CZ229594A3 - Peptides, process of their preparation, their use and pharmaceutical compositions based thereon - Google Patents

Abstract

Compounds of formula (I) and pharmaceutically acceptable salts thereof and bioprecursors therefore wherein R<1> is C1-C6 alkyl, C3-C8 cycloalkyl, aryl, heterocyclyl or CONR<9>R<10>; R<2> is C1-C6 alkyl, C3-C8 cycloalkyl(C1-C4)alkyl, aryl(C1-C4)alkyl or heterocyclyl(C1-C4)alkyl; R<3> is C1-C6 alkyl, C3-C8 cycloalkyl, C3-C8 cycloalkyl(C1-C4)alkyl, aryl(C1-C4)alkyl, aryl(C2-C4)-alkenyl, heterocyclyl(C1-C4)alkyl or heterocyclyl(C2-C4)-alkenyl; R<4> is C1-C6 alkyl, C3-C8 cycloalkyl, aryl or heterocyclyl; each of R<5>, R<6>, R<7> and R<8> is independently H, C1-C6 alkyl or C3-C8 cycloalkyl; or R<5> and R<6>, or R<7> and R<8> may be joined together to form a 3 to 8 membered carbocyclic ring; X is a 4-10 membered mono or bicyclic heterocyclic group containing carbon ring atoms and one ring nitrogen atom through which the group is attached to the adjacent carbonyl group; the group may be saturated or partially unsaturated and, in addition to the -(CR<7>R<8>)m-Het substituent, it may be substituted by up to 4 further substituents independently chosen from F, C1-C6 alkyl, C3-C8 cycloalkyl, OR<11> or NR<9>R<10>; Het is an imidazolyl or triazolyl group either of which may optionally be substituted by C1-C6 alkyl, C3-C7 cycloalkyl, NR<9>R<10> or CONR<9>R<10>, each of R<9> and R<10> is independently H, C1-C6 alkyl or C3-C8 cycloalkyl, or R<9> and R<10> may be joined together to form, with the nitrogen to which they are attached, a 4 to 8 membered nitrogen-containing heterocyclic group, R<11> is H, C1-C6 alkyl or C3-C8 cycloalkyl; n and m are each independently 0, 1 or 2; wherein any alkyl or cycloalkyl group included in the aforementioned definitions may optionally be fully or partially substituted by fluorine; are inhibitors of retroviral proteases of utility in the treatment and prophylaxis of human retroviral infections.

Description

The invention relates to certain heterocyclic group containing peptide derivatives useful in the treatment and prophylaxis of retroviral infections in humans. The invention further relates to a process for the production of these peptides, their use and pharmaceutical preparations based thereon.

BACKGROUND OF THE INVENTION

Human immunodeficiency virus (HIV) is a retrovirus that causes various clinical conditions, the most serious of which is usually called AIDS (acquired immune deficiency syndrome) and ARC (AIDS-related complex). HIV infection is characterized by a gradual breakdown of the immune system and central nervous system dysfunction. Patients who are severely affected by immunodeficiency suffer from a number of occasional infections (such as pneumocystis carinii, human cytoroegalovirus or Candida) and various cancers such as Kaposi's sarcoma. An important factor in the gradual deterioration of immune function after infection

AND

HIV is the loss of cells, especially CD4 - lymphocytes. Infection of monocyte and macrophage lineage cells also contributes to the observed pathology. The key step in the disease process is thus successful infection of cells with ^HIV-1- 004.

HIV is a retrovirus. It encodes its genetic information in RNA, which is converted to DNA after the virus enters the host cell. An important step in the retrovirus replication cycle is the processing of the initial polypeptide precursor into mature (mature) structural and replicable proteins. This processing is performed with a virus-encoded protease, and in the absence of this enzymatic activity, virus replication is blocked.

It has now been found in the context of the invention that certain heterocyclic-group-linked peptide derivatives are potent inhibitors of retroviral proteases in both out-of-cell and infected-cell experiments, and additionally exhibit antiviral activity in cell culture assays. This activity renders these compounds useful in the treatment and prophylaxis of retroviral infections, particularly HIV-induced infections.

SUMMARY OF THE INVENTION

The present invention relates to peptides of the formula I - (CFPR ⁵ ) -

O

HER

R ²

O

II

R ⁴

X- (CR ⁷ R ⁸ ) _m -Het (I)

HIM

R ³ O where

R ¹ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl, heterocyclyl or CONR ⁹ R ¹⁰ ;

R ² is C 1 -C 6 alkyl, C 3 -C 5 cycloalkylalkyl and C 1 -C 4 alkyl, C 1 -C 4 arylalkyl

Ί carbon in the alkyl moiety or heterocyclylalkyl of 1 to 4 carbon atoms in the alkyl moiety;

R ³ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkylalkyl and C 1 -C 4 alkyl, C 1 -C 4 arylalkyl, (C 2 -C 4) arylalkenyl, (C 1 -C 4) heterocyclylalkyl or C 2 -C 4 heterocyclylalkenyl;

R ⁴ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl or heterocyclyl;

R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently hydrogen, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl or R ⁵ and R ⁶ or R ⁷

O and R are joined together to form a 3- to 8-membered carbocyclic ring;

X represents a 4- to 10-membered monocyclic or bicyclic heterocyclic group containing ring carbon atoms and one ring nitrogen atom through which it is attached to an adjacent carbonyl group, the heterocyclic group being saturated or partially unsaturated and except for - (CR R °) _In -Het optionally contains up to four additional substituents each independently selected from fluoro, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl and OR ¹¹ and NR ⁹ R ¹⁰ ;

Het is imidazolyl or triazolyl, each of which is optionally substituted with C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl or NR ⁹ ; ¹⁰ or CONR ⁹ R ¹⁰ ;

R ⁹ and R ¹⁰ are each independently hydrogen, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl, or R ⁹ are linked and, together with the nitrogen to which they are attached, represent a 4 to 8 membered heterocyclic group containing nitrogen;

R-- is hydrogen, C alkyl-C alkyl alkyl or C 8-C cyk cycloalkyl;

n and m are each independently 0, 1 or 2;

wherein any of the alkyl or cycloalkyl groups mentioned in the above definitions is optionally fully or partially substituted by fluorine, and pharmaceutically acceptable salts and bioprecursors thereof.

In the above definitions R @ ^1, R ^2, R ^3, and

R ⁴ is a heterocyclyl group of 4- to 6-membered heterocyclic group containing up to four nitrogen or oxygen or sulfur atoms as heteroatoms, optionally in combination with one or two nitrogen atoms. The ring may be aromatic or partially or fully saturated and optionally may contain a fused benzo group or may be substituted by C1-C6 alkyl, C3-C8 cycloalkyl, C2-C5 alkanoyl, C1-C4 alkoxy carbon, halogen, hydroxy, oxo or aryl. Preferred heterocyclyl groups include pyridyl, pyrimidinyl, thienyl, isoquinolyl and tetrazolyl.

In the above definitions of R ¹ , R ² , R ² and R ⁴ , aryl means phenyl optionally substituted by one to three substituents independently selected from C 1 -C 6 alkyl, C 3 -C 3 cycloalkyl. C 1 -C 4 alkoxy, C 1 -C 4 alkoxy, C 2 -C 5 alkanoyl, hydroxy, halogen, C 1 -C 4 alkyl substituted in whole or in part, C 1 -C 4 alkoxy is completely or partially substituted by fluorine, phenyl, phenoxy, benzyl, benzoyl, phenylsulfonyl, pyridyl, tetrazolyl, phenyltetrazolyl and NR ⁹ R ¹⁰ and CONR ⁹ R ¹⁰ , wherein R ⁹ and R ¹⁰ are as defined above. The term halogen refers to fluorine, chlorine, bromine or iodine.

The imidazolyl or triazolyl group, Het, may be attached either to the ring carbon or ring nitrogen atom and may be unsubstituted or mono-, di- or trisubstituted. The term triazolyl includes both 1,2,3- and 1,2,4-triazolyl.

Alkyl and alkoxy groups containing three or more carbon atoms may be branched or straight chain. Any of the alkyl, alkoxy or cycloalkyl groups mentioned in the above definitions is optionally fully or partially substituted by fluorine.

Bioprecursor as defined above refers to a pharmaceutically acceptable biodegradable derivative of a compound of formula (I) which, when administered to an animal or human, is converted in the body to a compound of formula (I). And, for example, an amino acid (such as L-valine).

The compounds of formula I contain a number of asymmetric carbon atoms and all possible stereoisomers, whether separated or not, are within the scope of the present invention.

One preferred embodiment of the present invention are compounds having the stereochemistry shown in Formula Ia

Ό N

HO? ³ (1a) _NH ? -X- (CR ⁷ H ⁸ ) _m -Het 10 O

In the above formula, the thick bonds indicate that the attached groups lie above the plane of the molecule, while the broken bonds indicate that these groups lie below the plane of the molecule.

In the definition of R ¹ aryl is preferably phenyl and heterocyclyl is preferably oxetan-3-yl or 1, l-dioxothietan-3-yl. R ¹ preferably represents tert-butyl, isopropyl, oxetan-3-yl or 1,1-dioxothietan-3-yl and the group (CR ⁵ R ⁶ ) n is absent; or R ¹ is phenyl and (CR ⁵ R ⁶ ) n is methylene; or R ¹ represents a group H2NCO-, CH3NHCO- or (CH3) 2NCO- and (CR ⁵ R ⁶⁾ n is a methylene or CH (CH _3). Particularly preferred-are compounds in which R ¹ is t-butyl, isopropyl or oxetan-3-yl, n is 0, especially compounds wherein is a group of formula ^(CR> R ^O) _R For R ¹ tert-butyl.

In the definition of ^R2, aryl is preferably phenyl and heterocyclyl is for example pyridyl, pyrimidinyl or thienyl. R @ ^{2 is} preferably C1 -C4 arylalkyl, the benzyl group being particularly preferred.

In the definition of R ⁴ , aryl is preferably phenyl and heterocyclyl is preferably pyridyl, pyrimidinyl or thienyl. R ^{@ 4 is} preferably C1 -C6 alkyl, with isopropyl and sec-butyl (valine or isoleucine derivatives) being particularly preferred.

The heterocyclic group X is preferably a 4 to 6 membered saturated or monounsaturated group, most preferably a group derived from azetidine, pyrrolidine, tetrahydropyridine or piperidine, with a piperidine derived group being particularly preferred.

Preferably R ⁷ and R ⁸ are each hydrogen and m is preferably 0 or 1.

In the definition of R ³ , aryl is phenyl which is unsubstituted or substituted as described above for aryl, and heterocyclyl is, for example, pyridyl, pyrimidinyl, isoquinolyl or thienyl. R ³ is preferably arylC 1 -C 4 alkyl or arylalkenyl having 2 to 4 carbon uhlíku.v alkenyl moieties. Most preferably, R ³ is benzyl which is optionally substituted in the phenyl ring by fluorine, chlorine, iodine, methyl, trifluoromethyl or trifluoromethoxy, 3-phenylprop-2-enyl or 3-phenylpropyl.

Preferred specific compounds of formula (I) include

1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -3 (imidazole- 1-yl) azetidine,

1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -4 (imidazole- 1-yl) piperidine,

1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (3-phenylprop-2-en-1-yl) hexanoyl) - (S) valyl] -4- (imidazol-1-yl) piperidine,

1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) valyl] -4 - (imidazol-1-yl) piperidine,

1- [N - ((S) -5- (tert-butoxycarbonylamino) - (R) -2- (4-chlorobenzyl) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl) - 3- (imidazol-1-yl) azetidine a

1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -

4-Hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) -isoleucyl] -4- (imidazol-1-yl) piperidine.

The invention furthermore relates to compounds of the formula I or pharmaceutically acceptable salts or bioprecursors thereof for use as a medicament, in particular for use in the treatment or prophylaxis of human retroviral infections, in particular infections caused by HIV. The invention also provides the use of the compounds of formula (I) or pharmaceutically acceptable salts or bioprecursors thereof in the manufacture of a medicament for the prophylaxis or treatment of retroviral infections.

The present invention also provides a pharmaceutical composition comprising a compound of Formula I, a pharmaceutically acceptable salt or biopresor thereof, and a pharmaceutically acceptable diluent or carrier.

The antiviral activity of the compounds of formula I can be demonstrated using in vitro assay systems. For example, the compounds of Formula I are able to completely protect the human H9 T-cell line for 7 days from the progressive effects of HIV infection. Untreated virus-infected cells exhibit typical cytopathic effects such as syncytia formation and cell death. In addition, virus particles produced in virus-infected cells that have been treated with a compound of formula I are non-infectious.

Examples of infections that can be treated or prevented using the compounds of Formula I include infections caused by human or animal retroviruses, particularly HIV-1. Clinical conditions that can be treated or prevented by administration of the compounds of Formula I include AIDS, ARC, and HIV-associated dementia.

Λ. sj

The compounds of the invention may also be used to block the progression of disease in infected individuals who have not yet developed any symptoms.

Compounds of formula (I) may be prepared by coupling and protecting group techniques well known to those skilled in the art of peptide chemistry.

A suitable process is illustrated by the following methods for preparing the compounds of formula (Ia). These procedures are given as illustrative examples.

