.89 g, 6.76 mmol) and 1-hydroxy-7azabenzotriazole 0.92 g, 6.76 mmol) in dichloromethane (10 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of the crude hydrochloride salt of N-Dimethyl-N - (((2S) -pyrrolidin-2-yl) methyl) amine (1.36 g, 6.76 mmol) in dichloromethane (10 ml) and N, N-dimethylformamide ( 10 ml) and ethyldiisopropylamine (5.75 ml, 33. 8 mmol) was added

10 successively. The reaction mixture was stirred for 16 h, while heating to room temperature. It was diluted with ethyl acetate (100 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (100 ml). The aqueous phase was extracted with ethyl acetate (3 x 80 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to

15 give 2.26 g of N - [(1 R) -1-benzyl-2 - ((2 S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -Ncarbamic acid tert-butyl ester .

1 H-NMR (CDCl3, selected values): � 1.20, 1.33, and 1.37 (all s, together 9 H); 2.22 and 2.28 (both s, together 6 H);

2.82 and 2.84 (both s, together 3 H); 4.25 (m, 1 H); 4.80, 5.11, and 5.30 (dd, t, and m, together 1 H); 7.10 - 7.30 (m, 5 H).

C22H35N3O3 [389.5]

calc. C67.83 H9.06 N10.79

Found C67.39 H9.13 N10.73

(2R) -1 - ((2S) -2 - ((Dimethylamino) methyl) pyrrolidin-1-yl) -2-methylamino-3-phenylpropan-1-one

20 [0178]

image 1

[0179] At 0 ° C, trifluoroacetic acid (8 ml) was added to a solution of tert-butyl ester of N - [(1R) -1-benzyl-2 - ((2S) 2 - ((dimethylamino) methyl ) pyrrolidin-1-yl) -2-oxoethyl] -N-methylcarbamic (2.26 g, 5.80 mmol) in dichloromethane (8 ml). The reaction mixture was stirred for 20 min at 0 ° C. The solvent was removed in vacuo. Dichloromethane (70 ml) was added, and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 1.24 g of (2R) -1 - ((2S ) -2 - ((dimethyl-amino) methyl) pyrrolidin-1-yl) -2-methylamino-3-phenylpropane-1-one.

1 H-NMR (CDCl3, selected values): � 2.33 (s, 3 H); 2.43 (s, 6 H); 3.25 (m, 3 H); 4.17 (m, 1 H); 7.25 (m, 5 H).

acid tert-butyl ester N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((25) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2 -oxoethyl] -N30 methylcarbamoyl} -2- (2- naphthyl) ethyl) -N-methylcarbamic

[0180]

image 1

[0181] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (530 mg, 2.76 mmol) was added to a solution of (2R) -2- (N- (tert-butoxycarbonyl) acid -N-Methylamino) -3- (2-naphthyl) -propionic (911 mg, 2.76 mmol) and 1 hydroxy-7azabenzotriazole (376 mg, 2.76 mmol) in dichloromethane (5 ml). The reaction mixture was stirred for 20 min at 0 ° C. 5 A solution of (2R) -1 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-methylamino-3-phenylpropan-1-one (800 mg, 2.76 mmol) in dichloromethane (5 ml) and N, N-dimethylformamide (5 ml) and ethyldiisopropylamine (0.71 ml, 4.15 mmol) was added successively. The reaction mixture was stirred for 3 days, while this was heating to room temperature. It was diluted with ethyl acetate (100 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (100 ml). The aqueous solution was extracted with ethyl acetate (3 x 70 ml). The combined organic layers

10 were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (90 g), using dichloromethane / methanol / 25% aqueous ammonia (200: 10: 1) as eluent, to give 1.37 g of tert-butyl ester of N- ( (1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} - 2 - (2-Naphthyl) ethyl) -N-methylcarbamic.

1 H-NMR (CDCl3, selected values): � 0.64 (m, 1 H); 1.10, 1.29, 1.36, and 1.47 (all s, together 9 H); 4.99, 5.09, 5.45, 15 and 5.53 (t, t, m, and t, together 2 H); 7.10 - 7.90 (m, 12 H).

