SI8911201A - Amino acid derivatives - Google Patents

Abstract

Compounds of general formula .1 J, 'CH' b \ ch / BI, in which n represents the number 0 or 1; R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclyl-carbonyl, alkylsulfonyl, arylsulfonyl, monoaralkylcarbamoyl, cinamoyl or alpha-aralkoxycarbonyl-aminoalkanoyl and R2 represents hydrogen, or R1 and R2 together with the nitrogen atom at of which they are bound, are cyclically imidated a group of formula alkyl or alkoxycarbonylalkyl, or when n represents the number 0, R3 may also represent alkylthioalkyl or, where n denotes the number 1, R3 may also represent alkylsulfonylalkyl; R4 represents alkyl, cycloalkyl, cycloalkylalkyl, aryl or aralkyl; R5 represents hydrogen and R6 represents hydroxy, or R5 and R6 together represent oxo; R7 and R8 together represent the trimethylene group or tetramethylene, which may be substituted by hydroxy, alkoxycarbonylamino or acylamino or in one of the -CH2- groups replaced by -NH-, -N (alkoxycarbonyl) -, -N (acyl) - or -S-, or which one carries fused cycloalkane, aromatic or heteroaromatic ring; and R 9 represents alkoxycarbonyl, monoalkylcarbamoyl, monoarylcarbamoyl, monoarylcarbamoyl or a group of formula

Description

F.HOFFMANN-LA ROCHE AG

DERIVATIVE AMINO ACID

The present invention relates to amino acid derivatives. Amino acid derivatives, the preparation of which is possible with the present invention, are compounds of the general formula

4

R R (0) p

II t> ^R hzz

R ⁹ in which n represents a number 0 or 1; R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl, monoaralkylcarbamoyl, cinamoyl or α-aralkoxycarbonylaminoalkanoyl and R4 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, a cyclic imide group of formula

Oh

N— (a) in which P and Q together represent the aromatic system; R4 represents alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylsulfinylalkyl, carbamoylalkyl or alkoxycarbonylalkyl, or when n represents O, R1 may represent alkylthioalkyl, or when n represents 1, R4 may also represent alkylsulfonylalkyl; R4 represents alkyl, cycloalkyl, cycloalkylalkyl, aryl or aralkyl; R4 represents hydrogen and R4 represents hydroxy, or R4 and R4 together represent oxo;

R ⁷ and r ⁸ together represent a trimethylene or tetramethylene group which may be substituted by hydroxy, alkoxycarbonylamino or acylamino, or in which one of the groups -CH 2 'is replaced by -NH-, -N (alkoxycarbonyl) -, -N (acyl) - or -S-, or which carries a fused cycloalkane, aromatic or heteroaromatic ring; and R4 represents alkoxycarbonyl, monoalkylcarbamoyl, monoaralkylcarbamoyl, monoarylcarbamoyl or a group of the formula -CO '/ ^H \ with ^co \

CH

JO

NHJ 1 (b) wherein R 10 and R 4 each represent alkyl;

and their pharmaceutically acceptable acid addition salts.

The compounds of formula I and their pharmaceutically acceptable acid addition salts are novel and have valuable pharmacological properties. In particular, they inhibit proteases of viral origin, and may be used in prophylaxis or for the treatment of viral infections, in particular infections caused by HIV and other retroid viruses.

The subject of the present invention are the compounds of formula I and their abovementioned salts per se, as well as the use of the therapeutically active substances, the process for the manufacture of said compounds and salts, the intermediates used in said process, the medicinal preparations containing said compounds and salts, the use of said compounds and salts in the control or elimination of diseases, especially for the treatment or prophylaxis of viral infections, and the use of said compounds and salts for the preparation of medicaments for the treatment and prophylaxis of viral infections.

For the purposes of this specification, the term alkyl is used alone or in combination to denote a straight or branched alkyl group containing not more than 8, preferably not more than 4, carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl , sec-butyl, tert-butyl, pentyl, hexyl and the like. The term alkoxy, alone or in combination, represents a group of alkyl ether in which the term alkyl has a given meaning, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like . The term cycloalkylalkyl represents an alkyl group, as previously defined, which is substituted by a cycloalkyl group and contains 3-8, preferably 3-6 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. The term aryl, alone or in combination, represents a phenyl or naphthyl group which may optionally carry one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, hydroxy, amino and the like, such as phenyl, p-tolyl, 4-methoxyphenyl, 4-tert-butoxyphenyl, 4fluorophenyl, 4-chlorophenyl, 4-hydroxyphenyl, 1-naphthyl, 2-naphthyl, etc. The term aralkyl, alone or in combination, means an alkyl group as previously defined in which one hydrogen atom is replaced by an aryl group as previously defined, such as benzyl, 2-phenylethyl and the like. The term aralkoxycarbonyl, alone or in combination, means a group of formula -C (O) -O-aralkyl, wherein the term aralkyl has the aforementioned meaning, such as benzyloxycarbonyl, etc. The term alkanoyl, alone or in combination, means an ayl group which is derived from an alkanecarboxylic acid such as acetyl, propionyl, butyryl, valeryl, 4methylvaleryl, etc. The term cycloalkylcarbonyl means an acyl group derived from monocyclic or bridged cycloalkanecarboxylic acid, such as cyclopropanecarbonyl, cyclohexanecarbonyl, adamantanecarbonyl, etc., or from a benzcondensed monocyclic cycloalkanecarboxylic acid, which may be substituted with, for example, alkanoyl, substituted by, e.g. , 4-tetrahydro-2-naphthoyl, 2-acetamido-1,2,3,4tetrahydro-2-naphthoyl. The term aralkanoyl means an acyl group which is derived from an arylsubstituted alkanecarboxylic acid such as phenylacetyl, 3-phenylpropionyl (hydrocinamoyl), 4-phenylbutyryl, (2-naphthyl) acetyl, 4-chlorohydrocinamoyl, 4-aminohydrocinamoyl, 4-methoxyhydroxyhydrocyclo, etc. The term aroyl represents an acyl group derived from an aromatic carboxylic acid, e.g. of benzoic or naphthalenecarboxylic acid, which may be substituted, such as benzoyl, 4-chlorobenzoyl, 4-carboxybenzoyl, 4 (benzyloxycarbonyl) -benzoyl, 1-naphthoyl, 2-naphthoyl, 6-carboxy-2-naphthoyl, 6- (benzyloxycarbonyl) ) -2-naphthoyl, 3-benzyloxy-2-naphthoyl, 3-hydroxy-2-naphthoyl, 3- (benzyloxyformamido) -2-naphthoyl, etc. A heterocyclic moiety of a heterocyclylcarbonyl or heterocyclylalkyl group is a saturated, partially unsaturated or aromatic monocyclic, bicyclic or tricyclic heterocycle containing one or more hetero atoms selected from the group consisting of nitrogen, oxygen and sulfur, which may be substituted on one or more substituents carbon with halogen, alkyl, alkoxy, oxo etc. and / or on a secondary nitrogen atom (i.e. on -NH-) with alkyl, aralkoxycarbonyl, alkanoyl, phenyl or phenylalkyl, or on a tertiary nitrogen atom (i.e. on -N-) with an oxide and which is bonded via a carbon atom. Examples of such heterocyclic groups are pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiamorpholinyl, pyrrolyl, imidazolyl, (e.g. imidazol-4-yl, 1-benzyloxycarbonylimidazol-4-yl, etc.), pyrazolyl, pyridyl, pyrazinyl, pyrazinyl, pyrazinyl , triazolyl, oxazolyl, thiazolyl, indolyl (e.g. 2-indolyl, etc.), quinolyl (e.g. 2-quinolyl, 3-quinolyl, 1-oxido-2-quinolyl, etc.), isoquinolyl (e.g., 1-isoquinolyl, 3 -isoquinolyl, etc.), tetrahydroquinolyl (e.g., 1,2,3,4-tetrahydro-2-quinolyl, etc.) 1,2,3,4-tetrahydroisoquinolyl (e.g., 1,2,3,4-tetrahydro-1-oxo- isoquinolyl, etc.), quinoxalinyl, β-carbolinyl and the like.

The term halogen means fluorine, chlorine, bromine or iodine.

The cinnamyl group denoted by R1 may be unsubstituted or may bear on the phenyl ring one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, nitro and the like.

The aromatic system, denoted in the foregoing formula (a) together with P and Q, may be monocyclic (e.g. 1,2-phenylene or thienylene), or polycyclic (e.g. 1,2-naphthylene, 2,3 naphthylene, 1 , 8-naphthylene, 2,3-anthylene, etc.) and may be unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halogen and the like.

As previously mentioned, the group is trimethylene or tetramethylene, wherein R ^ and R ⁸ together may be substituted by a hydroxy group or an alkoxycarbonylamino group (eg. Tert-butoxycarbonylamino) or an acylamino group (eg alkanoylamino, cikloalkilkarbonilamino, aralkanoylamino or aroylamino). Alternatively, one of the -CH 2 - groups from the group is trimethylene or tetramethylene, denoted together with R 1 and R ⁸ , is replaced by -NH-, -N (alkoxycarbonyl) -, e.g. -N (tert-butoxycarbonyl) -, -N (acyl) - or -S-. Where the group is trimethylene or tetramethylene, wherein R ^ and R ⁸ together carry a fused cycloalkane ring, the latter may, for example. a fused cycloalkane ring containing 3-6 carbon atoms, such as e.g. a fused cyclopentane, cyclohexane or similar ring, and when the trimethylene or tetramethylene group bears a fused aromatic or heteroaromatic ring, it may be e.g. a fused benzene, indole or thiophene ring which may be substituted on one or more carbon atoms by halogen, alkyl, alkoxy, etc. Thus -N (R ^) - CH (R ⁸ ) (R ^) may represent e.g. one of the following groups:

where R ^ has the meaning described above, R ² represents hydrogen, hydroxy, alkoxycarbonylamino or acylamino, R 4 represents hydrogen, alkoxycarbonyl or acyl, m denotes i or 2 and p denotes 1 or 2.

The pharmaceutically acceptable acid addition salts of the compounds of formula I are salts formed with inorganic acids such as e.g. hydrohalic acids such as hydrochloric acid or hydrobromic acid, sulfuric acid, hydrochloric acid, phosphoric acid, etc., or with organic acids such as e.g. acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid, p-toluenesulfonic acid, etc.

The compounds of formula I contain at least three asymmetric carbon atoms and are therefore present in the form of optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates. This invention includes all of these forms.

A particular group of compounds of formula I comprises a compound, in which n denotes the number 0, R represents an alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylthioalkyl, carbamoylalkyl or alkoxycarbonylalkyl and R ^2, and R & together represent a group of trimethylene or tetramethylene, in which one group -CH2- may be replaced by -NH- or -S- or which may carry a fused cycloalkane, aromatic or heteroaromatic ring.

In the compounds of formula I previously shown, it is desirable that R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl or α-aralkoxycarbonylamino-alkanoyl, preferably benzyloxycarbonyl, 2-hydroxycarbonyl, 2-hydroxycarbonyl, 2-hydroxycarbonyl, 2-hydroxycarbonyl, naphthoyl, 3-hydroxy-2-naphthoyl, 3-benzyloxy-2-naphthoyl, 2quinolylcarbonyl or 3-quinolylcarbonyl and R ² represents hydrogen. Preferably, R ² represents alkyl, cyanoalkyl, alkylthioalkyl or carbamoylalkyl and preferably cyanomethyl, methylthiomethyl or carbamoylmethyl. It is desirable that R1 represents aralkyl, especially benzyl. It is desirable that R ^ represents hydrogen and that R ^d represents hydroxy. It is desirable that -N (R ² ) -CH (R 8) (R 9) represents one of the groups shown previously of formula (c) to (i), a special group of formula (c) in which R ² represents hydrogen and m denotes 2, or R ^ ² represents tert-butoxycarbonylamino and m denotes 1, a group of formula (d) in which R ^d2 represents tert-butoxycarbonyl, a group of formula (e), in which m denotes 1, group s formula (f), in which m and p both denote 1, or a group of formula (g), (i) or (j). R4 is preferably represented by alkoxycarbonyl, especially tert-butoxycarbonyl, monoalkylcarbamoyl, especially isobutylcarbamoyl, or

Ί tert-butylcarbamoyl or a group of formula (b), a special group in which β1θ represents sec-butyl and RH represents isobutyl.

From the foregoing, it is understood that compounds of formula I in which R1 represents benzyloxycarbonyl, 2-naphthoyl, 1-hydroxy-2-naphthoyl, 3hydroxy-2-naphthoyl, 3-benzyloxy-2-naphthoyl, 2-quinolylcarbonyl are particularly preferred or 3-quinolylcarbonyl and R 4 represents hydrogen, R 4 represents cyanomethyl, methylthiomethyl or carbamoylmethyl, R 4 represents benzyl, R 8 represents hydrogen and R 4 represents hydroxy, and

-N (r7) -CH (R8) (r9) represents a group of the aforementioned formula (c), wherein R12 is hydrogen and m is 2, or R12 is tert-butoxycarbonylamino and m is a group of formula (a) d) wherein R1 represents tert-butoxycarbonyl, a group of formula (e) above, in which m is 1, a group of formula (f), and m and p are 1, or a group of formulas ( g), (i) or (j), and R 4 represents tert-butoxycarbonyl, isobutylcarbamoyl, tert-butylcarbamoyl or a group of formula (b) in which R 4 represents sec-butyl and R 4 represents isobutyl.

The compounds of formula I having particular advantage are:

N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide,

N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] -Nl-isobutyl-L-prolinamide,

N- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -thiazolidine-carboxamide, N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] -2 (S) -piperidinecarboxamide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -Nert-butyl-octahydro- (3aS, 6aS) -cyclopenta [b] pyrrole-2 (S) -carboxamide,

1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -Nert-butyl-2 (S) -piperidine-carboxamide,

2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1,2,3,4-tetrahydropyrido ] 3,4-b] indole-1-carboxamide, N-tert-butyl-3- [2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino ] -4-phenylbutyl] 4 (R) -thiazolidinecarboxamide,

N1-tert-butyl-N2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] -L-prolinamide N-oxide, 1 - [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2- (R) -hydroxy-4-phenylbutylN-tert-butyl-2 (S) -piperidinecarboxamide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4 (tert-butoxycarbonyl) -N-tert-butyl- 2- (R or S) -piperazinecarboxamide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2- (R) -hydroxy-4-phenylbutyl] 4 - (tert-butoxycarbonyl) -N-tert-butyl-2 (R or S) -piperazinecarboxamide, N ^ - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] 4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide, 1- [3 (S) - [[N- (3-benzyloxy-2 -naphthoyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] N- (tert-butyl-2 (S) -piperidine-carboxamide,

N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] -2-piperidinecarboxamide 1-oxide,

N-tert-butyl-1- [3 (S) - [[N- (3-hydroxy-2-naphthoyl) -L-asparaginylamino] -2 (R) -hydroxy-4-phenylbutyl] -2 (S) -piperidinecarboxamide , trans-2- [3 (S) - [[N-benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro- (4aR, 8aS) -isoquinoline-3 (S) -carboxamide,

4- (tert-butoxycarbonyl) -N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2quinolylcarbonyl) -L-asparaginyl] amino] butyl ] -2 (R or Siperiperazinecarboxamide,

N-tert-butyl-1- [2 (R) -hydroxy-3 (S) - [[N- (1-hydroxy-2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] -2 (S) -piperidinecarboxamide, trans-N-tert-butyl-decahydro-2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] - (4aR, 8aS) -isoquinoline-3 (S) -carboxamide and N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lcysteinyl] amino] butyl] -2 (S) -piperidinecarboxamide.

The most preferred compounds of formula I are:

N-tert-butyl-1- [2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] 2 (S) -piperidinecarboxamide,

N-tert-butyl-octahydro-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] - (3aS, 6aS) -cyclopenta [b] pyrrole-2 (S) -carboxamide and N-tert-butyl-1,2,3,4-tetrahydro-2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) L-asparaginyl] amino] butyl] pyrido [3,4-b] indole-1 (R or S) -carboxamide.

On the basis of the process of the invention, the compounds of formula I and their pharmaceutically acceptable acid addition salts are prepared such that (a) for the preparation of a compound of formula I in which n is 0, a compound of general formula

P.

CH / \ Z h ₂ nc

R ⁵ R ⁶

CH_-NR ²

CH 'b

in which r £ R ³ , r6 r7 r8 j _n r9 p _re j are given meaning, reacts with an acid of the general formula

R '

R i

-N

CH3

-COOH in which R 1, R ² and R ^{3 have} the meaning given above, or with its reactive derivative, or (b) for the preparation of a compound of formula I wherein n is 0, R ³ represents hydrogen and R 4 represents hydroxy, a compound of formula I wherein n is 0 and R ³ and R 6 together represent oxo, or (c) for the preparation of a compound of formula I in which n is 0 and R 1 represents alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl , arylsulfonyl, cinamoyl or Cr-aralkoxycarbonylaminoalkanoyl, and R ² represents hydrogen, or R ³ and R ² together with the nitrogen atom to which they are attached represent a cyclic imide group of the formula (a) shown above, a compound of the general formula / ^{co 1} . CH NH

R

I

-CH, CH_-N z

in which R ³ , R £ R ³ , r6, r7 _? r8 j _n r9 p _re jd _an jp _Omen ,

IV is reacted with an agent which gives the group alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl, cinamoyl or α-aralkoxycarbonylaminoalkanoyl, or with an agent forming the cyclic imide group of formula (a) or (previously shown) d) for the preparation of a compound of formula I in which n is 0 and R1 represents monoaralkylcarbamoyl and R1 represents hydrogen, the compound of formula IV shown above is reacted with a compound of general formula r1'_N = C = OV in which R1 represents aralkyl, or (e) to prepare a compound of formula I in which R1 represents allylsulfinylalkyl and n denotes 0, oxidize a compound of formula I in which R4 represents alkylthioalkyl and n denotes 0, or (f) to prepare a compound of formula I, v wherein n denotes 1, oxidize a compound of formula I in which n denotes 0, or (g) to prepare a compound of formula I in which n denotes 1 and R 1 represents an aromatic N-heterocyclylcarbonyl N-oxide group, and R 4 represents hydrogen , we oxidize a compound of formula I in which n represents 1 and R 1 represents an aromatic N-heterocyclylcarbonyl group and R 1 represents hydrogen, or (h) to prepare a compound of formula I in which n represents I and R 1 represents alkylsulfonylalkyl, oxidize a compound of formula I, in which n represents 1 and represents alkylsulfinylalkyl, or (i) for the preparation of a compound of formula I, in which R 1 represents carboxy-substituted aroyl, hydroxy-substituted aroyl or hydrocinamoyl, and R 1 represents hydrogen, catalytically hydrogenating a compound of formula I in which R represents benziloksikarbonilsubstituiran aroyl, benzyloxy-substituted aroyl or cinnamoyl and R ^ represents hydrogen, or (j) for the preparation of a compound of formula I in which R represents an imidazol-4-yl and / or R ⁴ represents a hydroxy-substituted aryl or hydroxy-substituted aralkyl and / or and R ⁸ together represent a trimethylene or tetramethylene group in which one group CH 2 - is replaced by -NH-, is treated with a compound of formula I in which R 4 represents oil 111 (benzyl-oxycarbonyl) -imidazol-4-yl and / or R4 represents tert-butoxy-substituted aryl or tert-butoxy-substituted aralkyl and / or R? and R ⁸ together represent a trimethylene or tetramethylene group in which one group -CH 2 - is replaced by -N (tert-butoxycarbonyl) - a strong acid group, or (k) for the preparation of a compound of formula I in which R? and R ⁸ together represent the group trimethylene or tetramethylene which is substituted with acylamine, or in which one group -CH 2 - is replaced by -N (acyl) -, acylates a compound of the general formula ^{r 1} - \ / ^co \ / ^ch \ / ^ch ^ · ^X CH NH £ <2'n R ⁷ '

-V.

, 8 '

VI in which n, R, R R3, R4, R5, R6, j _n R9 p _re j given meaning, and R and R ⁸ 'together represent a group of trimethylene or tetramethylene which is substituted by amino or in which one group of -CH ^ - replaced by -NH-, and / or (l) if desired, separate the mixture of diastereoisomeric racemates into diastereoisomeric racemates or optically pure diastereoisomers and / or (m) optionally separate the mixture of diastereoisomers into optically pure diastereoisomers and / or ( n) if desired, the resulting compound of Formula I is converted into a pharmaceutically acceptable acid addition salt.

The reaction of a compound of formula II with an acid of formula III according to embodiment (a) of the process of the present invention can be carried out by methods known in the chemistry of peptides. Thus, when using an acid of formula III, it is desirable to carry out this reaction in the presence of a condensing agent such as hydroxybenzotriazole and dicyclohexylcarbodiimide. This reaction can be carried out in an inert organic solvent, such as e.g. ether (e.g. diethyl ether, tetrahydrofuran, etc.) or dimethylformamide, at low temperature, preferably at about -10 ° C to +5 ° C and preferably about 0 ° C. Suitable reactive acid derivatives of formula III which can be used are e.g. suitable acid halides (eg acid chlorides), acid anhydrides, mixed anhydrides, activated esters, etc. When using a reactive derivative, it is advisable to carry out the reaction in an inert organic solvent such as a halogenated aliphatic hydrocarbon (eg dichloromethane, etc.) or ether (eg, diethyl ether, tetrahydrofuran, etc.) and, where appropriate, in the presence of an organic base ( e.g., N-ethylmorpholine, di-isopropylethylamine, etc.), at low temperature, preferably at about 10 ° C to + 5 ° C and preferably about 0 ° C.

The reduction of a compound of formula I in which R1 and R4 together represent oxo can be carried out in accordance with embodiment (b) of the present invention by methods known to reduce the carbonyl group to a hydroxy group. So e.g. the reduction can be carried out using a complex metal hydride such as an alkali metal borohydride, especially sodium borohydride, in a suitable organic solvent such as acanol (eg methanol, ethanol, propanol, isopropanol, etc.). It is appropriate to carry out this reduction at about room temperature.

According to embodiment (c) of this process, suitable contributing agents are alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl, cinamoyl or aaralkoxycarbonylaminoalkanoyl, suitable acids or their reactive derivatives such as the corresponding acid halides (eg acid acids, anhydrides, anhydrides) , mixed anhydrides, activated esters, etc., and suitable agents that form the cyclic imide group of Formula (a) previously presented are compounds of the formula HOOC-PQ-COOAralkyl, in which P and Q have the foregoing meaning. The reaction of a compound of formula IV with the above-mentioned agents is carried out in the same manner as previously described in connection with embodiment (a) of the process of the present invention. In the reaction of a compound of formula IV with a compound of formula HOOC-PQ-COOAralkyl, the initially formed reaction product spontaneously loses one molecule of aralkanol (HO-Aralkyl) to form a cyclic imide group.

The reaction of a compound of formula IV with a compound of formula V according to embodiment (d) of the process of the present invention can be carried out in a manner known in the art. This reaction can be carried out in an inert organic solvent, such as a halogenated aliphatic hydrocarbon (e.g. dichloromethane, etc.), at a temperature between about 0 ° C and room temperature, preferably at room temperature.

Oxidation according to embodiments (e), (f), (g) and (h) of this process can be carried out by known methods. The oxidation is preferably carried out using an organic peracid such as peracetic acid, perbenzoic acid, haloperbenzoic acid such as m-chloroperbenzoic acid, perphthalic acid or the like, although it can also be carried out using hydrogen peroxide. The oxidation can be carried out in the presence of an organic solvent which is inert under the given reaction conditions, such as e.g.

alkanols such as methanol, ethanol, etc., a halogenated hydrocarbon such as methylene chloride, etc., and the like. Oxidation can be carried out over a wide temperature range, e.g. in the range of about -70 ° C to about room temperature.

The catalytic hydrogenation according to embodiment (i) of the process of the present invention can be carried out in a known manner. It is recommended that the catalytic hydrogenation be carried out in the presence of a precious metal as a catalyst, preferably a palladium catalyst such as palladium on charcoal, and in an inert organic solvent (eg alkanol such as ethanol, isopropanol, etc.) at about room temperature below atmospheric pressure. When a compound of formula I in which R1 represents nitro-substituted cinamoyl and R1 represents hydrogen is hydrogenated catalytically according to this embodiment, a compound of formula I is obtained wherein R1 represents amino-substituted hydrocinamoyl and R4 represents hydrogen.

Embodiment (j) of the process of the present invention can be performed using a strong inorganic acid, e.g. hydrohalic acids such as hydrochloric acid or hydrobromic acid or strong organic acids, e.g. halogenated alkanecarboxylic acids such as trifluoroacetic acid and the like. This embodiment may be carried out in accordance with known procedures; e.g. in the presence or absence of an inert organic solvent (e.g. an alkanecarboxylic acid ester such as ethyl acetate, etc.) and at a temperature between about 0 ° C and about room temperature, preferably at room temperature.

The acylation of a compound of formula VI according to an embodiment of the process (k) of the present invention can be carried out according to known methods. It is appropriate to perform the acylation using an acyl halide such as acyl chloride or acyl bromide in the presence of an inert organic solvent such as dimethylformamide etc. and at a temperature between about 0 ° C and room temperature. Alternatively, a reactive acid derivative such as acid anhydride or the like may be used in place of the acid halide.

Optional separations according to embodiments (1) and (m) of the process of the present invention can be achieved by conventional methods; e.g. by column chromatography, thin layer chromatography, high pressure liquid chromatography, etc.

Conversion of a compound of formula I into a pharmaceutically acceptable acid addition salt according to embodiment (n) of the process of the present invention can be accomplished by treating the compound in a conventional manner with an inorganic acid such as e.g. hydrohalic acid such as e.g. hydrochloric acid or hydrobromic acid, sulfuric acid, hydrochloric acid, phosphoric acid, etc., or with an organic acid such as acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid, ptoluensulfonic acid, ptoluensulfonic acid, ptoluenic acid.

The compounds of formula II used as starting materials in carrying out (a) of the process are novel and also form the object of the present invention.