The process shown in Scheme A is based on the protected lactone of formula II. This lactone is alkylated, using for example n-butyllithium or lithium hexamethyldisilazide and alkylated product is treated with "" a compound of formula R 3 R ^J and then separating the desired isomer, i.e. the product of formula III. The lactone ring of this compound is opened by treatment with dilute alkali metal hydroxide. This yields the corresponding hydroxy acid, the hydroxy group of which is subsequently protected, for example by forming a target. butyldimethylsilyl derivative. This protecting reaction is carried out using tert-butyldimethylsilyl chloride in Ν, Ν-dimethylformamide. The protected ester is then subjected to hydrolysis to provide the intermediate of formula IV.

The condensation of this intermediate with the second fragment of formula VIII, which leads to compounds of formula Ia, is illustrated in Scheme B. In this procedure, the protecting group of intermediate V is removed (in the case of tert-butoxycarbonyl by treatment with hydrogen chloride) and the resulting compound VI is reacted with N-protected amino acid of formula Boc-NK-CH (R ⁴ ) CO ₂ H using an amide coupling reagent. There was thus obtained intermediate VII. Thereafter, the N-protecting group is cleaved from intermediate VII and the amine product of formula VIII is condensed with an intermediate of formula IV to give an intermediate of formula IX. Thereafter, the hydroxy protecting group (X ') is cleaved from intermediate IX to give the desired product of formula Ia. In an alternative process, the intermediate of formula IX can be produced by condensation of intermediate X with intermediate of formula VI. An intermediate of formula X can be prepared from an intermediate of formula IX by reaction with an amino acid whose carboxyl group is protected, of formula H ₂ NCH (R ⁴ ) CO ₂ R and subsequent basic hydrolysis.

Scheme A

1. LiHMDS

2. R ³ -Br

AND

T o

1. OH

2. Protection of alcoholic and acidic functional groups

3. Deprotection of acidic functional group

R '- (CR ⁵ R ⁵ ) _n —θ'

OR ² O !! I II

flV)

Protecting group X ': preferably tert-butyldimethylsilyl

R ^{1 2 3} - (CR ⁵ R ⁶ ) _n : preference is given to tert-butyl or benzyl

OR ²

Diagram

PX- (CR ⁷ R ⁸ ) _m -Het (V) I deprotection

T

H-X-RCR ^^ - HeL (Ví)

Boc-NHCH (R ⁴ ) CO ₂ H amide coupling reagent

R ¹ (CR ⁵ R ⁵ )

R ¹ - (CR ⁵ R 0) _n .

Ό NH (IV) _x > 5 £ 2 ^l 3u.

X- (CR'R ^s) _mHet

The amine condensing agent

-¾ / n

rr

H ₂ N _p (ΜΠ) O .X {CR ^{7 R)} _m -Het

X- (CR ⁷ R 0) _m -Het deprotection i

The HX- (CR'R ⁵ ) -Het (VI) i amide condensation agent of the alcohol group

Ω!

(-and)

Ό NH ^(X) X'O Ø ² • NH, OH amide condensing agent

Base 2 (IV)

Amide coupling reagent: for example 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide

Protecting groups: P- preferably tert-butoxycarbonyl or benzyloxycarbonyl

X '- preferably a t-butyldimethylsilyl group

R @ 1 is preferably methyl or ethyl

In another alternative process as illustrated in Scheme C, cleavage of the tert-butoxycarbonyl group from intermediate XI, treatment with an acid such as trifluoroacetic acid and subsequent reaction with a carbonic acid derivative of formula XII yields products of formula XIII in which R skupina (CR) R R) _{n is} other than tert-butyl. For example, reaction of products of formula XI with 3-oxetanyloxycarbonyloxysuccinimide yields products of formula XIII wherein R ² represents

And n is 0.

Diagram

FT-ICFPR ⁵ ),

O '0 (XIII)

X 'is either hydrogen or a protecting group, e.g. butyldimethylsilyl

Y represents a group capable of undergoing a nucleophilic rearrangement; succinimidooxy group * σ is preferred

The starting materials of formula V required for the above process are in some cases compounds known from the literature or compounds which can be prepared by routine synthetic procedures from readily available starting materials. For example, reaction of 1- (N-tert-butoxycarbonyl) -3-hydroxyazetidine with methanesulfonyl chloride followed by reaction with a compound of formula Het-H in the presence of a base gives the desired compounds of formula V wherein X represents an azetidine ring. Alternatively, for example, 1- (N-benzyloxycarbonyl) -4-ketopiperidine can be converted to the 4-amino derivative by reaction with sodium cyanoborohydride in the presence of ammonium acetate. Subsequent treatment with dimethylformamidazine affords the 4- (1,2,4-triazol-4-yl) derivative.

Treatment of 1- (N-tert-butoxycarbonyl) -4-ketopiperidine with an anion obtained by reacting 1- (diethoxymethyl) imidazole with n-butyllithium followed by reaction of the product with methanesulfonyl chloride in the presence of a base affords N-protected-4-imidazole-2- yl- (1,2,5,6-tetrahydropyridine) intermediate. Catalytic hydrogenation affords the corresponding N-protected-4- (imidazol-2-yl) piperidine. Reaction of the above keto compound directly with a heterocyclic compound, for example imidazole, in the presence of thionyl chloride affords compounds of formula V wherein X is tetrahydropyridine. Reduction of this material yields the piperidine derivative again.

Intermediates of formula V in Scheme B, wherein m is not 0, can be prepared by standard transformation procedures from the appropriate precursors, for example using nucleophilic opening of the epoxy group to introduce the Het group. For example, reaction of 1- (N-tert-butoxycarbonyl) -4-ketopiperidine with trimethylsulfonium iodide in the presence of a base yields

4-Spiro-2'-oxyran. Reaction of this product with imidazole, followed by elimination of the 4-hydroxy group, is obtained

1- (N-tert-butoxycarbonyl) -4- (imidazol-1-yl) methyl-1,2,5,6-tetrahydropyridine from which the corresponding piperidine derivative is obtained by reduction.

The naturally occurring N-tert-butoxycarbonyl derivatives (Boc derivatives) used in the synthesis of compounds of formula VII are commercially available, as are their hydroxysuccinimidoesters. The corresponding intermediates derived from non-natural (i.e., non-natural) amino acids can be prepared by standard procedures (see, for example, M. J. O'Dellell et al., J. Amer. Chem. Soc., 1589, 1111, 2353). Compounds of formula II may be prepared from the corresponding tert-butoxycarbonyl-protected amino aldehydes (see DH Rich et al., J. Org. Chem., 1978, 43, 3624 and Y. Hamada et al., Chem. Pharm. Bull. 1982, 30 (5), 1521) by reaction with ethyl propionate (see AH Fray et al., J. Org. Chem.,

1086, 51, 4828), followed by reduction to form

5- tert. butoxycarbonylamino-4-hydroxy-6-phenylhexanoate. Cyclization by refluxing in toluene then affords the lactones of formula II as diastereomer mixtures which can be separated by standard procedures.

Certain of the hydroxy and amino protecting groups and certain carboxy activating groups need to be used in the above methods and in the procedures set forth in the specific examples below. It will be apparent to those skilled in the art that the described coupling and protection procedures can be carried out by any standard method known from peptide synthesis, and the use of all such methods is therefore within the scope of the invention. The choice of a specific protecting group in considerable. It depends on the availability of the required reagent, its effect on the solubility of the protected compounds, its ease of cleavage, and the presence of other groups that could be affected by its use. For example, in the above process, certain amino groups need to be protected and then deprotected to allow further reaction on the regenerated amino groups. The choice of protecting group for a given amino group was dependent on the role of the amino group in the overall reaction sequence. Amino protecting groups with varying levels of lability can be used. Such protecting groups are known in the art and in this context reference is made to Bodansky et al., Peptide Synthesis, 2nd Edition, John Wiley & Sons, New York (1976); Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York (1981); McOmie, Protective Groups in Organic Chemistry, Plenum Press, New York (1973) and Sheppard, Comprehensive Organic Chemistry, The Synthesis and Peactions of Organic Compounds, Pergamon Press, New York (1979), redig. E. Haslam, part 23.6, pages 321 to 339.

Non-limiting examples of representative amino acid protecting groups include aryloxycarbonyl such as benzyloxycarbonyl; substituted or unsubstituted aralkyl such as benzyl, trityl, benzhydryl and 4-nitrobenzyl; benzylidene; arylthio such as phenylthio, nitrophenylthio and trichlorophenylthio, phosphoryl derivatives such as dimethylphosphoryl and O, O-dibenzylphosphoryl; trialkylsilyl groups such as trimethylsilyl; and j. as disclosed in U.S. Patent No. 4,322,341. A preferred amino protecting group for use in the above reaction sequence is tert-butoxycarbonyl. Substitution procedures for introducing this group into a given amino group are well known. Generally, they include acylation of the corresponding amino compound with the corresponding 1a carbonyl chloride or anhydride in a solvent inert to the reaction, such as water, methylene chloride or tetrahydrofuran, in the presence of a base (acid acceptor) such as sodium or potassium hydroxide when water is used as solvent. and in the presence of a tertiary amine such as triethylamine or pyridine when an organic solvent is used as the solvent.

When an aqueous solvent is used, the pH of the reaction mixture is usually maintained at about 8 to 10, and preferably at about 9.

Protected amino groups are converted to unprotected amino groups by methods well known to those skilled in the art and suitable for the particular groups used. For example, tert-butoxycarbonyl is readily cleaved by treatment with dichloromethane saturated with hydrogen chloride gas.

Various hydroxy protecting groups are also known and are described in the references cited above. A preferred hydroxy protecting group is tert-butyldimethylsilyl. This group is introduced as above and is readily cleaved by treatment with tetra-n-butylammonium fluoride in tetrahydrofuran at room temperature.

Activation of carboxy groups as a means of carrying out a given acylation reaction is also a technique known to those skilled in the art. In particular, anhydrides and activated esters, especially esters derived from N-hydroxyphthalimide, N-hydroxysuccinimide or 1-hydroxybenzotriazole, are preferably used in the reaction sequence described herein. All of these substances are used in the art of peptide synthesis.

A dehydrating condensing agent is used to form the reactive ester. Representative examples of such condensing agents include 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide,

N, N'-dicyclohexylcarbodiimide, N, N'-carbonyldiimidazole,

1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, ethoxyacetylene, diphenylketene and N-ethyl-5-phenylisoxazoline-3'-sulfonate. The reaction conditions for the use of these condensing agents are well described in the literature. They typically involve the use of a solvent inert to the desired reaction and temperatures ranging from ambient temperature to 100 ^° C. The above carbodiimide reagents are preferred because they allow the use of reaction temperatures near ambient temperature and provide satisfactory yields of the desired esters. .

Upon completion of the condensation reactions leading to the end products, the various protecting groups may be cleaved by suitable techniques as described above and the compounds of formula I may be isolated and purified by conventional techniques such as recrystallization or column chromatography. The above sequence may be suitably adapted by appropriate selection of the starting materials in order to obtain any of the compounds with different variants of the meanings of R ¹ to R ⁸ , X and Het.

The present invention further provides a process for the preparation of a compound of formula (I) which comprises removing a protecting group from a compound of formula (IX).

O

RHCRSR ⁶ ) W o

NH

R *

ΧΌ

X- (CR ⁷ R ⁸ ) _m -Het (IX) wherein X 'represents a selectively removable hydroxy protecting group, whereupon the compound of formula I is isolated and optionally converted to its pharmaceutically acceptable salt.

A preferred protecting group X 'is tert-butyldimethylsilyl and this group is removed by treatment with tetra-n-butylammonium fluoride in an organic solvent, preferably tetrahydrofuran.

The novel intermediates of formulas VIII, IX and X are also an object of the present invention.

Examples of pharmaceutically acceptable salts of the compounds of formula (I) include acid addition salts such as sulfates, hydrogen sulfates, phosphates, lactates, mesylates, fumarates, citrates, succinates and gluconates.

For the treatment of patients suffering from retroviral infection, in particular HIV infection, the compounds of formula I will be administered by any suitable route, for example oral, parenteral (e.g. subcutaneous, intravenous, intramuscular or intratermal), rectal, nasal, topical (including buccal and sublingual) or vaginal route. Pharmaceutical compositions containing the antiviral agent of the invention and one or more pharmaceutically acceptable carriers and optionally other therapeutic agents may be prepared by conventional procedures well known in the pharmaceutical art. Oral dosage forms include, in particular, syrups, tablets and capsules, which may contain flavoring agents in addition to the inert carrier. Tablets may be made by conventional compression or molding processes, the starting raw material for these processes being a powder mix containing suitable additives, e.g., an antiviral agent in combination with a binder, diluent, lubricant and surfactant. Rectal preparations may be in the form of suppositories and vaginal preparations may be in the form of tampons, creams or foams. Parenteral formulations should be in sterile form, for example, packaged in vials for injection and contain aqueous or non-aqueous diluents, buffers, and antioxidants, the formulation being isotonic to the blood. A suitable dose of the anti-retroviral agents of formula (I) will range from 1 to 50, preferably 1 to 25 mg / kg / day, and may be administered in up to six divided doses over the course of the day. Of course, there are cases where higher or lower dosages are preferable, which is influenced by the age, weight, degree of disease and patient response. Appropriate treatment must be determined by a specialist.