(2R) -N - [(1R) -1-Benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino ) -3- (2-

naphthyl) propionamide

[0182]

image 1

[0183] At 0 ° C, trifluoroacetic acid (10 ml) was added to a solution of tert-butyl ester of N - ((1R) -1- {N - [(1R) -1-benzyl-2 - (( 2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} 2- (2-naphthyl) ethyl) -N-methylcarbamic (1.37 g,

2.28 mmol) in dichloromethane (10 ml). The reaction mixture was stirred for 75 min at 0 ° C. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (70 ml) and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (90 g), using

25 dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 692 mg of (2R) -N - [(1R) -1-benzyl-2 ((2S) -2 - ((dimethylamino ) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methyl-2- (methylamino) -3- (2-naphthyl) propionamide.

1 H-NMR (CDCl3, selected values): � 1.85 and 2.01 (both s, together 3 H); 2.20 and 2.31 (both s, together 6 H); 3.65 and

3.80 (both t, 1 H); 4.04 and 4.45 (both m, together 1 H); 5.60 and 5.91 (t and dd, together 1 H); 7.10 - 7.90 (m, 12 H).

acid tert-butyl ester {(3E) -4- [N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethyl) amino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N30 methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] -1,1-dimethylbut-3-enyl} carbamic

[0184]

image 1

5

10

fifteen

twenty

25

30

[0185] At 0 ° C, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (132 mg, 0.69 mmol) was added to a solution of (2E) -5- (tert-butoxycarbonylamino) -5- methylhex-2-enoic (168 mg, 0.69 mmol) and 1-hydroxy-7-azabenzotriazole (94 mg, 0.69 mmol) in dichloromethane (5 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of (2R) -N [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methyl-2- (methyl-amino) -3- (2-naphthyl) propionamide (345 mg, 0.69 mmol) in dichloromethane (10 ml) and N, N-dimethylformamide (5 ml) and ethyldiisopropylamine were added successively. The reaction mixture was stirred for 16 h, while heating to room temperature. It was diluted with ethyl acetate (70 ml) and washed with a saturated aqueous solution of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 491 mg of acid tert-butyl ester {( 3E) -4- [N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2oxoethyl ] -N- methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] -1,1-dimethylbut-3-enyl} carbamic.

one

H-NMR (CDCl3, selected values): 1.30 and 1.32 (both s, together 6 H); 1.45 (s, 9 H); 1.60 (m, 2 H); 4.00 (m, 1 H);

4.48 (m, 1 H); 5.48 (dd, 1 H); 5.92 (dd, 1 H); 6.11 and 6.20 (both d, together 1 H); 6.82 and 6.92 (both m, together 1 H); 7.10

7.90 (m, 12 H).

[0186] At 0 ° C, trifluoroacetic acid (7 ml) was added to a solution of acid tert-butyl ester {(3E) -4- [N - ((1R) -1- {N [(1R) - 1-Benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] 1,1-dimethylbut-3-enyl} carbamic (491 mg, 0.68 mmol) in dichloromethane (7 ml). The reaction mixture was stirred for 60 min at 0 ° C. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (100 ml) and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography on silica (40 g), using dichloromethane / methanol / 25% ammonia (100: 10: 1) as eluent, to give 285 mg of the title compound.

1 H-NMR (CDCl3, selected values): � 0.55 (m, 1 H); 1.11, 1.12, and 1.17 (all s, together 6 H); 2.25 (s, 6 H); 2.45 (s, 3 H); 2.85 (s, 3 H); 4.02 (m, 1 H); 5.48 (dd, 1 H); 5.80 and 5.93 (m, and dd, together 1 H); 6.10 and 6.18 (both d, together 1 H); 6.87 and 7.00 (both m, together 1 H); 7.10 - 7.90 (m, 12 H).

HPLC 27. 97 min (A1).

27. 80 min (B1).

MS: 626.4 [M + 1] +

[0187] To carry out biological tests, the title compound was transferred into its diacetate salt by lyophilization of 0.5 acetic acid (40 ml).

Example 5

N - ((1R) -1- {N - [(1R) -1-Benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N- methylcarbamoyl} -2- (2-naphthyl) ethyl) -Nmethyl-3 - ((methylamino) methyl) benzamide

[0188] N- {3- [N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl acid tert-butyl ester ) pyrrolidin-1-yl) -2-oxoethyl] -Nmethylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] benzyl} -N-methylcarbamic

image 1

[0189]

image10

[0190] At 0 ° C, N - (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (132 mg, 0.69 mmol) was added to a solution of 3- (N- (tert-butoxycarbonyl) -N-methylamino acid ) benzoic acid (183 mg, 0.69 mmol) and 1-hydroxy-7- azabenzotriazole (94 mg, 0.69 mmol) in dichloromethane (5 ml). The reaction mixture was stirred for 20 min at 0 ° C. A solution of (2R) -N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methyl-2 - (methylamino) -3- (2-naphthyl)

