SK114094A3 - Peptides, method of their producing, their using and pharmaceutical preparation on their base - 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

Peptides, process for their preparation, their use and pharmaceutical preparations based thereon

Technical field

The invention relates to certain peptide derivatives containing a heterocyclic group which are 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 commonly referred to as AIDS (Acquired Immune Deficiency Syndrome) and ARC (AIDS-related complex). HIV infection is characterized by progressive breakdown of the immune system and central nervous system dysfunction. Patients who are severely affected by immunodeficiency suffer from a number of occasional infections (for example pneumocystis carinii, human cytomegalovirus or Candida) and various cancers such as Kaposi's sarcoma. An important factor for the gradual deterioration of immune function after HIV infection is the loss of cells, especially CD4 ⁺ lymphocytes. Infection of monocyte and macrophage lineage cells also contributes to the observed pathology. Thus, a key step in the progression of the disease is the successful infection of CD4 ⁺ cells with HIV.

HIV is a retrovirus. It encodes its genetic information in RNA, which is converted into 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-linked peptide derivatives are potent inhibitors of retroviral proteases, both in experiments outside the cell and in experiments with infected cells, 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 invention relates to peptides of the general formula

X- (CR ⁷ R ⁰ ) _m -Het (I) wherein

R ¹ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl, heterocyclyl or q in 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

C 1 -C 4 alkyl or C 1 -C 4 heterocyclylalkyl;

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 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 ⁶ , R ⁷ and R ⁸ are each independently hydrogen, C 1 -C 6 alkyl or cycloalkyl-5 (C 3 -C 8 alkyl) or R and R or R and R ⁸ 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, said heterocyclic group being saturated or partially unsaturated and except for - (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 ;

R ⁹ and R ¹⁰ are each independently hydrogen, C 1 -C 6 alkyl or C 3 -C 8 cycloalkyl, or R ⁹ and R ^{10 are taken} together and together with the nitrogen atom to which they are attached are four to eight membered a nitrogen-containing heterocyclic group;

R ^1X is hydrogen, C 1 -C 6 alkyl or C 3 -C 8 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 of R ¹ , R ² , R ³ and R ⁴ , the term heterocyclyl refers to a four to six 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 of this group may be aromatic or partially or fully saturated, and optionally may contain pricondenzo5

or benzyl, or may be substituted by C1-C6 alkyl, C3-C8 cycloalkyl, C2-C5 alkanoyl, C1-C4 alkoxy, halogen, hydroxy, oxo or aryl. Preferred heterocyclyl groups include pyridyl, pyrimidinyl, thienyl, isoquinolyl and tetrazolyl.

3 p 3

In the above definitions of R ^x , R, R and R ⁴ , the term aryl refers to a phenyl group optionally substituted by one to three substituents independently selected from the group consisting of C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl. C 1 -C 4 -alkoxy, C 2 -C 5 -alkanoyl, hydroxy, halogen, C 1 -C 4 -alkyl, wholly or partially substituted by fluorine, C 1 -C 4 -alkoxy or partially substituted by fluorine, phenyl, phenoxy, benzyl, benzoyl, 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 as

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 ΐ ’

I | listed in the above definitions is optionally entirely

I or partially substituted by fluorine.

t Under the designation bio-precursor, see above

By definition, a pharmaceutically acceptable biologically means

I; a degradable derivative of a compound of formula (I) which, when administered to an animal or human, is converted in the body into a compound of formula (I). * possible to mention. ester derivatives formed between the free hydroxy group of the compound of formula I and, for example, an amino acid line (such as L-valine).

The compounds of formula I contain a series; asymmetric carbon atoms and within the scope of this invention! all possible stereoisomers are included, whether separated or not.

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

X- (CR ⁷ R0) _m -Het (Ia)

In the above formula, the strengthened 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 oxethan-3-yl or 1,1-dioxothietan-3-yl. R ^{1 is} preferably tert-butyl, isopropyl, oxethan-3-yl or 1,1-dioxothietan-3-yl, and (CR ⁵ R ⁶ ) n is absent; or R ¹ is phenyl and (CR ⁵ R ⁶ ) n is methylene; or R ¹ is H 2 NHCO-, CH 3 NHCO- or (CH 3) 2 NCO 3, and (CR ⁵ R ⁶ ) n is methylene or CH (CH ₃ ). Particular preference is given to compounds in which R- ¹ · is t-butyl, isopropyl or oxetan-3-yl and n is 0, preferably compounds in which the group of formula (CR ⁵ R ⁶ ) _{n is} represented by R ¹ tert-butyl.