The compound of formula II can be produced e.g. so that the compound of the general formula

ch ₂ —X

II o

VII in which the meaning is given above, R1 represents an amino-protecting group (e.g. tert-butoxycarbonyl or benzyloxycarbonyl) and X represents a chlorine or bromine atom, reacted with a compound of the general formula /

I «

VIII in which R1, R1, and R4 have the meanings given above, and either cleave the group R14 from the compound obtained by the general formula

CH

Oh

IX in which R ^, R? r8 r9 j _n r14 the aforementioned meaning, to give a compound of formula II in which they together represent oxo, or reduce a compound of formula IX and cleave the group R14 from the compound obtained with the general formula / \ / ^CH 2 r ¹⁴ -h / \ /

H OH

wherein R ^, R7, R8, R9 and R L4 is previously given meaning, with a compound of formula II, wherein R $ is hydrogen and R ⁶ is hydroxy.

The reaction of a compound of formula VII, preferably a compound in which R14 is benzyloxycarbonyl, with a compound of formula VIII can be carried out in a known manner, e.g. in an inert organic solvent, such as a halogenated aliphatic hydrocarbon (e.g. dichloromethane, etc.), and in the presence of a base (e.g., trialkylamine such as triethylamine, etc.); carrying out the reaction at room temperature is appropriate.

The cleavage of the group R14 _{on a} d compound of formula IX can also be carried out in a known manner, e.g. using a strong inorganic acid such as hydrochloric acid or a strong organic acid (e.g. trifluoroacetic acid, etc.), suitable at about 0 ° C to about room temperature. Alternatively, if the amino-protecting group Rl4 is cleaved by hydrogenolysis, it can be cleaved using hydrogen in the presence of a noble metal catalyst (e.g., palladium catalyst such as palladium on carbon) in an organic solvent or solvent mixture that is inert under given reaction conditions (e.g., in alkanol such as ethanol, isopropanol, etc., an alkanecarboxylic acid ester such as ethyl acetate, etc.); appropriately, the reaction is carried out at room temperature.

The reduction of a compound of Formula IX to give a compound of formula X is carried out as described earlier in connection with the reduction of a compound of formula I in which n denotes 0 and R and R ⁸ together represent oxo in accordance with the type (b ) of the process of the present invention.

The cleavage of the R14 group from a compound of formula X can be performed in a manner analogous to that previously described with respect to the cleavage of the R14 group from a compound of formula IX.

Another method for the preparation of a compound of formula II in which R 5 represents hydrogen and R 6 represents hydroxy involves first reacting a compound of the general formula

in which R4 and R14 are given the above meaning, with the compound of formula VIII already shown, conveniently in an inert organic solvent, such as e.g. alkanol (e.g. methanol, etc.), dimethylformamide or the like, at elevated temperature, suitable at a temperature of from about 60 ° C to about 120 ° C, then cleave the group R1 from the product of this reaction (compound of Formula X shown above) described way.

The compounds of formula IV used as starting material for embodiments (c) and (d) of the process of the present invention are novel and constitute another object of the invention.

The compounds of formula IV can be obtained e.g. by cleaving the benzyloxycarbonyl or tert-butoxycarbonyl R1 group from a compound of formula I wherein n is 0, R1 represents benzyloxycarbonyl or tert-butoxycarbonyl and R ² represents hydrogen. This cleavage is carried out in a manner analogous to the method described above with respect to the cleavage of the group R14 from a compound of formula IX.

Compounds of formula VI in which R ² and R 1 together represent a trimethylene or tetramethylene group in which one group -CH 2 - is replaced by -NH- used as starting material in the embodiment (k) of the process of the present invention are subgroups of compounds of the Formula I. Compounds of formula VI in which R ² 'and R ² ' together represent a trimethylene or tetramethylene group which is substituted by an amino, also used as starting material for carrying out (k) of this process, are novel and they are also an object of the present invention. They can be obtained e.g. by separating the alkoxycarbonyl group from a compound of formula I in which R4 and R4 together represent a trimethylene or tetramethylene group which is substituted by alkoxycarbonylamine. This cleavage can be accomplished by conventional procedures; e.g. by treatment with an acid such as e.g. halogens (e.g. hydrochloride) in an inert organic solvent (e.g. alkanecarboxylic acid ester such as ethyl acetate, etc.).

The starting materials of Formula III and their reactive derivatives, starting materials of Formula V as well as the compounds of Formulas VII, VIII and XI previously shown, in the absence of known compounds or analogs of known compounds, can be produced in a similar manner to that produced known compounds, or as described in the Examples given below, or in an analogous manner. In addition, the agents required to carry out process (c) of the present invention are generally known compounds.

As mentioned above, the compounds of Formula I and their pharmaceutically acceptable acid addition salts inhibit proteases of viral origin and are useful for the treatment or prophylaxis of viral infections, especially infections caused by HIV and other retroid viruses.

In vitro inhibition of HIV protease with the compounds obtainable by the present invention can be demonstrated by the following assay:

The HIV protease is expressed in E. coli and is partially purified by soluble bacterial extracts by ammonium sulfate fractionation (0-30%). Protease activity is analyzed using a protected hexapeptide as a substrate, i.e. succinyl-Ser-Leu-Asn-Tyr-Pro-Ile isobutylamide. The cleavage of the substrate was quantified by measuring the H-Pro-Ile-isobutylamide product by spectrophotometric analysis of the N-terminal proline.

Dissolve 1.25 mM of substrate in 125 mM citrate buffer (pH 5.5) containing 0.125 mg / ml Tween 20. In 80 μΐ of the above buffered substrate, 10 / d of solutions of various concentrations of the test compound (dissolved in methanol or dimethyl sulfoxide and diluted with water, added) contains 0.1% Tween 20) and 10 μ \ proteases. Digestion was performed at 37 ° C for a fixed time and terminated by the addition of 1 ml of coloring reagent (30 mg / ml isatin and 1.5 mg / ml 2- (4-chlorobenzoyl) -benzoic acid in 10% acetone in ethanol (vol./vol .)). This solution was heated in a water bath and then the pigmented residue was redissolved in 1 ml of 1% pyrogallol in 33% water in acetone (w / v / v). The optical density of the solution was measured by spectrophotometry at 599 nm. The treatment of H-Pro-Ile isobutylamide in the presence of the test compound is compared with controls and the concentration of the compound required for 50% inhibition (bo) is determined by graphically displaying the result for the various concentrations of the test compound used.

The results obtained by the above test using representative compounds of Formula I as tested compounds are shown in the following table.

Table

Compound Ι ₅₀ (μΜ) A 0.87 B 0.15 C 0.3 D 0.13 E 0.75 F 0.75 Mr 0.08 H 0.01 I 0.085

Compound A: N- [N - [[N- (benzyloxycarbonyl) -L-asparaginyl] -L-phenylalanyl] methyl] -L-proline tert-butyl ester.

Compound B: N [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester.

Compound C: N- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] 2 (S) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester.

Compound D: N2- [N- [3 (S) - [[N- (benzoxycarbonyl) -L-asparaginyl] amino] -2 (R and S) hydroxy-4-phenylbutyl] -L-prolyl] -Nl-isobutyl -L-isoleucinamide (isomer 1; Example 13).

Compound E: N2- [N- [3 (S) - [[N [N- (benzoxycarbonyl) -L-leucyl] -L-asparagmyl] -amino] 2 (R and S) -hydroxy-4-phenylbutyl] - L-prolyl] -Nl-isobutyl-L-iso-leucinamide (isomer 1; Example 14).

Compound F: N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R and S) hydroxy-4-phenylbutyl] -Nl-isopentyl-L-prolinamide (isomer 2; Example 17).

Compound G: N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R or S) hydroxy-4-phenylbutyl] -Nl-isobutyl-L-prolinamide (isomer 2; Example 21).

Compound H: N- [2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] -L-proline tert-butyl ester.

Compound I: 2- [3 (S) - [[N- (Benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1,2,3,4 -tetrahydro-3 (R, S) -isoquinolinecarboxamide.

The compounds of formula I and their pharmaceutically acceptable acid addition salts can be used as medicaments (e.g. in the form of pharmaceutical preparations). These pharmaceutical preparations can be administered enterally, e.g. orally (as tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions or suspensions), nasally (eg in the form of nasal sprays), or rectally (eg in the form of suppositories). The administration can also be performed parenterally, e.g. intramuscularly or intravenously (eg in the form of injection solutions).

For the preparation of tablets, coated tablets, dragees, and hard gelatin capsules, the compounds of formula I and their pharmaceutically acceptable acid addition salts may be treated together with pharmaceutically inert organic or inorganic binders. As binders for tablets, dragees and hard gelatin capsules, e.g. lactose, maize starch or derivatives thereof, talc, stearic acid or its salts, etc.

Suitable binders for soft gelatin capsules are e.g. vegetable oils, waxes, greases, semi-solid and liquid polyols, etc.

Suitable binders for the preparation of solutions and syrups are e.g. water, polyols, sucrose, invert sugar, glucose, etc.

Suitable binders for injection solutions are e.g. water, alcohols, polyols, glycerol, vegetable oils, etc.

Suitable binders for suppositories are e.g. natural or hydrogenated oils, waxes, greases, semi-solid or liquid polyols, etc.

In addition, these pharmaceutical preparations may contain preservatives, solvents, viscosity enhancers, stabilizers, humectants, emulsifiers, sweeteners, colorants, flavoring agents, salts of osmotic pressure, buffers, coating agents or antioxidants. They may also contain other therapeutically relevant substances.

According to the present invention, the compounds of formula I and their pharmaceutically acceptable acid addition salts may be used for the treatment or prophylaxis of viral infections, in particular infections caused by retroviruses. The dosage amount may vary within wide limits and it is understandable that it must be tailored to the individual requirements of each case. In principle, an oral dosage amount of from about 3 mg to about 3 g, preferably from about 10 mg to about 1 g (e.g., about 300 mg per person), preferably divided into 1-3 dosage units, should be sufficient for oral administration. The above dosage limit may be exceeded when deemed appropriate.

The following Examples illustrate the present invention. The solvent systems listed in the Examples are as follows:

System A: 5% methanol in chloroform

System B: 10% methanol in chloroform

System C: chloroform: methanol: acetic acid: water (120: 15: 3: 2)

System D: chloroform: methanol: acetic acid: water (90: 15: 3: 2)

System E: chloroform: methanol: acetic acid: water (60: 18: 2: 3)

System F: chloroform: methanol: acetic acid: water (240: 24: 3: 2)

System G: dichloromethane: methanol: acetic acid: water (120: 15: 3: 2)

System H: diethyl ether: n-hexane: methanol (47.5: 47.5: 5)

Systems: dichloromethane: methanol: acetic acid: water (60: 18: 2: 3)

System J: dichloromethane: methanol: acetic acid: water (120: 15: 2: 3)

Example 1

0.5 g (1.07 mmol) of N - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-propyl tert-butyl ester was dissolved in 50 ml of isopropanol and 5 ml of ethyl acetate and hydrogenated for 5 hours in the presence of 0.406 g (2.14 mmol) ) toluene-4-sulfonic acid and 50 mg 5% palladium on charcoal. The catalyst was removed by filtration and the filtrate was evaporated. The residue was collected with 5 ml of dichloromethane and 5 ml of dimethylformamide and cooled in an ice / salt bath. 0.285 g (1.07 mmol) of N- (benzyloxycarbonyl) -L-asparagine in 25 ml of dimethylformamide were added, followed by 0.145 g (1.07 mmol) of hydroxybenzotriazole, 0.221 g (1.07 mmol) of dicyclohexylcarbodiimide and 0.246 g (2.14 mmol) of N-ethylamine. The mixture was stirred overnight, the separated dicyclohexylurea was removed by filtration and the filtrate was evaporated to dryness in vacuo. The resulting dark brown gum was stratified between ethyl acetate and water. The organic phase was washed successively with 5% hydrogen carbonate solution and saturated sodium chloride solution and dried over sodium sulfate. The solvent was removed by evaporation and the solid was chromatographed on silica gel using 5% isopropanol in ethyl acetate to elute. 0.125 g of N- [N - [[N- (benzyloxycarbonyl) -Lasparaginyl] -L-phenylalanyl] methyl] -L-proline tert-butyl ester was obtained as a pale yellow solid.

Analysis for € 3ΐΗ4θΝ4θγ (580.69).

Calculated: C, 64.12; H, 6.94; N, 9.65%

Found: C, 63.18; H, 6.79; N, 9.8%: ash 1.2%

Ash free: C, 63.91; H, 6.87; N, 9.92%.

N - [[N- (Benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-proline tert-butyl ester, which was used as starting material, was obtained by stirring 0.77 g of [N (benzyloxycarbonyl) -L -phenylalanil] methyl bromide with 0.35 g of t-butyl ester of L-proline and 0.203 g of triethylamine in 15 ml of dichloromethane at room temperature overnight. The solvent was removed by evaporation, the crude product was chromatographed on silica gel, eluting with chloroform to give 0.65 g of N - [[N (benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-proline tert-butyl ester in solid. form, from the ground. 98-99 ° C.

Example 2

1.5 g (3'22 mmol) of N - [[N- (benzyloxycarbonyl) -L-phenyl-alanyl] methyl] L-proline tert-butyl ester was hydrogenated in 10 ml of ethyl acetate and 5 ml of isopropanol in the presence of 0.15 g of 5% palladium on charcoal and 1.223 g (6.44 mmol) of toluene-4-sulfonic acid. The product was combined with 1.24 g (3.27 mmol) of N- [N- (benzyloxycarbonyl) -L-leucyl] -Lasparagine in the presence of 0.442 g (3.27 mmol) of hydroxybenzotriazole, 0.675 g (3.27 mmol) of dicyclohexylcarbodiimide and 0.753 g (6.55 mmol) of 6.55 mm (6.55 mmol). -ethylmorpholine in an analogous manner to that described in Example 1. Further treatment gave 0.34 g of a solid material which was chromatographed on silica gel using 5% isopropanol in dichloromethane for elution. We obtained 310 mg of N - [[N- [N- [N- (benzyloxycarbonyl) -L-leucyl] -L-asparaginyl] -L-phenylalanyl] methyl-proline tert-butyl ester; MS: m / e 694 [M + H] &lt; + &gt;.

The N- [N- (benzyloxycarbonyl) -L-leucyl] -L-asparagine used as starting material was obtained as follows:

4.5 g (12.4 mmol) of succinimide ester of N- (benzyloxycarbonyl) -L-leucine were dissolved in 40 ml of dimethylformamide and the solution was then cooled in an ice / salt bath. A solution of 1.64 g (12.4 mmol) of L-asparagine in 3.1 ml (12.4 mmol) of 4M sodium hydroxide solution was added, then 2 g (24.8 mmol) of sodium hydrogen carbonate were added. The mixture was stirred for 18 hours at room temperature. The solvent was removed by evaporation and the residue was collected with 100 ml of water. The pH was adjusted to 9.5 using 2M sodium hydroxide solution and then the mixture was extracted twice with 25 ml of diethyl ether each. This aqueous solution was adjusted to pH 2.5 using 3M hydrochloric acid. The solid crystalline product which precipitated from the solution was separated by filtration, washed with diethyl ether and dried. 3.5 g of N- [N- (benzyloxycarbonyl) -L-leucyl] -L-asparagine were obtained.

Analysis for (379.42).

Calculated: C, 56.98; H, 6.64; N, 11.07%

Found: C, 56.76; H, 6.62; N, 11.05%.

Example 3

0.71 g (1.52 mmol) of tert-butyl ester of N - [[Nbenzyloxycarbonyl) -L-phenyl-alanyl] methylL-proline was hydrogenated in the presence of 0.58 g (3.05 mmol) of toluene-4-sulfonic acid over 5% palladium on charcoal. The product was combined with 0.35 g (1.52 mmol) of N- (4methylvaleryl) -L-asparagine in an analogous manner as described in Example 1. The crude product was purified by chromatography on silica gel using isopropanol in ethyl acetate to elute (gradient 5% - 8%). 0.23 g of N - [[- N- [N- (4-methylvaleryl) -L-asparaginyl] -L-phenylalanyl] -methyl] -L-proline tert-butyl ester are obtained in solid form; MS: m / e 545 [M + H] &lt; + &gt;.

The N- (4-methylvaleryl) -L-asparagine used as starting material was obtained as follows:

5.35 g (25 mmol) of 4-methyl valeric acid succinimide ester was dissolved in 40 ml of dimethylformamide, a solution was added to a solution of 3.3 g (25 mmol) of L-asparagine in 6.25 ml (25 mmol) of 4M sodium hydroxide solution and 5 ml of dimethylformamide pre-cooled on ice. 5.5 g (65 mmol) of sodium hydrogen carbonate were added and the mixture was stirred for 18 hours at room temperature. The solvent was removed by evaporation and the residue was collected with 100 ml of water. The pH was adjusted to 3 with 4M hydrochloric acid. The solvent was then removed by evaporation and the residue was collected in methanol. The insoluble material was removed by filtration and the solvent was removed by evaporation. The crude product was chromatographed on silica gel using system D. Elution gave 900 mg of N- (4-methylvaleryl) -L-asparagine in solid form; Rf (system D): 0.25.

Example 4

In the analogous manner as described in Example 1, but using N- (tert-butoxycarbonyl) -L-asparagine instead of N- (benzyloxycarbonyl) -L-asparagine, N- [N - [[ N- (tert-butoxy) -L-asparaginyl] -L-phenyl-alanyl] -methyl] -L-proline in solid form; MS: m / e 546 [M] &lt; ^{+ &gt};.

Example 5

1.3 g (2.79 mmol) of N - [[N- (benzyloxycarbonyl) -L-phenyl-alanyl] methyl] L-proline tert-butyl ester was hydrogenated in a mixture of 10 ml of ethyl acetate and 10 ml of isopropyl alcohol in the presence of 0.39 g 5 of palladium-on-carbon and 1.06 g (5.58 mmol) of toluene-4-sulfonic acid and the resulting product was combined with 0.62 g (2.78 mmol) of N- (benzyloxycarbonyl) -alanine in the presence of 0.572 g (2.78 mmol) of dicyclohexylcarbodiimide, 0.375 g (2.78 mmol) ) hydroxybenzotriazole and 0.64 g (5.57 mmol) of N-ethylmorpholine in the analogous manner as described in Example 1. After further treatment and chromatography on silica gel using 3% isopropanol in dichloromethane for elution, 0.55 g of tert-butyl ester was obtained N- [N - [[N- (benzyloxycarbonyl) -L-alanyl] -L-phenylalanyl] methyl] -L-proline in solid form; MS: m / e 538 [M + H] &lt; ⁺ &gt;.

Example 6

In the analogous manner as described in Example 1, tert-butyl was obtained from the tert-butyl ester of N - [[N (benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-proline and N- (benzyloxycarbonyl) -Lglutamine. N- [N - [[N- (benzyloxycarbonyl) -L-glutaminyl] -L-phenylalanyl] methyl] -L-proline ester; MS: m / e 595 [M + H] &lt; + &gt;.

Example 7

0.45 g (0.77 mmol) of N- [N - [[N- (benzyloxycarbonyl) -L-asparaginyl] L-phenylalanyl] -methyl] -L-proline tert-butyl ester obtained as described in Example 1, was dissolved in 10 ml of isopropanol, stirring this solution for 45 minutes at room temperature in the presence of 0.075 g (1.98 mmol) of sodium borohydride. The solvent was removed by evaporation and the residue was collected with 50 ml of ethyl acetate and washed first with water and then with saturated sodium chloride solution. After drying over sodium sulfate and evaporation, 0.43 g of a solid material was chromatographed on silica gel using 2% methanol in dichloromethane for elution. 80 mg of N- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] 2 (S) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester was obtained in solid form; Rf (system Β): 0.38;

MS: m / e 583 [M + H] &lt; + &gt;.

Further elution of the column with 5% methanol in dichloromethane gave 70 mg of N- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] 2- (R) hydroxy-4- tert-butyl ester. Phenylbutyl] -L-proline in solid form: Rf (system Β): 0.23;

MS: m / e 583 [M + H] +

Example 8

0.3 g (0.45 mmol) of N - [[N- [N- [N- (benzyloxycarbonyl) -L-leucyl]] - Lasparaginyl] -L-phenylalanyl] -methyl] -L-proline tert-butyl ester obtained as was described in Example 2, in 10 ml of isopropanol, was treated at room temperature with 40 mg (1.06 mmol) of sodium borohydride. After 1 hour, the mixture was treated as described in Example 7. 0.32 g of a solid material were obtained which was chromatographed on silica gel using 3% methanol in dichloromethane for elution. 90 mg of N- [3 (S) - [[N- [N- (benzyloxycarbonyl) -L-leucyl] -L-asparaginyl] -amino] -2 (R or S) hydroxy-4 mg were obtained -phenylbutyl] -L-proline (isomer 1) in solid form; Rf (system A): 0.26;

MS: m / e 696 [M + H] &lt; + &gt;.

Further elution of the column with 5% methanol in dichloromethane gave 70 mg of N- [3 (S) - [[N- [N- (benzyloxycarbonyl) -L-leucyl] -L-asparaginyl] amino] - tert-butyl ester - 2 (R or S) -hydroxy-4-phenylbutyl] -L-proline (isomer 2) in solid form;

Rf (system A): 0.19; MS: m / e 696 [M + H] &lt; + &gt;.

Example 9

0.18 g (0.33 mmol) of N - [[N- [N- (4-methylvaleryl) -L-asparaginyl] -L-phenylalanyl] -methyl] -L-proline tert-butyl ester obtained as described in Example 3 was obtained. was reduced with 30 mg (0.80 mmol) of sodium borohydride in 10 ml of isopropanol as described in Example 7. The two isomers of tert-butyl ester 2 (R or S) -hydroxy-3 (S) [[N- (4- methyl valeryl) -L-asparaginyl] amino] -4-phenylbutyl] -L-proline was separated by chromatography on silica gel using methanol in chloroform (gradient 5% to 8%) for elution. 35 mg of isomer 1 are obtained in solid form; Rf (system Β): 0.2; MS: m / e 547 [M + H] +, and 27 mg of isomer 2 in solid form; Rf (system Β): 0.15; MS: m / e 547 [M + H] + 25

Example 10

0.6 g (1.1 mmol) of N- [N - [[N- (tert-butoxycarbonyl) L-asparaginyl] -L-phenylalanyl] -methyl] -L-proline tert-butyl ester obtained as described in Example 4 was obtained. was reduced with 0.1 g (2.65 mmol) of sodium borohydride in 10 ml of isopropanol as described in Example 7. The two isomers obtained were separated by chromatography on silica gel using an elution system. 115 mg of N- [3 (S) - [[N (tert-butoxycarbonyl) -L-asparaginyl] amino] -2 (R or S) -hydroxy-4-phenyl-butyl isomer 1 mg isomer is obtained. ] -Lprolin in solid form; Rf (system D): 0.23; MS: m / e 549 [M + H] + 'and 100 mg of isomer 2 in solid form; Rf (system D): 0.15; MS: m / e 549 [M + H] &lt; + &gt;.

Example 11

0.45 g (0.84 mmol) of N - [[N- [N- (benzyloxycarbonyl) -L-alanyl] -L-phenylalanyl] -methyl] -L-proline tert-butyl ester obtained as described in Example 5 was reduced to 15 ml of isopropanol in the presence of 80 mg of sodium borohydride as described in Example 7. After treatment, the two isomers obtained were separated by chromatography on silica gel, using elution system A. 70 mg of N [3 (S) - [[N- (benzioxycarbonyl) -L-alanyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester (isomer 1) was obtained in the form of rubber; Rf (system A): 0.38; MS: m / e 540 [M + H] ⁺ and 50 mg of isomer 2 in gum form; Rf (system A): 0.21; MS: m / e 540 [M + H] &lt; ⁺ &gt;.

Example 12

0.55 g (0.93 mmol) of N - [[N- [N- (benzyloxycarbonyl) -L-glutaminyl] -L-phenylalanyl] methyl] -L-proline tert-butyl ester obtained as described in Example 6 was reduced to 15 ml of isopropanol in the presence of 90 mg of sodium borohydride as described in Example 7. After treatment, the product was chromatographed on silica gel using 2% methanol in chloroform for elution. 60 mg of N [3 (S) - [[N- (benzyloxycarbonyl) -L-glutaminyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] L-proline isomer 1 mg isomer ( isomer 1) in solid form; Rf (system Β): 0.37; MS: m / e 597 [M + H] + and 65 mg of isomer 2 in solid form; Rf (system Β): 0.25; MS: m / e 597 [M + H] &lt; ⁺ &gt;.

Example 13 g (5.2 mmol) of N ² - [N- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] L-prolyl] -Nl-isobutyl-L-isoleucinamide was given in 25 ml of ethyl acetate and 10 ml of isopropanol and hydrogenated over 0.3 g of 5% palladium on charcoal for 5 hours in the presence of

1.97 g (10.36 mmol) of toluene-4-sulfonic acid. The catalyst was removed by filtration and the filtrate was evaporated. The resulting solid was dried over phosphorus pentoxide in high vacuum and combined with 1.48 g (5.57 mmol) of N-benzyloxycarbonyl) -L-asparagine in a manner analogous to that described in Example 1. 3.3 g of crude N ² - [ N- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R and S) -hydroxy-4-phenylbutyl] -L-prolyl] -Nl-isobutyl-L-isoleucinamide in solid form. These two isomers were separated by chromatography on silica gel, using system F for elution. 90 mg of isomer 1 were obtained as a solid material; Rf (system F): 0.23; MS: m / e 695 [M + H] + and 0.55 g of isomer 2 in solid form; Rf (system F): 0.11; MS: m / e 695 [M + H] &lt; ⁺ &gt;.

N ² - [N- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -L-prolyl] -Nisobutyl-L-isoleucinamide, which was used as starting material, was obtained as follows:

(i) A solution of 20.0 g of N- (benzyloxycarbonyl) -L-isoleucine and 9.6 ml of N-ethylmorpholine in 400 ml of tetrahydrofuran was cooled to -20 ° C and 9.8 ml of isobutylchloroformiate was added dropwise. This mixture was stirred for 5 minutes and 5.52 g of isobutylamine was added dropwise. The mixture was stirred for 20 minutes at -15 ° C and warmed to room temperature. 20 ml of water were added and the solution was evaporated to dryness. The residue was layered between 200 ml of water and 800 ml of ethyl acetate, the organic solution was washed with 200 ml of 5% citric acid solution, 200 ml of saturated sodium hydrogen carbonate solution and 200 ml of water, dried over sodium sulfate and evaporated to dryness. The residue was triturated with diethyl ether to give 18.2 g of N ² (benzyloxycarbonyl) -Nl-isobutyl-L-isoleucinamide, which was used in the next step without further purification.