10 propionamide (345 mg, 0.69 mmol) in dichloromethane (10 ml) and N, N-dimethylformamide (5 ml) and ethyldiisopropylamine (0.118 ml) was added successively. The reaction mixture was stirred for 16 h, while heating to room temperature. It was diluted with ethyl acetate and washed with a saturated aqueous solution of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted with ethyl acetate (3 x 50 ml). The combined organic layers were dried over magnesium sulfate. The solvent was removed in vacuo. The crude product was purified by chromatography on

15 quick column on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 524 mg of tert-butyl ester of N- {3- [N - (( 1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2oxoethyl] -N-methylcarbamoyl} -2- ( 2-naphthyl) ethyl) -N-methylcarbamoyl] benzyl} -N-methylcarbamic.

1 H-NMR (CDCl3, selected values): � 0.72 (m, 1 H); 1.45 (br, 9 H); 3.18 (br, 6 H); 4.05 (m, 1 H); 4.32 and 4.40 (both br, together 2 H); 5.60 (dd, 1 H); 5.95 (m, 1 H); 6.80 - 6.90 (m, 16 H).

[0191] At 0 ° C, trifluoroacetic acid (7 ml) was added to a solution of tert-butyl ester of N- {3- [N - ((1R) -1- {N - [(1R) 1 -benzyl-2 - ((25) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylcarbamoyl] benzyl } -N-methylcarbamic (523 mg, 0.70 mmol) in dichloromethane (7 ml). The reaction mixture was stirred for 25 min at 0 ° C. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (90 ml), and the solvent was removed in vacuo. This procedure was repeated twice. The crude product was purified by flash column chromatography.

25 on silica (40 g), using dichloromethane / methanol / 25% aqueous ammonia (100: 10: 1) as eluent, to give 436 mg of the title compound.

1 H-NMR (CDCl3, selected values): � 0.87 (m, 1 H); 1.22 (m, 1 H); 1.45 (m, 1 H); 1.67 (m, 1); 4.09 (m, 1 H); 5.53 and

5.90 (dd and m, together 2 H); 6.80 - 7.90 (m, 16 H).

HPLC 28. 43 min (A1).

30. 63 min (B1).

MS: 648.4 [M + 1] +

[0192] To carry out biological tests, the title compound was transferred into its diacetate salt by lyophilization of 0.5 M acetic acid (40 ml).

Example 6

N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4- (dimethyl-amino) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2E) -5-Amino-5-methylhex-2- enoic acid (2-naphthyl) ethyl) -N-methylamide

[0193]

image10

[0194] The title compound was prepared as in Example 1 using 4- (dimethylamino) piperidine hydrochloric salt, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino) -3-phenylpropionic acid, (2R) -2- (N- (tert-butoxycarbonyl) -Nmethylamino-3- (2-naphthyl)) propionic acid, and (2E) -5- (butoxycarbonylamino) -5-methylhex-2-enoic acid.

1 H-NMR: (CDCl3; selected values) � 1.40 (s, 6 H); 2.00 (s, 6 H); 4.42-4.85 (2 H); 5.45-5.90 (m, 2 H); 6.28 (dd, 1 10 H); 6.85 (m, 1 H); 7.10 - 7.85 (m, 12 H) MS (ES): m / Z 626.2 (M + H) +

Example 7 N-methyl-N - [(1R) -1- (N-methyl-N - {(1R) -1- [N-methyl-N- (1-methylpiperidin-4-yl) carbamoyl] -2- phenylethyl} carbamoyl) -2- (2-naphthyl) ethyl] (2E) -5-amino-5-methylhex-2-enoic acid

15 [0195]

image 1

[0196] The title compound was prepared as in Example 1 using 1-methyl-4- (methylamino) piperidine acid, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino) -3- phenylpropionic, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino-3- (2naphthyl)) propionic acid, and (2E) -5- (butoxycarbonylamino) -5-methylhex-2- enic acid.

20 1 H-NMR (CDCl3; selected values)? 5.50 - 6.08 (m, 2 H); 6.20-6.70 (m, 2 H); 7.10 - 7.85 (m, 12 H) Example 8 3-Aminomethyl-N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-methylpiperazin-1-yl) -2- oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N

methylbenzamide [0197]

image 1

[0198] The title compound was prepared as in Example 1 using N-methylpiperazine, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino) -3-phenylpropionic acid, (2R) -2- ( N- (tert-butoxycarbonyl) -N-methylamino-3- (2-naphthyl)) propionic acid, and 3- ((tert-butoxycarbonylamino) methyl) benzoic acid.