The compounds of formula I may be used in combination with other drugs, some of which may potentiate their efficacy. Such drugs include:

(a) reverse transcriptase inhibitors such as AZT, dd1, ddC, foscarnet, TIBO compounds, dipyridodiazepinones or 6-substituted acyclopyrimidine derivatives (HEPT);

b) gp120-CD4 blockers such as dextran sulfate and soluble CD4, including a combination thereof with toxic agents such as pseudomonas toxin;

c) tat antagonists such as D-penicillamine;

d) other retroviral protease inhibitors such as Ro

31-8959 a

e) biological response modifiers such as interferons, interleukins or colony stimulating factors, for example GM-CSF.

The compounds of the invention are evaluated for their antiviral activity by dissolving the test compound in 50 μΐ DMSO, diluting the solution with RPMI 1640 and adding a complex salt solution of pH 7.2 to a concentration of 1 mg / ml. Assays are performed at concentrations of 0.001, 0.01, 0.1, 1 and 10 pg / ml against HIV-1 (strain IIIB) in the human T-cell line (H9). At the same time, untreated control infections are also initiated.

Seven days after infection, the supernatant from each tissue culture is titrated for the presence of infectious virus on CS166 cells (human T-cell lines). On days 3, 5 and 7, the cultures are examined to determine the presence or absence of syncytia. In control infections without drug treatment, typical viral cytopathic effects such as syncytia formation and cell death are evident. The reported IC ₁₀₀ value is the lowest concentration of test substance that provides 100% culture protection. Using this test method, the compounds of the invention exhibit IC ₁₀₀ values in the range of 0.1 to 10.0 µg / ml.

The preparation of certain starting materials and compounds of formula (I) is further illustrated in the following examples. These examples are illustrative only, and are not intended to limit the scope of the invention in any way. The purity of the compounds is routinely monitored by thin layer chromatography using Merck Kieselgel 60 F ₂₅₄ silica gel plates. 1 H-NMR spectra are recorded on either a Nicolet QE-300 or Bruker AC-300 spectrometer and are consistent with the proposed structure in all cases. Chemical shift values are reported in parts per million parts (ppm) downstream of tetramethylsilane. Conventional abbreviations are used to denote the principal peaks, i.e. s = singlet, d = doublet, t = triplet, m = multiplet and b = broad. Optical values

- 2 3 rotation is measured at degrees Celsius. AND

0.1% concentration in methanol at 25 ° C, otherwise. All temperatures are in

R. = infrared spectrum.

DETAILED DESCRIPTION OF THE INVENTION

Preparation of starting materials and intermediates

Preparation 1 (S) -5 - [(S) -1-tert-butoxycarbonylamino-2-phenylethyl] gamma-butyrolactone)

a) Ethyl (4S, 5S) - and (4R, 5S) -5-tert-butoxycarbonylamino-4-hydroxy-6-phenylhex-2-ynoate

A solution of diisopropylamine (6.4 mL) in dry tetrahydrofuran (25 mL) was stirred under a nitrogen atmosphere at

- 25 ° C. A 1.6 M solution of n-butyllithium in hexane (24.4 ml) was added over 5 minutes while maintaining the temperature below - 20 ° C. After an additional 15 minutes at -20 ° C, the solution is cooled to -70 ° C and ethyl propiolate (3.8 g) is added dropwise over 10 minutes, keeping the temperature below

- 60 'C. The resulting yellow suspension was stirred for an additional 20 minutes at -70 ° C and then a solution of N-tert-butoxycarbonyl-L-phenylalaninal (6.5g, see J R. Luly et al., J. Org) was added dropwise over 10 minutes. Chem., 1987, 52, 1487) in dry tetrahydrofuran (15 mL), keeping the temperature again below -65 ° C. The clear yellow solution formed was stirred at -70 ° C for 2 hours and then acetic acid (4 mL) was added. The cooling bath was removed, the mixture was allowed to warm to -30 ° C and when this was reached, water (100 mL) and ethyl acetate (100 mL) were added with vigorous stirring. The organic layer was separated and washed sequentially with 1M hydrochloric acid (50 mL), saturated aqueous sodium bicarbonate solution (50 mL), and saturated brine (50 mL). The organic solution was then dried over sodium sulfate and the solvent was evaporated. The crude product obtained as an oily residue was purified by chromatography on silica gel (ethyl acetate / hexane 1: 4). Product containing fractions were evaporated to give an oily residue which solidified overnight upon standing. This product was recrystallized from a mixture of ether and hexane to give the title compounds as a mixture wherein the diastereomer ratio of 4S, 5S: 4R, 5S was about 2: 1 (4.3 g), mp 98-99 ^° C.

Analysis calculated for ^C 19 H 25 ^KO 5: C, 65.70; H, 7.20; N, 4.03% found: C, 65.62; K, 7.42; N, 4.33

NMR (CDCl ₃ ) δ = 1.30-1.39 (m, 3H); 1.43

2.90-3.11 (m, 2 H); 3.37-3.38 and 4.16-4.19 1H); 3.93-4.04 (m, IH); 4.22-4.33 (m, 2 H); (LH); 4.77-4.79 and 4.87-4.90 (2 x brm, 7.35 (m, 5H)).

(s, SH); (2x brm, 4.51-4.56 1H); 7,24b) Ethyl (4S, 5S) - and (4R, 5S) -5-tert-butoxycarbonylamino-4-hydroxy-6-phenylhexanoate

The above product (1.17 g) was dissolved in ethanol (50 mg) and 5% palladium on barium sulfate was added as a catalyst. The mixture is hydrogenated at 50 psi for 2 hours. Then the mixture was filtered, the solvent was evaporated under reduced pressure to give the title compounds as a mixture of diastereomers in an approximate 2: 1 ratio. The product was a white solid (1.18 g) with a shade temperature of 125-126 ° C.

Analysis calculated for ^C 19 ^H 29 N 5 5: _{C 18 A 19} = _; , 25; N, 3.98% found: c, 64.91; H, 3.40; N, 2.98

N.M.R. (CDCl) δ = 1.24-1.22 (m, 3H); 1.25 and x s, SH); 1.72-1.95 (m, 2 H); 2.38-2.62 (m, 2H);

2.58 (m, 2 H); 3.02-2.04 and

D, O); 3.55-2.51 (m, 2 H);

., 40-3.42 (:

r— - 3--4 \ “· '--->' t

1,42 (2

2.77change.

t

c) (S) -5 - [(S) -1-tert-butoxycarbonylamino-2-phenynyl] -gamma-butyrolactone)

The gamma-hydroxyester of (b) was dissolved in a mixture of 2.5% acetic acid and toluene (35 mL) and the solution was refluxed for 2 hours. The solution was then cooled and evaporated to dryness under reduced pressure. The residue was purified by silica gel chromatography (40:60 diethyl ether / hexane). The title compound (0.4 g) is obtained, m.p. 98-99 ° C.

Analysis for ^C 17 ^H 23 ^NO 4 calculated: c, soy, SS-, 7.54; N, 4.59. Found: C, 55.77; H, 7.7 £; N, 4.3o m / e = 306 (MH +).

NMR (CDCl ₃ ) δ = 1.42 (s, SH); 2.11-2.19 (m, 2 H);

2.51-2.58 (m) 2H); 2.87-3.02 (m, 2 H); 4.00-4.07 (m,

1H); 4.47-4.52 (m, IH); 4.63 (d, J = 10, NH); 7.26-7.36 (m, 5H).

[α] ²⁵ D -22.6 ° (c = 1, MsOK).

D

I-R. (KBr) 1775, 1690, 1 = 25 cm ^-1 .

Preparation 2 (R) -2-Benzyl- (S) -5 - [(S) -5-tert-butoxycarbonylamino- (S) -4 (tert-butyldimethylsilyloxy) -6-phenylhexanoic acid

a) (R) -3-Benzyl- (S) -5- [(S) -1-tert. butoxycarbonylamino-2-phenylethyl] gamma-butyrolactone

To a cold (-10 ° C) solution of hexamethyldisilazane (7.9 mL) in tetrahydrofuran (15 mL) was added a 1.6 M solution of n-butyllithium in hexane (23 mL) over 3 minutes, keeping the temperature below 0 ° C. After another 5 minutes at 0 ^e C, the solution was cooled to - 70 ^C C and treated with a solution of (S) -5- [(S) -1-terc.butcxykarbonylamino-2-phenylethyl-j gammabutyrolaktonu (5 g) in tetrahydrofuran (38 ml), keeping the temperature below - 65 ^e C. the solution was stirred for 15 minutes at - 70 ^{° C} C and then to it over 1 minute was added benzyl bromide (1.95 ml) in tetrahydrofuran (12.5 ml) and the solution was stirred for an additional 10 minutes at - 70 ^C C. to the solution was added acetic acid (6.5 mL) and the cooling bath removed. Water (50 mL) and ethyl acetate (50 mL) were added and the mixture was allowed to warm to room temperature. The organic layer was separated, dried (MgSO4) and the solvent was evaporated under reduced pressure. The crude product is obtained as an oil. The oil was purified by silica gel chromatography (50:50 diethyl ether / hexane). This afforded the title compound (3.52 g) as a clear oil.

Analysis calculated for ^C 24 ^H 29 ^NO 4: C 72.91; found: _r .

7.34; N, 3.54%

7,5 0; N, 3.50 [α] _D ²⁵ -14 ° (c = 0.1%, MeOH)

N.M.R. (CDCl) < 5 = 1.37 (s, 9H); 1.9-5.2 (m, 2H);

3.20 (m, 5H); 3.92-4.01 (m, 1H); 4 _t 21-4.25 (m, 1H); Δ / 56 (m, 1H); 7.21-7.3 δ (m, 10H).

2.7952b) (R) -2-Benzyl- (S) -5-tert. butoxycarbonylamino (S) -4- (tert-butyldimethylsilyloxy) -6-phenylhexanoic acid

To a suspension of the product of (a) (17.63 g) in dioxane (120 mL) and water (60 mL) was added sodium hydroxide (1 N, 53.5 mL) at room temperature. The reaction mixture was stirred for 3 hours and then acidified to pH 5 with acetic acid. The mixture was allowed to stand for 30 minutes and the precipitate formed was filtered off and washed with water. The solid was washed with ethyl acetate, dried over magnesium sulfate and evaporated under reduced pressure. The white solid thus obtained was triturated with hexane, filtered and dried to give the intermediate hydroxy acid (17.85 g). To a solution of this hydroxy acid in Ν, Ν-dimethylformamide was added imidazole (29.38 g) and tert-butyldimethylsilyl chloride (32.53 g) at room temperature. The mixture was stirred for 18 hours, the solvent was evaporated under reduced pressure, and a mixture of water and ice, 10% citric acid was added to the residue to pH 4, and the resulting mixture was extracted with two 400 ml portions of ethyl acetate. The combined extracts were dried (MgSO 4) and evaporated under reduced pressure to give a light oil (29.2 g). To a solution of this oil in tetrahydrofuran (240 mL) was added acetic acid (240 mL) and water (80 mL) at room temperature. The mixture was stirred at room temperature for 2 hours and at 4 ^° C for 18 hours. The solution was then evaporated under reduced pressure and the residue partitioned between water (400 mL) and ethyl acetate (400 mL). The separated organic layer was washed with water (2 x 400 mL), saturated brine (100 mL), dried (MgSO 4) and concentrated in vacuo to a light oil. The oil was purified by chromatography on silica gel using diethyl ether / hexane (70:30) as eluent. The title compound was obtained as a glass (22.4 g). m / e 528 (MH) ^{&lt; + &gt;}

N.M.R. (DMSO-d6) S = 0.10 (s, 6H); 0.95 (s, 9H);

1.30 (n, 10H); 1.35 (m, 1H); 1.9 (m, 1H); 2.40 (ra, 1H);

2.72 (m, 2 H); 2.85 (m, 1H); 3.60 (m, 1H); 3.75 (m, 1H);

6.88 (d, IH); 7.22 (m, 10H).

Preparation 3

5-bromomethylisoquinoline

To a solution of an isomeric mixture of 5- and 7-bromoisoquinoline (5: 7, 40:60 → 1.0 g; see Glyde and Taylor, J. Chem. Soc. Perkin Trans 2, 1975, 1783) in solid tetrahydrofuran (15 mL) a 1.6 M solution of n-butyllithium in hexane (3.3 mL) at -70 ° C was added. The reaction mixture was maintained at this temperature for 30 minutes and then a solution of dry dimethylformamide (0.74 mL) in dry tetrahydrofuran (5 mL) was added. After an additional 15 minutes, the reaction was quenched by the addition of ethanol (5 mL) and allowed to warm to room temperature. Saturated ammonium chloride solution (10 ml) and diethyl ether (15 ml) were added sequentially and the organic phase was separated, washed with saturated sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by silica gel chromatography eluting with hexane / ethyl acetate (50:50). The faster progressing aldehyde was isolated to give 5-formylisoquinoline as an unstable yellow solid (0.05 g). Rf 0.3 (hexane: ethyl acetate, 50:50).

To a solution of this product (0.88 g) in dry methanol (20 mL) was added sodium borohydride (0.53 g) at 5 ° C and the resulting mixture was allowed to warm to room temperature and maintained at this temperature for 1 hour. The solution was then diluted with diethyl ether (20 mL) and water (10 mL) was added.

The aqueous layer was separated and extracted with diethyl ether (20 mL). The two organic phases were combined, washed with saturated sodium chloride solution (20 ml), dried over magnesium sulphate and the solvent was evaporated under reduced pressure. It is obtained

5-hydroxymethylisoquinoline as a pale yellow powder (0.37 g), m.p. 71-72 ° C.