(ii) A solution of 18.0 g of the above product in 200 ml of ethanol was hydrogenated over 1.0 g of 10% palladium on charcoal for 5 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 9.8 g of N, N-isobutyl-L-isoleucinamide as a colorless oil, which was used directly in the next step.

(iii) A solution of 17.3 g of succinimide ester of N- (benzyloxycarbonyl) -L-proline and 9.35 g of N1-isobutyl-L-isoleucinamide in 120 ml of tetrahydrofuran was stirred overnight at room temperature. The solvent was removed by evaporation and the residue was layered between 200 ml ethyl acetate and 250 ml 5% citric acid solution. The organic solvent was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was recrystallized from a mixture of ethyl acetate and n-hexane to give 13.6 g of N ² - [N27 (benzyloxycarbonyl) -L-prolyl] -Nl-isobutyl-L-isoleucinamide as a white solid from the ground. 87-88 ° C.

(iv) A solution of 13.6 g of the above product in 500 ml of ethanol was hydrogenated over 10% palladium on charcoal for 2 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 9.10 g of N, N -L-prolyl-NI-isobutyl-L-isoleucinamide as a colorless oil, which was used without further purification.

(v) A solution of 8.86 g of [N- (benzyloxycarbonyl) -L-phenylalanyl] -methyl bromide, 6.71 g of N-L-prolyl-N-isobutyl-L-isoleucinamide and 2.60 g of triethylamine in 400 ml of dichloromethane was stirred at room temperature overnight. The solvent was removed by evaporation and the crude product was chromatographed on silica gel using ethyl acetate for elution. The resulting product was triturated with a mixture of petroleum ether (boiling point 40-60 ° C) and ethyl acetate to give 8.70 g of N ^ - [N - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-prolyl] - Nl-isobutyl-L-isoleucinamide in the form of a dirty white solid material from the ground. 80-81 ° C.

(vi) A solution of 3.5 g of N2 - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-prolyl] -Nisobutyl-L-isoleucinamide in 200 ml of ethanol was treated with 1.0 g of sodium borohydride in the manner analogous to Example 7, yielding 7.1 g of N ^ [N- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -L-prolyl] -N ^4- isobutylL-isoleucinamide.

Analysis for: C33H48N4O5 x 0.5 H2O.

Calculated: C, 67.21; H, 8.37; N, 9.50%

Found: C, 67.29; H, 8.31; N, 9.47%.

Example 14

0.78 g (1.34 mmol) of N, N - [N- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -L-prolyl] -Nl-isobutyl-L-isoleucinamide was hydrogenated and the resulting product was combined with 0.53 g (1.39 mmol) of N- [N- (benzyloxycarbonyl) -L-leucyl] -L-asparagine in a manner analogous to that described in Example 1. 1.1 g of crude N2- [N- [3 ( S) - [[N [N- (benzyloxycarbonyl) -L-leucyl] -L-asparaginyl] amino] -2 (R and S) -hydroxy-4-phenylbutyl] -L-prolyl] -Nl-isobutyl-L- isoleucinamide in solid form. The two isomers were separated by chromatography on silica gel using system F for elution. 60 mg of isomer 1 are obtained in solid form; Rf (system C): 0.34; MS: m / e 808 [M] + and 50 mg of isomer 2 in solid form; Rf (system C): 0.24; MS: m / e 808 [M] &lt; ^{+ &gt};.

Example 15

0.275 g (0.473 mmol) of N- [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester was collected by 10 ml of isopropanol and hydrogenated for 18 hours over 50 mg of 5% palladium on charcoal in the presence of 0.18 g (0.94 mmol) of toluene-4-sulfonic acid. The catalyst was removed by filtration and the filtrate was evaporated. The resulting solid was dissolved in 10 ml of dichloromethane and cooled in an ice bath. 50 mg (0.49 mmol) of acetic anhydride were added followed by 0.10 g (0.99 mmol) of triethylamine and 0.2 ml of pyridine. The mixture was stirred for 1 hour at room temperature. The solvent was removed by evaporation and the residue was layered between ethyl acetate and water. The organic phase was washed with 5% sodium hydrogen carbonate solution and saturated sodium chloride solution and then dried over anhydrous sodium sulfate. The solvent was removed by evaporation to give 35 mg of N- [3 (S) - [(N-acetyl-L-asparaginyl) amino] -2 (R) hydroxy-4-phenylbutyl] -L tert-butyl ester. -proline; Rf (system E): 0.28; MS: m / e 491 [M + H] &lt; ⁺ &gt;.

Example 16 g (2.14 mmol) of N- [3 (S) - (benzyloxyformamido) -2- (R and S) hydroxy-4-phenylbutyl] -L-proline tert-butyl ester was collected with 25 ml of methanol and hydrogenated over 250 mg of 5% palladium on charcoal for 3 hours in the presence of 0.81 g (4.26 mmol) of toluene-4-sulfonic acid. The catalyst was removed by filtration and the filtrate was evaporated. The resulting solid was dried over phosphorus pentoxide under high vacuum and then dissolved in 5 ml of tetrahydrofuran.

0.566 g (2.134 mmol) of N- (benzyloxycarbonyl) -L-leucine were collected in 15 ml of tetrahydrofuran and cooled to -15 ° C. 0.245 g (2.134 mmol) of Nethylmorpholine and 0.291 g (2.134 mmol) of isobutyl chloroformate were added. After 5 minutes, a solution of tetrahydrofuran was prepared, prepared as described in the previous paragraph, and then 0.451 g (4.27 mmol) of N-ethylmorpholine was added. The mixture was stirred for 10 hours at room temperature, the solvent was removed by evaporation and the residue was layered between ethyl acetate and water. The ethyl acetate solution was washed with saturated sodium hydrogen carbonate solution and saturated sodium chloride solution and then dried over anhydrous sodium sulfate. The solvent was removed in vacuo to give 1.5 g of N- [3 (S) - [[N- (benzyloxycarbonyl) -leucyl] amino-2- (R and S) -hydroxy-4- crude tert-butyl ester. phenylbutyl] -L-proline in the form of an oil. The two isomers were separated by chromatography on silica gel using% n-hexane in ethyl acetate for elution. 180 mg of N- [3 (S) [[N- (benzyloxycarbonyl) -L-leucyl] amino] -2- (R or S) -hydroxy-4-phenylbutyl] -L- isomer 1 isomer is obtained. proline in the form of crystalline material; Rf (system A): 0.34; MS: m / e 582 [M + H] + and 70 mg of isomer 2 as crystalline material; Rf (system A): 0.28; MS: m / e 582 [M + H] &lt; + &gt;.

The N- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester used as starting material was obtained as follows:

2g (4.3 mmole) of N - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] -L-proline tert-butyl ester (obtained as described in Example 1) was dissolved in 25 ml of isopropanol, 25 ml of ethanol and 25 ml. of methanol, then this solution was stirred for 4 hours at room temperature in the presence of 0.4 g (10.7 mmol) of sodium borohydride. The solvent was removed by evaporation and the residue was layered between 50 ml of ethyl acetate and 25 ml of water. The organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed by evaporation to give 2 g of N- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester in the form of a transparent rubber; Rf (system C): 0.68 and 0.57.

Example 17

1.5 g (3.12 mmol) of N2- [3 (S) - (benzyloxyformamido) -2- (R and S) -hydroxy-4-phenylbutyl] -Nl-isopentyl-L-prolinamide were hydrogenated in 25 ml of methanol in the presence of 1.19 g (6.24 mmol) of toluene-4-sulfonic acid and 0.25 g of 5% palladium on charcoal. After 3 hours, the catalyst was removed by filtration and the filtrate was evaporated, yielding 2.1 g of solid material. This material was dried over phosphorus pentoxide under high vacuum and combined with 0.8 g (3.12 mmol) of n- (benzyloxycarbonyl) -Lasparagine in the analogous manner as described in Example 1. 1.45 g of N ^ [3 (S) - was obtained. [[N- (Benzyloxycarbonyl) -L-asparaginyl] amino-2- (R and S) -hydroxy-4-phenylbutyl] N1-isopentyl-L-prolinamide as a crude product chromatographed on silica gel using a system C for elution. 60 mg of isomer 1 are obtained in solid form; Rf (system E): 0.58; MS: 596 [M + H] ⁺ and 0.25 g of isomer 2 in solid form;

Rf (system E): 0.41; MS: m / e 596 [M + H] +.

The N2- [3 (S) - (benzyloxyformamamido) -2- (R and S) -hydroxy-4-phenylbutyl] -N 1-isopentyl-Lprolinamide used as starting material was obtained as follows:

(i) A solution of 15.0 g of succinimide ester of N (benzyloxycarbonyl) -L-proline and 4.15 g of isopentylamine in 100 ml of tetrahydrofuran was stirred at room temperature overnight. The solvent was removed by evaporation and the residue was collected with 250 ml of ethyl acetate. This solution was washed with 250 ml of 5% citric acid solution, twice with 250 ml of saturated sodium hydrogen carbonate solution and 150 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to dryness. The residue was recrystallized from a mixture of ethyl acetate and n-hexane to give 11.1 g of N2- (benzyloxycarbonyl) -Nl-isopentyl-L-prolinamide as a white solid with a melting point of 110-112 ° C.

(ii) A solution of 5.73 g of N ^ - (benzyloxycarbonyl) -Nl-isopentyl-L-prolinamide in 600 ml of ethanol was hydrogenated over 0.8 g of 10% palladium on carbon for 3.75 hours. The catalyst was removed by filtration and the filtrate was evaporated to give 3.4 g of N1-isopentyl-L-prolinamide in the form of an oil, which was used without further purification.

(iii) A solution of 3.4 g of N1-isopentyl-L-prolinamide, 6.76 g of [N- (benzyloxycarbonyl) -L-phenylalanyl] methyl bromide and 2.0 g of triethylamine in 360 ml of dichloromethane was stirred at room temperature overnight. The crude product obtained was isolated in an analogous manner as described in Example 1 and recrystallized from a mixture of ethyl acetate and nhexane to give 3.3 g of N2 - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] N1-isopentyl- L-prolinamide in the form of a white solid material from the ground. 82-84 ° C.

(iv) A solution of 0.96 g of N2 - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl-N1-isopentyl-L-prolinamide in 40 ml of ethanol was treated with 0.17 g of sodium borohydride in an analogous manner as described in Example 2. Obtained the crude product was recrystallized from a mixture of diethyl ether and n-hexane to give 0.65 g of N2- [3 (S) - (benzyloxyformamido) -2- (R and S) -hydroxy-4-phenylbutyl] -Nl-isopentyl-L -Prolinamide in the form of a white solid material from the ground. 73-82 ° C.

Example 18

0.58 g (1.24 mmol) of N2- [3 (S) - (benzyloxyformamido) -2- (R or S) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide was dissolved in 25 ml of methanol and hydrogenated for 5 hours at room temperature in the presence of 0.472 g (2.48 mmol) of toluene-4-sulfonic acid and 0.1 g of 5% palladium on charcoal. The catalyst was removed by filtration and the filtrate was evaporated. 0.82 g of a solid material were obtained which was dried over phosphorus pentoxide in high vacuum and combined with 0.322 g (1.21 mmol) of N- (benzyl 31 oxycarbonyl) -L-asparagine in dimethylformamide in the analogous manner as described in Example 1. After treatment in an analogous manner as described in Example 10, 0.6 g of crude product was obtained which was chromatographed on silica gel using an elution system C. We obtained 225 mg of N ² - [3 (S) - (benzyloxycarbonyl) -Lasparaginil] -amino] -2- (R or S) -hydroxy-4-phenylbutyl] -nl-tert-butyl-L-prolinamide in the solid form; MS: m / e 582 [M + H] &lt; ⁺ &gt;.

N ² - [3 (S) - (benzyloxyformamido) -2- (R or S) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-pyrrolinamide, which was used as starting material, was obtained as follows :

(i) 5.2 g (15 mmol) of succinimide ester of N- (benzyloxycarbonyl) -L-proline and 1.63 g (22.3 mmol) of tert-butyl amine were collected with 50 ml of dichloromethane and the mixture was stirred at -8 ° C for 1 hour and then 18 hours at room temperature. This solution was then washed with 5% citric acid solution, saturated sodium hydrogen carbonate solution and saturated sodium chloride solution and dried over anhydrous sodium sulfate. After evaporation, a solid material from ethyl acetate / n-hexane crystallized to give 3 g of N ² - (benzyloxycarbonyl) -Nl-tert-butyl-L-prolinamide;

MS: m / e 305 [M + H] &lt; + &gt;.

(ii) 2.5 g (8.22 mmol) of N ² - (benzyloxycarbonyl) -Nl-tert-butyl-L-prolinamide in 25 ml of methanol were hydrogenated for 5 hours over 0.5 g of 5% palladium on charcoal. The catalyst was removed by filtration and the solvent was removed by evaporation to give 1.4 g of oil which crystallized in a few hours while stored in the refrigerator. 0.35 g (2.05 mmol) of this solid was added to a solution of 0.77 g (2.05 mmol) of [N- (benzyloxycarbonyl) -L-phenylalanyl] methyl bromide in 15 ml of dichloromethane and then 0.207 g (2.05 mmol) of triethylamine were added. The mixture was stirred at room temperature for 2 hours. The solvent was removed by evaporation and the residue was collected with 25 ml of ethyl acetate. The resulting solid was filtered off and discarded. The mother liquor was washed with water, saturated sodium hydrogen carbonate solution and saturated sodium chloride solution and then dried over anhydrous sodium sulfate. Evaporation and crystallization from ether / n-hexane gave 0.65 g of N ² - [[N (benzyloxycarbonyl) -L-phenylalanyl] methyl] -Nl-tert-butyl-L-prolinamide in solid form; MS: m / e 466 [M + H] +.

(iii) 1.25 g (2.7 mmol) of N ² - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] -Nl-tert-butyl-L-prolinamide was collected with 25 ml of isopropanol and stirred for 4 hours at room temperature in of 0.255 g (6.7 mmol) of sodium borohydride. The solvent was removed by evaporation and the residue was layered between ethyl acetate and water. The organic layer was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. 1.2 g of N2- [3 (S) - (benzyloxyformamido) -2 (R and S) hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide were obtained, which was chromatographed on silica gel using the system F for elution. 15 mg of isomer 1 are obtained in the form of a rubber; Rf (system F): 0.38; MS: m / e 468 [M + H] ⁺ and 0.225 g of isomer 2 as rubber; Rf (system F); 0.27; MS: m / e 468 [M + H] &lt; + &gt;.

Example 19

A solution of 474 mg of N- [3 (S) - (benzyloxyformamido) -4-cyclohexyl-2 (R or S) -hydroxybutyl] -L-proline tert-butyl ester in 100 ml of ethanol was hydrogenated over 50 mg of 10% palladium catalyst on charcoal for 3 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 380 mg of oil. This oil was combined with 285 mg of N- (benzyloxycarbonyl) -L-asparagine in an analogous manner as described in Example 1. The crude product obtained was chromatographed on silica gel using 7% methanol in dichloromethane for elution to give 30 mg of N- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl-] amino-4-cyclohexyl-2 (R or S) hydroxybutyl] -L-proline tert-butyl ester in solid form, with m.p. 135-136 ° C (isomer 1).

Similarly, 474 mg of the second N- [3 (S) (benzyloxyformamido) -4-cyclohexyl-2 (R or S) -hydroxybutyl] -L-proline tert-butyl ester is hydrogenated and evaporated with 285 mg of N- ( benzyloxycarbonyl) -L-asparagine to give 90 mg of isomer 2; MS: m / e 589 [Μ + Η] ⁺ .

The N- [3 (S) - (benzyloxyformamido) -4-cyclohexyl-2 (R and S) -hydroxybutyl] -L-proline tert-butyl ester used as starting material was obtained as follows:

(i) A solution of 19.6 g of N- (benzyloxycarbonyl) -3-cyclohexyl-L-alanine in 60 ml of tetrahydrofuran was cooled to -10 ° C and 8.9 ml of N-ethylmorpholine was added. 11.6 ml of isobutyl chloroformate were added dropwise and stirred at -10 ° C for 15 minutes. 250 ml of diethyl ether were added and the mixture filtered. The filtrate was added to a cold solution of diazomethane in diethyl ether (obtained from 21.5 g of N-methyl-N-nitroso-4-toluenesulfonamide) and the mixture was stirred for 1.5 hours at room temperature. The resulting solution was then washed with water and 200 ml of saturated sodium hydrogen carbonate solution, dried over anhydrous sodium sulfate and evaporated, yielding 6.6 g of benzyl [233 cyclohexyl-1 (S) - (2-diazoacetyl) ethyl] carbamate in a form of yellow oil that was used without further purification.

(ii) A solution of 6.6 g of the above product in 60 ml of diethyl ether was stirred at room temperature and bubbled through the hydrogen chloride solution for 30 minutes throughout the stirring. The solvent was removed and the resulting oil was crystallized from a mixture of diethyl ether and n-hexane to give 5.8 g of [N- (benzyloxycarbonyl) -3-cyclohexyl-L-alanyl] methyl chloride, which was used without further purification.

(iii) A solution of 4.4 g of the above product in a mixture of 90 ml of tetrahydrofuran and 10 ml of water was cooled to 0 ° C and 0.75 g of sodium borohydride was added. This mixture was stirred for 1 hour at 0 ° C and evaporated to dryness. The residue was collected with 100 ml of dichloromethane and 100 ml of water, the aqueous layer was adjusted to pH 3 with concentrated hydrochloric acid and the organic layer was separated, dried over anhydrous sodium sulfate and evaporated. The residue was triturated with 200 ml of hot n-hexane and separated by filtration to give 2.65 g of 3 (S) - (benzyloxyformamido) -1chloro-4-cyclohexyl-2 (R and S) -butanol as a white solid;

MS: 339. 341 [M] &lt; + &gt;.

(iv) 14 ml of 0.71 M ethanolic potassium hydroxide solution was added to the solution

2.65 g of the above product in 60 ml of ethanol. This mixture was stirred for 1 hour at room temperature and then evaporated to dryness. The residue was layered between 100 ml of dichloromethane and 100 ml of water, the organic layer was dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on silica gel using 3% methanol in dichloromethane for elution. 1.84 g of 3 (S) - (benzyloxyformamido) -4-cyclohexyl-1,2 (R and S) -epoxybutane were obtained in the form of an oil;

MS: m / e 303 [M] &lt; + &gt;.

(v) A solution of 1.84 g of the above product and 1.21 g of t-butyl ester of L-proline in 65 ml of methanol was refluxed for 24 hours. The mixture was evaporated to dryness and the crude product was chromatographed on silica gel using% methanol in dichloromethane to elute. 0.41 g of N, N - [3 (S) (benzyloxyformamido) -4-cyclohexyl-2 (R or S) -hydroxybutyl] -L-proline (isomer 1) tert-butyl ester (colorless oil) was obtained; NMR (CDCl3, 250 MHz) δ 0.78-1.0 (2H, m) 1.1-1.4 (5H, m), 1.43 (9H, s) 1.55-1.7 (5H, m), 1.8-1.95 (4H, m), 2.03 -2.15 (1H, m), 2.3-2.4 (1H, m), 2.5 (1H, dd), 2.66 (1H, t), 3.16-3.26 (2H, m), 3.55-3.7 (2H, m), 4.08 (1H, broad, s), 5.02-5.15 (3H, m) 7.25-7.4 (5H, m).

Further elution with the same solvent gave 1.18 g of isomer 2 as a colorless oil; NMR (CDCl3, 300 MHz) δ 0.75-0.85 (1H, m), 0.9-1.0 (1H, m), 1.1-1.4 (5H, m), 1.43 (9H, s), 1.58-1.70 (5H, m) , 1.75-1.9 (5H, m) 2.05-2.18 (1H, m), 2.54-2.75 (3H, m), 3.05-3.15 (2H, m) 3.45-3.52 (1H, m), 3.7-3.8 (1H. m), 4.96-5.15 (3H, m), 7.3-7.38 (5H, m).

Example 20

0.663 g (1.42 mmol) of N - [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -Nl-isobutyl-L-prolinamide was hydrogenated in 30 ml of methanol in the presence of 0.539 g (2.84 mmol) of toluene-4-sulfonic acid and 0.1 g of 5% palladium on charcoal. This product was then combined with 0.378 g (1.42 mmol) of N- (benzyloxycarbonyl) -Lasparagine and further treated as described in Example 1. The crude product obtained was subjected to flash chromatography on silica gel using System F to give 80 mg of N2 - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] -Nl-isobutyl-L-prolinamide (isomer 1) in solid form; Rf (system E): 0.46; MS: m / e 582 [M + H] &lt; + &gt;.

The upper column was then eluted with System C to give 70 mg of isomer 2 in solid form; Rf (system E): 0.33; MS: m / e 582 [M + H] &lt; ⁺ &gt;.

The N2- [3 (S) - (benzyloxyformamido) -2 (R and S) -hydroxy-4-phenylbutyl] -Nl-isobutyl-Lprolinamide was used as starting material as follows:

(i) N1-isobutyl-L-prolinamide was obtained as described in Example 17 (i) using isobutylamine instead of isopentylamine.

(ii) A solution of 1.5 g (3.98 mmol) of [N- (benzyloxycarbonyl) -L-phenylalanyl] methyl bromide, 0.678 g (3.98 mmol) of N1-isobutyl-L-prolinamide and 0.402 g (3.98 mmol) of triethylamine in 30 ml of dichloromethane was obtained. stirred for 4 hours at room temperature. The solvent was removed by evaporation and 50 ml of ethyl acetate were added. The resulting solid material was removed by filtration and discarded. The filtrate was washed with water, sodium hydrogen carbonate solution and saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed by evaporation and the crude product was chromatographed on silica gel using chloroform for elution to give 1.6 g of N2 - [[N- (benzyloxycarbonyl) -L-phenylalanyl] methyl] -Nl-isobutyl-L-proline amide in solid. form; MS: m / e 466 [M + H] ⁺ · (iii) A solution of 0.7 g (1.5 mmol) of N2 - [[N- (benzyloxycarbonyl) -L-phenylalanyl] -methyl] N1-isobutyl-L-prolinamide in 20 ml. ethanol was treated with 0.143 g (3.75 mmol) of sodium borohydride at room temperature for 2 hours. The solvent was removed by evaporation and the residue was layered between ethyl acetate and water. The ethyl acetate phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated, yielding 0.67 g of N ^ - [3 (S) - (benzyloxyformamido) -2 (R and S) hydroxy-4-phenylbutyl] -Nl-isobutyl-L-prolinamide in solid form; Rf (system A) 0.39 and 0.33; MS: m / e 467 [M] + ·

Example 21

A solution of 249 mg of N- [3 (S) - [[L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester, 106 mg of 2-naphthoyl chloride and 72 mg of diisopropylethylamine in 10 ml of dry dichloromethane was stirred for 20 hours at 20 ° C. The gelatin reaction mixture was stratified between dichloromethane and water. The organic phase was evaporated and the resulting oil was crystallized from ethyl acetate / n-hexane to give 215 mg of N [2 (R) -hydroxy-3 (S) [[N- (2-naphthoyl) tert-butyl ester - L-asparaginylamino] -4-phenylbutyl] -L-proline in the form of a white solid; MS: m / e 603 [M + H] &lt; ⁺ &gt;. N- [3 (S) - [[Lasparaginyl-amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester, which was used as starting material, was obtained as follows:

A solution of 1.75 g of N- [3 (S) - [[(N- (benzyloxycarbonyl) -L-asparaginylamino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester) in 100 ml of ethanol was hydrogenated over 10% palladium on charcoal for 64 hours The catalyst was removed by filtration and the filtrate was evaporated The residue in the form of a rubber was stratified between ethyl acetate and 2M hydrochloric acid The aqueous phase was basified by dilute sodium hydroxide solution and extracted with Ethyl acetate The ethyl acetate extract was evaporated, yielding 1.14 g of N- [3 (S) - [[L-asparaginyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -L- tert-butyl ester. white solid foam proline;

MS: m / e 449 [M + H] +.

Example 22

In an analogous manner to that described in Example 21, N [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L- was extracted from 223 mg of tert-butyl ester. proline, 95 mg of toluene-4-sulfonyl chloride and 65 mg of di-isopropylethylamine gave 170 mg of N- [2 (R) hydroxy-4-phenyl-3 (S) - [[N- (p-toluenesulfonyl) -L] tert-butyl ester -asparaginyl] amino] butyl] -L-proline as a white solid (from ethyl acetate / n-hexane); MS: m / e 603 [M + H] &lt; ⁺ &gt;.

Example 23

In an analogous manner to that described in Example 21, N [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline was obtained from 223 mg of tert-butyl ester. , 77 mg of phenylacetyl chloride and 65 mg of diisopropylethylamine gave 190 mg of N- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (phenylacetyl) -L-asparaginyl] amino] butyl] tert-butyl ester - L-proline as a white solid (from ethyl acetate / n-hexane); MS: m / e 567 [Μ + Η] +.

Example 24

In an analogous manner to that described in Example 21, N [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L- was obtained from 249 mg of tert-butyl ester. proline, 94 mg hydrocinamoyl chloride and 72 mg diisopropylethylamine gave 214 mg of N-3 (S) [(N-hydrocinamoyl-L-asparaginyl) amino] -2 (R) -hydroxy-4-phenylbutyl] tert-butyl ester - L-proline as a white solid (from ethyl acetate / n-hexane); MS: m / e 581 [M + H] &lt; ⁺ &gt;.

Example 25

In an analogous manner to that described in Example 21, N [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L- was recovered from 270 mg of tert-butyl ester. proline, 114 mg of 1-naphthoyl chloride and 77 mg of diisopropylamine were obtained by chromatography on silica gel using 10% methanol in dichloromethane for elution, 130 mg of N- [2 (R) hydroxy-3 (S) [[N - (1-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] -L-proline as a white solid; MS: m / e 603 [M + H] &lt; ⁺ &gt;.