5 1 H-NMR (CDCl3; selected values) � 3.30 (m, 1 H); 3.50 (dd, 1 H); 3.75 (m, 1 H); 3.95 (s, 2 H); 5.78 (t, 1 H); 3.88 (m, 1 H); 7.00 - 7.80 (16 H).

HPLC: 24. 55 min (A1).

26. 52 min (B1). MS (ES): m / Z = 606.4 [M + H] +. Example 9 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2- acid naphthyl) ethyl-N-methylamide

10 (2E) -5-amino-5-methylhex-2- oenic [0199]

image 1

[0200] The title compound was prepared as in Example 1 using N-methylpiperazine, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino) -3-phenylpropionic acid, (2R) -2- ( N- (tert-butoxycarbonyl) -N-methylamino-3- (2-naphthyl)) propionic acid, and (2E) -5- (butoxycarbonylamino) -5-methylhex-2-oenic acid.

one

H-NMR (CDCl3; selected values) � 1.24 (s, 6 H); 1.65 (s, 3 H); 2.35 (s, 3 H); 2.80 (s, 3 H); 5.68 (dd, 1 H); 5.78 (dd, 1 H); 6.18 (dd, 1 H); 6.95 (m, 1 H); 7.15 - 7.80 (m, 12 H).

HPLC: 25. 03 min (A1).

27. 50 min (B1). MS (ES): m / Z = 598.4 [M + H] +. Example 10

20 N-methyl-N - ((1R) -1- {N-methyl-N - [(1R) -2-phenyl-1 - ((2,2,6,6-tetramethylpiperidin-4-yl) carbamoyl) ethyl] carbamoyl} -2- (2naphthyl) ethyl) (2E) -5-Amino-5-methylhex-2-enoic acid [0201]

image 1

[0202] The title compound was prepared as in Example 1 using 4-amino-2,2,6,6-tetramethyl-piperidine, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino acid ) -3-phenylpropionic, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino-3- (2naphthyl)) propionic acid, and (2E) -5- (butoxycarbonylamino) -5-methylhex- 2-mad.

5 1 H-NMR (CDCl3 selected values) � 1.25 (s, 6 H); 1.40 (two s, 6 H); 1.52 (two s, 6 H); 2.92 (s, 3 H); 3.02 (two s, 3 H); 5.10 (dd, 1 H); 5.50 (dd, 1 H); 6.15 (d, 1 H); 6.75 (m, 1 H); 7.00 - 8.00 (m, 12 H).

HPLC: 29. 27min (A1).

31. 67min (B1).

MS (ES): m / Z = 654.8 [M + H] +. Example 11

10 3-Aminomethyl-N-methyl-N - ((1R) 1- {N-methyl-N - [(1R) -2-phenyl-1 - ((2,2,6,6-tetramethylpiperidin-4-yl ) carbamoyl) ethyl] carbamoyl} -2 (2-naphthyl) ethyl) benzamide [0203]

image 1

[0204] The title compound was prepared as in Example 1 using 4-amino-2,2,6,6-tetramethyl-piperidine, 15 (2R) -2- (N- (tert-butoxycarbonyl) -N- methylamino) -3-phenylpropionic, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino-3- (2naphthyl)) propionic acid, and 3 - ((tert-butoxycarbonylamino) methyl) benzoic acid.

1 H-NMR (CDCl3; selected values) � 3.60 - 3.85 (m, 2 H); 3.90-4.30 (m, 1 H); 5.25-5.95 (m, 2 H); 6.70 - 7.90 (m, 16 H).

HPLC: 29. 27 min (A1).

31. 55 min (B1). MS (ES): m / Z = 662.4 [M + H] +. 20 Example 12

N-methyl-N - ((1R) -1- {N-methyl-N - [(1R) -2-phenyl-1 - ((2,2,6,6-tetramethylpiperidin-4-yl) carbamoyl) ethyl ] carbamoyl} -2- (2naphthyl) ethyl) (2E) -5-amino-3,5-dimethylhex-2-enoic acid

[0205]

image 1

[0206] The title compound was prepared as in Example 1 using 4-amino-2,2,6,6-tetramethyl-piperidine, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino acid ) -3-phenylpropionic acid (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino-3- (2

5

naphthyl)) propionic, and (2E) -5- (butoxycarbonylamino) -3,5-methylhex-2-enoic acid.