/in

Analysis for C ₁₀ H _g NO requires: C, 75.45; H, 5.70; N, S, 80% Found: C, 75.06; K, 5.72; N, S, 66.

A solution of the above product (1.11 g) in glacial acetic acid (15 mL) was treated with 49% aqueous hydrobromic acid (30 mL), and the resulting mixture was refluxed for 2 hours. The reaction mixture was concentrated under reduced pressure, the residue was suspended in methylene chloride, and the mixture basified with saturated aqueous sodium bicarbonate. The phases were separated and the aqueous phase was extracted with methylene chloride. The combined organic phases were washed with a saturated aqueous sodium bicarbonate solution, dried over magnesium sulfate for 0.5 hours and then evaporated at room temperature under reduced pressure. There was thus obtained the title compound as a colorless solid. This solid was azeotroped with toluene and then used directly for the next reaction (1.28 g).

N.M.R. (CDCl3)? -7.90 (m, 2H); 8.65 (

4.90 (s, 2 H); 7.50 (t, 1H), 1H); 9.25 (s, 1H).

7.70 (d, IH);

Preparation 4

7-bromomethylisoquinoline

7-Formylisoquinoline was obtained from the initial step of Preparation 3 as a slower running product and was isolated as a yellow solid (0.08 g); Rf 0.25 (hexane: ethyl acetate, 50:50). Reaction of this material with sodium borohydride as described above gives 7-hydroxymethylisoquinoline, m.p. 129-130 ° C.

A solution of the hydrochloride of the above product (0.05 g) in thionyl bromide (0.5 ml) was heated to 60 ° C and maintained at this temperature for 45 minutes. The reaction mixture was ice-cooled, cautiously treated with excess water and then diethyl ether (15 mL). Concentrated aqueous ammonia is then added to pH 9 and the ether phase is separated, washed with water and dried over magnesium sulfate. A solution of hydrogen chloride in isopropyl alcohol (5.9N, 0.06 mL) was added and the resulting cloudy suspension was evaporated under reduced residue, azeotroped with the product as the hydrochloride solid (0.04 g).

m / e (MH +) 222

NMR (DMSO-d _e) = 5.05 (s, 2H), 8.25 (d, lH); S, 30 (d, 1H); S, 45 (s 9.70 (s, 1H).

pressure at room temperature, with toluene to give a colorless form

Preparations 5 to 13

The following compounds of formula (IV) are prepared as described in Preparation 2, but using the appropriate substituted benzyl bromide or bromomethylisoquinoline for the alkylation of gamma-butyrolactone in step (a), followed by ring opening and reaction with tert-butyldimethylsilyl chloride as described in step (b).

(IV)

preparation font 1 R ³ KH ’ Analysis (%) C Η N (theoretical values in brackets) 5 and CH ₂ F 2 54 6, 1 66.06 3.34 2.51 (66.02 5.31 2.56) t € _CH2-C! 562.2 and ⁷ _ 564.6 ''! _ / 'ii: W \ _from i: i * L \>' i! and X Σ 'Ί ! - - t - 1 AND 4,5 5,75 2,07 ' <from χ η x 7 c ό η '\ i (.., _., 7 and 1 ί C 'ch ^ J-cf 55 6 and 52.2 7.71 2.25! (62.5 7.44 2.35) and 10 and t and AND and CK ₂ N 2 -OCF ₃ c _ 2, 61.20 7.30 2.30 (60.90 7.30 2.30) 11 ^X CH ₂ CH = CH- ^ y » 452 69.53 3.59 2.52 (69.40 3.55 2.53) 12 ₂ kA ^ N 579 67.92 7.92 4.72 (63.26 3.02 4.32) ’ 13 'chAA4 579 67.77 7.85 4.64 (67.78 3.04 4.79) ^:

· 0.1 mol Η-Ό

0.33 mol of H 2 O

->

Preparation 14 1- (N-t-butoxycarbonyl) -3-methanesulfonyloxyazetidine

a) Azetidin-3-ol hydrochloride (2.10 g) was stirred in methylene chloride (40 ml) and diisopropylethylamine (2.59 g) was added followed by di-tert-butyl dicarbonate (4.36 g). The mixture was stirred at room temperature for 4 hours and then the solvent was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (200 mL) and the solution was washed with 1.5 M hydrochloric acid (50 mL), saturated sodium bicarbonate solution (25 mL), and brine (25 mL). The organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (50:50 ethyl acetate / hexane). 1- (N-tert-butoxycarbonyl) -3-hydroxyazetidine was obtained as a white solid (2.57 g), m.p. 51-53 ° C.

Analysis for c ₈ Hi ₅ no ₃ calculated: C, o 5 _f 4 7; found: 0.55.29;

09% 9S.

b) A solution of the above product (1.0 g) in methylene chloride (35 ml) was mixed with methanesulfonyl chloride (0.75 ml) and pyridine (1.5 ml) and the resulting mixture was stirred at room temperature for 3 days. The resulting solution was diluted with methylene chloride (75 mL), washed with aqueous citric acid solution (5%, 100 mL), saturated aqueous sodium bicarbonate solution (100 mL), dried over magnesium sulfate and evaporated under reduced pressure. The title product was obtained as a colorless oil (1.4 g).

Calcd for C ₉ H ₁₇ NO ₅ S: c, 43, C3; H, 5.77; N, 5.5S% found: 0.43.00; H, 6.80; N, 5.50

r./e 2 69 (Mis. ".)

(CDCl 3) δ = 1.27 (δ, δ :); 2.24 (s, 3H); 2.383, δ (m, 2H); 4.17-4.2 S (m, 2H); ); 5.25 (m, 1H).

Preparation 15

N, 4.53% N, 4.3%; 2.5 ° (m, 1H); 3.0 (s, 3h);

7.30 (m, 5H).

1- (N-benzyloxycarbonyl) - (R) -3-methanesulfonyloxypyrrolidine

The title compound was prepared from 1- (N-benzyloxycarbonyl) - (R) -2-hydroxypyrrolidine (J. Med. Chem., 1992, 35, 1764) using the procedure described in Preparation 14, except that pyridine used as the base of triethylamine. The product is obtained as an oil.

Analysis for C 0,5^H H iN NOO -S calculated: 0.51.53; h, 5.7S; Found: C, 51.60; Δ, 5, SO;

m / e MH + 300.

N.M.P. (CDCl3) δ = 2.15 (m, 1H)

3.45-3.30 (r, 4H); 5.10 (s. 2H);

Preparation 16

1- (N-Benzyloxycarbonyl) - (S) -3- (para-toluene) sulfonyloxy-pyrrolidine

To a solution of 1- (N-benzyloxycarbonyl) - (R) -3 (hydroxypyrrolidine (7.90 g) in dry tetrahydrofuran (100 mL) was added triphenylphosphine (13.58 g) under nitrogen and the resulting solution was cooled to -30 '. C 1 and methyl tosylate (10.06 g), and diethyl azodicarboxylate (10.41 g) were added over 0.5 hour, after which the reaction mixture was allowed to warm to room temperature and maintained at this temperature for 65 hours. The reaction mixture is evaporated under reduced pressure,

The residue was dissolved in dichloromethane, washed with water, dried over magnesium sulphate and evaporated under reduced pressure to an oil. The residue was purified by chromatography on silica gel using methylene chloride / methanol (58: 2 to 96: 4) as eluent. A second chromatography is then performed using a 80:20 to 50:50 mixture of hexane and ethyl acetate as eluent. Obtained as a golden oil (10.22 g). Analysis of the C- H ₀ - NO ,, ₌ S vypočreno: 2,50,78; H, 5.54;

.rezeno:

-.eur. ;

(2D2 4 y.);

• in \ * · i

Preparation 17

1- (N-tert-butoxycarbonyl) -4-methanesulfonyloxypiperidine

The title compound was prepared from 1- (N-tert-butoxycarbonyl) -4-hydroxypiperidine as described in Preparation

14. The product obtained has a melting point of S5 to 86 ° C.

Analysis for C ₁₁ H ₂₁ NO-S calculated: C, 4 7.3; 7, 7, 58; N, 5, 02¾ Found: C, 47.2; H, 7, 66; N, 4, c χ N.N.P. (CDC1;) <? = 1,5 ( S, 5H); 1 , S5 (m j, 0 o (s, j), 3, 2.-5. ; 3.7 5 (m , 2H);

Preparation

8

1- (N-tert-butoxycarbonyl) -3- (imidazol-1-yl) azetidine

To a solution of imidazole (0.41 g) in Ν, Ν-dimethylformamide (30 mL) was added sodium hydride (60%, 0.24 g) and the mixture was stirred at room temperature for 1 hour. 1- (N-butoxycarbonyl) -3- (methanesulfonyloxyxazetidine (Preparation 14) (1.4 g)) was added and the mixture was heated to 75 ° C for 3 days. The solvent was evaporated under reduced pressure, the residue was dissolved in ethyl acetate (50 mL). The residue was purified by chromatography on silica gel using methylene chloride / methanol / conc. aqueous ammonia (93: 7: 1) as the eluent. The title compound was obtained as an oil (0.72 g).

m / e 22 (MH) +

NMR (DMSO-d ₆ ) δ 4.2 ° -4.4 - (δ, 95-4.05)

2H);

(s, 1 H); 7.43 (s, (5, SH); 3H); 5

-I ·> f - · * / ´ 'J v' in - f - * * 7

Preparations 19 to 25

The following compounds of formula V are prepared as described in Preparation 18, but using the appropriate starting materials from Preparations 14 to 17, which are reacted with an appropriately substituted or unsubstituted imidazole or triazole.

Preparation number 1 PX- (CR ^{7 R)} _m -Het (MH) * Analysis (%) C Η N (theoretical values in brackets) 19 Dec NH; SOC-N ^ N N N X / '_! 239.3 55.1 7.50 22.9 (oo, 4 / _f /, 5) 1 - * ·, r \ ώΟΟ-Ν— —N ^X N NMR: 1.33 (=, 1H) 4.05 (m, 2H); 4.2 (m, 2H); o, _ i, j ..,,, ·, - ~ pcs; ; ^C t - ~ '· - ----' CH2-N ^ - N ^ N c. z _t z _ o, ϋ -τ. (6 o, 60 0 z, z - _o, 2oj ? ? / \ Rl Cbz-N y-N N 272 55.69 6.33 15.25 (55.60 5.25 15.25) ^: Boc-1H-N-N for 24 Boc-N \ -N ^ N _N = / 253 25 ch ₃ Boc-N ^ y — N ^ N 265.9 61.0 S, S2 15.6 (61.3 S, S2 15.3) ^:

1. 0.05 mol of methylene chloride

2. Hemihydrate

Preparation

6

a) 1- (N-tert-butoxycarbonyl) -4- (4-methylimidazol-1-yl) piperidine

Reaction of 4-methylimidazole with 1- (N-tert-butoxycarbonyl) -4-methanesulfonyloxypiperidine affords two regioisomeric products, which are separated by chromatography on silica gel using dichloromethane / methanol / concentrated aqueous ammonia 96: 3.5: 0.5 as eluent. The major isomer has an Rf of 0.47; m / e 265.9 (MH) &^lt; - &^gt;;

NKR (DMSO-d _6)? = 1.43 (s, 9H); 1.7 (dc, 2K); 1.93 (bd, 2 H); 2.07 (s. 3H); 2 _of E3 (m, 2n); 4, Co (^, jr.),

6.95 (s, 1 H); and _t 5 (s, -C-) b) 1- (N-tert. butoxycarbonyl) -4- (5-methylimidazol-1-yl) piperidine

Minor isomer, Rf 0.52; m / e 265.9 (K H)

NKR (DKSO _d-e) (bd, 2H); 2.05 (s, (s, 1H); 7.65 (s,

Preparation 27

1- (N-tert-butoxycarbonyl) -4- (imidazol-2-yl) - (1,2,5,6-tetrahydropyridine)

a) 1- (Diethoxymethyl) imidazole (6.8 g) was obtained in dry tetrahydrofuran (50 ml) under nitrogen at -40 ° C. Add n-butyllithium (25 mL, 1.6 N in hexane) to the mixture at a rate such that the reaction temperature remains below -3.5 ° C. The mixture was then added dropwise over 10 minutes

1- (N-tert-butoxycarbonyl) -4-ketopiperidine (2.65 g) in dry tetrahydrofuran (10 mL), keeping the temperature below -40 ° C and the resulting mixture was stirred at -40 ° C for 2 hours. Stir with hydrochloric acid (50 mL, 0.1N) for minutes, then add ethyl acetate (50 mL) and stir the resulting mixture for 5 minutes. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (1 x 50 mL). The combined organic extracts were washed with saturated sodium bicarbonate solution (1 x 50 mL) followed by saturated sodium chloride solution. The organic layer was dried over magnesium sulfate and evaporated to a yellow oil (8.0 g). The residue was purified by silica gel chromatography eluting with ethyl acetate: methanol: concentrated aqueous ammonia (90: 10: 1). 1- (N-tert-butoxycarbonyl) -4-hydroxy-4-imidazol-2-ylpiperidine was obtained as a cream-colored solid (2.2 g). m / e 268.0 (M + H) &lt; + & gt ^;.

b) This product (267 mg) was mixed with diisopropylamine (3 50 ml) in dry dimethylformamide (2 ml) at 0 ° C. Methanesulfonyl chloride (150 mL) was added in one portion, and the reaction mixture was stirred at 0 ° C for 2 hours. Additional portions of diisopropylamine (350 mL) and methanesulfonyl chloride (155 mL) were then added to the reaction mixture, and the reaction mixture was stirred at room temperature for 1 hour. The resulting mixture was diluted with water (4 mL), adjusted to pH 9 with 1M sodium hydroxide solution, and extracted with ethyl acetate (3 x 10 mL).