Example 26

In an analogous manner to that described in Example 21, N [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L- was recovered from 270 mg of tert-butyl ester. proline, 100 mg of cinamoyl chloride and 77 mg of di-isopropylethylamine were obtained after complete chromatography on silica gel using 10% methanol in dichloromethane for elution, 99 mg of N [3 (S) - [(N-cinamoyl-L- asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline in solid form; MS: m / e 579 [M + H] +

Example 27

A solution of 270 mg of N-3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester and 98 mg of 4-phenylbutyric acid in 10 ml of dry tetrahydrofuran cooled in ice / salt mixture. 81 mg of hydroxybenzotriazole, 69 mg of Nethylmorpholine and 136 mg of dicyclohexylcarbodiimide were added and the mixture was stirred for 64 hours. The mixture was then diluted with ethyl acetate and filtered. The filtrate was washed with aqueous sodium hydrogen carbonate solution and sodium chloride solution. The solvent was removed by evaporation and the residue was chromatographed on silica gel using system G for elution to give 79 mg of N- [2 (R) hydroxy-4-phenyl-3 (S) tert-butyl ester [[ N- (4-phenylbutyryl) -L-asparaginyl] amino] butyl] -L-proline as a white solid; MS: m / e 595 [M + H] &lt; ⁺ &gt;.

Example 28

In the analogous manner as described in Example 27, N- [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] - L-proline, 104 mg of 2-naphthylacetic acid, 75 mg of hydroxybenzotriazole, 64 mg of N-ethylmorpholine and 126 mg of dicyclohexylcarbodiimide were obtained after chromatography on silica gel using 10% methanol in dichloromethane, 62 mg of N- [2-tert-butyl ester] (R) -hydroxy-3 (S) - [[N (2-naphthyl) acetyl] -Lasparaginyl] amino] -4-phenylbutyl] -L-proline, as a white solid; MS: m / e 617 [M + H] ⁺ .

Example 29

In an analogous manner, as described in Example 27, N [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L- was recovered from 270 mg of tert-butyl ester. proline, 108 mg of 4-methoxyhydrocycic acid, 81 mg of hydroxybenzotriazole, 69 mg of N-ethylmorpholine and 136 mg of dicyclohexylcarbodiimide were obtained after chromatography on silica gel using an elution system G, 113 mg of N- [2 (R) tert-butyl ester -hydroxy-3 (S) - [[N- (4methoxyhydrocinamoyl) -L-asparaginyl] amino] -4-phenylbutyl] -L-proline, as a white solid; MS: m / e 611 [M + H] +.

Example 30

0.179 g of dicyclohexylcarbodiimide, 0.016 g of hydroxybenzotriazole and 0.09 g of Nethylmorpholine were added to a stirred solution of 0.238 g of N-3 (S) amino-2 (R or S) -hydroxy-5-methylhexyl] -L-proline tert-butyl ester and 0.210 g N- (benzyl-oxycarbonyl) 38

L-asparagine in 10 ml of tetrahydrofuran at 0 ° C. This mixture was stirred for 16 hours and then filtered. The filtrate was stripped between ethyl acetate and water, then the organic phase was washed with sodium hydrogen carbonate solution and sodium chloride solution. The organic phase was evaporated and the residue was chromatographed on silica gel using ethyl acetate / methanol (9: 1) as elution to give, after recrystallization from ethyl acetate / n-hexane, 0.13 g of N-3 tert-butyl ester. (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino-2 (R or S) -hydroxy-5-methylhexyl] -L-proline as a white solid; MS: m / e 549 [M + H] &lt; ⁺ &gt;.

The N- [3 (S) -amino-2 (R or S) -hydroxy-5-methylhexyl] -L-proline tert-butyl ester used as starting material was obtained as follows:

(i) A solution of 1.31 g of 3 (S) - (benzyloxyformamido) -1,2 (R or S) -epoxy-5-methyl-hexane and 0.855 g of proline tert-butyl ester in 10 ml of dimethylformamide was stirred for 16 hours at

100 ° C and then the solvent was removed by evaporation. The residue was layered between ethyl acetate and water. The organic phase was washed with sodium chloride solution and evaporated. The residue was chromatographed on silica gel using ethyl acetate / acetic acid / ethanol (4: 1: 1) for elution. The material having an Rf of about 0.24 was re-chromatographed on silica gel using H elution system to give 0.358 g of N- [3 (S) - (benzyloxyformamido) -2 (R or S) tert-butyl ester. hydroxy-5-methylhexyl] -L-proline as a colorless oil;

MS: m / e 435 [M + H] +.

(ii) A solution of 0.35 g of N- [3 (S) - (benzyloxyformamido) -2 (R or S) hydroxy-5-methylhexyl] -L-proline tert-butyl ester in 20 ml of ethanol was hydrogenated over 10% palladium on charcoal for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 0.238 g of N- [3 (S) -amino-2 (R or S) hydroxy-5-methylhexyl] -L-proline tert-butyl ester as a colorless gum. ; MS: m / e 301 [M + H] ⁺ .

Example 31

A solution of 0.28 g of N- [3 (S) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester and 0.083 g of benzyl isocyanate in 5 ml of dichloromethane were stirred for 2 hours at 25 ° C. The solvent was then removed by evaporation and the residue triturated with diethyl ether to give 0.174 g of N- [3 (S) - [[N (benzylcarbamoyl) -L-asparaginyl] amino] -2 (R) tert-butyl ester. hydroxy-4-phenylbutyl] -L-proline as a white solid; MS: m / e 582 [M + H] &lt; + &gt;.

Example 32

Solution of 0.28 g of N- [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline tert-butyl ester, 0.124 g of 1-adamantanecarbonyl chloride and 0.081 g of diisopropylethylamine in 5 ml of dichloromethane was stirred for 8 hours at 20 ° C. 0.124 g of 1adamantanecarbonyl chloride and 0.081 g of di-isopropylethylamine were added and stirred at 20 ° C for 16 hours. The resulting solution was diluted with 20 ml of dichloromethane and washed sequentially with water, sodium carbonate solution, potassium bisulphate solution and sodium chloride solution. The organic solution was then evaporated and the residue triturated with diethyl ether to give 0.2 g of N [3 (S) - [[N- (1-adamantylcarbonyl) -L-asparaginyl] amino] -2 (R) tert-butyl ester. ) -hydroxy-4-phenylbutyl] -Lprolin as a white solid; MS: m / e 611 [M + H] ⁺ .

Example 33

0.208 g of dicyclohexylcarbodiimide, 0.124 g of hydroxybenzotriazole and 0.106 g of Nethylmorpholine were added to a stirred solution of 0.315 g of N2-3 (S) -amino-2 (R) hydroxy-4-phenylbutyl] -Nl-phenyl-L-prolinamide and 0.244 g of N-prolineamide (benzyloxycarbonyl) -Lasparagine in 10 ml of tetrahydrofuran at 0 ° C. This mixture was stirred for 16 hours and then diluted with ethyl acetate and filtered. The filtrate was washed with sodium hydrogen carbonate solution and sodium chloride solution and the solvent was removed by evaporation. The residue was chromatographed on silica gel, eluting with dichloromethane / methanol (9: 1) to give 0.22 g of N ^ -3 (S) - [[N (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-phenyl-L-pyrrolinamide as a white solid; with a melting point of 169-171 ° C (from methanol).

The N2-3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-phenyl-L-prolinamide used as starting material was obtained as follows:

(i) 3.5 g of sodium borohydride was added to an ice and stirred solution of 21 g of [N (benzyloxycarbonyl) -L-phenylalanylmethyl chloride in 500 ml of aqueous tetrahydrofuran. Stirring was continued for a further 0.5 h and then the solvent was removed by evaporation. The residue was decomposed into dichloromethane and water and then carefully acidified with concentrated hydrochloric acid. After further treatment of the organic phase, a white solid material was obtained which gave two spots on chromatography; Rf 0.4 and 0.5 (5% methanol / chloroform). The solid material was extracted with boiling hexane until a faster component was extracted. The combined n-hexane extracts were evaporated and the residue was re-extracted with boiling N-hexane to separate a smaller amount (0.5 g) of the slower component. 11.5 g of 3 (S) - (benzyloxyformamido) 1-chloro-4-phenyl-2 (S) -butanol were obtained, m.p. 151-153 ° C (from ethyl acetate / n-hexane).

(ii) A solution of 11.4 g of 3 (S) - (benzyloxyformamido) -1-chloro-4-phenyl-2 (S) -butanol in 300 ml of ethanol containing 2.24 g of potassium hydroxide was allowed to stand for 15 minutes. The solvent was removed by evaporation and the residue was layered between water and dichloromethane. Further treatment gave 10.1 g of crude 3 (S) - (benzyloxyformamido) -1,2 (S) epoxy-4-phenylbutane as a off-white solid which was used without further purification.

(iii) A solution of 0.594 g of 3 (S) - (benzyloxyformamido) -1,2 (S) -epoxy-4-phenylbutane and 0.38 g of N1-phenyl-L-prolinamide in 5 ml of dimethylformamide was heated to 90 ° C for 16 hours, and then at 120 ° C for 8 hours. The solvent was removed by evaporation and the residue was chromatographed on silica gel using diethyl ether / methanol (96: 4) for elution to give 0.48 g of N - [3 (S) - (benzyloxyformamido) -2 (R) hydroxy-4 -phenylbutyl] -Nl-phenyl-L-prolinamide as a white solid, mp 133-135 ° C (from diethyl ether).

(iv) A solution of 0.45 g of tert-butyl ester N ² - [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nl-phenyl-L-prolinamide in 20 ml of ethanol was hydrogenated for 4 hours over 10% palladium on charcoal to give 0.315 g of N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-phenyl-L-prolinamide as a white solid;

MS: m / e 354 [M + H] +.

Example 34

In an analogous manner as described in Example 33, 0.987 mg of 2- [3 (S) (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1,2,3 , 4-tetrahydro3 (R, S) -isoquinolinecarboxamide, 0.665 g of N- (benzyloxycarbonyl) -L-asparagine, 566 mg of dicyclohexylcarbodiimide, 0.337 g of hydroxybenzotriazole and 0.287 g of N-ethyl-morpholine gave 0.5 g of 2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1,2,3,4-tetrahydro-3 (R, S) - isoquinoline-carboxamide in the form of pale yellow foam; MS: m / e 644 [M + H] +.

Example 35

In the analogous manner as described in Example 33, 0.225 dicyclohexylcarbodiimide, 0.135 g hydroxybenzotriazole, 0.115 g N-ethylmorpholine, 0.38 g N ^ 41 [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] are obtained -Nl-benzyl-L-prolinamide and 0.266 g of N (benzyloxycarbonyl) -L-asparagine gave 0.056 g of Nl-benzyl-N2-3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-prolinamide as a white solid; MS: m / e 616 [M + H] ⁺ .

Example 36

A solution of 270 mg of N- [3 (S) - (benzyloxyformamido) -4- (4-fluorophenyl) 2 (R) -hydroxybutyl] -L-proline tert-butyl ester in 50 ml of ethanol was hydrogenated for 2 hours over 30 mg 10 % palladium on charcoal. The catalyst was removed by filtration, the filtrate was evaporated to give an oil which was combined with 158 mg of N- (benzyloxycarbonyl) -L-asparagine in a manner analogous to that described in Example 1. The crude product obtained was chromatographed. on silica gel, using 10% methanol in dichloromethane for elution. We obtained 120 mg of N- [3 (S) - [[N- (benzyloxycarbonyl) L-asparaginyl] amino] -4- (4-fluorophenyl) -2 (R) -hydroxybutyl] -L-proline tert-butyl ester. in the form of a white solid material with a melting point of 163-164 ° C.

The N- [3 (S) - (benzyloxyformamido) -4- (4-fluorophenyl) -2 (R) -hydroxybutyl] -L-proline tert-butyl ester used as starting material was obtained as follows :

(i) In an analogous manner to that described in Example 19 (i), 7.70 g of N (benzyloxycarbonyl) -3- (4-fluorophenyl) -DL-alanine were treated with isobutyl chloroformate and then reacted with diazomethane, whereby 7.15 g of benzyl [2- (4fluorophenyl) -1 (RS) - (2-diazoacetyl) ethyl] carbamate were obtained as a yellow solid, mp 97 ° C.

(ii) Treatment of the above product with hydrogen chloride in a manner analogous to that described in Example 19 (ii) yielded 6.70 g of [N- (benzyloxycarbonyl) -3- (4fluorophenyl) -DL-alanyl] methyl chloride, which used without further purification.

(iii) Reduction of 6.30 g of the above product with sodium borohydride in an analogous manner as described in Example 19 (iii) gave 5.83 g of a mixture of alcohols. This mixture was chromatographed on a silica gel column using 2% methanol in dichloromethane to elute to give 0.95 g of threo-3 (benzyloxyformamido) -1-chloro-4- (4-fluorophenyl) -2-butanol in the form of white solid material. Further elution of the upper column with the same solvent system was obtained

3.25 g of erythro-3- (benzyloxyformamido) -1-chloro-4- (4-fluorophenyl) -2-butanol as a white solid, mp 148-149 ° C.

(iv) Treatment of 3.20 g of erythro-3- (benzyloxyformamido) -1-chloro-4- (4-fluorophenyl) -2-butanol with an ethanolic potassium hydroxide solution in an analogous manner as described in Example 19 (iv) gave 2.03 g erythro-3- (benzyloxyformamido) -1,2-epoxy4- (4-fluorophenyl) butane as a white solid; MS: m / e 315 [M] &lt; ^{+ &gt};.

(v) 0.63 g of the above product was treated with the t-butyl ester of L-proline in an analogous manner as described in Example 19 (v). The crude product obtained was chromatographed on a silica gel column using 5% methanol in dichloromethane for elution. 0.26 g of N- [3 (R) - (benzyloxyformamido) -4- (4-fluorophenyl) -2 (S) -hydroxybutyl] -L-proline tert-butyl ester was obtained as a colorless oil; NMR (CDCl3, 300 MHz) δ 1.47 (9H, s), 1.75-1.95 (4H, m), 2.05-2.15 (1H, m), 2.25-2.35 (1H, m), 2.50-3.00 (4H, m) , 3.20 (2H, m), 3.58 (1H, m), 3.85 (1H, m), 4.88 (1H, d), 5.00 (2H, dd), 6.94 (2H, t), 7.13-7.37 (7H, m ).

Further elution with the same solvent system gave 0.275 g of N- [3 (S) - (benzyloxyformamido) -4- (4-fluorophenyl) -2 (R) -hydroxybutyl] -L-proline tert-butyl ester as a colorless oils; NMR (CDCl3), 300 MHz) δ 1.42 (9H, s), 1.75-1.95 (4H, m), 2.05-2.20 (1H, m), 2.54-2.80 (3H, m), 2.85-2.98 (2H, m ), 3.03-3.24 (2H, m), 3.44 (1H, m), 3.92 (1H, m) 4.88-5.06 (3H, m), 6.93 (2H, t), 7.10-7.40 (7H, m).

Example 37

A solution of 730 mg of N- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4 (4-methoxyphenyl) butyl] -L-proline tert-butyl ester in 20 ml of ethanol was hydrogenated for 2 hours over 50 mg 10% palladium on charcoal. The catalyst was removed by filtration, the filtrate was evaporated to give an oil which was combined with 340 mg of N- (benzyloxycarbonyl) -Lasparagine in a manner analogous to that described in Example 1. The crude product obtained was chromatographed on silica gel. , using 10% methanol in dichloromethane for elution. 82 mg of N- [3 (S) - [[N (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4- (4-methoxyphenyl) butyl] -L-proline 82 mg was obtained in white solid material; MS: m / e 613 [M + H] ⁺ .

The N- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4- (4-methoxyphenyl) butyl] -L-proline tert-butyl ester used as starting material was obtained as follows way:

(i) In an analogous manner as described in Example 19 (i), 3.10 g of N (benzyloxycarbonyl) -3- (4-methoxyphenyl) -L-alanine were treated with isobutyl chloroformate and then diazomethane to give 2.83 g benzyl- [2- (4-methoxyphenyl) -1 (S) (2-diazoacetyl) ethyl] carbamate as a pale yellow solid with a melting point of 88-90 ° C.

(ii) 2.80 g of the above product was treated with hydrogen chloride in a manner analogous to that described in Example 19 (ii) to give 2.85 g of [N- (benzyloxycarbonyl) -3 (4-methoxyphenyl) -L-alanyl] methyl chloride, in the form of a white solid, mp 107 ° C.

(iii) The above product was treated with sodium borohydride in an analogous manner as described in Example 19 (iii). The crude product obtained was triturated with 30 ml of 5% methanol in dichloromethane to give 1.15 g of 3 (S) (benzyloxyformamido) -1-chloro-4- (4-methoxyphenyl) -2 (S) -butanol as a white solid material. MS: m / e 363. 365 [M] +.

(iv) The above product was treated with an ethanolic potassium hydroxide solution in an analogous manner as described in Example 19 (iv) to give 0.86 g of 3- (S) (benzyloxyformamido) -1,2 (S) -epoxy- 4- (4-Methoxyphenyl) butane as a white solid, mp 86-87 ° C.

(v) 0.51 g of the above product was treated with the L-proline tert-butyl ester in an analogous manner as described in Example 19 (v) to give 0.63 g of N- [3 (S) t-butyl ester ) - (Benzyloxyformamido) -2 (R) -hydroxy-4- (4-methoxyphenyl) -butyl] -L-proline as a colorless oil; MS: m / e 498 [M] &lt; + &gt;.

Example 38

A solution of 1.58 g of N2- [3 (S) - (benzyloxyformamido) -4- (4-tert-butoxyphenyl) -2 (R) hydroxybutyl] -Nl-tert-butyl-L-prolinamide in 200 ml of ethanol was hydrogenated 4 hours above 100 mg 10% palladium on charcoal. The catalyst was removed by filtration, the filtrate was evaporated to give an oil which was combined with 0.74 g of N- (benzyloxycarbonyl) L-asparagine in a manner analogous to that described in Example 1. The crude product obtained was chromatographed on silica gel columns, using 10% methanol in dichloromethane for elution. 0.59 g of N - [3 (S) - [[N (benzyloxycarbonyl) -L-asparaginyl] amino] -4- (4-tert-butoxyphenyl) -2 (R) -hydroxybutyl] -N ^ -terc was obtained .-butyl-L-prolinamide as a white solid;

MS: m / e 654 [M + H] &lt; + &gt;.

N2- [3 (S) - (benzyloxyformamido) -4- (4-tert-butoxyphenyl) -2 (R) -hydroxybutyl] -Nlterc-butyl-L-prolinamide was used as starting material as follows:

(i) In an analogous manner to that described in Example 19 (i), N- (benzyloxycarbonyl) -3- (4-tert-butoxyphenyl) -L-alanine (obtained from 7.45 g of dicyclohexylammonium salt) was treated with isobutyl chloroformate and then reacted with diazomethane to give 3.58 g of benzyl [2- (4-tert-butoxyphenyl) -1 (S) - (2diazoacetyl) ethyl] carbamate as a yellow solid with a melting point of 80-82 ° C.

(ii) Treatment of the above carbamate with hydrogen chloride in a manner analogous to that described in Example 19 (ii) yielded 3.6 g of [N- (benzyloxycarbonyl) -3- (4-tert-butoxyphenyl) -L-alanyl] of methyl chloride which was used without further purification.

(iii) Reduction of the above chloride with sodium borohydride in an analogous manner as described in Example 19 (iii) yielded a mixture of alcohols triturated with hot n-hexane to give 3.14 g of a white solid. . This material was recrystallized from a mixture of ethyl acetate and n-hexane to give 1.88 g of 3- (S) - (benzyloxyformamido) -4- (4-tert-butoxyphenyl) -1-chloro-2 (S) butanol. in the form of a white solid material; NMR (CDCl3, 250 MHz) δ 1.35 (9H, s), 2.85-3.05 (3H, m), 3.50-3.70 (2H, m), 3.80-3.90 (1H, m), 3.92-4.04 (1H, m) , 4.85 (1H, d), 5.00-5.10 (2H, m), 6.93 (2H, d), 7.10 (2H, d), 7.25-7.40 (5H, m).

(iv) 5.52 g of the above product was treated with an ethanolic potassium hydroxide solution in an analogous manner as described in Example 19 (iv) to give 4.98 g of 3- (S) - (benzyloxyformamido) -4- (4-tert) .-butoxyphenyl) -1,2 (S) -epoxy-butane as a colorless gum; NMR (CDCl3), 250 MHz) δ 1.36 (9H, s), 2.75-3.05 (5H, m), 3.76 (1H, m), 4.73 (1H, d), 5.07 (2H, s), 6.95 (2H. d), 7.10 (2H, d), 7.25-7.40 (5H, m).

(v) 2.50 g of the above product was treated with N1-tert-butyl-L-prolinamide in an analogous manner as described in Example 19 (v) to give 1.58 g of N ^ [3 (S) - ( benzyloxyformamido) -4- (4-tert-butoxyphenyl) -2 (R) -hydroxybutyl] -Nl-tert-butyl-L-prolinamide as a colorless oil; MS: m / e 539 [M] &lt; ^{+ &gt};.

Example 39

200 mg solution of N ² - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] 4- (4-tert-butoxyphenyl) -2 (R) -hydroxybutyl] -Nl-tert-butyl -L-prolinamide in 5 ml of trifluoroacetic acid was stirred for 20 hours at room temperature. The solution was poured into a saturated solution of hydrogen carbonate and the mixture was extracted three times with 30 ml of dichloromethane and three times with 30 ml of ethyl acetate each. The combined extracts were dried over anhydrous sodium sulfate, evaporated to give 100 mg of a colorless gum. This was chromatographed on a silica gel column using 10% methanol in dichloromethane to elute to give 42 mg of a glassy solid. This material was triturated with a mixture of 1 ml diethyl ether and 3 ml n-hexane to give 28 mg trifluoroacetate N ² - [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] -2 (R) -hydroxyphenyl) -butyl] N1-tert-butyl-L-prolinamide as a glassy solid; MS: m / e 598 [M-CF 3 CO 2] ⁺ .

Example 40

In the analogous manner as described in Example 1, N ² - [3 (S) (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L- is obtained from 4-methyl ester. of prolinamide and N (benzyloxycarbonyl) -L-aspartic acid gave N ² - [3 (S) - [[N- (benzyloxycarbonyl) 4-O-methyl-La-aspartyl] amino] -2 (R) -hydroxy-4- phenylbutyl] -Nl-tert-butyl-L-prolinamide in the form of a glassy solid material with a melting point of 55-60 ° C.

MS: m / e 597 [M + H] &lt; + &gt;.

Example 41

In the analogous manner as described in Example 1, N ² - [3 (S) (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N 1-tert-butyl-L-prolinamide and N (benzyloxycarbonyl) -L-methionine gave N ² - [3 (S) - [[N- (benzyloxycarbonyl) -Lmethionyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L -Prolinamide as a white solid with a melting point of 132-133 ° C.

Example 42

N ² - [3 (S) - [[N, 3-bis (benzyloxycarbonyl) -L-histidyl] amino] -2 (R) -hydroxy-4-phenyl-butyl] N1-tert-butyl-L-prolinamide was produced from N ² - [3 (S) - (benzyloxyformamido) -2 (R) hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide and N, 3-bis (benzyloxycarbonyl) -Lhistidine per analog the method as described in Example 1; it was obtained in the form of a colorless glassy material; MS: m / e 739 [M + H] &lt; ⁺ &gt;.

Example 43

30 mg solution of N2- [3 (S) - [[N, 3-bis (benzyloxycarbonyl) -L-histidyl] amino] -2 (R) hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide in 6 ml of 2M hydrochloric acid was stirred at room temperature overnight. The mixture was neutralized by the addition of 2M sodium hydroxide solution and extracted twice with 10 ml of dichloromethane each. The combined extracts were dried over anhydrous sodium sulfate and evaporated. The crude product obtained was triturated with diethyl ether to give 14 mg of N ² - [3 (S) - [[N- (benzyloxycarbonyl) -L-histidyl] amino] -2 (R) -hydroxy- [4-phenylbutyl ] N1-tert-butyl-L-prolinamide in the form of a white solid, melting point 100 ° C (decomposition); MS: m / e 604 [M] &lt; + &gt;.

Example 44

In an analogous manner to that described in Example 21, N [3 (S) - [[L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L- was extracted from 224 mg of tert-butyl ester. of proline, 70 mg of benzoyl chloride and 65 mg of di-isopropylethylamine were obtained after complete chromatography on silica gel using 10% methanol in dichloromethane for elution, 20 mg of N [3 (S) - [N- (benzoyl- L-asparaginyl) amino] -2 (R) -hydroxy- [4-phenylbutyl] -L-proline as a white solid methanol / diethyl ether material; MS: m / e 553 [M + H] &lt; ⁺ &gt;.

Example 45

In an analogous manner to that described in Example 27, N3 (S) - [[L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline was extracted from 224 mg of tert-butyl ester. 93 mg of 1-naphthylacetic acid, 68 mg of hydroxybenzotriazole, 58 mg of N-ethylmorpholine and 113 mg of dicyclohexylcarbodiimide after complete chromatography on silica gel using 10% methanol in dichloromethane for elution gave 106 mg of N- [2 ( R) hydroxy-3 (S) - [[N- [2- (1-naphthyl) acetyl] -L-asparaginyl] amino] -4-phenylbutyl] -L-proline as a white solid methanol / diethyl ether material; MS: m / e 617 [M + H] ⁺ .

Example 46

In an analogous manner to that described in Example 27, N3 (S) - [[L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline was obtained from 493 mg of tert-butyl ester. 190 mg of kinaldic acid, 149 mg of hydroxybenzotriazole, 127 mg of N-ethylmorpholine and 249 mg of dicyclohexylcarbodiimide after complete chromatography on silica gel using 10% methanol in dichloromethane for elution gave 167 mg of N- [2 (R) tert-butyl ester hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] butyl] -L-proline as a white solid methanol / diethyl ether material; MS: m / e 604 [M + H] ⁺ .