1 H-NMR (CDCl3; selected values) � 3.92 - 4.30 (m, 1 H); 5.05-5.88 (m, 3H); 7.00 - 7.80 (m, 12 H).

HPLC:

29. 80 min (A1).

32. 43 min (B1).

MS (ES): m / Z = 668.4 [M + H] +.

Example 13

N - ((1R) 1- {N - [(1R) 1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

from acid

10

(2E) -4- (1-aminocyclobutyl) but-2-enoic

[0207]

image 1

[0208] The title compound was prepared as in Example 1 using N-methylpiperazine, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino) -3-phenylpropionic acid, (2R) -2- ( N- (tert-butoxycarbonyl) -N-methylamino-3- (2-naphthyl)) propionic acid, and (2E) -4- (1- (butoxycarbonylamino) cyclobutyl) but-2-enoic acid.

1 H-NMR (CDCl3; selected peaks) � 1.62 (s, 3 H); 2.35 (s, 3 H); 2.80 (s, 3 H); 5.70 (dd, 1 H); 5.80 (dd, 1 H); 6.22 (d, 1 H); 6.98 (m, 1 H); 7.15 - 7.80 (m, 12 H).

HPLC: 25. 88 min (A1).

28. 65 min (B1). MS (ES): m / Z = 610.4 [M + H] +. Example 14

20 N - ((1R) 1- {N - [(1R) 1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl ) -N-Methylamide (2E) -5-amino-3,5-dimethylhex-2-enoic acid [0209]

image 1

[0210] The title compound was prepared as in Example 1 using N-methylpiperazine, (2R) -2- (N- (tert-butoxycarbonyl) -N-methylamino) -3-phenylpropionic acid, (2R) -2- ( N- (tert-butoxycarbonyl) -N-methylamino-3- (2-naphthyl)) 5 propionic acid, and (2E) -5- (butoxycarbonylamino) -3,5-methyl-hex-2-oenic acid.

1 H-NMR (CDCl3; selected values) � 1.18 (s, 6 H); 1.68 (s, 3 H); 1.95 (s, 3 H); 2.30 (s, 3 H); 2.85 (s, 3 H); 3.40 (dd, 1 H); 3.54-3.75 (m, 2 H); 5.68-5.85 (m, 3 H); 7.15 - 7.80 (m, 12 H).

HPLC: 25. 70 min (A1).

28. 27 min (B1). MS (ES): m / Z = 612.4 [M + H] +. Example 15

10 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4 -yl) ethyl) -N-Methylamide (2E) -4- (1-aminocyclobutyl) but-2-enoic acid [0211]

image 1

[0212] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) - (N-tert-15-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- (N-tert- Butoxycarbonyl-N-methylamino) -3- (biphenyl-4-yl) propionic acid and (2E) -4- (1- (tert-butoxycarbonylamino) cyclobutyl) but-2-enoic acid as raw materials. ESMS: 637. 4 (M + H) + HPLC: rt = 33.58 min. (A1) HPLC: rt = 34.95 min. (B1)

Example 16 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl -4-yl) ethyl) -N-Methylamide (2E) -5-amino-5-methylhex-2-enoic acid [0213]

image 1

[0214] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- (N-tert) -butyloxycarbonyl-N-methylamino) -3- (biphenyl-4-yl) 5 propionic acid and (2E) -5-tert-butoxycarbonylamino-5-methylhex-2-enoic acid as ESMS raw materials: 625.4 (M + H) + HPLC: rt = 32.65 min. (A1) HPLC: rt = 34.02 min. (B1) Example 17 10 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxy-piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (Biphenyl-4-yl) ethyl) -N-methylamide (2E) -5-amino-3,5-dimethylhex-2-enoic acid [0215]

image 1

[0216] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-15-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- (N- tert-butoxycarbonyl-N-methylamino) -3- (biphenyl-4-yl) propionic acid and (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid as raw materials. ESMS: 639.4 (M + H) + HPLC: rt = 33.29 min. (A1) HPLC: rt = 36.40 min. (B1)

Example 18 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2 -naphthyl) ethyl) -N-methylamide (2E) -5-amino-5-methylhex-2-enoic acid

[0217] [0218] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- ( N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -5-tert-butoxycarbonylamino-5-methyl-hex-2-enoic acid as raw materials.

image 1

5 ESMS: 599.4 (M + H) + HPLC: rt = 29.88 min. (A1) Example 19 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-Naphthyl) ethyl) -N-methylamide acid

(2E) -5-amino-3,5-dimethylhex-2-enoic 10 [0219]

image 1

[0220] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- (N-tert) -butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid as raw materials.