The combined organic extracts were dried over magnesium sulfate and evaporated to a gum (260 mg). The residue was purified by silica gel chromatography eluting with dichloromethane: methanol: concentrated aqueous ammonia (95: 5: 1). The title compound is obtained as a yellow resin (144 mg), m / e 250.1 (MH) ⁺ NKR (CDCl ₃ ) δ = 1.46 (s, 9K); 2.6 (bs, 2 H); 3.53 (t,

2H); 4.0 (bs, 2 H); 6.3 (m, IH); 7.0 (s. 2H); 9.4 (bs,

1H).

- i ο Preparation

1- (N-tert-butoxycarbonyl) -4- (imidazol-2-yl) piperidine

The product of Preparation 27 (0.55 g) was dissolved in ethanol (30 mL) and hydrogenated at 50 psi using 10% palladium on carbon catalyst. The catalyst was filtered off from the reaction mixture and the solvent was evaporated to give the product as a foam (0.55 g), m / e 252.1 (MH) ⁺

N.M.R. (CDCl3) <5 = 1, (bd, 2H); 2.75 (bR,:

(s, 9H); 1.68 (cd); 2.52 (bR, 1H);

2H); 1.55, 1 (bd, 2K);

3.7 ~ (bs,

Preparation 29

1- (N-tert-butoxycarbonyl) -4- (imidazol-1-yl) methyl-1,2,5,6-tetrahydropyridine

a) A 60% dispersion of sodium hydride in oil (4 g) in dry dimethylsulfoxide (100 mL) was washed with hexane to remove the oil and then heated to 70 ^° C with stirring for 1 hour. Dry tetrahydrofuran (100 mL) was added thereto. ) and the resulting mixture was cooled to - 20 ^e C. to this mixture was added a solution of trimethylsulfonium iodide (20.4 g) in dimethyl sulfoxide (80 ml) and a solution of l- (N-t-butoxycarbonyl) -4-ketopiperidine (19 9 g) in dry tetrahydrofuran (100 ml) and the resulting reaction mixture was stirred for 0.5 hour at -10 ° C and then for 1 hour at room temperature. Water (500 ml) was added and the mixture was extracted with ethyl acetate (3 x 250 ml). The combined extracts were washed with brine, dried (MgSO 4) and evaporated under reduced pressure. The residue was purified by silica gel chromatography eluting with cyclohexane / diethyl ether / isopropyl alcohol (60: 40: 1). 1- (N-tert.

butoxycarbonyl) piperidin-4-spiro-2'-oxirane as a colorless solid (19.6 g), mp 65-66 ^e C.

Analysis calculated for ^C 11 ^H 19 ^NO 3: 0.51.97; H, 3.92; N, 5.70% found: ^c , 62.11; 9.06; N, 6.55.

b) To a solution of imidazole (1.91 g) in dry acetic acid. nitrile (30 mL) was added under stirring under nitrogen atmosphere a 30% oil dispersion of sodium hydride (0.84 g) and the resulting mixture was heated to 60 ^° C until a solution was formed. After 15 minutes, the product of step a) (2.0 g) was added thereto, and the reaction mixture was maintained for 5 hours. The reaction mixture was allowed to stand overnight at room temperature, the solvent was evaporated under reduced pressure and the oily residue was partitioned between methylene chloride (40 mL) and water (20 mL). The organic phase is separated and washed with water, dried over magnesium sulphate and evaporated under reduced pressure. The residue was purified by silica gel chromatography using a 95: 4: 1 mixture of methylene chloride, methanol and concentrated (0.88 density) aqueous ammonia as eluent. There was thus obtained 1- (N-tert-butoxycarbonyl) -4-hydroxy-4- (imidazol-1-yl) methylpiperidine as a colorless powder (2.13 g).

Analysis calculated for: C, 8 8.43; K, found: ⁵ ° ^{/ 25;} m / e 282 (MH +).

1/10 ch ₂ or ₂ 8.07; N, 14.50% 8.30; N, 14.52

c) To a solution of the product of step b) (2.0 g) and triethylamine (5.44 ml) in dry methylene chloride (80 ml) was added a solution of methanesulfonyl chloride (2.20 ml) in dry methylene chloride (10 ml) with stirring; the resulting mixture was allowed to warm to room temperature and maintained at this temperature for 14 hours. The reaction mixture was washed with water, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using methylene chloride / methanol / concentrated (0.88 density) aqueous ammonia (96: 4: 0 to 95: 4: 1) as the eluent. Obtained as a gold-colored oil (1.36 g).

Analysis for C ₁₃ H ₂₁ N 3 ° 2 · 1/4 ch ₂ or ₂ calculated: C, 60.14 l! 'í ^z - / N i.e. * »f / i '© found: C, 59.93 t / ', 0 0 / N i - 4/9 m / e (MH) - 254 N.M.P .. (CDCl3) δ - 1 / X (WITH, SH); 1.57 (m, 2 H); 3.50 (-, 2H); 3.90 (s, 2 H); 4.50 (with, 2.-); o, 50 (s, 1K); 6.9 0 (s, 1H), 10.10 (s, J); 7.55 (with, 1H). At p ^r and v a 3 0

1- (N-tert-butoxycarbonyl) -4- (imidazol-1-yl) methylpiperidine

A solution of the product of Preparation 29 (1.33 g) in absolute ethanol (25 mL) was hydrogenated with stirring in the presence of 10% palladium on activated carbon (0.3 g) as a catalyst for 4 hours at room temperature and 50 psi. . The reaction mixture was filtered and evaporated under reduced pressure and the residue azeotroped with methylene chloride. The residue was purified by silica gel chromatography eluting with methylene chloride: methanol (96: 4). Obtained as a colorless oil (1.09 g).

Analysis for ^C 13 H ₂₃ N ₃ O ₂ · 1/4 CH ₂ Cl ₂ calculated: ^C, 63.36; H, S, 74; N, 15.84% found: ^C ' ⁶³ '^08; H, 8.60; N, 15.50 m / e (MH) + 266

v 2 - N.M.R. (CDClj) S = 1.15 (m, 2H); 1.45 (s, 9H); 2H); 1.85 (m, 1H); 2.65 (t, 2K); 3.80 (d, 2H); 2H); 6.90 (s, 1H); 7.10 (with, 1H); 7.45 (s, 1 H). At P r and v and 3 1

1- (N-tert-butoxycarbonyl) -4- (imidazol-1-yl)

1,2,5,6-tetrahydropyridine

Imidazole (8.20 g) was stirred in dry methylene chloride (30 mL) at -10 ° C and thionyl chloride (4.8 mL) in dry methylene chloride (30 mL) was added. The resulting suspension was allowed to warm to room temperature and after 2 hours a solution of 1- (N-tert-butoxycarbonyl) -4-ketopiperidine (6.0 g) in dry methylene chloride (50 mL) was added dropwise.

The reaction mixture was stirred overnight and then evaporated under reduced pressure. To the oily residue was added potassium carbonate (6.0 g) in water (30 mL) and the product was extracted with methylene chloride (2 x 60 mL). United extra ?. The reaction mixture was washed with water (40 mL), dried (MgSO 4) and evaporated under reduced pressure. The residue was purified by silica gel chromatography eluting with ethyl acetate / methanol (100: 0 to 90:10). Obtained as an oil (1.0 g).

N.M.R. (CDCl 3) δ = 1.50 (s: -H); 2.55 (s, 2 H); 3.10

2H); 4.05 (s, 2 H); 5.80 (s, IH); 7.10 (s, 2H); 7.65 (s,

4?

Preparation 32 1- (N-Benzyloxycarbonyl) -4- (1,2,4-triazol-4-yl) piperidine

To a solution of 1- (N-benzyloxycarbonyl) -4-ketopiperidine (5.0 g) in methanol (25 mL) was added ammonium acetate (16.5 g) and sodium cyanoborohydride (0.94 g), and the mixture was stirred at room temperature. .for 24 hours. The solvent was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and 1M sodium hydroxide solution. The ethyl acetate layer was separated, dried over magnesium sulfate and evaporated under reduced pressure to a yellow oil. The residue was purified by silica gel chromatography eluting with methylene chloride / methanol / concentrated aqueous ammonia (95: 5: 1). There was thus obtained 4-amino-1- (N-benzyloxycarbonyl) piperidine as a yellow oil. A solution of this product (1.5 g) in toluene (20 ml) was mixed with dimethylformamidazine (1.0 g) and p-toluenesulfonic acid (0.1 g) and the mixture was heated to reflux for 24 hours. Then, the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography eluting with methylene chloride / methanol / concentrated aqueous ammonia (93: 7: 1). The title compound was obtained as a colorless oil, m / e (MH +) 287;

N.K.R. (DMSO-d6) 3.09 (m, 2H); 4.12 7.25-7.45 (m, 5H);

= 1.52 (m, 2H); 2.01 (m, 2 H); 2.53 (m, 2 H); 4.40 (m, IH); 5.11 (s, 2H);

5.65 (s, 2 H).

Λ Λ

Preparation 33

1- (N-tert-butoxycarbonyl- (S) -valyl) -3- (imidazol-1-yl) azetidine

A solution of 1- (N-tert-butoxycarbonyl) -3- (imidazol-1-yl) azetiaine from Preparation 18) (0.72 g) in methylene chloride (30 mL) was saturated with hydrogen chloride at 0 ° C and kept at this temperature for an additional hour. The solvent was evaporated under reduced pressure and the amine hydrochloride obtained was dissolved in Ν, Ν-dimethylformamide (25 ml). To the resulting solution was added the N-tert ester at room temperature. butoxycarbonyl- (S) -valine and N-hydroxysuccinimide (1.0 g) and N, N-diisopropylethylamine (1.7 mL). The reaction mixture was stirred at room temperature for 1 hour and then freed from the solvent by evaporation under reduced pressure. The residue was purified by silica gel chromatography eluting with methylene chloride / methanol / concentrated aqueous ammonia (97: 7: 1). The title compound was obtained as a colorless foam (0.79 g). m / e 323 (MH) ^&lt; ^{+ &gt};;

NMR (DMSO-d ₆ )? = 0.39 (m, 6H); 1.38 (s, 9H); 1.90 (m, 1H); 3.61-3.78 (m, 1H); 3.92-4.10 (m, IH); 4.244.80 (m, 3H); 5.19 (m, IH); 6.98 (s, IH); 7.06 (dd,

1H); 7.38 (s, 1 H); 7.77 (s, 1H).

Preparations 34 to 43

The following compounds of formula (VII) wherein R ⁴ is (S) -isopropyl and R ⁷ and R ⁸ are each hydrogen are prepared as described in Preparation 33 using the appropriate intermediates from Preparations 18 to 32, which are condensed with an N ester tert-butoxycarbonyl- (S) valine and N-hydroxysuccinimide.

t3u

rs \ Z 'ρ »t_ ´ Z, v--' · Z \ '1.-Het (VII)

L. Preparation font -X- (CH3) 'Het MH * Analysis (%) C (theoretical 1 H odnoty v N brackets) í 3 4 ί NH- —N ^ -N ^ N 338 í 1 5 _N ^Z / —N ^ N 3 91 52.09 6.73 14.03 \ / í = / C =, (52.32 6.46 14.36) —N - N N oc,.-c c, 3.46 ic, cc 16,22) '

2 ~ ί - N N N (59.54 ! ^of ! C ⁴ _R z τ - from * ~ 15.24) - - C j and and and -N - N N j AND 1 - C * 3 9! _CH. ^? 3 £ 5; £ Ί 1 ~ “T - C 9 2 14,9 1 , t ; and - N - N N (€ 1.1 ^c i - - 15.0) ¹ i and i 1 1 \ _ / __ ’ 4 u • 1 and —N) - N N \ _ / j 364.9 £ l _r 3 {ί Ί C k - - Z * r _t -L3 cct 14.9 15.2) ⁵ ; (and </ Γ11 and and i 41. and and and H / - \, N “ ^N / ~ <) 2oi, 1 and and ....... ‘f 42 - / - - CH 2 N 4 N 3 £ 5 59.29 (59.44 8.15 8.43 14.29 14.37) ⁴ III 4 3 —T / Vn ^ n \ _ / \ = J 34 9 51 5 ^Ί (61.46 8.11 8, C3 15.63 15.89) ⁵

1 · Hemihydrator 3. 0.25 mol of HCl 57 0.05 mol of CK _; NO. 1,

0.1 mol CH _? NO. 1; 4. 0.3 mol of CK _: C1 _?

- 47 Preparations 44

The following compounds of formula VII wherein R ⁴ is sec-butyl and m is 0 are prepared as described in Preparation 33 using the N-tert-butoxycarbonyl- (S) -isoleucine ester and N-hydroxysuccinimide in the condensation step.