Example 47

In an analogous manner to that described in Example 27, N3 (S) - [[L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline was obtained from 234 mg of tert-butyl ester, 96 mg of 4-chlorohydrocycic acid, 70 mg of hydroxybenzotriazole, 60 mg of N-ethylmorpholine and 118 mg of dicyclohexylcarbodiimide, after completion of the chromatography on silica gel using elution system G, gave 171 mg of N- [3 (S) - tert-butyl ester. [[N- (4-chlorohydrocinamoyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline as a white solid; MS: m / e 615 [M-1H] ⁺ .

Example 48

In an analogous manner to that described in Example 27, N3 (S) - [[L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline was obtained from 234 mg of tert-butyl ester, 90 mg of 1-isoquinolinecarboxylic acid, 70 mg of hydroxybenzotriazole, 60 mg of N-ethylmorpholine and 118 mg of dicyclohexylcarbodiimide, after complete chromatography on silica gel using 10% methanol in dichloromethane for elution, yielded 146 mg of N- [2-butyl ester (2-) R) hydroxy-3 (S) - [[N- (1-isoquinolylcarbonyl) -L-asparaginyl] amino] -4-phenylbutyl] -L-proline as a pale yellow solid from ethyl acetate / n-hexane;

MS: m / e 604 [M + H] +.

Example 49

In an analogous manner as described in Example 21, 209 mg of N ^ -3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L -prolineamide, 89 mg of 2-naphthoyl chloride and 61 mg of di-isopropylethylamine gave 191 mg of N1-tert-butyl-N ^ - [2 (R) hydroxy-3 (S) - [[N- (2-naphthoyl) -L- asparaginyl] amino] -4-phenylbutyl] -L-prolinamide as a white solid; MS: m / e 602 [M + H] +.

The starting material was obtained by hydrogenation of N2- [3 (S) - [[N- (benzyloxycarbonyl) L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide ;

MS: m / e 582 [M + H] ⁺ , This last compound was obtained in an analogous manner as described in Example 33 only using N1-tert-butyl-L-prolinamide in paragraph (iii) instead of N1- of phenyl-L-prolinamide.

Example 50

Solution 2.798 g of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-octahydrocyclopent [b] pyrrole-2-carboxamide (four diastereomers) and 2.72 g of succinimide ester N- (benzyloxycarbonyl) -L-asparagine in 50 ml of dimethoxyethane was stirred for 2 hours at 20 ° C and then the solvent was removed by evaporation. The residue was collected in ethyl acetate and the solution was washed with aqueous sodium carbonate solution and sodium chloride solution. The organic solution was evaporated and the residue was chromatographed on silica gel using a G elution system. 434 mg of 1- [3 (S) - [[N-benzyloxycarbonyl) -Lasparaginyl] amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-octahydro- (3aS, 6aS) cyclopenta was obtained [b] pyrrole-2 (S) -carboxamide as a white solid material from ethyl acetate; MS: m / e 622 [M + H] +; Rf 0.22.

1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-octahydro-cyclopenta- [b] pyrrole 2-carboxamide, which was used as starting material, was obtained as follows.

(i) A solution of 2.98 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenylbutane and 2.1 g of N-tert-butyl-octahydro-cyclopenta [b] pyrrole-2-carboxamide in 50 ml of ethanol was heated at reflux for 10 hours and then the solvent was removed by evaporation. Chromatography on silica gel using 4.7 g of 1- [3 (S) (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-octahydrocyclopent was obtained by chromatography on silica gel [b ] pyrrole-2-carboxamide in the form of a mixture of four diastereomers,

MS: m / e 508 [M + H] +.

(ii) A solution of 4.6 g of 1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-octahydro-cyclopenta- [b] pyrrole-2-carboxamide (four diastereomers ) in 90 ml of ethanol was hydrogenated above 10% palladium on charcoal at 20 ° C and at atmospheric pressure for 72 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 2.958 g of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-octahydro-cyclopenta- [b] pyrrole -2-carboxamide (four diastereomers) in the form of a brownish oil; MS: m / e 374 [M + H] ⁺

Example 51

In an analogous manner to that described in Example 27, 310 mg of 1-3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-octahydro- ( 3aS, 6aS) cyclopenta [b] pyrrole-2 (S) -carboxamide, 111 mg quinaldic acid, 86 mg hydroxy49 benzotriazole, 74 mg N-ethylmorpholine and 132 mg dicyclohexylcarbodiimide were obtained after chromatography on silica gel, eluting dichloromethane / methanol (9: 1), 200 mg of N-tert-butyl-octahydro-1- [2 (R) -hydroxy-4-phenyl-3 (S) [[N- (2-quinolylcarbonyl) -L] -asparaginyl] amino] butyl- (3aS, 6aS) -cyclopenta- [b] pyrrole-2 (S) carboxamide, in the form of a white solid; MS: m / e 643 [M + H] ⁺ 1- 3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-octahydro (3aS, 6aS ) -cyclopenta- [b] pyrrole-2 (S) -carboxamide, which was used as starting material, was obtained as follows:

A solution of 397 mg of 1- [3 (S) - [[N-benzyloxycarbonyl) -L-asparaginyl] amino-2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-octahydro- (3aS, 6aS) - of cyclopenta [b] pyrrole-2 (S) carboxamide in 20 ml of ethanol was hydrogenated for 4 hours over 10% palladium on charcoal at 20 ° C and under atmospheric pressure. The catalyst was removed by filtration and the filtrate was evaporated, yielding 312 mg of 1-3 (S) - [[Lasparaginyl] amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-octahydro- ( 3aS, 6aS) -cyclopenta- [b] pyrrole-2carboxamide in the form of pale yellow gum; MS: m / e 488 [M + H] &lt; + &gt;.

Example 52

In the analogous manner as described in Example 27, 295 mg of N ² - [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl -L-prolinamide, 204 mg of 6- (benzyloxycarbonyl) -2-naphthalenecarboxylic acid, 20 mg of hydroxybenzo-triazole, 77 mg of N-ethylmorpholine and 151 mg of dicyclohexylcarbodiimide were obtained after complete chromatography on silica gel using G elution system, 340 mg N ^2- [3 (S) - [[N [6-benzyloxycarbonyl) -2-naphthoyl] -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nlert-butyl-L-prolinamide in dirty-white solid ethyl acetate / nhexane material; MS: m / e 736 [M + H] &lt; + &gt;.

Example 53

294 mg N ² - [3 (S) - [[N- [6- (benzyloxycarbonyl) -2-naphthoyl] -L-asparaginyl] amino] -2 (R) hydroxy-4-phenylbutyl] -Nl-tert.- of butyl L-prolinamide in 100 ml of isopropanol was hydrogenated over 10% palladium on charcoal at 20 ° C under atmospheric pressure for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 145 mg of N ² - [3 (S) - [[N- (6-carboxy-2-naphthoyl) -L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide as a white solid from methanol / -ethyl acetate; MS: m / e 646 [M + H] +.

Example 54

In an analogous manner as described in Example 27, 295 mg of N ² - [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl- Of L-prolinamide, 171 mg of 4- (benzyloxycarbonyl) benzoic acid, 90 mg of hydroxybenzotriazole, 77 mg of Nethylmorpholine and 151 mg of dicyclohexylcarbodiimide were obtained after chromatography on silica gel using a G elution system, 390 mg of N ² - [3 (S) - [[N- [4 (Benzyloxycarbonyl) benzoyl-L-asparaginyl] amino] -2 (R) -hydroxy-4-phenyl-butyl] -Nlterc-butyl-L-prolinamide as a dirty white solid from ethyl acetate / n-hexane; MS: m / e 686 [M + H] &lt; + &gt;.

Example 55

274 mg N ² - [3 (S) - [[N- [4- (benzyloxycarbonyl) benzoyl] -L-asparaginyl] amino] -2 (R) hydroxy-4-phenylbutyl] -Nl-tert-butyl-L -prolineamide in 100 ml of isopropanol was hydrogenated over 10% palladium on charcoal at 20 ° C under atmospheric pressure for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 193 mg of N ² - [3 (S) - [[N- (4-carboxybenzoyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4- phenylbutyl] -Nl-tert-butyl-L-prolinamide as a white solid methanol / ethyl acetate solid; MS: m / e 596 [M + H] &lt; ⁺ &gt;.

Example 56

In an analogous manner as described in Example 27, from 228 mg of N ² - [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl- L-prolinamide, mg of 4-nitrocyclic acid, 69 mg of hydroxybenzotriazole, 59 mg of N-ethylmorpholine and 116 mg of dicyclohexylcarbodiimide were obtained after chromatography on silica gel using dichloromethane / methanol (9: 1) for elution, 68 mg of N1-tert.- butyl-N ² - [2 (R) hydroxy-3 (S) - [[N- (4-nitrocinamoyl) -L-asparaginyl] amino] -4-phenylbutyl] -L-prolinamide as a white solid methanol material / diethyl ether; MS: m / e 623 [M + H] &lt; + &gt;.

Example 57

A solution of 200 mg of N1-tert-butyl-N ² - [2 (R) -hydroxy-3 (S) - [[N- (4-nitrocinamoyl) -Lasparaginyl] amino] -4-phenylbutyl] -L-prolinamide in 20 ml of ethanol were hydrogenated over 10% palladium on charcoal at 20 ° C under atmospheric pressure for 5 hours. The catalyst was removed by filtration and the filtrate was evaporated. The residue was chromatographed on silica gel using dichloromethane / methanol (9: 1) to elute to give mg of N2- [3 (S) - [[N- (4-aminohydrocinamoyl) -L-asparaginyl] amino] -2 ( R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide as a white solid methanol / ethyl acetate solid; MS: m / e 595 [M + H] &lt; ⁺ &gt;.

Example 58

In an analogous manner as described in Example 27, from 418 mg of N2- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L -prolineamide,

205 mg of l-acetyl-1,2,3,4-tetrahydro-2 (R, S) -quinolinecarboxylic acid, 127 mg of hydroxybenzotriazole, 108 mg of N-ethylmorpholine and 212 mg of dicyclohexylcarbodiimide after chromatography on silica gel using dichloromethane / methanol (9: 1) for elution to give 220 mg of N2-3 (S) - [[N - [(1-acetyl-1,2,3,4-tetrahydro-2 (R, S) -quinolyl) carbonyl] - L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide (two diastereomers) as a white solid material from dichloromethane / diethyl ether; MS: m / e 649 [M + H] ⁺ .

Example 59

In the analogous manner as described in Example 27, 418 mg of N ^ - [3 (S) - [(Lasparaginyl) amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl -L-prolinamide, 179 mg 1,2,3,4-tetrahydro-1-oxo-3 (R, S) -isoquinolinecarboxylic acid, 127 mg hydroxybenzotriazole, 108 mg N-ethylmorpholine and 212 mg dicyclohexylcarbodiimide, after silica gel chromatography , using system I for elution, 132 mg of N1-tert-butyl-N2- [3 (S) - [[N - [(1,2,3,4-tetrahydro-1-oxo-3 (R or S) -isoquinolyl) carbonyl] -L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -L-prolinamide (diastereomer A) as a white solid methanol / diethyl ether material,

MS: m / e 621 [M + H] ⁺ , and 66 mg N1-tert-butyl-N2- [3 (S) - [[N - [(1,2,3,4-tetrahydro-loco-3) (R or S) -isoquinolyl) carbonyl] -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] L-prolinamide (diastereomer B) as a white solid methanol / diethyl ether material; MS: m / e 621 [M + H] ⁺ .

Example 60

In an analogous manner as described in Example 27, from 418 mg of n2- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L -prolineamide,

218 mg of 2-acetamido-1,2,3,4-tetrahydro-2-naphthalenecarboxylic acid, 127 mg of hydroxybenzotriazole, 108 mg of N-ethylmorpholine and 212 mg of dicyclohexylcarbodiimide after chromatography on silica gel using dichloromethane / methanol (9: 1 ) for elution to give 142 mg of N ^ - [3 (S) - [[N- [2 (R or S) -acetamido-1,2,3,4-tetrahydro-2 naphthoyl] -L-asparaginyl] amino] - 2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl] -L-prolinamide (diastereomer A) as a white solid MS: m / e 663 [M + H] ⁺ and 80 mg N2- [ 3 (S) - [[N- [2 (R or S) -acetamido-1,2,3,4-tetrahydro-2-naphthoyl] -L-asparaginyl] amino] -2 (R) -hydroxy-4- phenylbutyl] -N-tert-butyl-L-prolinamide (diastereomer B) as a white solid; MS: m / e 663 [M + H] &lt; ⁺ &gt;.

Example 61

In the analogous manner as described in Example 27, 209 mg of N, N - [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl -L-prolinamide, 81 mg quinaldic acid, 63 mg hydroxybenzotriazole, 54 mg N-ethylmorpholine and 106 mg dicyclohexylcarbodiimide, after complete chromatography on silica gel using dichloromethane / methanol (9: 1) to give 88 mg N2- [2 ( R) -hydroxy-4-phenyl3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] butyl] -Nl-tert-butyl-L-prolinamide as a white solid MS: m / e 603 [M + H] ⁺ .

Example 62

A solution of 844 mg of 1- [3 (S) -amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) piperidinecarboxamide and 882 mg of N- (benzyloxycarbonyl) -Lasparagine succinimide ester in 25 ml of tetrahydrofuran was stirred for 16 hours at 20 ° C and then evaporated. The residue was dissolved in 50 ml of dichloromethane and the organic solution was washed with saturated sodium chloride solution and water. The solvent was removed by evaporation and the residue was chromatographed on silica gel using dichloromethane / methanol (9: 1) for elution. 461 mg of 1- [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide was obtained in the form of a pale yellow solid; MS: m / e 596 [M + H] &lt; ⁺ &gt;.

1- [3 (S) -amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide, which was used as starting material, was obtained as follows :

(i) 2.576 g of N-tert-butyl-2 (S) -piperidinecarboxamide and 4.158 g of 3 (S) - (benzyloxyformamido) -1,2 (S) -epoxy-4-phenylbutane in 70 ml of ethanol were heated for 16 hours at reflux. The solvent was removed by evaporation and the residue was dissolved in 100 ml of diethyl ether and treated with 10 g of activated magnesium silicate. The solvent was then removed by evaporation to give 6.16 g of 1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-453 phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide in the form colorless glassy material, MS: m / e 482 [M + H] ⁺ (ii) A solution of 1.25 g of 1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl -2 (S) -piperidinecarboxamide in 80 ml of ethanol was hydrogenated over 10% palladium on charcoal at 20 ° C and under atmospheric pressure for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 0.844 g of 1- [3 (S) amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide in pale yellow rubber; MS: m / e 348 [M + H] ⁺

Example 63

In an analogous manner as described in Example 21, 226 mg of 1- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide, 93 mg of 2-naphthoyl chloride and 63 mg of di-isopropylethylamine, after complete chromatography on silica gel using dichloromethane / methanol (9: 1) for elution to give 76 mg of N-tert-butyl-1- [2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] -2 (S) -piperidinecarboxamide as a white solid; MS: m / e616 [M + H] ⁺ 1- [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S ) The piperidinecarboxamide used as starting material was obtained by hydrogenation of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) piperidinecarboxamide.

Example 64

In an analogous manner as described in Example 27, from 226 mg of 1- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide, 85 mg quinaldic acid, 56 mg N-ethylmorpholine, 66 mg hydroxybenzotriazole and 111 mg dicyclohexylcarbodiimide, after chromatography on silica gel using dichloromethane / methanol (9: 1), eluted to give 100 mg N-teryl .-Butyl-1 [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] -amino] butyl] -2 (S) piperidinecarboxamide in white form solid material; MS: m / e 617 [M + H] +.

Example 65

In an analogous manner to that described in Example 27, 228 mg of N- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl- Of L-prolinamide, 88 mg of 3-isoquinolinecarboxylic acid, 69 mg of hydroxybenzotriazole, 59 mg of N-ethylmorpholine and 116 mg of dicyclohexylcarbodiimide, after chromatography on silica gel using dichloromethane / methanol (9: 1), eluted to give 88 mg of N -terter -butyl-N ^ [2 (R) -hydroxy-3 (S) - [[N- (3-isoquinolylcarbonyl) -L-asparaginyl] amino] -4-phenylbutyl] -L-propylamino as a white solid; MS: m / e 603 [M + H] &lt; ⁺ &gt;.

Example 66

In an analogous manner to that described in Example 27, 228 mg of N- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl- L-prolinamide, 88 mg of 3quinolinecarboxylic acid, 69 mg of hydroxybenzotriazole, 59 mg of N-ethylmorpholine and 116 mg of dicyclohexylcarbodiimide, after chromatography on silica gel using dichloromethane / methanol (9: 1), were eluted to give 86 mg of N-tert. butyl-N2- [2 (R) hydroxy-4-phenyl-3 (S) - [[N- (3-quinolylcarbonyl) -L-asparaginyl] amino] butyl] -L-prolinamide as a white solid; MS: m / e 603 [M + H] &lt; + &gt;.

Example 67

In an analogous manner to that described in Example 27, 295 mg of N ^ - [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L -prolineamide, 214 mg of 3- (benzyloxyformamido) -2-naphthalenecarboxylic acid, 90 mg of hydroxybenzotriazole, 77 mg of N-ethylmorpholine and 151 mg of dicyclohexylcarbodiimide, after complete chromatography on silica gel, using G eluting system, 380 mg of N ⁷ [3 (S) - [[N- (3-benzyloxyformamido) -2-naphthoyl] -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide, white solid methanol material; MS: m / e 751 [M + H] ⁺ .

Example 68

Solution 216 mg of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2,3-dihydro-1H-indole-2 (R or S) -carboxamide (diastereomer A), 151 mg of N- (benzyloxycarbonyl) -L-asparagine, 76 mg of hydroxybenzotriazole, 65 mg of N-ethylmorpholine and 128 mg of dicyclohexylcarbodiimide in 10 ml of tetrahydrofuran were stirred for 16 hours at 20 ° C. The solvent was removed by evaporation and the residue was chromatographed on silica gel using dichloromethane / methanol (92: 8) for elution. 70 mg of 1 [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-2,3-dihydro-1H is obtained -indole-2 (R or S) -carboxamide (diastereomer A) in the form of a pale yellow solid; MS: m / e 630 [M + H] &lt; ⁺ &gt;.

1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2,3-dihydro-1H-indole-2 (R or S) -carboxamide which is it was used as starting material, as follows:

A solution of 891 mg of N-tert-butyl-2,3-dihydro-1H-indole-2 (R or S) -carboxamide and 654 mg of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy- 4-Phenylbutane in 5 ml of dimethylformamide was heated at 140 ° C for 40 hours. The solvent was removed by evaporation and the residue was chromatographed on silica gel using diethyl ether / n-hexane (2: 1) for elution. 300 mg of 1- [3 (S) - (benzyloxyformamido) -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-2,3-dihydro-1H-indole-2 (R or S) was obtained -carboxamide (diastereomer A) in the form of a pale yellow oil; m / e 516 [M + H] ⁺ , and 290 mg of 1 [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2,3-dihydro- lindol-2 (R or S) -carboxamide (diastereomer B) as a pale yellow oil;

MS: m / e 516 [M + H] +.

(ii) A solution of 300 mg of 1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-2,3-dihydro-1H-indole-2 (R or S ) -carboxamide (diastereomer A) in 20 ml of ethanol was hydrogenated above 10% palladium on charcoal at 20 ° C and pressurized for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 216 mg of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-2,3-dihydro-1H- indole-2 (R or S) -carboxamide (diastereomer B) as a brownish oil.

Example 69

In an analogous manner as described in Example 68, from 223 mg of 1- [3 (S) -amino-2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-2,3-dihydro-1H -indole-2 (R or S) -carboxamide (diastereomer B), 151 mg N- (benzyloxycarbonyl) -L-asparagine, 76 mg hydroxybenzotriazole, 65 mg N-ethylmorpholine and 128 mg dicyclohexylcarbodiimide gave 210 mg l- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2,3-dihydro-1H-indole-2 (R or S) -carboxamide (diastereomer B), in the form of a pale yellow solid; MS: m / e 630 [M + H] &lt; ⁺ &gt;.

Example 70

A solution of 0.422 g of N2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] -Nl-tert-butyl-L-prolinamide and 142 mg of 3-chloroperbenzoic acid in 10 ml of dichloromethane was stirred for 1 hour at 20 ° C. The solvent was removed by evaporation and the residue was chromatographed on silica gel using dichloromethane / methanol (9: 1) to elute to give 289 mg of N, N-tert-butyl-N 2 - [2 (R) - hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] -butyl] -L-prolinamide as a white solid;

MS: m / e 619 [M + H] +.

Example 71

0.123 mg solution of N ^ -oxide N1-tert-butyl-N2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2quinolylcarbonyl) -L-asparaginyl] amino] butyl] - L-prolinamide and 35 mg of 3-chloroperbenzoic acid in 5 ml of dichloromethane were stirred for 24 hours at 25 ° C. Another 70 mg of 3-chloroperbenzoic acid was added and stirred for 48 hours at 20 ° C. The solution was washed with aqueous sodium carbonate solution and sodium chloride solution, then the solvent was removed by evaporation. The residue was chromatographed on silica gel using elution system J to give 70 mg of N1-tert-butyl N2- [2 (R) -hydroxy-3 (S) - [[N- (1-oxide) -2-quinolylcarbonyl) -L-asparaginyl] -amino] -4-phenylbutyl] -L-prolinamide as an off-white solid ethyl acetate material; MS: m / e 635 [M + H] &lt; + &gt;.

Example 72

A solution of 98 mg of 3- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-4 (R) -thiazolidinecarboxamide dihydrobromide in 0.5 ml of dry dimethylformamide was stirred and cooled to -10 ° C in an ice / salt bath adding 44 mg of N-ethylmorpholine and then 76 mg of N- (benzyloxycarbonyl) -L-asparagine succinimide ester. The mixture was allowed to warm to room temperature, then stirred overnight. Dimethylformamide was removed by evaporation under reduced pressure and the residue dissolved in chloroform. The solution was washed with water, then dried over sodium sulfate and evaporated. The rubber residue was purified by flash chromatography on silica gel using 4% methanol in dichloromethane to elute. 66 mg of 3- [3 (S) - [[N (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl 4 (R) -thiazolidinecarboxamide was obtained in form of colorless foam. The analytically pure product was obtained by recrystallization from ethyl acetate; MS: m / e 600 [Μ + Η] ⁺ .

3- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert.-butyl-4 (R) thiazolidine-carboxamide dichlorobromide, which was used as starting material, was obtained as follows the way;

(i) A solution of 2.67 g of N-benzyloxycarbonyl-4 (R) -thiazolidinecarboxylic acid in 42 ml of dry tetrahydrofuran was stirred and cooled to -15 ° C in an ice / salt bath with the addition of 1.15 g of N-ethylmorpholine, and 2 minutes later was added 1.87 g of isobutyl chloroformate. After 3 minutes, 0.73 g of tert-butyl amine was added dropwise and the mixture was allowed to warm to room temperature overnight. The tetrahydrofuran was removed by evaporation under reduced pressure and the residue was dissolved in dichloromethane. The solution was washed successively with water, 10% citric acid solution, water, saturated sodium hydrogen carbonate solution and water, then dried over anhydrous sodium sulfate, filtered off and evaporated to give 2.85 g of 3 (benzyloxycarbonyl) -N-tert.- butyl-4 (R) -thiazolidinecarboxamide in solid form, having a melting point at 96-98 ° C after recrystallization from diethyl ether / n-hexane.

(ii) A mixture of 3.2 g of 3- (benzyloxycarbonyl) -N-tert-butyl-4 (R) -thiazolidinecarboxamide and 10 ml of 32% (w / w) hydrobromic hydrogen in ice was stirred for 2 hours at room temperature. The resulting solution was poured into diethyl ether, the separated hydrobromide salt was separated by filtration, washed with diethyl ether and then dissolved in water. This solution was basified by the addition of IM sodium hydroxide solution and extracted twice with dichloromethane. The combined dichloromethane extracts were washed with water, dried over anhydrous sodium sulfate, filtered and evaporated, yielding 1.57 g of N-tert-butyl-4 (R) -thiazolidinecarboxamide in crystalline form with a melting point of 68-71 ° C.

(iii) A solution of 3.2 g of N-tert-butyl-4 (R) -thiazolidinecarboxamide and 5.0 g of 3 (S) - (benzyloxyformamido) -1,2 (S) -epoxy-4-phenylbutane in 130 ml of isopropanol was stirred in argon and heated to 90 ° C under reflux for 3 days. The solution was evaporated under reduced pressure and the residue was purified by flash chromatography on silica gel using diethyl ether / n-hexane (2: 1) for elution. 3.47 g of 3- [3 (S) (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-4 (R) -thiazolidinecarboxamide were obtained. Recrystallization from ethyl acetate gave an analytically pure material with a melting point of 62-65 ° C.

(iv) A mixture of 2 g of 3- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl4 (R) -thiazolidinecarboxamide and 4.1 ml of 32% (w / mass) of hydrogen bromide in ice was stirred for 1 hour at room temperature. The resulting solution was diluted with anhydrous diethyl ether and the product extracted was quickly filtered off and washed with fresh diethyl ether. 2.01 g of 3- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl 4 (R) -thiazolidinecarboxamide dihydrobromide was obtained as a white solid; NMR (300 MHz): δ (D ₂ O) 1.37 (9H, s), 3.0 (2H, m), 3.09 (2H, dq), 3.29 (2H, d), 3.85 (2H, m), 4.16 (1H , m), 4.3 (2H, q) and 7.42 (5H, m) ppm.

Example 73

In the analogous manner as described in Example 21, from 110 mg of 3- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -thiazolidine-carboxamide, 46 mg of 2-naphthoyl chloride and 31 mg of di-isopropylethylamine were obtained by flash chromatography on silica gel using 2% methanol in dichloromethane for elution, 83 mg of N-tert-butyl-3- [2 ( R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] -amino] -4-phenylbutyl] -4- (R) -thiazolidinecarboxamide; MS: m / e 620 [M + H] &lt; ⁺ &gt;.