15 ESMS: 613.4 (M + H) + HPLC: rt = 30.58 min. (A1) Example 20 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxy-piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} - 2- (2-Naphthyl) ethyl) -N-methylamide acid

(2E) -4- (1-aminocyclobutyl) but-2-enoic 20 [0221]

image 1

[0222] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- (N-tert) acid -butyloxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -4- (1- (tert-butoxycarbonylamino) cyclobutyl) but-2-enoic acid.

5 ESMS: 611.4 (M + H) + HPLC: rt = 30.82 min. (A1) Example 21 N - ((1R) -1- {N - [(1R) -1- (4-fluorobenzyl) -2- (4-hydroxy-piperidin-1-yl) -2-oxoethyl] -N -methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

of (2E) -5-amino-5-methylhex-2-enoic acid [0223]

image 1

[0224] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (4-fluorophenyl) propionic acid and (2R) -2 - (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -5-tert-butoxycarbonylamino-5-methylhex-2-enoic acid as raw materials.

15 ESMS: 617.4 (M + H) + HPLC: rt = 30.27 min. (A1) HPLC: rt = 31.60 min. (B1) Example 22 N - ((1R) -1- {N - [(1R) -1- (4-fluorobenzyl) -2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl } -2- (2-Naphthyl) ethyl) -N-methylamide

20 (2E) -5-amino-3,5-dimethylhex-2-enoic acid [0225]

image 1

[0226] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (4-fluorophenyl) propionic acid and (2R) -2 - (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid as raw materials.

5 ESMS: 631.4 (M + H) + HPLC: rt = 30.98 min. (A1) HPLC: rt = 32.38 min. (B1) Example 23 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4- (dimethyl-amino) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-Naphthyl) ethyl) -N-methylamide

10 (2E) 4- (1-aminocyclobutyl) but-2-enoic acid [0227]

image 1

[0228] The title compound was prepared as in Example 1 using 4-N, N-dimethylpiperazine, (2R) -2- (N (tert-butoxycarbonyl) -N-methylamino) -3-phenylpropionic acid, (2R ) -2- (N- (tert-butoxycarbonyl) -N-methylamino-3- (2-naphthyl))

Propionic, and (2E) -4- (1- (butoxycarbonylamino) cyclobutyl) but-2-enoic acid. 1 H-NMR (CDCl3; selected maximum values) d 1.90 (s, 3 H); 2.38 (s, 3 H); 2.45 and 2.47 (two s, 3 H) 2.78 and 2.80 (two s, 3 H); 6.32 (dd, 1 H); 6.90 (m, 1 H); 7.15 - 7.84 (m, 12 H).

HPLC: 26.72 min (A1). MS (ES): m / z = 638.4 [M + H] +. 20 Example 24

N - ((1R) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} -2- (2-Naphthyl) ethyl) -N-Methylamide (2E) -5-amino-5-methylhex-2-enoic acid [0229]

image 1

[0230] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-thienyl) propionic acid and (2R) -2 - (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -5-tert-butoxycarbonylamino-5-methylhex-2-enoic acid as raw materials.

5 ESMS: 605.4 (M + H) + HPLC: rt = 29.07 min. (A1) Example 25 N - ((1R) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-methylamide

of (2E) -5-amino-3,5-dimethylhex-2-enoic acid [0231]

image 1

[0232] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-thienyl) propionic acid and (2R) -2 - (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid and (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic acid as raw materials.

15 ESMS: 619.4 (M + H) + HPLC: rt = 29.76 min. (A1) Example 26 N - ((1R) -2- (biphenyl-4-yl) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1- ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} ethyl) -N

(2E) -5-amino-5-methylhex-2-enoic acid methylamide 20 [0233]

image 1

[0234] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-thienyl) propionic acid and (2R) -2 - (N-tert-butoxycarbonyl-N-methylamino) -3- (biphenyl-4-yl) propionic acid and (2E) -5-tert-butoxycarbonylamino-5-methylhex-2-enoic as raw materials.