1 preparation yoga number -X-He " MH * Analysis (%) C H (theoretical values in N brackets)! 4 4 1 - N \ _ / \ _ 3 6 5 62.01 (61.99 S / 2S 8.37 14.94 15.22) ^{1 45} ! ií / -N γ \ n n - 'N' 365 _ř 3 and 1 II II and 4 6 and —N ch ₃ OJ.,. (62.0 9.22 9.10 | 4 —N ) - n / ^ N 379.9 52.2 (52.0 3 ₍ 9 3 5.10 'J.S, JJ i 14.5) - i CK ₃ ! 43 Γ - N _ / \ _; 379.9 and 1 ch /

1. 0.2 mol H _; O. 0.5 mol H ₃ O

Preparation 49

1- (N-tert-butoxycarbonyl- (S) -isoleucyl) -4-ketopiperidine

The title compound was prepared according to the procedure in Preparation 33, except that 4-ketopiperidine hydrochloride hydrate was used in place of 3- (imidazol-1-yl) azetidine hydrochloride and (S) -isoleucine ester with N-hydroxysuccinimide. m / e 313 (MH) -] - -16 ^e (c = 0.34%, MeOH)

Preparation 50

N- ((R) -2-benzyl- (S) -5-tert-butoxycarbonylamino- (S) -4 (tert-butyldimethylsilyloxy) -6-phenylhexanoyl) - (S) -valine

The title compound was prepared as described in

S. J. deSolms et al., J. Med. Chem., 34, 2852 (1991).

Preparation 51 1-Isocyano-3-methyl-1- (p-toluenesulfonyl) -but-1-ene

The title compound was prepared from p-toluenesulfonylmethylisocyanide and isobutyraldehyde by the method

described in Van Leusen, Schaart and Van Leusen Recueil, 98, No 5, 258 (1979). IR. (nujol) 2100 cm ^-1 m / e 257 (M + H +) 3) * N.M.R. (CDC1) <S = 1.15 (d, 6H); 2.5 (s. 3H); 2.84 (m, 1H); 6.88 (d, 1H); 7.4 (d, 2H); 7.85 (d, 2 H).

Preparation

2

3-oxetanyloxycarbonyloxysuccinimide

Oxetan-3-ol (2.0 g) and N, N-diisopropylethylamine (7.76 g) were dissolved in methylene chloride (50 mL) and the solution was added dropwise to a solution of bis (tri-chloromethyl) carbonate over 15 minutes under nitrogen. (2.69 g) in methylene chloride (100 mL) was maintained at -20 ° C. The solution was stirred at 20 ° C for a further 15 min. N-hydroxysuccinimide (3.45 g) was added all at once. The resulting solution was allowed to warm to room temperature over 2 hours and then washed with water (50 mL), saturated aqueous sodium bicarbonate (50 mL), and brine (25 mL). The organic layer was dried over magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The title compound was obtained as a light brown oil (4.65 g).

NMR (CDCl ₃ ) δ = 2.83 (s, 4H); 4.75 (m, 2 H); 4.90 (m, 2 H);

5.58 (m, IH).

Example 1

1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -3 (imidazole-1) -yl) azetidine

a) 1- [N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-tert-butyldimethylsilyloxy-6-phenylhexanoyl) - (S) valyl] -3 - (imidazol-1-yl) azetidine

A solution of 1- [N-tert-butyloxycarbonyl- (S) -valyl) 3- (imidazol-1-yl) azetidine (from Preparation 33, 0.79 g) in methylene chloride (50 mL) was saturated with hydrogen chloride at temperature

-c and held at this temperature for an additional 1 hour. The solvent was evaporated under reduced pressure to give the amine as a colorless solid. A solution of this product in dimethylformamide (20 mL) was added to a solution of the reactive ester, which was prepared previously by intermixing (R) -2-benzyl- (S) -5-tert. Lamino- (S) -4- (tert-butyldimethylsilyloxy) -6-phenylhexanoic acid butoxycarbones (from Preparation 2,

1.32 g), 1-hydroxybenzotriazole (0.36 g), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.58 g) and Ν, N-diisopropylethylamine (1.75 ml) in dimethylformamide (75 g) ml) (20 minutes). After stirring for an additional 24 hours, the solvent was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and water. The ethyl acetate layer was separated, dried over magnesium sulfate and the solvent was evaporated under reduced pressure. The crude product was obtained as a colorless oil. The oil was purified by silica gel chromatography eluting with ethyl acetate: methanol: concentrated aqueous ammonia (90: 10: 1). The title compound was obtained as a colorless foam (1.1 g).

Analysis for ^{C H} 4 R 6 R 5 ^N ° 5 and ^a · / ^2H 2 ° Calculated: C, 66.39; H, 8.36; N, 9.44%. Found: C, 65.04; H, 8.26; N, 9.61 m / e 733 (MH) +.

[α] ²⁵ -6 ° (c = 0.29%, MeOK)

D

NKR _(DKSO-d) 6 = 0, 12 (m, 6H); 0.90 (m, 15H); 1,251.29 (2 xs, 9H); 1.96 (m, 1H); 2.32-2.77 (m, 4H);

2.89 (n, 1H); 3.48-3.74 (m, 2 H); 3.8S-4.06 (m, 3H); 4,184.66 (m, 4H); 5.12 (m, 1H); 6.77 (t, 1H); 6.98 (s, 1H); 7.077.32 (m, 10H); 7.40-7.82 (2.times.s, 1H); 7.76-7.96 (2.times.s, 1H); 8.0-8.20 (2 × d, 1H).

b) 1- [N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-hydroxy-6-phenylhexanoyl) - (S) valyl] -3- (imidazole 1-yl) azetidine

The product of step a) (1.19 g) was dissolved in tetrahydrofuran and a 1M solution of tetra-n-butylammonium fluoride in tetrahydrofuran was added at room temperature. After 48 hours, the solvent was evaporated under reduced pressure, the product was taken up in ethyl acetate, washed with saturated aqueous sodium bicarbonate solution and water, dried over magnesium sulfate and the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography eluting with methylene chloride / methanol / concentrated aqueous ammonia (93: 7: 1). The product obtained is then recrystallized from a mixture of ethyl acetate and hexane. The product was a colorless solid (0.52 g), mp 124-126 C. ^e

^Calcd for ^C 35 ^H 47 ^N 5 ^O 5: C, 63.05; H, 7.67; N, 11.1, 34% found: C, S7.99; H, S, O; N, 11.06 m / e 613 (MH) &lt; + &gt; -3 = (c = 0.14%, MeOH)

D

NMR _(DMSC-d) o = 0.30 (m, 5H); 1.21-1.47 (m, 1H);

1.27 (s, 9H); 2.62 (m, 2 H); 4.02 (m, 2 H); 5.13 (m, IH); 7.26-7.77 (2

58-1.71 (m, IH); 1,31-1 63-2.92 (m. 3H); 3,37-3 13-4.36 (m, 2 H); 4,41-4

43 (d, 1H); 6.98 (s, IH);

<- 1 in \ · 7 7? - “7? v o, - .. - / from J G> <f O Λ.

98 (m, 1 H); 2.44.61 (m, 2H); 3.85.56 (m, 2H);

7.05-7.30 (m, 10K); S, 1H); 7.38-7.98 (2µ, 1H).

Examples 2 to 10

The following compounds were prepared according to the general procedure described in Example 1, but employing, in condensation step a), a suitably protected intermediate carboxylic acid of formula IV and a suitable amine of formula VIII, and then removing the tert-butyldiroethylsilyl protecting group as described in step b) .

H

O

c e;

Example 11 - [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl) - (R) -2- (4-trifluoromethoxy) benzyl) hexanoyl) (S) - valyl] - (S) -3- (imidazol-1-yl) pyrrolidine

The procedure described in Example 1 was repeated using 1- (N-tert-butoxycarbonyl- (S) -valyl) -3- (S) -imidazol-1-yl) pyrrolidine (Preparation 36) in the condensation step, whereupon the tert. the butyldimethylsilyl protecting group is removed as described in step 1b). The title compound is obtained, m.p. 111-112 ° C.

Analysis for ^C 37 H ₄ FF ₃ N ₅ O ₅ Calcd: C, 62.08; K, 6.76; N, 9.78% found: _c , 61.90; H, 6.85; N, 9.61 m / e · (MH) - 716 [α] 7-2 ° (c = 0.1%, MeOH) [α] ~ 714 ° (c = 0.1%, MeOK)

D 3 65

N.M.R. (DMSO-δ) δ = 0.65-0.90 (m, 6K); 1.15-1.35 (m,

10H); 1.60 (m, IH); 1.85 (m, IH); 2.05-2.90 (m, 7H);

3.20-4.10 (m, 6H); 4.20 (m, IH); 4.55 (m, IH); 4.85 (m, 1H); 6.40 (d, 1H); 6.90-6.95 (2X p , 1H); 7,10—7,30 (m, -On); z, z 0 - /, 80 (2 X s, 1H); 7.90-8.0 (2 × d, 1H). Example 1 2

1- [N- ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl) - (R) -2- (4-trifluoromethoxy) benzyl) hexanoyl) (S) -valyl 1 - (R) -3- (Imidazol-1-yl) pyrrolidine

The procedure described in Example 1 was repeated using 1- (N-tert-butoxycarbonyl-1- (S) -valyl) -3- (R) - (imidazol-1-yl) pyrrolidine. The title product of melting point 109 ^DEG C. is obtained.

- Analysis for ^C 30 3 ₇ H ₄₈ F ₃ N ₅ O _sixth 2/5 H ₂ 0 calculated: found: m / e (KH) ”fa] - -23»

D

N.M.R. (DMSO-C 6) δ = 0.70-0.55 (m,

C, 61.47; H, 6.80; N, 9.55% C, 61.44; H, 7.05; N, 9.71 715 (c = 0.1%, MeOH)., 15-1.40 (-,

1.60 (m, 1H); 1.5 (m, 1H); 2.00-2.90 (m, 7H); 3.25-2.70 (m, 5H); 3.95 (m, 2 H); 4.20 (1H,); 4,454.65 (m, IH); 4.75-4.95 (2 × m, 1H); 5.40 (m, IH); 6.90 (s, 1 H); 7.05-7.25 (m, 10H); 7.70 - /, / = (2 x s,;

7.90-3.05 (2 × d, 1H).

Example

1- [N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -

4-Hydroxy-6-phenylhexanoyl) - (S) -valyl-4- (imidazol-1-yl) piperidine

a) 1- [N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-tert-butyldimethylsilyloxy-6-phenylhexanoyl) (S) -valyl] -4- (imidazol-1-yl) piperidine

The title compound was prepared from 1- (N-tert-butoxycarbonyl- (S) -valyl-4-imidazol-1-ylpiperidine and (S) -5-tert-butoxycarbonylamino- (S) -4-tert-butyldimethylsilyloxy- (R). (2-Benzyl) -6-phenylhexanoic acid using the same procedure as described in Example 1, step a). The crude product was purified by silica gel chromatography eluting with ethyl acetate: methanol: concentrated aqueous ammonia (90: 10: 1). Obtained as a colorless foam (0.71 g).

Analysis for C ₄₃ H ₆₅ N ₅ O ₅ Si. 1/4 H ₂ O calculated: c, 67.54; H, S, 56; N, 9.15%

[delta] found: c, 67.22; H, S, 44; N, 9.22 m / e 760 (MH) ^{&lt; + &gt;}

N. K. R.

1.30 (m 2.0c (rr.

(DMSO-d ₆ ) δ = 0.12 (rc, 6H) 2H); 1.26 (5.9H); 1.47-1 4K); 2.3 6-2.97 (m, 5H); 3; 0.9l (m, 15H);

.75 (m, 2 π); 1c, 50-3.75 (m, 2H);

1,134.61 (m, 4K); 6.79 (m, 1H); 6.90 (d, 1H); 7.07-7.29 (m, 11H); 7.68-7.73 (2 × S, 1K); 7.93-7.99 (2 × d, ln).

b) 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -4 (imidazole 1-yl) piperidine

The title compound was prepared from the product of step a) by the same procedure as described in Example 1 step b). The crude product was purified by silica gel chromatography eluting with methylene chloride / methanol / concentrated aqueous ammonia followed by recrystallization from ethyl acetate / hexane. The title compound (0.48 g) is obtained as a colorless solid, m.p. 121-123 ° C.

Analysis calculated for ^C 37 ^H 51 ^N 5 ^O 5 · 1/4 calculated: C, 6 S, 30; X, Ί, 9 2; found: .-., /, 9ο;

m / e 646 (KH) H ₂ °

N, 10.77% N, 10.5 9 ^{[Q] d} -10 ° (c = 0.1, MeOH)

NMR (DMSO-d ₆ ) δ = 0.77 (m, 6H); 1.19-1.37 (m, 1H); 1.28 (s, 9H); 1.48-1.72 (m, 3H); 1.82-2.04 (m, 3H); 2.422.94 (m, 7H); 3.38-3.60 (m, 2H)); 3.96-4.15 (m, IH); 4.28 (m, 1K); 4.37-4.58 (m, 3H); 6.43 (d, 1H); 6.83 (s, 1H);

7.04-7.29 (m, 11H); 7.65-7.69 (2 × S, 1H); 7.84-7.88 (2 × d, 1H).

YEAH

Examples 14 to 30

The following compounds were prepared as described in Example 13, but using a suitable protected intermediate carboxylic acid of formula IV and a suitable valine or isoleucine derivative of formula VIII in condensation step a) and then deprotecting the tert-butyldimethylsilyl protecting group as described in step b ).