3- [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-4 (R) thiazolidine-carboxamide used as starting material We have obtained the material as follows:

A mixture of 170 mg of 3- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-4 (R) -thiazolidinecarboxamide and 0.3 ml of 32% (w / w) hydrobromide in ice was stirred for 1 hour at room temperature. The solution was diluted with diethyl ether and the solids recovered filtered off rapidly, washed with diethyl ether and dissolved in water. The solution was basified by the addition of potassium carbonate and extracted three times with chloroform. The combined chloroform extracts were dried over anhydrous sodium sulfate, filtered, and evaporated to give 110 mg of 3- [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N -tert.-butyl4 (R) -thiazolidinecarboxamide; MS: m / e 466 [M + H] +.

Example 74

330 mg solution of N ^ - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -4- (4-tert-butoxyphenyl) -2 (R) -hydroxybutyl] -Nl-tert-butyl -L-prolinamide in 30 ml of ethanol was hydrogenated over 50 mg of 10% palladium-on-carbon catalyst for 3 hours. The catalyst was removed by filtration and the filtrate was evaporated. The residue was dissolved in dichloromethane and treated with 72 mg di-isopropylethylamine and 106 mg 2-naphthoyl chloride in an analogous manner as described in Example 21 to give 165 mg N ^ ^_ [4- (459 tert-butoxyphenyl) - 2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino] butyl] -Nyl-tert-butyl-L-prolinamide as a off-white solid.

Example 75

90 mg solution of N '- [4- (4-tert-butoxyphenyl) -2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -Lasparaginyl] amino] butyl] -Nl-tert .butyl-L-prolinamide in 40 ml of 3.5M hydrogen chloride in ethyl acetate was stirred for 30 minutes at room temperature. The solvent was removed by evaporation and the residue triturated with diethyl ether and filtered to give 80 mg of N2-2 (R) -hydroxy-4 (4-hydroxyphenyl) -3 (S) - [[N- (2-naphthoyl) hydrochloride ) -Lasparaginyl] -amino] butyl] -Nl-tert-butyl-L-prolinamide as a white solid with a melting point of 171-174 ° C.

Example 76

Solution 59 mg 2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1,2,3,4tetrahydropyrido [3,4-b] indole-1- of carboxamide (isomer A) and 71 mg of succinimide ester of N- (benzyloxycarbonyl) -L-asparagine in 3 ml of dry tetrahydrofuran was stirred for 16 hours at room temperature and then evaporated. The residue was dissolved in dichloromethane and the solution was washed twice with water and twice with saturated aqueous sodium hydrogen carbonate solution. The solvent was removed by evaporation under reduced pressure to give 103 mg of crude product. 32 mg of this product was purified by high pressure liquid liquid chromatography and reversed phase using 55% 0.05M ammonium formate in acetonitrile for elution. 9.9 mg of 2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1,2 was obtained. 3,4-tetrahydro-pyrido [3,4-b] -indole-1 (R or S) -carboxamide (isomer A) as a colorless oil; MS: m / e 683 [M + H] &lt; ⁺ &gt;.

2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1,2,3,4-tetrahydro- Pyrido [3,4-b] -indole-1-carboxamide (isomer B) was obtained from 2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl-N-tert-butyl-1 , 2,3,4-tetrahydropyrido [3,4-b] indole-1 (R or S) -carboxamide (isomer B) in an analogous manner to the preparation of isomer A described above with the same purification method, whereby get a colorless oil; MS: m / e 683 [M + H] &lt; ⁺ &gt;.

2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl-N-tert-butyl-1,2,3,4-tetrahydropyrido [3,4b] indole-1-carboxamide, which We used them as starting materials as follows:

(i) Ice-cooled solution consisting of 3.0 g of 1,2,3,4-tetrahydropyrido [3,4-b] indole-1-carboxylic acid in 7 ml of 2M sodium hydroxide solution, 17 ml of water and 17 ml of dioxane, 12.5 ml of 2M sodium hydroxide solution and a solution of 3.6 ml of benzylchloroformiate in 9 ml of dioxane were added alternately and in small quantities. After the addition was complete, the mixture was stirred at room temperature overnight, then the dioxane was removed by evaporation under reduced pressure. The resulting solution was diluted with water, washed twice with diethyl ether, acidified with 25 ml of IM sulfuric acid and extracted three times with ethyl acetate. The organic extracts were combined, washed twice with water and evaporated, yielding 4.54 g of 2-benzyloxycarbonyl-1,2,3,4-tetrahydropyrido [3,4-b] indole-1-carboxylic acid;

MS: m / e 351 [M + H] +, which was used without further purification.

(ii) A solution of 4.54 g of 2-benzyloxycarbonyl-1,2,3,4-tetrahydropyrido [3,4-b] indole-carboxylic acid in 45 ml of anhydrous tetrahydrofuran was cooled in an ice / acetone bath while protecting with a drying tube. . 1.82 ml of isobutyl chloroformate and 1.82 ml of N-ethylmorpholine were added and the mixture was stirred for 10 minutes then 2.1 ml of tert-butylamine was added. The mixture was then stirred for 40 minutes at 0 ° C and 45 minutes at room temperature, then diluted with ethyl acetate and washed twice with water, twice with 0.5M sodium hydroxide solution and again with water. The solution was dried over magnesium sulfate and evaporated to dryness. Recrystallization from ethyl acetate and nhexane gave 1.21 g of tert-butyl 2-benzyloxycarbonyl-1,2,3,4-tetrahydropyrido [3,4-b] indole-1-carboxamide; MS: m / e 406 [M + H] ⁺ . 0.76 g of identical material was obtained by subjecting the mother liquor to chromatography on silica gel using ethyl acetate / n-hexane (1: 1) for elution.

(iii) 1.62 g of tert-butyl 2-benzyloxycarbonyl-1,2,3,4-tetrahydro-pyrido [3,4-b] indole-carboxamide were dissolved in 10 ml of 45% hydrobromic acid solution in acetic acid. After 30 minutes, the solution was evaporated and the residue was collected in water. After filtration, the filtrate was washed three times with diethyl ether and then neutralized by the addition of saturated sodium hydrogen carbonate solution in water. The resulting product was extracted from a cloudy aqueous layer using ethyl acetate. The combined organic extracts were dried over anhydrous magnesium sulfate and evaporated to give 0.84 g of tert-butyl 1,2,3,4tetrahydro-pyrido [3,4-b] indole-1-carboxamide as a white, crystalline material with a melting point 144 ° C.

(iv) Solution of 0.60 g of tert-butyl 1,2,3,4-tetrahydro-pyrido [3,4-b] indole-1-carboxamide and 0.66 g of 3 (S) - (benzyloxyformamido) -1,2- ( S) -Epoxy-4-phenylbutane in 20 ml of methanol was refluxed in argon for 16 hours and then evaporated to give a clear oil. These two diastereomeric products were determined by flash chromatography on silica gel using n-hexane / ethyl acetate (3: 1) for elution. We obtained 268 mg of isomer A 2- [3 (S) - (benzyl-oxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1,2,3,4-tetrahydropyrido- [3. 4-b] indole-1-carboxamide;

MS: m / e 569 [M + H] + and 65 mg of isomer B of the same compound; MS: m / e 569 [M + H] + (v) 150 mg solution of 2- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1,2 , 3,4-Tetrahydro-pyrido [3,4-b] indole-1-carboxamide (isomer A) in 5 ml of ethanol was hydrogenated at a pressure of 3.4 bar above 10% palladium on charcoal at 20 ° C for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 120 mg of 2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl-N-tert-butyl-1,2,3,4tetrahydropyrido [3,4-b] indole-1-carboxamide (isomer A); MS: m / e 435 [M + H] ⁺ 2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl-N-tert-butyl-1,2,3,4-tetrahydropyrido [ 3,4bindol-1-carboxamide (isomer B), MS: m / e 435 [M + H] ⁺ , was produced in an analogous manner from 2- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy- 4-Phenylbutyl] -N-tert-butyl-1,2,3,4-tetrahydropyrido [3,4-b] indole-1-carboxamide (isomer B).

Example 77

In an analogous manner as described in Example 70, 154 mg of N-tert-butyl-1-2 (R) hydroxy-3 (S) - [[N- (quinolylcarbonyl) -L-asparaginyl] amino-4 was obtained -phenylbutyl] -2 (S) -piperidinecarboxamide and 51 mg of 3-chloroperbenzoic acid gave 114 mg of 1-oxide of N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[ N- (2-quinolylcarbonyl) -L-asparaginyl] -amino] butyl] -2-piperidinecarboxamide as a white solid;

MS: m / e 633 [M + H] + ·

Example 78

In the analogous manner as described in Example 27, 230 mg of 1- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl- 2 (S) -piperidinecarboxamide, 139 mg of 3-benzyloxy-2-naphthalenecarboxylic acid, 113 mg of dicyclohexylcarbodiimide, 68 mg of hydroxybenzotriazole and 58 mg of N-ethyl-morpholine were obtained after chromatography on silica gel using G elution system, 242 mg 1 [3 (S) '[[N- (3-benzyloxy-2-naphthoyl) -L-asparaginyl] amino] -2 (R) -4-phenyl-butyl] -N-tert-butyl-2 (S) - piperidinecarboxamide in foam form; MS: m / e 722 [M + H] ⁺ ·

Example 79

A solution of 181 mg of 1- [3 (S) - [[N- (3-benzyloxy-2-naphthoyl) -L-asparaginyl] amino] -2 (R) hydroxy-4-phenyl-butyl] -N-tert.- butyl-2 (S) -piperidinecarboxamide in 5 ml of ethanol was hydrogenated over 10% palladium on charcoal at 20 ° C under atmospheric pressure for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated. Trituration with diethyl ether gave 110 mg of N-tert-butyl-1- [3 (S) - [[N- (3-hydroxy-2-naphthoyl) -Lasparaginyl] amino] -2 (R) -hydroxy- 4-Phenylbutyl] -2 (S) -piperidine-carboxamide, in the form of a pale yellow solid, MS: m / e 632 [M + H] + ·

Example 80

In an analogous manner as described in Example 27, 400 mg of cis-1- [3 (S) -amino2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro-2 (R or S) -quinolinecarboxamide, 226 mg of N- (benzyloxycarbonyl) -L-asparagine, 226 mg of dicyclohexylcarbodiimide, 135 mg of hydroxybenzotriazole and 115 mg of N-ethylmorpholine, obtained after chromatography on silica gel using dichloromethane / methanol (94: 6: e): 94: 225 mg cis-1 [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-decahydro-2 (R or S ) -quinolinecarboxamide, in the form of a white, solid material; MS: m / e 650 [M + H] ⁺ .

Cis-1- [3 (S) -amino] 2 (R) -hydroxy-4-phenyl-butyl] -N-tert-butyl-decahydro-2 (R or S) quinolinecarboxamide, which was used as starting material , we prepared as follows:

(i) A solution of 2.376 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenylbutane and 1.904 g of cis-N-tert-butyl-decahydro-2 (R, S) -quinolinecarboxamide in 32 ml of ethanol was stirred for 24 hours at 80 ° C. 0.474 g of 3 (S) - (benzyloxyformamido) -1,2- (S) epoxy-4-phenylbutane in two portions were added and the mixture stirred at 80 ° C for 5 hours. The solvent was removed by evaporation and the residue was chromatographed on silica gel using dichloromethane / methanol (97.5: 2.5) to elute to give 1.17 g of cis-1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy- 4-Phenylbutyl] N-tert-butyl decahydro-2 (R or S) quinolinecarboxamide (isomer A) as white solid diethyl ether / n-hexane, MS: m / e 536 [M + H] ⁺ and 1.146 g cis-1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro-2 (R or S) -quinolinecarboxamide (isomer B) in the form of pale- yellow tires; MS: m / e 536 [M + H] ⁺ .

(ii) A solution of 0.535 g of cis-1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] N-tert-butyl-decahydro-2 (R or S) -quinoline-carboxamide (isomer B) in 25 ml of ethanol was hydrogenated above 10% palladium on charcoal at 20 ° C and under atmospheric pressure for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 400 mg of cis-1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro2 (R or S) -quinoline-carboxamide in the form of a colorless gum.

Example 81

Solution 561 mg trans-2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyldecahydro- (4aR, 8aS) -isoquinoline-3 (S) -carboxamide and 372 mg of N- (benzyloxycarbonyl) -L-asparagine in 20 ml of dry tetrahydrofuran was cooled using an ice / salt mixture. 189 mg of hydroxybenzotriazole, 161 mg of N-ethylmorpholine and 317 mg of dicyclohexylcarbodiimide were added and the mixture was stirred for 16 hours. The mixture was then diluted with ethyl acetate and filtered. The filtrate was washed with aqueous sodium hydrogen carbonate solution and sodium chloride solution. The solvent was removed and the residue chromatographed on silica gel using dichloromethane / methanol (9: 1) to elute to give 434 mg of trans-2- [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro- (4aR, 8aS) isoquinoline-3 (S) -carboxamide as a white solid methanol / diethyl ether material; MS: m / e 650 [M + H] ⁺ .

Trans-2- [3 (S) -amino] 2 (R) -hydroxy-4-phenyl-butyl] -N-tert-butyl-decahydro- (4aR, 8aS) isoquinoline-3 (S) -carboxamide, which we used it as starting material, we obtained as follows:

(i) A solution of 440 mg of trans-N-tert-butyl-decahydro- (4aR, 8aS) -isoquinoline-3 (S) carboxamide and 549 mg 3 (S) - (benzyloxyformamido) -1,2- (S) - of epoxy-4-phenylbutane in 6 ml of ethanol was stirred for 7 hours at 60 ° C. 54 mg of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenylbutane was added and the solution was stirred at 20 ° C for 16 hours. The solvent was removed by evaporation and the residue was chromatographed on silica gel using an H elution system to give 771 mg of trans-2- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] - N-tert-butyl-decahydro- (4aR, 8aS) -isoquinoline-3 (S) -carboxamide, as a white solid;

MS: m / e 536 [M + H] +.

(ii) A solution of 747 mg of trans-2- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro- (4aR, 8aS) -isoquinoline-3 The (S) -carboxamide in 40 ml of ethanol was hydrogenated over 10% palladium on charcoal at 20 ° C and pressurized for 5 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 561 mg of trans-2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] N-tert-butyl-decahydro-4aR, 8aS ) -isoquinoline-3 (S) -carboxamide as a pale-brown solid.

Example 82

In an analogous manner as described in Example 27, 276 mg of 1- [3 (S) - [[Lasparaginyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 ( S) -piperidinecarboxamide, 167 mg of 1-benzyloxy-2-naphthalenecarboxylic acid, 81 mg of hydroxybenzotriazole, 69 mg of N-ethylmorpholine and 136 mg of dicyclohexylcarbodiimide were obtained after chromatography on silica gel using dichloromethane / methanol (9: 1) 1- [3 (S) - [[N- (1-benzyloxy-2-naphthoyl) -L-asparaginyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S ) -piperidinecarboxamide as a white solid methanol / diethyl ether material; MS: m / e 722 [M + H] &lt; ⁺ &gt;.

Example 83

In the analogous manner as described in Example 79, from 119 mg of 1- [3 (S) - [[N (benzyloxy-2-naphthoyl) -L-asparaginyl] amino] 2 (R) -hydroxy-4-phenyl -butyl] -N-tert-butyl2 (S) -piperidinecarboxamide obtained by chromatography on activated magnesium silicate using dichloromethane / methanol (9: 1) for elution, 67 mg of N-tert-butyl-1 [2 (R) -hydroxy-3 (S) - [[N- (1-hydroxy-2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] 2 (S) -piperidine-carboxamide as a white solid;

MS: m / e 631 [M + H] +.

Example 84

In the analogous manner as described in Example 27, from 276 mg of 1- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide, 183 mg of 3- (benzyloxycarbonyl) -2-naphthalenecarboxylic acid, 81 mg of hydroxybenzotriazole, 138 mg of N-ethylmorpholine and 136 mg of dicyclohexylcarbodiimide, after chromatography on silica gel using system J for elution, gave 98 mg , butyl-1- (S) - [1 (S) - (2,3-dihydro-1,3-dioxo-1H-benz [f] isoindol-2-yl) -3-carbamoylpropionamido] -2 (R ) hydroxy-4-phenylbutyl] -2 (S) -piperidinecarboxamide;

MS: m / e 642 [M + H] +

Example 85

A solution of 650 mg of N- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-L-propylamino and 538 mg of N- (benzyloxycarbonyl) -S-methyl-L-cysteine in 20 ml of dry tetrahydrofuran was cooled in an ice / salt mixture. 270 mg of hydroxybenzotriazole, 230 mg of N-ethylmorpholine and 412 mg of dicyclohexylcarbodiimide were added and the mixture was stirred for 16 hours. The mixture was diluted with ethyl acetate and filtered. The filtrate was washed with aqueous sodium hydrogen carbonate solution and sodium chloride solution. The solvent was removed by evaporation and the residue was chromatographed on silica gel using an elution system G to give 800 mg of N2- [3 (S) - [[N (benzyloxycarbonyl) -S-methyl-L-cysteinyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide; MS: m / e 585 [M + H] &lt; + &gt;.

The N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide used as starting material was obtained in an analogous manner as starting material in Example 33, only using N1-tert-butyl-L-prolinamide in paragraph (iii) instead of N ^ -phenyl-L-prolinamide.

Example 86

193 mg solution of N2- [3 (S) - [[N- (benzyloxycarbonyl) -S-methyl-L-cysteinyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L- of prolinamide in 2 ml of methanol was cooled to -70 ° C. A solution of 62 mg of 3-chloroperbenzoic acid in 5 ml of methanol was added and the mixture was stirred for 30 minutes at -70 ° C. The solvent was then removed by evaporation and the residue chromatographed on silica gel using an elution system G to give 62 mg of N2- [3 (S) - [[N- (benzyloxycarbonyl) -3- (methylsulfinyl) -L-alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide as a mixture of 1: 1 diastereomers; MS: m / e 601 [M + H] &lt; ⁺ &gt;.

Example 87

A solution of 450 mg of N2- [3 (S) - [[N- (benzyloxycarbonyl) -S-methyl-L-cysteinyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide in 4 ml of methanol was cooled to -70 ° C. 166 mg of 3-chloroperbenzoic acid were added portionwise over 10 minutes, the solution was stirred for 15 minutes at -70 ° C and then allowed to warm to 20 ° C. The solution was cooled again to -70 ° C, 33 mg of 3-chloroperbenzoic acid was added and the mixture was stirred at -70 ° C for 30 minutes. The solvent was removed by evaporation and the residue was layered between dichloromethane and 2N sodium hydroxide solution. The organic phase was evaporated and the residue was chromatographed on silica gel using an elution system G to give 100 mg of N ^{2 -} oxide N ² - [3 (S) - [[N- (benzyloxycarbonyl) -3 (methylsulfinyl) -L- alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide (diastereomer A), MS: m / e 617 [M + H] ⁺ and 154 mg N ² - of N ² - [3 (S) - [[N (benzyloxycarbonyl) -3- (methylsulfinyl) -L-alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nlert-butyl-L-prolinamide oxide (diastereomer B); MS: m / e 617 [M + H] ⁺ .

Example 88

A solution of 154 mg of N ² -oxide N ² - [3 (S) - [[N- (benzyloxycarbonyl) -3- (methylsulfinyl) -Lanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert .butyl-L-prolinamide (two diastereomers) in 5 ml of methanol was treated with 84 mg of 3-chloroperbenzoic acid and the solution was stirred for 16 hours at 20 ° C. The solvent was removed by evaporation and the residue was layered between dichloromethane and 2M sodium hydroxide solution. The organic phase was evaporated and the residue was crystallized from ethyl acetate / n-hexane to give 28 mg of N ² -oxide N ² - [3 (S) - [[N- (benzyloxycarbonyl) -3- (methylsulfonyl) -alanil] monohydrate. ] -amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide;

MS: m / e 633 [M + H] +.

Example 89

In an analogous manner as described in Example 27, 400 mg of cis-1- [3 (S) -amino2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro-2 (R or S) -quinolinecarboxamide, 266 mg of N- (benzyloxycarbonyl) -L-asparagine, 226 mg of dicyclohexylcarbodiimide, 135 mg of hydroxybenzotriazole and 115 mg of N-ethylmorpholine were obtained after chromatography on silica gel using dichloromethane / methanol (94: 6: e): 94: 225 mg cis-1 [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-decahydro-2 (R or S ) -quinolinecarboxamide as a white solid; MS: m / e 650 [M + H] ⁺ .

The cis-1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro-2 (R or S) quinolinecarboxamide used as starting material obtained as follows:

(i) A solution of 2.376 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenylbutane and 1.904 g of cis-N-tert-butyl-decahydro-2 (R, S) -quinolinecarboxamide in 32 ml of ethanol was stirred for 24 hours at 80 ° C. 0.474 g of 3 (S) - (benzyloxyformamido) 1,2- (S) -epoxy-4-phenylbutane were added in two portions and the mixture was stirred at 80 ° C for 5 hours. The solvent was removed by evaporation and the residue chromatographed on silica gel using dichloromethane / methanol (97.5: 2.5) to elute to give 1.17 g of cis-1- [3 (S) 67 (benzyloxyformamido) -2 (R ) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro-2 (R or S) quinolinecarboxamide (isomer A) as a white solid diethyl ether / nhexane; MS: m / e 536 [M + H] ⁺ and 1.146 g of cis-1- [3 (S) - (benzyloxyformamido) -2 (R) hydroxy-4-phenylbuty] -N-tert-butyl-decahydro-2 (R or S) -quinolinecarboxamide (isomer B) in the form of pale yellow gum; MS: m / e 536 [M + H] ⁺ .

(ii) A solution of 0.535 g of cis-1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] N-tert-butyl-decahydro-2 (R or S) -quinolinecarboxamide (isomer B) was hydrogenated in 25 ml of ethanol over 10% palladium on charcoal at 20 ° C and under atmospheric pressure for 16 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 400 mg of cis-1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro2 (R or S) -quinolinecarboxamide as a colorless gum.

Example 90

In the analogous manner as described in Example 27, 27 mg of N3 (S) - [[- L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -L-proline was obtained from 27 mg and 11.3 mg of 2-indolecarboxylic acid gave 15 mg of N- [3 (S) - [[N- (2-indolylcarbonyl) -Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] tert-butyl ester - L-proline; MS: m / e 592 [M + H] &lt; ⁺ &gt;.

Example 91

In an analogous manner as described in Example 72, from 240 mg of 3- [3 (S) -amino-2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-tetrahydro-2H-1,3 -thiazine-4 (R and S) -carboxamide and 234 mg of N- (benzyloxycarbonyl) -L-asparagine succinimide ester, 162 mg of 3- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino were obtained ] -2 (R) -hydroxy-4-phenylbutyl] -Nert-t-butyl-tetrahydro-2H-1,3-thiazine-4 (R and S) -carboxamide as a mixture of two diastereomers. Flash chromatography on silica gel using 3% methanol in dichloromethane for elution gave 20 mg less of the polar diastereomer (isomer A); MS: m / e 614 [M + H] ⁺ , and using 5% methanol in dichloromethane 32 mg more polar diastereomer (isomer B); MS: m / e 614 [M + H] ⁺ .

3- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-tetrahydro-2H-1,3-thiazine-4 (R and S) -carboxamide used as starting material was obtained from the known tetrahydro-2H-1,3-thiazine-4 (R and S) -carboxylic acid compound by N-benzyloxycarbonylation in a known manner and then by reaction in an analogous manner as described in 72 (i) - (iv), and by alkalinizing the dihydrobromide obtained with sodium hydrogen carbonate solution.

Example 92

0.122 g of N- (benzyloxycarbonyl) -3-cyano-L-alanine was dissolved in 2 ml of dry dimethylformamide. The solution was stirred and cooled in an ice / salt bath and treated with 0.066 g of hydroxybenzotriazole and 0.1 g of dicyclohexylcarbodiimide. The mixture was stirred for 5 minutes and then treated with 0.163 g of N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nlterc-butyl-L-prolinamide in 2.5 ml of dry dichloromethane. The mixture was allowed to warm to room temperature and then stirred overnight. The resulting dicyclohexylurea was determined by filtration and washed with methylene chloride. The combined filtrate and washings were evaporated at 40 ° C in vacuo to give an oil which was triturated between ethyl acetate and water. The organic phase was washed successively with aqueous sodium hydrogen carbonate solution and saturated sodium chloride solution and dried over sodium sulfate. The solvent was removed by evaporation to give an oil which was chromatographed on silica gel using 14% methanol in dichloromethane for elution. In this way 50 mg of product was recrystallized from diethyl ether / n-hexane to give 0.045 g of N ^ - [3 (S) - [[N (benzyloxycarbonyl) -3-cyano-L-alanyl hemihydrate ] amino] -2 (R) -hydroxy-4-phenyl-butyl-N1-tert, butyl-L-prolinamide in solid form, m.p. 65-70 ° C (softening).

The N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide used as starting material was obtained as follows:

(i) A solution of 0.425 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenylbutane and 0.244 g of tert-butylamide L-proline in 10 ml of dry isopropanol was heated at 80 ° for 20 hours. C. The solvent was removed by evaporation in vacuo and the residue was chromatographed on silica gel using 5% methanol in dichloromethane for elution. 0.44 g of N- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N1-tert-butyl-L-pyrrolinamide was obtained in the form of a white foam.

(ii) A solution of 0.46 g of N ^ '[3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nlterc.-butyl-L-prolinamide in 40 ml of ethanol was hydrogenated over 40 mg of 10% palladium-on-charcoal at room temperature and atmospheric pressure for 1.5 hours to give 0.33 g of N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl- Lprolinamide in the form of a rubber that crystallizes during the state.

Example 93

0.167 g of N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide was dissolved in 10 ml of dry dimethylformamide and the solution was stirred at 0 ° C while adding 0.191 g of N- (benzyloxycarbonyl) -L-phenyl-glycine succinimide ester. The resulting solution was stirred for 1 hour at 0 ° C and then stored overnight at 4 ° C. The solution was then evaporated in vacuo and the residue was layered between ethyl acetate and water. The organic phase was washed with saturated aqueous sodium hydrogen carbonate solution and saturated sodium chloride solution. The aqueous phases were back-extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate and evaporated to give a gum which was chromatographed on silica gel using acetone / adichloromethane (1: 1) for elution. The combined fractions containing the product were evaporated to give a rubber which was re-evaporated with diethyl ether to give 0.127 g of solid material. This material was extracted with dichloromethane and the combined organic phases were evaporated to give a solid material which was triturated with diethyl ether. 0.05 g of N2- [3 (S) - [[N (benzyloxycarbonyl) -L-phenylglycyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-pyrrolinamide are obtained in solid form with a melting point of 101-103 ° C.