5 ESMS: 631.2 (M + H) + HPLC: rt = 32.20 min. (A1) Example 27 N - ((1R) -2- (biphenyl-4-yl) -1- {N - [(1R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl ) methyl) ethyl] -N-methylcarbamoyl} ethyl) -N

(2E) -5-amino-3,5-dimethylhex-2- oenic acid methylamide 10 [0235]

image 1

[0236] This compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-thienyl) propionic acid and (2R) -2 - (N-tert-butoxycarbonyl-N-methylamino) -3- (biphenyl-4-yl) propionic acid and (2E) -5-tert-butoxycarbonylamino-3,5-dimethylhex-2-enoic as raw materials.

15 ESMS: 465.4 (M + H) + HPLC: rt = 32.89 min. (A1) Example 28 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4-yl) ethyl) -N-methylamide

(2E) -5-methyl-5- (methylamino) hex-2-enoic acid [0237]

image 1

[0238] The title compound was prepared as in Example 1 but using 4-hydroxypiperidine, (2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3-phenylpropionic acid and (2R) -2- (N -terc-butoxycarbonyl-N-methylamino) -3- (biphenyl-4-yl) propionic acid and (2E) -5- (N- (tert-butoxycarbonyl) -N-methylamino) -5-methylhex-2- enic acid as materials cousins

5 MS: m / z: 639.4 (M + H) + HPLC: method A1: Rt = 32.94 min Example 29 ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin -1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4-yl) ethyl) acid amide (2E) -4

(1-aminocyclobutyl) but-2-enoic 10 [0239]

image 1

HPLC: Rt = 31.55 min. (A1) Rt = 33.11 min. (B1) LC-MS: 623.6 [M + 1] +

Claims (5)

1. Compound of the general formula I image 1  where R1 is hydrogen or C1-6 alkyl; R2 is C1-6 alkyl;  L is image 1 where q, s, t and u are independently of each other 0 or 1; 10 r is 0 or 1 the sum q + r + s + t + u is 2 or 3; R9, R10, R11 and R12 are independently from each other hydrogen or C1-6 alkyl; Q is> N-R13 or image 1 where or is 0 or 1; T is -N (R15) (R16) or hydroxyl;  R13,  R15 and R16 are independently from each other hydrogen or C1-6 alkyl; R14   it is hydrogen; G is image 1  where R17, R18, R20 and R21 are independently from each other hydrogen, where R19 is independently hydrogen or aryl; J is image2  where R22, R23 R25 and R26 independently of each other are hydrogen;  where R24 is hydrogen or halogen; a is 1; b is 1; 10 c is 0; d is 0 and is 0 or 1; fis1; R5  it is hydrogen or C1-6 alkyl optionally substituted with one or more hydroxyl, aryl or heteroaryl; R6 and R7 are independently from each other hydrogen or C1-6 alkyl; or R6 and R7 form - (CH2) i-U- (CH2) j-, 15 where i and j independently of each other are 1 or 2, and U is a Valencia bond; R8 is hydrogen or C1-6 alkyl; and M is arylene or -CR27 = CR28-; where R27 and R28 are independently from each other hydrogen or C1-6 alkyl; or a pharmaceutically acceptable derivative salt.

2. A pharmaceutically acceptable compound or salt thereof according to claim 1, wherein R1 is C1-6 alkyl.

3.

Pharmaceutically acceptable compound or salt thereof according to claim 1 or 2, wherein R6 and R7 independently of one another are hydrogen or C1-6 alkyl.

Four.

Compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R6 and R7 form - (CH2) i-U (CH2) j-, wherein i and j independently of one another are 1 or 2, and U is a chemical bond.