O

OH Ø ² 6

Tartrate salt, hemihydrate

Example 31

1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl) -4 (1,2) (4-triazol-4-yl) piperidine

The benzyloxycarbonyl protecting group is cleaved from 1- [N- ((R) -2-benzyloxycarbonyl) -4- (1,2,4-triazol-4-yl) piperidine (Preparation 32) by catalytic hydrogenation and the resulting amine product is condensed with N - ((R) -2-benzyl- (S) -5-tert-butoxycarbonylamino- (S) -4-tert-butyldimethylsilyloxy) -6-phenylhexanoyl) - (S) -valine as in Example 1, step a), after to form 1- [N - ((R) -2-benzyl- (S) -5-tert-butoxycarbonylamino- (S) -4-tert-butyldimethylsilyloxy) -6-phenylhexanoyl) - (S) -valyl] -4- (1) 2,4-triazol-4-yl) piperidine, m / e 761 (MH ^&lt; ^{+ &gt;} ).

This product was treated with tetran-n-butylammonium fluoride as described in Example 1 step b). This gives the title compound.

Analysis for C ₃₆ H ₆ O _5Q N _5th H ₂ O calculated: <2.65, 06; E, 7.83; N, 12.55. Found: C, 64.50; K, 7.93; N, 12.43

L2] ~ -9 ° (c = 0.12 MeOK)

D

N.lí.R. (D.MSO-d ₆₎

1.46-1.76 (m, 3H);

3.33-3.64 (m, 2 H);

5.36-6.44 (m, 1H);

3.54 (s, 1H); 3.60 (s, 1H).

δ = 0.70-0.92 (m, SK); 1.1c -1.41 (m, 10r); 1.31-2, 15 (m, 3K); 2.53-2.96 (m, 7K);

3.95-4.15 (m, 1H); 4.27-4.51 (m, 4H);

7.01-7.12 (m, 10K); 7.73-7.91 (m, 1K);

Example 32

1- [Ν- ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) valyl] -4- (imidazol-1-yl) -1,2,5,6-tetrahydropyridine; tartrate

a) Reaction of (S) -5-tert-butoxycarbonylamino- (S) -4terc. butyldimethylsilyloxy) - (R) -2- (4-trifluoromethoxybenzyl) -6-phenylhexanoic acid with amine prepared by deprotection of 1- (N- (tert-butoxycarbonyl) -4-imidazol-2-yl (1,2,5) 6-tetrahydropyridine) as described in Example 1a) affords 1- (N- (S) -5-tert-butyloxycarbonylamino) - (S) 4-tert. butyldimethylsilyloxy) -6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl 1- (S) -valyl] -4- (imidazol-1-yl) 1,2,5,6-tetrahydropyridine.

For C ₃₃ ^H 8 ^F 3 ^N 5 ^O 6 ^s · 1/10 CH ₂ Cl ₂ calculated: C, 62.27; H, 7.37; N, 8.23% Found: C, 6 2.19; H, 7.39; N, 8.3S m / e 842 (K) * [α-10 ^C (c = 0.1%, MeOH)]

TO. E.R. (CDClj) = 0.10 (m - · π - Π t - · J i v, u u (d, 5K); ° W 90 (s. 9H); 1, 35 (s. 9H); *) - i 20-1.95 m - - -; / - / - 9 * - / ~ o (.5 n), 2, 70 (m. 2H); T - / 41-4, 100 (m, 6H); 4.55 (m, 1H); ⁴ z 65 (d, 1K); 5, 60-5.80 (2 X S, ii *), 6 25 (d, 1 H); 6, 90- ⁷ z 30 (m, 1H); 7 65 (s, 1 H)

b) The title compound as the free base is obtained by cleavage of the protecting group with tetra-n-butylammonium fluoride as described in Example 1b). This material was dissolved in absolute ethanol, and a solution of 1-tartaric acid (0.14 g) in absolute ethanol was added. Ether was added to the mixture and the resulting precipitate was filtered off and dried. The acid salt is obtained

1-tartaric acid as a colorless solid, mp 92-152 C ^e (0.61 g).

Analysis for ^c 3 ^H 3 ^F 3 ^N 5 ° 6. CH ₁₆ O ₆ . 1/3 H ₂ O requires C, 57.11; H, 6.23; N, 7.93% found: 57.40; Η, 6.64; N, 7.51 m / e (MH +) 728 [oj- -r5,7 ^e (c = 0.1%, MeOH) - - Γ) ICJ ² - - 6.7 ^(c = 0.1% , MeOH)

6 5

N. M . R. (DMSO-d 6) c = O, 7 o (m, 5H); 1,2 5 (s, 9H); 0,95- 1 i - 1 " 0 1H); 1.50 (m OF 1H) ; 1.30 (m, IH) ; 2.35-2.95 (m 7H) • 7 t t 0 -, c 5 (m, 4 π) of 4.0 (and, (m, 1 H); 4.25 (with, 2ΗΪ · - * * ; 4.40-4.60 (m of [0100] 2H ); 6.0 (m, IH) ; 5.40 (d, IH) 7.0 (5, 1H); 7,10-7,3 0 (m, 9H); 7.50 (s, 1H); 7.95 (m,

2H).

Example 33

1- (N - ((S) -5- (tert-butyloxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S) isoleucyl) -4- (5-Isopropyl-imidazol-1-yl) -piperidine

a) Deprotection of 1- (N-tert-butoxycarbonyl- (Ξ) isoleucyl) -4-ketopiperidine (Preparation 49) followed by reaction with (S) -5-tert. butoxycarbonylamino- (S) -4- (tert-butyldimethylsilyloxy) -6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoic acid (Preparation 10) gives 1- (N- ((S) -5) -5- - (tert-butyloxycarbonylamino) - (S) -4- (tert-butyldimethylsilyloxy) -6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S) -isoleucyl] -4-ketopiperidine.

<4

- Analysis for bo ^ 42 ^ 62 ^ * ^ 33θ7 ¹ 'Calculated: C, 62.60; H, 7.80; N, 5.20 Found: C, 62.50; H, 3.00; N, 5.20 m / e 806.5 (MH) +. [α] ²³ -5.30 (c = 0.1%, MeOH).

D

b) The above product (2.0 g) and aqueous ammonia (density 0.88) in ethanol (25 ml) were hydrogenated for 4 hours at 0.2 MPa in the presence of palladium on carbon (5,100 mg). The catalyst was filtered off and the solvent was evaporated. The colorless foam obtained (2.0 g) was purified by chromatography on silica gel using methylene chloride / methanol / concentrated aqueous ammonia (90: 10: 1) as eluent. Concentration of the appropriate fractions gave 1- (N - ((S) -5-tert-butyloxycarbonylamino- (S) -4-tert-butyldimethylsilyloxy-6-phenyl-1- (R) 2- (4-)). trifluoromethoxybenzyl) hexanoyl- (S) -isoleucyl] -4-aminopiperidine as a colorless glass (1.53 g).

c) The product of step b) (404 mg) was stirred with 1-isocyano-3-methyl-1- (p-toluenesulfonyl) but-1-ene (from preparation 51, 150 mg) in methanol (15 ml) with diisopropylethylamine (100 mg) for 16 hours. The solvent was evaporated at 40 ^° C. The residue was purified by silica gel chromatography using a mixture of methylene chloride, methanol and concentrated aqueous. 98: 2: 0.4 ammonia. It is obtained

1- (N - ((S) -5- (tert-butyloxycarbonylamino) - (S) -4-tert-butyldimethylsilyloxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S) -isoleucyl] -4- (5-isopropylimidazolyl-1-yl) piperidine as a colorless glass (380 mg), m / e 900 (MH ⁺ ).

d) The product of step c) is deprotected by treatment with tetra-butylammonium fluoride as described in Example 1b). Obtained the title compound.

Analysis for ^C 42 ^H 58 ^F 3 ^N 5 ° 6 '- / ^{2 H} 2 ° calculated: C, 63.5; K, 7.45; K, 8.81% found: ^C '= 3.5; H, 7.70; K, 8.70 m / e 785.9 (MH) *

NKK- (DKSO-de) 6 = 0. S (m, 6? :); 1.05 (m, 1H); 1.2 (m, -, x ( ^Ξ , 9y); x, 3 - 2.0 (m, i -); 2, O - 3.0 (m, 6, z>K);

x, 2 (- -, 0, c- ;; χ, χ (m, xr.); x,

-ir.} ·,, 4-4, z

4.12 (t, f) (s, 1H);

(s, s, 1 H); 7.55 (b R, 1H).

Example

1- [N - ((S) -4-hydroxy- (s) -5- (oxetan-3-yloxycarbonylamino) 6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S) -valyl] -4- (imidazol-1-yl) piperidine

a) The product of Example 16 (4.0 g) was dissolved in methylene chloride (40 ml) and the resulting solution was cooled in an ice bath. Trifluoroacetic acid (10 ml) was added dropwise over 5 minutes and the solution was stirred at 0 ° C for 1.5 hours. The solvent was evaporated under reduced pressure and the residue was taken up in ethyl acetate (250 mL). The organic phase was washed with 1M sodium hydroxide solution (50 ml) and brine (50 ml). The organic solution was dried (MgSO4), filtered and the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography using a 90: 10: 1 mixture of ethyl acetate, methanol and concentrated aqueous ammonia as eluent. 1- [N- (S) -5-amino- (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl (S) -valyl] -4- (imidazole-) was obtained. 1-yl piperidine as a white solid (2.98 g) m / e 630 (MH) ^{&lt; + &gt};

NMR (DMSO-d ₆ ) δ = 0.78 (m, 6H); 1.34-1.75 (m, 4H); 1.812.07 (m, 3H); 2.37-2.93 (m, 8K); 3.05-3.29 (m, 3H); 4 054,56 _r (m, 5H); 6.88 (s, 1H); 7.10-7.27 (m, 10H); 7.67 (d, 1H); 7.95 (m, 1H); . _ ..

b) The product from step a) (0.818 g) was dissolved in methylene chloride (30 ml) and added to the resulting solution.

3-Oxetanylcarbonyloxysuccinimide (0.344 g) was then stirred at room temperature for 1 hour. The solution was washed with 0.5M sodium hydroxide solution (15 ml) and brine (15 ml), dried over magnesium sulfate, filtered and evaporated under reduced pressure. The resulting solid was recrystallized from ethyl acetate to give the title compound as a white solid, mp 201-203 ° C.

Analysis for ^C 37 ^H 46 ^F 3 ^N 5 ° 7 1/2 H ₂ O calculated: C, 6 0.15; H, 6.41; N, 9.4% Found: C, 60.32; K, 6.22; N ^c 15 m / e 730 (MH) ’ [ ^α ] ^{1 D} 43 '(c = 0.1%, MeOH), [α] ~ 4-32 ° (c = 0.1, MéOK) D 3 6 5 NMR (DMSO-d ₆ ) δ - 0.77 (m 6H); 1.22-2.05 (m, 7H); 2,:

2.93 (m, 5H); 3.42-3.60 (m, 2K); 4.03 (d, 1K); 4.224.73 (m, 7H); 5.12 (m, 1H); 6.89 (s, 1H); 7.11-7.2S (m, 11H); 7.6 (d, 1H); 7.86 (d, 1H).

Ό C

Example 35

1- [N- (S) -4-hydroxy- (S) -5- (oxetan-3-yloxycarbonylamino) -6-phenyl- (R) -2- (3-phenylprop-2-enyl) hexanoyl- (S)] Isoleucyl] -4- (imidazol-1-yl) piperidine

To a solution of the product of Example 18 (0.45 g) in methylene chloride (20 mL) was added anhydrous trifluoroacetic acid (4 mL) at 0-5 ° C over 5 hours. The reaction mixture was concentrated under reduced pressure and the remaining resin was azeotroped 3 times with toluene and dried. A solution of the crude amine product and diisopropylethylarcin (0.82 mL) in methylene chloride (25 mL) was cooled to 5 ° C and a solution of 3-oxetanyloxycarbonyloxysuccinimide (0.22 g) in methylene chloride was added dropwise. The reaction mixture was allowed to warm to room temperature for 17 hours and then washed with water, dried (MgSO 4) and evaporated under reduced pressure. The residue was purified by silica gel chromatography eluting with methylene chloride / methanol / concentrated aqueous ammonia (95: 4: 1). Obtained as a colorless foam (0.26 g), mp 157-160 C. ^e

Analysis for ^C 38 ^H 49 ^N 5 ° 6 · ^{3/4 H} 2 ° calculated: C, 6.60; Η, 7.43; N, 10.22. Found: C, 66.45; H, 7.49; N, 10.22 cm @ -1

D (c = 0.1%, MeOH)

NMP. _(DKSO-d6) c 3.05 to 3.65 (m, 4H); 6.10 (m, IH); 6.30 6.95-7.35 (m, 11H)

0.75 (m, OH) ; 1,25-2,95 (m, 12H); , 95-4.75 (m, 10H); 5, 05 (m, 1 H); c, 1H); 6, 8 0-6, 8 5 (2 x p (1H);

[Delta] =: 5-7.6 [deg.] (2 * s, 1H); 7.95 (d,

-H)

Example 36

1- [N- (S) -4-hydroxy- (S) -5- (isopropyloxycarbonylamino) 6-phenyl- (R) -2- (3-phenylprop-2-enyl) hexanoyl- (S) isoleucyl] -4- - (imidazol-1-yl) piperidine; tartrate

The title compound was prepared according to the procedure in Example 35, except that isopropyl chloride, formate, was used to react with the intermediate amine. The crude product was purified by chromatography on silica gel eluting with methylene chloride: methanol: concentrated aqueous ammonia (97: 2: 1 to 95: 4: 1). Obtained as a colorless powder, which was recrystallized from ethyl acetate-hexane. The free base was dissolved in ethanol and a solution of 1-tartaric acid was added. Addition of diethyl ether gives the tartaric acid salt as a colorless powder, m.p. 156-157 ° C.