Example 94

In an analogous manner to that described in Example 92, 0.15 g of N- (benzyloxycarbonyl) -L-phenylalanine, 0.066 g of hydroxybenzotriazole, 0.1 g of dicyclohexylcarbodiimide and 0.166 g of N - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N ^4- tert-butyl-L-pyrrolinamide, yielding 0.1 g of N 2 - [3 (S) - [[N- (benzyloxycarbonyl) -L-phenylalanyl] amino] 2 (R) -hydroxy- 4-Phenylbutyl] -Nl-tert-butyl-L-prolinamide as a white solid with a melting point of 78-80 ° C.

Example 95

In an analogous manner as described in Example 92, 0.152 g of N- (benzyloxycarbonyl) -3-cyclohexyl-L-alanine, 0.066 g of hydroxybenzotriazole, 0.1 g of dicyclohexylcarbodiimide and 0.166 g of N - [3 (S) -amino- 2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide gave 0.1 g of N 2 - [3 (S) - [[- N- (benzyloxycarbonyl) -3-cyclohexyl-lanyl] amino] - 2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide, m.p. 71-75 ° C.

Example 96

In an analogous manner as described in Example 92, 0.1 g of N- (benzyloxycarbonyl) -L-asparagine, 0.05 g of hydroxybenzotriazole, 0.078 g of dicyclohexylcarbodiimide and 0.17 g of 1- [3 (S) -amino-2 (R) - hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R and S) -piperazinecarboxamide, after trituration with diethyl ether, gave 0.11 g of a white solid. The resulting material was purified by flash chromatography on silica gel using 10% methanol in dichloromethane for elution. The product first eluted (isomer A) was 0.043 g of 1- [3 (S) - [[N (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] - 4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R or S) -piperazinecarboxamide; MS: m / e 697 [M + H] &lt; ⁺ &gt;. The other eluted product (isomer B) was 0.007 g 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4- (tert .-butoxycarbonyl) -N-tert-butyl-2 (R or S) -piperazinecarboxamide; MS: m / e 697 [M + H] &lt; ⁺ &gt;.

1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R and S) -piperazinecarboxamide used as starting material, we obtained as follows:

(i) 0.65 g of 2-piperazinecarboxylic acid was dissolved in a mixture of 5 ml of water and 5 ml of dioxane, treated with 0.42 g of sodium hydrogen carbonate and stirred for 5 minutes. 1.09 g of di (tert-butyl) dicarbonate was added and the mixture was stirred overnight. The mixture was concentrated by evaporation and the residue was thoroughly extracted with ethyl acetate. The process was repeated with pH 6 and pH 4. The aqueous layer with pH 4 was then extracted with nbutanol. The combined organic extracts were dried over anhydrous sodium sulfate and evaporated to give 0.34 g of 4- (tert-butoxycarbonyl) -2-piperazinecarboxylic acid as a pale yellow solid, mp 226-229 ° C.

(ii) 0.1 g of 4- (tert-butoxycarbonyl) -piperazine-2-carboxylic acid was dissolved in 10 ml of 1N sodium hydroxide solution, cooled to 0 ° C and treated with 0.2 g of benzyl chloroformate. The mixture was allowed to warm to room temperature and then stirred overnight. The mixture was extracted with diethyl ether. The aqueous phase was then acidified to pH 4 with 2M hydrochloric acid and extracted with ethyl acetate to give 0.06 g of 1- (benzyloxycarbonyl) -4- (tert-butoxycarbonyl) -2-piperazinecarboxylic acid as a white solid; MS: m / e 365 [M + H] &lt; ⁺ &gt;.

(iii) 0.285 g of 1- (benzyloxycarbonyl) -4- (tert-butoxycarbonyl) -2-piperazinecarboxylic acid was dissolved in 10 ml of dry tetrahydrofuran and cooled to -15 ° C with stirring. Then 0.09 g of N-ethylmorpholine and then 0.107 g of isobutyl chloroformate were added. This mixture was stirred for 5 minutes and then 0.2 g of tert.butylamine was added dropwise. The stirring was continued overnight and the mixture was allowed to reach room temperature for some time. The solvent was removed by evaporation to give a light brown oil which was stripped to ethyl acetate and water. The organic phase was washed successively with 10% citric acid solution, sodium hydrogen carbonate solution and saturated sodium chloride solution, then dried over anhydrous sodium sulfate. The solvent was removed by evaporation to give 0.185 g of 1- (benzyloxycarbonyl) -4- (tert-butoxycarbonyl) -N-tert-butyl-2-piperazinecarboxamide as a light brown oil; MS: m / e 420 [M + H] &lt; ⁺ &gt;.

(iv) 1.1 g of 1- (benzyloxycarbonyl) -4- (tert-butoxycarbonyl) -N-tert-butyl-2-piperazinecarboxamide were dissolved in 40 ml of ethanol. 0.1 g of 10% palladium on charcoal was added and the mixture was hydrogenated at room temperature and atmospheric pressure for 2 hours. The catalyst was removed by filtration. The filtrate was evaporated, yielding 0.74 g of crude product, which was purified by flash chromatography on silica gel using 5% methanol in dichloromethane for elution. Evaporation of the solvents gave 0.44 g of 4- (tert-butoxycarbonyl) -N-tert-butyl-2-piperazinecarboxamide as an oil; MS: m / e 286 [M + H] +.

(v) 0.395 g of 4- (tert-butoxycarbonyl) -N-tert-butyl-2-piperazinecarboxamide was dissolved in 50 ml of dry isopropanol and treated with 0.413 g of 3 (S) - (benzyloxyformamido) -1, 2- ( S) -Epoxy-4-phenylbutane. The mixture was stirred for 72 hours at room temperature. The solvent was removed by evaporation to give a brown semi-liquid substance which was purified by flash chromatography on silica gel using 5% methanol in dichloromethane for elution. 0.234 g of 1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R or S) was obtained -piperazinecarboxamide as a mixture of diastereomers.

(vi) 0.234 g of 1- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R and S) -piperazinecarboxamide was dissolved in 20 ml of ethanol and treated with 100 mg of 10% palladium on charcoal. The mixture was hydrogenated at room temperature and at atmospheric pressure for 2.5 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 0.17 g of 1- [3 (S) -amino2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl -2 (R and S) of piperazinecarboxamide, which was used without further purification.

Example 97

0.035 g 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert. -butyl-2 (R or S) -piperazinecarboxamide (isomer A) was dissolved in 5 ml of ethyl acetate and treated with 5 drops of saturated hydrogen chloride solution in ethyl acetate. The mixture was allowed to stand at room temperature for 1 hour and then treated to give, after recrystallization from ethanol / diethyl ether, 0.024 g of 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] - hydrochloride. 2 (R) hydroxy-4-phenyl-butyl] -N-tert-butyl-2 (R or S) -piperazinecarboxamide (isomer A) with a melting point of 175 to 180 ° C.

Example 98

0.01 g of 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert. -butyl-2 (R or S) -piperazinecarboxamide (isomer B) was treated with a saturated solution of hydrogen chloride in ethyl acetate as described in Example 97 to give 0.007 g of hydrochloride 1- [3 (S) - [[N (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl (2R or S) -piperazinecarboxamide (isomer A) as a very hygroscopic solid; MS: m / e 597 [M + H] &lt; ⁺ &gt;.

Example 99

In an analogous manner as described in Example 92, 0.091 g of N- (benzyloxycarbonyl) -3-cyano-L-alanine, 0.05 g of hydroxybenzotriazole, 0.076 g of dicyclohexylcarbodiimide and 0.164 g of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R, S) -piperazinecarboxamide gave 0.075 g of a mixture of 60:40 diastereomers 1- [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2- piperazine carboxamide in the form of a light brown solid with a melting point of 80-85 ° C.

Example 100

In an analogous manner as described in Example 92, 0.108 g of 3-cyano-N- (2-naphthoyl) -L-alanine, 0.054 g of hydroxybenzotriazole, 0.083 g of dicyclohexylcarbodiimide and 0.18 g of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R, S) -piperazinecarboxamide gave 0.015 g of a mixture of 1: 1: 1: 1 isomers of 4- (tert .73 Butoxycarbonyl) -N-tert-butyl-1- [3 (S) - [[3-cyano-N- (2-naphthoyl) -L-alanyl] -amino] -2 (R) hydroxy-4- phenylbutyl] -2 (R, S) -piperazinecarboxamide in solid form;

MS: m / e 699 [M + H] &lt; + &gt;.

The 3-cyano-N- (2-naphthoyl) -L-alanine used as starting material was obtained as follows:

0.114 g of 3-cyano-L-alanine was dissolved in 5 ml of 1N sodium hydroxide solution and treated with 0.285 g of 2-naphthoyl chloride at 0 ° C. Acidification with 2M hydrochloric acid and flash chromatography on silica gel using 25% methanol in dichloromethane for elution gave 0.049 g of 3-cyano-N- (2-naphthoyl) -L-alanine, m.p. 95-100 ° C.

Example 101

In the analogous manner as described in Example 92, 0.048 g of 1- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -4-tert-butyl2 (R and S) -piperazinecarboxamide, 0.015 g of quinaldic acid, 0.012 g of hydroxybenzotriazole and 0.018 g of dicyclohexylcarbodiimide were obtained by flash chromatography on silica gel using 10% methanol in dichloromethane for elution, 0.004 g of pure ega, i.e. 4- (tert-butoxycarbonyl) -N-tert-butyl-1- [2 (R) hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino ] butyl] -2 (R or S) piperazinecarboxamide, MS: m / e 718 [M + H] ⁺ and 0.003 g of isomer B, i.e. 4- (tert-butoxycarbonyl) -N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] -2 (R or S) -piperazinecarboxamide, MS: m / e 718 [M + H] ⁺ .

1- [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -4 tert-butyl-2 (R and S) - The piperazinecarboxamide used as starting material was prepared as follows:

0.06 g 1- [3 (S) - [[- N- (benzyloxycarbonyl) -L-asparaginyl] amino-2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -N-tert. -butyl-2 (R, S) -piperazinecarboxamide (i.e.

The product of Example 96 was dissolved in 20 ml of ethanol before deciding the isomers A and B). 0.03 g of palladium on charcoal was added and the mixture was hydrogenated at room temperature and atmospheric pressure for 2 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 0.048 g of 1- [3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4- (tert-butoxycarbonyl) -4-tert-butyl 2-piperazinecarboxamide, which was used without further purification.

Example 102

In an analogous manner as described in Example 92, 0.151 g of N- (benzyloxycarbonyl) -3-cyano-L-alanine, 0.082 g of hydroxybenzotriazole, 0.126 g of dicyclohexylcarbodiimide and 0.212 g of 1- [3 (S) -amino-2 are obtained (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) piperidinecarboxamide, obtained after recrystallization from diethyl ether / n-hexane 0.085 g 1 [3 (S) - [[- N- (benzyloxycarbonyl)] -3-cyano-L-alanyl] amino-2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-2 (S) -piperidinecarboxamide, in the form of a white solid, mp 74-77 ° C.

Example 103

In an analogous manner to that described in Example 92, 0.372 g of N- (benzyloxycarbonyl) -L-aspartic acid, 0.189 g of hydroxybenzotriazole, 0.288 g of dicyclohexylcarbodiimide and 0.54 g of N2- [3 (S) -amino-2 (R) -hydroxy-4- (2-naphthyl) -butyl] N1-tert-butyl-L-prolinamide obtained after recrystallization from isopropanol / n-hexane (1: 4) 0.105 g of N2 [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy- (2-naphthyl) butyl] -Nl-tert-butyl-L-prolinamide, mp 149-151 ° C.

The N2- [3 (S) -amino-2 (R) -hydroxy-4- (2-naphthyl) -butyl] -Nl-tert-butyl-L-prolinamide used as starting material was obtained as follows:

(i) 5 g of 3- (2-naphthyl) -L-alanine and 0.93 g of sodium hydroxide in 12 ml of water were cooled to 0 ° C and stirred while adding a solution of 1.4 g of sodium hydroxide in 9 ml of water and 5 ml of benzyl chloroformate in 10 minutes. Stirring was continued for 2 hours, then the mixture was allowed to warm to room temperature. The mixture was diluted with water and extracted with diethyl ether. The aqueous layer was acidified with 4 ml of concentrated hydrochloric acid and extracted with ethyl acetate. The ethyl acetate extracts were washed again with water. The combined ethyl acetate extracts were dried over sodium sulfate, filtered, evaporated and triturated with petroleum ether (boiling point 40-60 ° C) to give 7.5 g of N- (benzyloxycarbonyl) -3- (2-naphthyl) -L-alanine with melting point 109-111 OC.

(ii) A solution of 7.5 g of N- (benzyloxycarbonyl) -3- (2-naphthyl) -L-alanine in 20 ml of dry tetrahydrofuran was stirred at -8 ° C and treated with 3.5 ml of N-ethylmorpholine and then with 4 ml of isobutyl chloroformate. which was added dropwise over the course of 10 minutes. The mixture was stirred at -8 ° C for another 20 minutes and cold (0 ° C) anhydrous diethyl ether was added. The resulting white precipitate was filtered off, and the cold filtrate was added dropwise to 100 ml of a stirred, cool (-8 ° C) solution of diazomethane in diethyl ether. We stopped cooling and stirred the solution for 3 hours. Water was added while stirring. The organic phase was washed successively with water, sodium hydrogen carbonate solution and saturated sodium chloride solution and then dried over sodium sulfate. The solvent was removed by evaporation to give a yellow oil, from which, after evaporation with light petroleum (boiling point 40-60 ° C), 9.4 g of a solid material was obtained. The material was triturated with diethyl ether and refrigerated overnight at 4 ° C, yielding 3.2 g of benzyl [3-diazo-1 (S) - [(2-naphthyl) methyl] -2-oxopropyl] carbamate.

(iii) 3.13 g of benzyl [3-diazo-1 (S) - [(2-naphthyl) methyl] -2-oxopropyl] carbamate were dissolved in 200 ml of anhydrous diethyl ether and stirred while the hydrogen chloride gas was bubbled through the solution. After 1 hour, the original sediment became granular, leaving excess hydrogen chloride. The solvent was removed by evaporation at room temperature and the resulting white solid was dried and the hydrogen chloride was removed in vacuo over sodium hydroxide for 2 hours. The obtained benzyl [3-chloro-1 (S) - [(2-naphthyl) -methyl] -2oxopropylcarbamate was immediately used in the next step.

(iv) The above benzyl [3-chloro-1 (S) - [(2-naphthyl) -methyl] -2-oxopropyl] carbamate was dissolved in 100 ml of 10% aqueous tetrahydrofuran, cooled to 0 ° C and treated with 0.456 g sodium borohydride, which was carefully added in solid form. The mixture was stirred for 1 hour at 0 ° C and then at room temperature overnight. The mixture was evaporated to give a white solid, which was stripped between dichloromethane and water. The stirred mixture was carefully acidified to pH 1 with concentrated hydrochloric acid. The phases were separated and the aqueous phase was re-extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate and evaporated, yielding 4.325 g of crude product with a melting point of 155-160 ° C. The crude product was extracted with boiling n-hexane. After removal of the solvent by evaporation, the residue was recrystallized from ethyl acetate / n-hexane to give 1.046 g of pure benzyl [3-chloro-2 (S) -hydroxy-1 (S) - [(2-naphthyl) methyl] propyl ] of carbamate with a melting point of 173-174 ° C.

(v) 1.02 g of benzyl [3-chloro-2 (S) -hydroxypropyl-1 (S) - [(2-naphthyl) methyl] propyl] carbamate was stirred in 40 ml of ethanol with 4 ml of 0.7M potassium hydroxide solution in ethanol within 0.75 hours. The solvent was removed by evaporation and the residue was layered between dichloromethane and water. The organic phase was dried over anhydrous sodium sulfate and evaporated to give a white solid which was recrystallized from ethyl acetate / n-hexane. 0.879 g of 3 (S) - (benzyloxyformamido) -1,2 (S) epoxy-4- (2-naphthyl) butane was obtained as a white solid with a melting point of 115-116 ° C.

(vi) 0.465 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4- (2-naphthyl) butane and 0.251 g of tert-butylamide L-proline in 10 ml of dry isopropanol were heated 23 hours at 80 ° C and further treated as described in Example 92 (i) to give 0.483 g of N ^ [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4- (2-naphthyl) ) -butyl] -Nl-tert-butyl-L-prolinamide in the form of a white foam with a melting point of about 75-85 ° C.

(vii) A solution of 0.725 g of N2- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4- (2-naphthyl) butyl] -Nl-tert-butyl-L-prolinamide was dissolved in 25 ml. ethanol and hydrogenated at room temperature and at atmospheric pressure above 0.5 g of 10% palladium on charcoal for 20 hours. The catalyst was filtered off and the filtrate was evaporated, yielding 0.54 g of N- [3 (S) -amino-2 (R) -hydroxy-L- (2-naphthyl) -butyl] -Nl-tert-butyl-L -Prolinamide in the form of white foam, which was used without further purification.

Example 104

In an analogous manner to that described in Example 92, 0.133 g of N- (benzyloxycarbonyl) -L-asparagine, 0.068 g of hydroxybenzotriazole, 0.103 g of dicyclohexylcarbodiimide and 0.16 g of N2- [3 (S) -amino-2 (R) - hydroxy-4-phenylbutyl] -Nl-tert-butyl4 (R) -hydroxy-L-prolinamide, only using dichloromethane instead of ethyl acetate as the separating agent and 20% methanol in dichloromethane for chromatography and re-evaporation with diethyl ether 0.1 g of N2- [3 (S) - (benzyloxycarbonyl) -Lasparaginyl-amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -hydroxy-L-prolinamide with melting point 115 ° C.

N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -hydroxy-L-prolinamide, which was used as starting material, was obtained as follows:

(i) 2.65 g of N- (benzyloxycarbonyl) -4 (R) -hydroxy-L-proline was dissolved in 10 ml of dry tetrahydrofuran and cooled to -10 ° C by stirring with a magnetic stirrer. 1.15 g of N-ethylmorpholine were added followed by 1.36 g of isobutylchloroformate immediately. The mixture was stirred for 30 minutes at -10 ° C and then 2.19 g of tert-butylamine was added. Stirring at -10 ° C was continued for 1 hour and then the mixture was allowed to stand for 2 hours to warm to room temperature and allowed to stand for another 2 hours. The solvent was removed by evaporation in vacuo and the residue was layered between ethyl acetate and water. The organic layer was washed with 10% citric acid solution and sodium hydrogen carbonate solution and then dried over sodium sulfate. The solvent was removed by evaporation to give a solid material which was triturated with diethyl ether and filtered off. 2.23 g of the crude product were recrystallized from ethyl acetate / diethyl ether to give 1.57 g of N2- (benzyloxycarbonyl) -Nl-tert-butyl-4 (R) -hydroxy-L-prolinamide with a melting point of 128- 130 ° C.

(ii) 0.224 g of N2- (benzyloxycarbonyl) -Nl-tert-butyl-4 (R) -hydroxy-L-prolinamide was dissolved in 20 ml of ethanol and hydrogenated over 50 mg of 10% palladium on carbon at room temperature and at atmospheric pressure for 2 hours. The catalyst was filtered off and the filtrate was evaporated to give 0.13 g of N1-tert-butyl-4 (R) -hydroxy-L-prolinamide, which was used in the next step without further purification.

(iii) 0.13 g of N1-tert-butyl-4 (R) -hydroxy-L-prolinamide and 0.208 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenylbutane in 10 ml of dry isopropanol was heated at 80 ° C for 24 hours. Further treatment in an analogous manner as described in Example 33 (iii), after trituration with diethyl ether, 0.236 g of N2- [3 (S) (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] was obtained - N1-tert-butyl-4 (R) -hydroxy-Lprolinamide in the form of a white gelatinous solid with a melting point at 135 ° C.

(iv) 0.226 g of N - [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -hydroxy-L-prolinamide was dissolved in 20 ml of ethanol and hydrogenated above 40 mg of 10% palladium on charcoal at room temperature and atmospheric pressure for 2 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 0.16 g of N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -hydroxyL -Prolinamide in the form of rubber, which was used without further purification.

Example 105

In an analogous manner to that described in Example 92, only using 0.067 g of N- (benzyloxycarbonyl) was obtained using dichloromethane instead of ethyl acetate as a delamination agent, using 20% methanol in dichloromethane for chromatography and re-evaporation with diethyl ether. Of L-asparagine, 0.034 g of hydroxybenzotriazole, 0.052 g of dicyclohexylcarbodiimide and 0.08 g of N - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (S) -hydroxy -L-prolinamide gave 0.03 g of N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl- 4 (S) hydroxy-L-prolinamide in the form of a pale yellow solid with a melting point of about 140 ° C.

N ^ - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (S) -hydroxy-L-prolinamide, which was used as starting material, was obtained as follows:

(i) 1.7 g of N- (benzyloxycarbonyl) -4 (S) -hydroxy-L-proline were dissolved in 15 ml of dry dimethylformamide with stirring and the solution was cooled to 0 ° C. 0.817 g of Nhydroxysuccinimide was added and the mixture was stirred until it reached 20 ° C. The mixture was then stirred overnight at room temperature. The resulting dicyclohexylurea was filtered off and the filtrate cooled to -10 ° C. Then 2 ml of tert.butylamine was added with stirring. Stirring was continued and at the same time allowed to warm to room temperature and continued stirring overnight. The solids recovered were filtered off, the filtrate was evaporated in vacuo to give a gum which was stripped between ethyl acetate and water. The organic layer was washed with 10% citric acid solution and then with saturated sodium hydrogen carbonate solution. The aqueous phases were back-extracted twice with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate and evaporated to give a solid material which was purified by flash chromatography on silica gel using 5% methanol in dichloromethane for elution. 1.36 g of crude product was obtained, which was recrystallized from ethyl acetate / diethyl ether / petroleum ether (boiling point 40-60 ° C) (1: 4: 4) and then stored at 4 ° C in a refrigerator overnight. Thus, 1.127 g of N, N * (benzyloxycarbonyl) -N1-tert-butyl-4 (S) -hydroxy-L-prolinamide were obtained with a melting point of 131-132 ° C.

(ii) 0.224 g of N2 '(benzyloxycarbonyl) -Nl-tert-butyl-4 (S) -hydroxy-L-prolinamide were hydrogenated in an analogous manner as described in Example 104 (ii) to give 0.135 g N1-tert-butyl-4 (S) -hydroxy-L-prolinamide as a gum-like solid, which was used in the next step without prior purification.

(iii) 0.135 g of N1-tert-butyl-4 (S) -hydroxy-L-prolinamide and 0.208 g of 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4-phenyl-butane in 10 ml of dry ethanol was stirred for 4 days at room temperature. The mixture was further treated as described in Example 92 (i), except that chromatography was performed using 10% methanol in dichloromethane for elution. 0.11 g of N2- [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N1-tert-butyl-4 (S) -hydroxy-L-prolinamide are obtained in the form of a foam.

(iv) 0.11 g of N - [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl4 (S) -hydroxy-L-prolinamide was dissolved in 10 ml. ethanol and hydrogenated above 20 mg 10% palladium on charcoal at room temperature and atmospheric pressure for 2 hours. The catalyst was filtered off and the filtrate was evaporated to give 0.08 g of N2- [3 (S) -amino2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (S) -hydroxy-L-prolinamide in a form of rubber that was used without further purification.

Example 106

In an analogous manner to that described in Example 92, but using dichloromethane instead of ethyl acetate as a delamination agent, using 20% methanol in dichloromethane for chromatography and re-evaporation with diethyl ether, 0.105 g of N- (benzyloxycarbonyl) is obtained -L-asparagine, 0.054 g of hydroxybenzotriazole, 0.082 g of dicyclohexylcarbodiimide and 0.17 g of N ^ - [3 (S) -amino-2 (R) hydroxy-4-phenylbutyl] -4 (R) -tert.-butoxyformamido-N1-tert .-Butyl-L-prolinamide gave 0.045 g of N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl-] amino] -2 (R) hydroxy-4-phenylbutyl] -4 (R) - tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide in the form of a dirty white solid with a melting point of 170-175 ° C.

N2- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -4 (R) -tert-butoxyformamido-N1-tert-butyl-L-prolinamide, which was used as starting material, we have obtained as follows:

(i) 0.32 g of N2- (benzyloxycarbonyl) -Nl-tert-butyl-4 (S) -hydroxy-L-prolinamide was dissolved in 5 ml of dry pyridine, cooled to 0 ° C, stirred and treated dropwise with 0.82 ml of methanesulfonyl chloride . The solution was stirred at 0 ° C for another 2 hours. The resulting mixture was poured into a mixture of ice and water and then extracted with ethyl acetate. The combined organic extracts were washed with 2M hydrochloric acid and saturated sodium hydrogen carbonate solution, then dried over anhydrous sodium sulfate. Evaporation gave 0.5 g of N, N - (benzyloxycarbonyl) -N1-tert-butyl-4 (S) - (methanesulfonyloxy) -L-prolinamide as an oil, which was used without further purification.

(ii) 0.5 g of N2- (benzyloxycarbonyl) -Nl-tert, -butyl-4 (S) - (methanesulfonyloxy) -L-prolinamide were dissolved in 10 ml of dry dimethylformamide and treated with 0.330 g of sodium azide. This heterogeneous mixture was stirred and heated at 75 ° C for 18 hours. The mixture was evaporated in an oil vacuum pump to give a solid material which was decomposed into ethyl acetate and water. The ethyl acetate phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed by evaporation to give 0.345 g of N, N - (benzyloxycarbonyl) -4 (R) -azido-N, N-butyl-L-prolinamide as a rubber.