5. A compound or pharmaceutically acceptable salt thereof according to any of claims 1-4, selected from:

 N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4 - ((dimethylamino) methyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} 2- (2-Naphthyl) ethyl) -N-Methylamide (2E) -5-amino-5-methylhex-2-enoic acid N - ((1R) -1- {N - [(1R) -1-benzyl) -2- ((3S) -3- (dimethylaminomethyl) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2naphthyl) ethyl) -N-methylamide acid (2E) -5-amino-5- methylhex-2-oenic image 1 image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((3S) -3- (dimethylaminomethyl) piperidin-1-yl-2-oxoethyl] -N-methylcarbamoyl} -2 - (2naphthyl) ethyl) -N-methylamide (2E) -4- (1-aminocyclobutyl) but-2-enoic acid image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N- methylcarbamoyl} -2- (2naphthyl) ethyl) -N-methylamide (2E) -5-amino-5-methylhex-2-enoic acid image 1  N - ((1R) -1- {N - [(1R) -1-Benzyl-2 - ((2S) -2 - ((dimethylamino) methyl) pyrrolidin-1-yl) -2-oxoethyl] -N- methylcarbamoyl} -2- (2 naphthyl) ethyl) -N-methyl-3 - ((methylamino) methyl) benzamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4- (dimethylamino) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2 -naphthyl) (2E) -5-amino-5-methylhex-2-enoic acid ethyl-N-methylamide image 1  3-Aminomethyl-N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- ( 2 naphthyl) ethyl) -N-methylbenzamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-Methylamide (2E) -5-amino-5-methylhex-2-enoic acid image 1  N - ((1R) 1- {N - [(1R) 1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) (2E) -4- (1-Aminocyclobutyl) but-2-enoic acid-N-methylamide image 1  N - ((1R) 1- {N - [(1R) 1-benzyl-2- (4-methylpiperazin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-Methylamide (2E) -5-amino-3,5-dimethylhex-2-enoic acid image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4- il) ethyl) (2E) -4- (1-aminocyclobutyl) but-2-enoic acid methylamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4- il) (2E) -5-amino-5-methylhex-2-enoic acid ethyl-N-methylamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4- il) ethyl) (2E) -5-amino-3,5-dimethylhex-2-enoic acid (2E) methylamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-Methylamide (2E) -5-amino-5-methylhex-2-enoic acid image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) -N-Methylamide (2E) -5-amino-3,5-dimethylhex-2-enoic acid image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2-naphthyl) ethyl) (2E) -4- (1-aminocyclobutyl) but-2-enoic acid N-methylamide image 1  N - ((1R) -1- {N - [(1R) -1- (4-fluorobenzyl) 2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2 -naphthyl) ethyl) -N-methylamide (2E) -5-amino-5-methylhex-2-enoic acid image 1  N - ((1R) -1- {N - [(1R) -1- (4-fluorobenzyl) -2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- ( 2-Naphthyl) ethyl) (2E) -5-amino-3,5-dimethylhex-2-enoic acid nmethylamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4- (dimethylamino) piperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (2 -naphthyl) ethyl) -N-Methylamide (2E) 4- (1-aminocyclobutyl) but-2-enoic acid image 1  N - ((1R) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} -2- (2-Naphthyl) ethyl) -N-Methylamide (2E) -5-amino-5-methylhex-2-enoic acid image 1  N - ((1R) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-i1) -2-oxo-1 - ((2-thienyl) methyl) ethyl] -N-methylcarbamoyl} -2- (2-Naphthyl) ethyl) -N-Methylamide (2E) -5-amino-3,5-dimethylhex-2-enoic acid image 1  N - ((1R) -2- (biphenyl-4-yl) -1- {N - [(2R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl ) methyl) ethyl] -N-methylcarbamoyl} ethyl) -N-methylamide (2E) -5-amino-5-methylhex-2-enoic acid image 1  N - ((1R) -2- (biphenyl-4-yl) -1- {N - [(1R) -2- (4-hydroxypiperidin-1-yl) -2-oxo-1 - ((2-thienyl ) methyl) ethyl] -N-methylcarbamoyl} ethyl) -N (2E) -5-amino-3,5-dimethylhex-2-enoic acid methylamide image 1  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4- il) ethyl) (2E) -5-methyl-5- (methylamino) hex-2- oenic acid methylamide image 1 ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4-yl) ethyl) (2E) -4- (1-aminocyclobutyl) but-2-enoic acid amide image 1 and pharmaceutically acceptable salts thereof. A compound or pharmaceutically acceptable salt thereof according to any one of claims 1-5, which is  N - ((1R) -1- {N - [(1R) -1-benzyl-2- (4-hydroxypiperidin-1-yl) -2-oxoethyl] -N-methylcarbamoyl} -2- (biphenyl-4- il) ethyl) (2E) -4- (1-aminocyclobutyl) but-2-enoic acid methylamide image 1 or a pharmaceutically acceptable derivative salt.

7.

Pharmaceutical composition comprising, as active substance, a pharmaceutically acceptable compound or salt thereof according to any one of claims 1-6, together with a pharmaceutically acceptable carrier or diluent.

8.

Use of a pharmaceutically acceptable compound or salt thereof according to any of claims 1-6 for the preparation of a medicament for stimulating the release of growth hormone from the pituitary gland of a mammal.