For ^C 35 ^H 51 ^N 5 ^O 5 · ^{2 H} 2 ° calculated: C, 61.75; H, 7.15; N, 3.57% Found: C, 61.60; H, 6. S7; N, S, 3S m / e 553 (NH) + [α] D 25 ^C (c = 0.1%, MeOH)

D

N.M.R. (DMSO-d6 < J = 0.75 (m, 6H); 1.0 (m, SK); 1.25, 3.65 (m, 14H); 3.95-4.70 (m, 10K)). 6.10 (m, 1H); 6.30 (d, 1H); 6.65 (d, 1H); 6.85-6.90 (2 xs, 1H); 7.05-7.35 * (m, 11H), 7.65-7.75 (2.times.s, 1H), 7.90 (d, 1H).

Example

7

1- [N- (S) -5- (tert-butoxycarbonylamino) -6-phenyl- (R) 2- (4-trifluoromethoxybenzyl) -4 - ((S) -oxyl) hexanoyl- (S) valyl] -4- (imidazol-1-yl) piperidine; tartrate

a) Dissolve N-benzyloxycarbonyl-L-valine (1.03 g) and dicyclohexylcarbodiimide (0.51 g) in dichloromethane. (25 mL) and stirred for 3 h. The precipitated dicyclohexylurea is filtered off and the filtrate is evaporated to a white foam. This white foam was mixed with the product of Example 16 in N, N-dimethylformamide (20 mL) and 4-dimethylaminopyridine (0.25 g) was added. The mixture was stirred at room temperature for 5 days and then partitioned between ethyl acetate and water. The organic extract was dried (MgSO4) and the solvent was evaporated. The residue was purified by silica gel chromatography eluting with ethyl acetate / methanol (0-10%). 1- [N- (S) -4- (N-Benzyloxycarbonyl (S) -valyloxy) - (S) -5-tert-butoxycarbonylamino) -6-phenyl- (R) 2- (4-trifluoromethoxy) was obtained. benzyl) hexanoyl- (S) -valyl 3-4- (imidazol-yl) piperidine as a white foam.

Analysis for ^C 51 ^H 65 ^F 3 ^N 6 ° 9 'calculated: c, 62.43; H,?,? S found: c, 62.63; : - :, 6.7 3 h ₂ °

b) The product from step a) (0.95 g) was dissolved in absolute ethanol (50 ml) and 10% palladium on activated carbon (0.1 g) was added, followed by hydrogenation for 4 hours under pressure. 0.41 MPa at room temperature. The catalyst was removed and the filtrate was evaporated to dryness. The residue was purified by silica gel chromatography eluting with methylene chloride: methanol: concentrated aqueous ammonia (97: 3: 0.5).

The product is obtained in the form of a foam. This foam was dissolved in ethyl acetate (6 mL) and a solution of tartaric acid (0.089 g) in a mixture of methanol and ethyl acetate (10% methanol, 10 mL of mixture) was added. The solvent was evaporated and the residue was triturated with diethyl ether. The product was obtained as a white glass (0.48 g), m.p. 122 ° C.

· Analysis for ^C 43 ^H 59 ^F 3 ^N 6 ° 7 · ^C 4 ^H 6 ° 6 '2 ° ^H Calc'd: C, 56.61; H, 6.77; N, 8.4% Found: C, 56.74; H, 6.76; N, 8.56 K. Ϊ · .. r .. (DMSO-d _s) 6 - 0,8 - ^, 93 (m, -2. ~); 1.2 (S,

z., - i .., 4.-./ ', 2 ₍ 4 -, z. (~, 4, _ (m, _r.); 4, _ · (m, \ yr)), 6, cj (s, 7.73-8.0 (m, 1H)).

Claims (23)

1. Peptides of the general formula I -X- {CR ^{7 R)} _m ^-Het! (and) OR ^{2 R} '- (C ^{R 5 R 6} ) -O ^A N _H ^ ho where ¹ - / R ¹ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl, heterocyclyl or CONR ⁵ R ¹⁰ ; R ² is C 1 -C 6 alkyl, C 3 -C 8 cycloalkylalkyl and C 1 -C 4 alkyl, C 1 -C 4 arylalkyl, or C 1 -C 4 heterocyclylalkyl alkyl moieties; R ³ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkylalkyl and C 1 -C 4 alkyl, C 1 -C 4 arylalkyl, (C 2 -C 4) arylalkenyl, (C 1 -C 4) heterocyclylalkyl or C 2 -C 4 heterocyclylalkenyl; i u R ⁴ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl or heterocyclyl; R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently hydrogen, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl or R ⁵ and R ⁶ or R ⁷ and R ⁸ are taken together to form a three to three membered alkyl group; an eight membered carbocyclic ring; X represents a four to ten membered monocyclic or bicyclic heterocyclic group containing ring carbon atoms and one ring nitrogen atom through which it is attached to an adjacent carbonyl group, the heterocyclic group being saturated or partially unsaturated and in addition to the substituent - (CR ⁷ R ⁸ ) _m 'Het optionally contains up to four additional substituents each independently selected from fluoro, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl and OR ¹¹ ; Het is imidazolyl or triazolyl, each of which is optionally substituted »alkyl group having 1 to 6 carbon atoms, cycloalkyl of 3-8 carbon atoms or a group of formula NR s R ⁹ or CONR ¹⁰ R ^{9 'L0;} R ⁹ and R ¹⁰ are each independently H, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl q η λ or R 8 and R 8 are taken together with the nitrogen atom to which they are attached to form z and t; a four to eight membered nitrogen-containing heterocyclic group; R ¹¹ is hydrogen, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl; n and n are each independently 0, 1 or 2; _with any of the alkyl groups or 'cycloalkyl, ·> which are mentioned in the above definitions are optionally fully or partially substituted by fluorine, and pharmaceutically acceptable salts and bioprecursors thereof. Peptides according to claim 1 having the stereochemistry corresponding to the general formula Ia R ¹ - (CR ⁵ F ¹⁵ ) _n X- (CR ⁷ R ⁸ ) _rr -Het (Ia) Wherein R ^A to R 0, n, m, X and Het are as defined in claim 1. Peptides according to claim 1 or 2, wherein R ¹ 'is tert-butyl, isopropyl or oxetanyl and n is 0. The peptides of claim 3, wherein R ² is benzyl. Peptides according to claim 3 or 4, wherein R ⁴ is isopropyl or sec-butyl. and Peptides according to any one of claims 3 to 5, wherein R ³ is benzyl which is optionally substituted in the phenyl ring by methyl, fluorine, chlorine, iodine, trifluoromethyl or trifluoromethoxy or wherein R ³ is 3-phenylpropyl or 3-phenylprop -2-enyl. 7. Peptides as claimed in any one of claims 3-6, wherein each of R ^5, R ^6, R ⁷ and R ^s is hydrogen. Peptides according to any one of claims 3 to 7, wherein m is 0 or 1. The peptides of claim 1 which are 1- [N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -3 (imidazole- 1-yl) azetidine, 1- [N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -4 (imidazole- 1-yl) piperidine, 1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (3-phenylprop-2-en-1-yl)] hexanoyl) - (S) valyl] -4- (imidazol-1-yl) piperidine, 1- [N- ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) valyl] -4 - (imidazol-1-yl) piperidine, 1- [N- ((S) -5- (tert-butoxycarbonylamino) - (R) -2 (4-chlorobenzyl) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) valyl] - 3- (imidazol-1-yl) azetidine; 1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) 4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) -isoleucyl] -4- (imidazol-1-yl) piperidine. A process for the production of peptides of the formula I according to claim 1, characterized in that the protecting groups are removed from a compound of the formula IX wherein X 'is a selectively cleavable hydroxy protecting group and R ¹ to R ⁸ , X and Het are as defined in claim 1, isolating the compound of formula I and optionally converting it to a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a peptide of formula (I) or (Ia), or a pharmaceutically acceptable salt or bioprecursor thereof, according to any one of claims 1 to 9, together with a pharmaceutically acceptable diluent or carrier. Peptides of formula I or Ia or pharmaceutically acceptable salts or bioprecursors thereof according to any one of claims 1 to 9 for use in medicine, in particular for use in the treatment or prophylaxis of human retroviral infections. 13. The use of peptides of the formula I or Ia or a pharmaceutically acceptable salt thereof according to any of> = ² 7 7 · <O '. of claims 1 to 9 for the manufacture of a medicament for use in the treatment or prophylaxis of human retroviral infections. 14. A process for producing peptides of formula (I) OR R'- _(C ^ rF _'<0 and _NH X- (CR ⁷ R ^B ) _m -Het HO R ³ (I) where R ¹ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl, heterocyclyl or qi η of CONR R; R ² is C 1 -C 6 alkyl, C 3 -C 8 cycloalkylalkyl and C 1 -C 4 alkyl, C 1 -C 4 arylalkyl or C 1 -C 4 heterocyclylalkyl parts; O R is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkylalkyl and C 1 -C 4 alkyl, arylalkyl C 1 -C 4 alkyl, arylalkenyl C 2 -C 4 alkenyl, C 1 -C 4 heterocyclylalkyl or C 2 -C 4 heterocyclylalkenyl; R ⁴ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl or heterocyclyl; R ^5, R ^5, R ⁷ and R ⁸ are each independently hydrogen, alkyl of 1 to 6 carbon atoms or cycloalkyl of 3-8 carbon atoms or R ⁵ and R ⁶ or R ⁷ and R ⁸ are joined together to form a three- up to an eight-membered carbocyclic ring? X represents a four to ten membered monocyclic or bicyclic heterocyclic group containing ring carbon atoms and one ring nitrogen atom through which it is attached to an adjacent carbonyl group, the heterocyclic group being saturated or partially unsaturated and in addition to the substituent - (CR ⁷ R ⁸ ) _m -Het optionally contains up to four additional substituents each independently selected from fluoro, C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl and OR ¹¹ and NR ⁹ R ¹⁰ ; Het is imidazolyl or triazolyl, each of which is optionally substituted with C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl or NR ⁹ R ¹⁰ or CONR ⁹ R ¹⁰ ; q 10, R ¹ and R ¹ are each independently hydrogen, C 1 -C 6 alkyl or cycloalkyl I 2 having from 3 to 8 carbon atoms, or R ⁹ and R ¹⁰ are linked and together with the nitrogen atom to which they are attached represent a 4- to 8-membered nitrogen-containing heterocyclic group; R ¹¹ is hydrogen, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl; . t n and m independently represent 0, 1 or 2; wherein any of the alkyl or cycloalkyl groups mentioned in the above definitions is optionally fully or partially substituted by fluorine, or a pharmaceutically acceptable salt thereof, characterized in that the protecting group is removed from the compound of formula IX X- (CR ⁷ R ⁸ ) _m -Het (ix) wherein X 'represents a selectively cleavable _Λ hydroxy protecting group and R ¹ to R ⁸ , X and Het are as defined in claim 1, isolating the compound of formula I and optionally converted to a pharmaceutically acceptable salt thereof. 15. The process of claim 14 wherein the selectively cleaved hydroxy protecting group X ' is t-butyldimethylsilyl and is cleaved by reaction with tetra-n-butylammonium fluoride in an organic solvent. AND - ου 16. The method of claim 14, wherein the compound of formula I exhibits a stereochemistry corresponding to formula Ib About NH X- (C? HO (Ib) wherein R ¹ to R ⁸ , n, m, X and Het have the meaning A method according to claim 14, 15 as recited in claim 15. or 16 wherein R ¹ is tert-butyl, isopropyl or oxetanyl and n is 0. The process of claim 17, wherein R ² is benzyl. The method of claim 17 or 18, wherein R ⁴ is isopropyl or sec-butyl. The method of any one of claims 17 to 19, wherein R ³ is benzyl optionally substituted in the phenyl ring with methyl, fluoro, chloro, iodo, trifluoromethyl or trifluoromethoxy or wherein R ³ is 3-phenylpropyl or 3-phenylprop-2- enyl. A method according to any one of claims 17 to 20, wherein each of R ⁵ , R ⁶ , R * ⁷ and R ⁸ represents a hydrogen atom. The method of any one of claims 17 to 21, wherein m is 0 or 1. The process of claim 14, wherein the compound of Formula I is 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -3 ( imidazol-1-yl) azetidine, 1- C N- ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -4 (imidazole (1-yl) piperidine; 1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (3-phenylprop-2-en-1-yl) hexanoyl) - (S) valyl-4- (imidazol-1-yl) piperidine, 1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) valyl] -4 - (imidazol-1-yl) piperidine, 1- [N- ((S) -5- (tert-butoxycarbonylamino) - (R) -2 (4-chlorobenzyl) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) valyl] -3 - (imidazol-1-yl) azetidine, or 1- [N - ((S) -5- (tert-butoxycarbonylamino) - (S) 4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl) - (S) -isoleucyl] -4- (imidazol-1-yl) piperidine. MP-1635-94