(iii) 0.345 g of N- (benzyloxycarbonyl) -4 (R) -azido-N1-tert-butyl-L-prolinamide were dissolved in 5 ml of dry tetrahydrofuran and the solution was evaporated. The rubber residue was dissolved in 10 ml of dry tetrahydrofuran and treated with 0.262 g of triphenylphosphine in a nitrogen atmosphere. The resulting solution was allowed to stand in a nitrogen atmosphere at room temperature for 18 hours. 0.027 g of water was added and the solution was allowed to stand at room temperature for 24 hours. The solvent was then removed by evaporation and the rubber-shaped residue was layered between water and diethyl ether. The aqueous phase was back-extracted with diethyl ether. The aqueous phase was then evaporated to give 0.09 g of N2- (benzyloxycarbonyl) 4 (R) -amino-N1-tert-butyl-L-prolinamide as a gum. The mixture was allowed to stand overnight at room temperature and the diethyl ether extracts were combined and evaporated to give 0.7 g of oil, which was chromatographed on silica gel using 10% methanol in dichloromethane for elution to give 0.16 g of N ^ - ( benzyloxycarbonyl) 4 (R) -amino-N1-tert-butyl-L-prolinamide in the form of a rubber.

(iv) 0.21 g of N2- (benzyloxycarbonyl) -4 (R) -amino-N1-tert-butyl-L-prolinamide were dissolved in a mixture of 5 ml of dioxane and 5 ml of water. 0.056 g of sodium hydrogen carbonate was added and a solution was added to which 0.144 g of di (tert-butyl) dicarbonate was added. The mixture thus obtained was stirred at room temperature overnight. The solvents were removed by evaporation and the residue was layered between water and diethyl ether. The aqueous phase was back-extracted with diethyl ether and then with ethyl acetate. The combined organic extracts were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The dried extracts were combined and evaporated to give 0.27 g of N ^ (benzyloxycarbonyl) -4 (R) - (tert-butoxy-formamido) -Nl-tert-butyl-L-prolinamide as a dirty white foam.

(v) 0.25 g of N2- (benzyloxycarbonyl) -4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide was dissolved in ethanol and hydrogenated over 0.1 g of 10% palladium on carbon at room temperature and below atmospheric pressure for 4 hours. The catalyst was filtered off and the filtrate was evaporated, yielding 0.17 g of 4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide as a vitreous material, which was used without further purification.

(vi) 0.17 g of 4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide and 0.178 g 3 (S) - (benzyloxyformamido) -1,2- (S) -epoxy-4 -phenylbutane in 15 ml of dry ethanol was stirred and the resulting solution was allowed to stand at room temperature for 4.5 days. The solution was further heated at 40 ° C for 96 hours. Further treatment, as described in Example 92 (i), except that 10% methanol in dichloromethane was used for eluting with chromatography, 0.230 g of N2 - [3 (S) - (benzyloxyformamido) -2 (R) was obtained -hydroxy-4-phenylbutyl] -4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide in gum form.

(vii) 0.22 g of N ² - [3 (S) - (benzyloxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L- of prolinamide was dissolved in 10 ml of ethanol and hydrogenated over 0.05 g of 10% palladium on charcoal at room temperature and atmospheric pressure for 2 hours. The catalyst was removed by filtration and the filtrate was evaporated, yielding 0.17 g of N ^ - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -4 (R) (tert-butoxyformamido) -Nl -terc.-butyl-L-prolinamide in the form of a gum, which was used without further purification.

Example 107

In an analogous manner as described in Example 27, 162 mg of 2 - [[3 (S) - [[Lasparaginyl] amino] -2 (R) -hydroxy-4-phenyl] butyl] -N-tert.- butyl-1,2,3,4-tetrahydro-pyrido [3,4-b] -indole-1 (R or S) -carboxamide (isomer B), 55 mg quinaldic acid, 43 mg 1hydroxybenzotriazole, 0.04 ml N-ethylmorpholine and 66 mg of dicyclohexylcarbodiimide were obtained by chromatography on silica gel using 3% methanol in ethyl acetate for elution, 95 mg of N-tert-butyl-1,2,3,4-tetrahydro-2- [2 (R) -hydroxy-4 -phenyl-3 (S) - [[N- (2quinolylcarbonyl) -L-asparaginyl] amino] butyl] pyrido [3,4-b] indole-1 (R or S) -carboxamide; MS: m / e 707 [M + H] &lt; ⁺ &gt;.

2 - [[3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenyl] butyl] -N-tert-butyl-1,2,3,4-tetrahydropyrido [3 , 4-b] indole-1 (R or S) -carboxamide, which was used as starting material, was obtained by hydrogenation of 2 [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] -2 ( R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1,2,3,4-tetrahydro-pyrido [3,4-b] indole-1 (R or S) -carboxamide.

Example 108

A solution of 154 mg of trans-2- [3 (S) - [(L-asparaginyl) amino] -2 (R) -hydroxy-4-phenyl-butyl] N-tert-butyl-decahydro- (4aR, 8aS) - isoquinoline-3 (S) -carboxamide and 52 mg quinaldic acid in 6 ml of dry tetrahydrofuran were cooled in an ice / salt mixture. 41 mg of hydroxybenzotriazole, 35 mg of N-ethylmorpholine and 68 mg of dicyclohexylcarbodiimide were added and the mixture was stirred for 64 hours. The mixture was diluted with ethyl acetate and filtered. The filtrate was washed with aqueous sodium hydrogen carbonate solution and sodium chloride solution and then evaporated. The residue was chromatographed on silica gel using dichloromethane / methanol (9: 1) to elute to give 50 mg of transN-tert-butyl-decahydro-2- [2 (R) -hydroxy-4-phenyl- 3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] - (4aR, 8aS) -isoquinoline-3 (S) -carboxamide as a white solid; MS: m / e 671 [M + H] ⁺ .

Trans-2- [3 (S) - [(L-asparaginyl) amino] -2 (R) -hydroxy-4-phenyl-butyl] -N-tert-butyldecahydro- (4aR, 8aS) -isoquinoline-3 ( The S) -carboxamide used as starting material was obtained by hydrogenation of trans-2- [3 (S) - [[N- (benzyloxycarbonyl) -Lasparagenyl] amino] -2 (R) -hydroxy-4-phenylbutyl ] -N-tert-butyl-decahydro- (4aR, 8aS) isoquinoline-3 (S) -carboxamide.

Example 109

A solution of 1.02 g of 1- [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) piperidinecarboxamide and 685 mg of N- (tert-butoxycarbonyl) -S -methyl-L-cysteine in 7 ml of dry tetrahydrofuran was cooled in an ice / salt mixture. 394 mg of hydroxybenzotriazole, 335 mg of N-ethylmorpholine and 661 mg of dicyclohexylcarbo-diimide were added and the mixture was stirred for 3 hours. The mixture was diluted with ethyl acetate and filtered. The filtrate was washed with aqueous sodium hydrogen carbonate solution and sodium chloride solution and evaporated. The solvent was removed by evaporation and the residue was chromatographed on silica gel using dichloromethane / methanol (96: 4) for elution to give 630 mg of 1- [3 (S) - [[N- (tert-butoxycarbonyl) -L -cysteinyl] amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide as a white solid; MS: m / e 565 [M + H] &lt; ⁺ &gt;.

Example 110

A solution of 650 mg of N-tert-butyl-1- [3 (S) - (L-cysteynyl) -amino-2 (R) -hydroxy-4-phenylbutyl] -2 (S) -piperidinecarboxamide and 242 mg of quinaldic acid cooled in ice / salt mixture. 189 mg of hydroxybenzotriazole, 161 mg of N-ethyl-morpholine and 317 mg of dicyclohexylcarbodiimide were added and the mixture was stirred for 64 hours. The mixture was diluted with ethyl acetate, filtered and the filtrate was evaporated. The residue was layered between dichloromethane and aqueous sodium hydrogen carbonate solution. The organic phase was washed with sodium chloride solution and then evaporated. The residue was chromatographed on silica gel using dichloromethane / methanol (19: 1) for elution to give 350 mg of N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lcysteinyl] amino] butyl] -2 (S) -piperidinecarboxamide as a white solid; MS: m / e 620 [M + H] &lt; + &gt;.

N-tert-butyl-1- [3 (S) - [(L-cysteinyl) amino] -2 (R) -hydroxy-4-phenylbutyl] -2 (S) -piperidinecarboxamide, which was used as starting material , we obtained as follows:

Solution 930 mg 1-3 (S) - [[N- (tert-butoxycarbonyl) -L-cysteinyl] amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-2 (S) -piperidinecarboxamide in 7 ml of trifluoroacetic acid was stirred at 20 ° C for 1 hour. The mixture was then evaporated to dryness and the residue was layered between dichloromethane and aqueous sodium hydrogen carbonate solution. The organic phase was evaporated to give 650 mg of N-tert-butyl-1- [3 (S) - [(Lc-phenyl) -amino] -2 (R) -hydroxy-4-phenylbutyl] -2 (S) -piperidinecarboxamide in the form of a colorless gum; MS: m / e [M + H] +.

Example 111 mg of freshly distilled acetyl chloride was added to a pre-cooled and stirred solution of 12 mg of hydrochloride 4 (R) -amino-N ² - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 ( R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide and 21 mg of sodium hydrogen carbonate in 0.5 ml of water and 0.25 ml of dimethylformamide. The mixture was stirred vigorously for 5 hours at 0 ° C and then allowed to stand at room temperature overnight. The mixture was diluted with water and extracted with dichloromethane. The combined dichloromethane extracts were evaporated to give a gum which was purified by flash chromatography on silica gel using 20% methanol in dichloromethane for elution. 2 mg of 4 (R) -acetylamino-N ² - [3 (S) [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert was obtained. butyl-L-prolinamide in semi-solid form; MS: m / e 639 [M + H] &lt; ⁺ &gt;.

Hydrochloride 4 (R) -amino-N ² - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] -amino] 2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl The -L-prolinamide used as starting material was prepared as follows:

mg N ² - [3 (S) - [[N ² - (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4 (R) - (tert-butoxyformamido) -N 1 of tert-butyl-L-prolinamide was dissolved in 0.5 ml of saturated hydrogen chloride solution in ethyl acetate and allowed to stand for 1 hour at room temperature. The solution was evaporated and the residue triturated with diethyl ether and stored at 0 ° C overnight. The recovered solid was filtered off and washed with diethyl ether to give 19 mg of 4 (R) -amino-N ² - [3 (S) - [[N- (benzyloxycarbonyl) L-asparaginyl] amino] -2 (2) hydrochloride. R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide as a solid material, m.p. 206-210 ° C.

The following Example illustrates the preparation of a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable acid addition salt thereof as the active ingredient:

Example A

The aqueous solution of the active ingredient was sterile filtered and stirred with heating with a sterile gelatin solution containing phenol as a preservative, using a quantity such that 1.00 ml of the solution obtained contained 3.0 mg of the active ingredient, 150.0 mg of gelatin, 4.7 mg of phenol and distilled water to 1.0 ml. Bottles of 1.0 ml were filled with this mixture under aseptic conditions.

Claims (20)

PATENT APPLICATIONS 1. Compounds of general formula R ² R ⁴ (0) _B 7 and II t> V / '“Χ / ^C \ / ^CH - ^N CH NH C R 'in which n represents a number 0 or 1; R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl, monoaralkylcarbamoyl, cinamoyl or α-aralkoxycarbonylaminoalkanoyl and R4 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, a cyclic imide group of formula Oh N— (a) O in which P and Q together represent the aromatic system; R ^ represents alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylsulfinylalkyl, carbamoylalkyl or alkoxycarbonylalkyl, or when n represents 0, R ^ represents alkylthioalkyl, or when n denotes number 1, R ^ may also represent alkylsulfonylalkyl; R ⁴ represents alkyl, cycloalkyl, cycloalkylalkyl, aryl or aralkyl; R ^ represents hydrogen and R ^ represents hydroxy, or R ^ and R ^ together represent oxo; R 6 and R ⁸ together represent a trimethylene or tetramethylene group which may be substituted by hydroxy, alkoxycarbonylamino or acylamino, or in which one of the -CH 2 - groups is replaced by -NH-, -N (alkoxycarbonyl) -, -N (acyl ) - or -S-, or which carries a fused cycloalkane, aromatic or heteroaromatic ring; and R4 represents alkoxycarbonyl, monoalkylcarbamoyl, monoaralkylcarbamoyl, monoarylcarbamoyl or a group of the formula _C / ^H \ / ^co \ --co CH l · ⁰ NH-R (b) wherein R1 and R4 each represent alkyl; and their pharmaceutically acceptable acid addition salts. Compounds according to claim 1, in which n is 0 (zero), R 4 represents alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylthioalkyl, carbamoylalkyl or alkoxycarbonylalkyl and R? and R4 together represent a trimethylene or tetramethylene group in which one of the -CH2- groups is replaced by -NH- or -S-, or which may carry a fused cycloalkane, aromatic or heteroaromatic ring. Compounds according to claim 1 or 2, in which R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl or α-aralkoxycarbonylaminoalkanoyl, preferably benzyloxycarbonyl, 2-naphthoyl, 1-hydroxyyl -2-naphthoyl, 3-benzyloxy-2-naphthoyl, 2-quinolylcarbonyl or 3quinolylcarbonyl, and R 4 represents hydrogen. Compounds according to any one of claims 1 to 3, in which R1 represents alkyl, cyanolakyl, alkylthioalkyl or carbamoylalkyl, preferably cyanomethyl, methylthiomethyl or carbamoylmethyl. Compounds according to any one of claims 1 to 4, wherein R 1 represents aralkyl, preferably benzyl. Compounds according to any one of claims 1 to 5, in which R 8 represents hydrogen and R 4 represents hydroxy. Compounds according to any one of claims 1 to 6, in which -N (R 7) -CH (R 8) (R 9) represents one of the following groups wherein R 1 has the meaning given in claim 1, R 12 represents hydrogen, hydroxy, alkoxycarbonylamino or acylamino, Rl3 represents hydrogen, alkoxycarbonyl or acyl, m denotes 1 or 2 and p denotes 1 or 2, preferably a group of formula (c) wherein Rl2 represents hydrogen and m denotes 2, or Rl2 represents tert-butoxycarbonylamino and m denotes 1, a group of formula (d) in which R 1 represents tert-butoxycarbonyl, a group of formula (e) in which m denotes 1, a group of formula (f) in which m and p both denote 1, or a group of formula (g), (i) or (j). Compounds according to any one of claims 1 to 7, in which R 1 represents alkoxycarbonyl, monoalkylcarbamoyl or a group of formula (b) as claimed in claim 1, preferably tert-butoxycarbonyl, isobutylcarbamoyl, tert-butylcarbamoyl or a group of formula ( b) as claimed in claim 1, wherein R1 represents sec-butyl and R4 represents isobutyl. Compounds according to any one of claims 1 to 8, in which R 1 represents benzyloxycarbonyl, 2-naphthoyl, 1-hydroxy-2-naphthoyl, 3-hydroxy-2-naphthoyl, 3-benzyloxy-2 naphthoyl, 2-quinolylcarbonyl or 3 -quinolylcarbonyl and R4 represents hydrogen, R4 represents cyanomethyl, methylthiomethyl or carbamoylmethyl, R4 represents benzyl, R8 represents hydrogen and R4 represents hydroxy, and -N (R7) -CH (R8) (R9) represents a group of the formula (c) stated in claim 7, wherein R12 is hydrogen and m is 2, or R12 is tert-butoxycarbonylamino and m is a group of formula (d) as claimed in claim 7, wherein tert-butoxycarbonyl is a group with formula (e) given in claim 7, in which m denotes 1, a group of formula (f) shown in claim 7, in which m and p denote 1, or a group of formulas (g), (i) or (j ) as claimed in claim 7, and R4 represents tert-butoxycarbonyl, isobutylcarbamoyl, tert-butylcarbamoyl, or a group of formula (b) as claimed in claim 1, in which R ^ represents sec-butyl and R ^ represents isobutyl. 10. N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-L-prolinamide or N2- [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] -2 (R or S) -hydroxy-4-phenylbutyl] -N 1-isobutyl-L-prolinamide. Compounds according to claim 1, selected from the group consisting of: N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nl-tert-butyl-4 (R) -thiazolidine-carboxamide, N-tert-butyl-1- [2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] 2 (S) -piperidinecarboxamide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -Nert-butyl-octahydro- (3aS, 6aS) -cyc openta [b] pyrrole-2 (S) -carboxamide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -Nert-butyl-2 (S) -piperidine-carboxamide, 2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1,2,3,4-tetrahydropyrido [3,4-b] indole-1-carboxamide, N-tert-butyl-3- [2 (R) -hydroxy-3 (S) - [[N- (2-naphthoyl) -L-asparaginyl] amino ] -4-phenylbutyl] 4 (R) -thiazolidinecarboxamide, N1-tert-butyl-N ² - [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] -L-prolinamide N ^ -oxide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2- (R) -hydroxy-4-phenylbutylN-tert-butyl-2 (S) -piperidinecarboxamide , 1- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -4 (tert-butoxycarbonyl) -N-tert-butyl -2- (R or S) -piperazinecarboxamide, 1- [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L * alanyl] amino] -2- (R) -hydroxy-4-phenylbutyl] 4- (tert-butoxycarbonyl) -N-tert-butyl-2 (R or S) -piperazinecarboxamide, N2- [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] 4 (R) - (tert-butoxyformamido) -Nl-tert-butyl-L-prolinamide, 1- [3 (S) - [[N- (3-benzyloxy- 2-naphthoyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] N- (tert-butyl-2 (S) -piperidine-carboxamide, N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] -2-piperidinecarboxamide 1-oxide, N-tert-butyl-1- [3 (S) - [[N- (3-hydroxy-2-naphthoyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -2 (S) -piperidinecarboxamide, trans-2- [3 (S) - [[N-benzyloxycarbonyl) -L-asparaginyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-decahydro- (4aR, 8aS) -isoquinoline-3 (S) -carboxamide, 4- (tert-butoxycarbonyl) -N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2quinolylcarbonyl) -L-asparaginyl] amino] butyl ] -2 (R or S) piperazinecarboxamide, N-tert-butyl-1- [2 (R) -hydroxy-3 (S) - [[N- (1-hydroxy-2-naphthoyl) -L-asparaginyl] amino] -4-phenylbutyl] -2 (S) -piperidinecarboxamide, trans-N-tert-butyl-decahydro-2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] - (4aR, 8aS) -isoquinoline-3 (S) -carboxamide and N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) ~ [[N- (2-quinolylcarbonyl) -Lcysteinyl] amino] butyl] -2 (S) -piperidinecarboxamide. 12. N-tert-butyl-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -Lasparaginyl] amino] butyl] -2 (S) - piperidinecarboxamide. 13. N-tert-butyl-octahydro-1- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) L-asparaginyl] amino] butyl] - (3aS , 6aS) -cyclopenta [b] pyrrole-2 (S) -carboxamide. 14. N-tert-butyl-1,2,3,4-tetrahydro-2- [2 (R) -hydroxy-4-phenyl-3 (S) - [[N- (2-quinolylcarbonyl) -L- asparaginyl] amino] butyl] pyrido [3,4-b] indole-1 (R or S) -carboxamide. 15. Compounds of general formula, CH '-N' R 6 and? 5 Λ in which R represents hydrogen or a group CH R4 represents alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylthioalkyl, alkylsulfinylalkyl, carbamoylalkyl or alkoxycarbonylalkyl; R4 represents alkyl, cycloalkyl, cycloalkylalkyl, aryl or aralkyl; R ^ represents hydrogen and R ^ represents hydroxy, or R ^ and R ^ together represent oxo; R? and R &apos; together represent a trimethylene or tetramethylene group which may be substituted by hydroxy, alkoxycarbonylamino or acylamino, or in which one of the -CH2- groups may be replaced by -NH-, * N (alkoxycarbonyl) -, N (acyl) - or -S-, or which bears a fused cycloalkane, aromatic or heteroaromatic ring; and R4 represents alkoxycarbonyl, monoalkylcarbamoyl, mono-aralkylcarbamoyl, monoarylcarbamoyl or a group of formula NH CO / \ z \ —CO CH NHi ¹⁰ .11 (b); VI in which each of and represents alkyl. 16. Compounds of the general formula ^r1 - \ ^X CH NH .C CH ₂ -N z \ in which n represents a number 0 or 1; R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl, monoaralkylcarbamoyl, cinamoyl or α-aralkoxycarbonylaminoalkanoyl and R4 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, or R3 represents hydrogen, a cyclic imide group of formula Oh O in which P and Q together represent the aromatic system; R 4 represents alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylsulfinylalkyl, carbamoylalkyl or alkoxycarbonylalkyl, or when n represents 0, R 4 may also represent alkylthioalkyl, or when n represents 1, R 4 may also represent alkylsulfonylalkyl ; R4 represents alkyl, cycloalkyl, cycloalkylalkyl, aryl or aralkyl; R 4 represents hydrogen and R 4 represents hydroxy or R 8 and R 4 together represent oxo; R? 'And R ’together represent a trimethylene or tetramethylene group which may be substituted by amino; and R4 represents alkoxycarbonyl, monoalkylcarbamoyl, monoaralkylcarbamoyl, monoarylcarbamoyl or a group of the formula NH —CO with ^co \ CH NH · (b), 11 in which each of βΐθ represents alkyl. Amino acid derivatives according to any one of claims 1 to 14 for use as a therapeutically active substance. Amino acid derivatives according to any one of claims 1 to 14 for use in the treatment or prophylaxis of viral infections. A process for the manufacture of a compound according to any one of claims 1 to 14, characterized in that it comprises (a) for the preparation of compounds of formula I, in which n is 0, a compound of general formula R I CH R ' CH -N / \ z ² H ₂ N c R ⁵ V Z / 'CH' .9 in which R4, R- ', R ^, R ^, R ^ and R ^ have the meaning indicated in claim 1, reacts with an acid of the general formula R ¹ —N CH.3 -COOH III in which R 1, R 2 and R 4 have the meaning indicated in claim 1, or with its reactive derivative, or (b) for the preparation of a compound of formula I, in which n is 0, R ³ represents hydrogen and R 4 represents hydroxy, reducing a compound of formula I in which n is 0 and R ³ and r 6 together represent oxo, or (c) to prepare a compound of formula I in which n is 0 and R ³ represents alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl , alkylsulfonyl, arylsulfonyl, cinamoyl or α-aralkoxycarbonylaminoalkanoyl, and R ² represents hydrogen, or R 1 and R ² together with the nitrogen atom to which they are attached represent a cyclic imide group of formula (a) as claimed in claim 1, a compound of the general formula IV in which R ³ , R 4, R ³ , r ⁸ , ⁷ , r 8 and _n 9 have the meaning shown _in claim 1, reacted with an agent which gives the group alkanoyl, cycloalkylcarbonyl, aralkanoyl, aroyl, heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl, cinamoyl or α-aralkoxycarbonylaminoalkanoyl, or by an agent forming the cyclic imide group of formula (a) shown in claim 1, or (d) for the preparation of a compound of formula I, in which n is 0 and R1 represents monoaralkylcarbamoyl and R ² represents hydrogen, a compound of Formula IV shown above is reacted with a compound of the general formula r1'-N = C = OV in which R1 represents aralkyl, or (e) to prepare a compound of formula I in which R ³ represents allylsulfinylalkyl and n is 0. oxidize a compound of formula I in which R ³ represents alkylthioalkyl and n denotes 0, or (f) to prepare a compound of formula I in which n denotes 1, oxidize a compound of formula I in which n denotes 0, or (g) for the preparation of a compound of formula I, in which n is 1 and R4 is representative a is an aromatic N-heterocyclylcarbonyl N-oxide group, and R ² represents hydrogen, oxidizing a compound of formula I in which n denotes 1 and R ^d represents an aromatic N-heterocyclylcarbonyl group and R ² represents hydrogen, or (h) to prepare a compound with of formula I in which n represents 1 and R 4 represents alkylsulfonylalkyl, oxidize a compound of formula I in which n represents 1 and R 4 represents alkylsulfinylalkyl, or (i) to prepare a compound of formula I in which R ^d represents carboxy-substituted aroyl, hydroxy-substituted aroyl or hydrocinamoyl, and R ² represents hydrogen, a catalytically hydrogenated compound of formula I in which R 1 represents benzyloxycarbonylsubstituted aroyl, benzyloxy-substituted aroyl or cinamoyl, and R ² represents hydrogen, or (j) for the preparation of a compound of formula I in which R 4 represents imidazol-4-yl and / or R 4 represents hydroxy-substituted aryl or hydroxy-substituted aralkyl and / or R ² and R 4 together represent a trimethylene or tetrameth group In which one group -CH2- is replaced by -NH-, a compound of formula I is treated in which R1 represents 1 (benzyloxycarbonyl) -imidazol-4-yl and / or R4 represents tert-butoxy-substituted aryl or tert-butoxy-substituted aralkyl and / or R ² and R 1 together represent a trimethylene or tetramethylene group in which one group -CH 2 - is replaced by a -N (tert-butoxycarbonyl) - group with a strong acid, or (k) for the preparation of a compound of formula I in which R ² and R 1 together represent a trimethylene or tetramethylene group which is substituted with acylamine, or in which one group -CH 2 - is substituted with -N (acyl) -, acylates a compound of the general formula H ^{2 R} / J _Z o A ^X CH NH C hz \ «<°> n R ⁷ ' V, VI in which n, R 1, R ⁷ , r ³ have _? r4 r5 _? r6 j _n r9 the meaning given _in claim 1 and R ⁷ 'and R &apos; together represent a trimethylene or tetramethylene group which is substituted with amino or in which one group -CH2- is replaced by -NH-, and / or (l) if desired , separate the mixture of diastereoisomeric racemates into diastereoisomeric racemates or optically pure diastereoisomers and / or (m) optionally separate the mixture of diastereoisomers into optically pure diastereoisomers and / or (n) optionally convert the compounds of formula I into pharmaceutically acceptable salts. Process according to claim 19, characterized in that compounds of formula I in which n is 0, R ⁷ represents alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, cyanoalkyl, alkylthioalkyl, carbamoylalkyl or alkoxycarbonylalkyl and R ⁷ and R 6 together represent a trimethylene or tetramethylene group in which one group -CH2- may be replaced by -NH- or -S-, or which may bear a fused cycloalkane, aromatic or heteroaromatic ring, and their pharmaceutically acceptable acid addition salts are prepared on the basis of embodiments (a ), (b), (c), (d) and / or (n) of the process.