In the definition of R @ ² is aryl, 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 is aryl, preferably phenyl and heterocyclyl is preferably "- pyridyl, pyrimidinyl or thienyl. R ⁴ f is preferably C 1 -C 6 alkyl, with isopropyl being particularly preferred ύ | and sec-butyl (derivatives of valine or isoleucine).

t '

The heterocyclic group X is preferably four-membered

i) ·

B to 6-membered saturated or monounsaturated group,

Even more preferably, a group derived from azetidine, pyrrolidine, [tetrahydropyridine or piperidine, with a specific | a group derived from piperidine is preferred.

R @ 1 and R @ 1 are each preferably hydrogen and m @ 1 is preferably 0 or 1.

In the definition of R ³ , aryl is phenyl which is unsubstituted or substituted as described above with the meaning of aryl and heterocyclyl is, for example, pyridyl, pyrimidinyl,

O isoquinolyl or thienyl. Preferably, R is C 1 -C 4 arylalkyl or C 2 -C 4 arylalkenyl. 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 (imidazol-1-yl) azetidine, 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) - valyl] -4- (imidazol-l-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-l-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. It is a further object of the present invention to provide the use of the compounds of formula I or their pharmaceutically acceptable salts or bio-precursors in the manufacture of medicaments for the prophylaxis or treatment of retroviral infections.

The present invention further provides a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof or a bioprecursor 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 cells infected with the virus exhibit typical cytopathic effects such as syncytia formation and cell death. In addition, viral particles produced in virus-infected cells, which? have been treated with a compound of formula I, are non-infectious.

As examples of infections that can be treated or treated can be prevented by the use of compounds of formula I, i infections caused by human or animal diseases may be mentioned

K | retroviruses, particularly HIV-1. As clinical conditions that can be treated or prevented by the administration of the compounds

I

AIDS, ARC, and dementia can be mentioned for Formula I

I [1] associated with HIV. The compounds of the invention can also be used to block disease progression in infected individuals who have not yet developed any symptoms.

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

A suitable process is illustrated by the following processes for the preparation of compounds of formula Ia. These procedures are given as illustrative examples.

The process shown in Scheme A starts from the protected lactone of formula II. This lactone is alkylated, for example using n-butyllithium or lithiumhexamethyldisilazide, and a compound of formula R ³ Br is added to the alkylated product, then the desired isomer, ie. 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 the formation of a tert-butyldimethylsilyl derivative. This protecting reaction is carried out using tert-butyldimethylsilyl chloride in N, N-dimethylformamide. The protected ester is then subjected to hydrolysis to give an 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-NH-CH (R 6) CO ₂ H using an amide coupling reagent. There was thus obtained intermediate VII. The N-protecting group and the amine are then cleaved from intermediate VII

the product of formula VIII is condensed with an intermediate of formula IV to form an intermediate of formula IX. Thereafter, the hydroxy protecting group (X ¹ ) 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 an 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 H2NCH (R ⁴ ) CO 2 R and subsequent basic hydrolysis.

Scheme A

R ¹ - (CR ⁵ R ⁶ ) _n -

(Π)

1. LiHMDS

2. R ³ -Br

R ¹ - (CR ⁵ R ⁶ ) _n —O

ABOUT

(ΙΠ) alcoholic acid functionality

3. Deprotection of the acid functional group

R ^{@ 1} - (CR ^{@ 5} R ^{@ 6} ) _n --O

OH (IV)

Protecting group X ': preferably tert-butyldimethylsilyl

R ¹ - (CR ⁵ R ⁶ ) _n : tert-butyl or benzyl is preferred

B

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

HX - (CR ⁷ R ⁸ ) _{m -} Het (VI)

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

ABOUT

R ¹ - (CR ⁵ R ⁵⁾ _n _A

X- (CR ⁷ R ⁸ ) _m -Het

R ^J

HX- (CR'R ⁸ ) -Het (VI) amide coupling reagent other base deprotection

--------------------------- 0 ».

alcohol group

X- (CR ⁷ R ⁸ ) _m -Het

Base 2

1. ^H 2 ^N

O-amide condensing agent (IV) amide condensing agent

ABOUT

R ⁴

OR

X- (CR ⁷ R ⁸ ) _m -Het (Ia)

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

Protecting groups: P - preferably tert-butoxycarbonyl or benzyloxycarbonyl

X'- preferably tert-butyldimethylsilyl

R is preferably methyl or ethyl

In another alternative process as illustrated in Scheme C, cleavage of tert-butoxycarbonyl 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 the group of formula R ^{1 2 3} (CR ⁵ R ⁶ ) _{n is} other than tert-butyl. For example, reaction of products of formula XI with 3-oxetanyloxycarbonyloxysuccinimide yields products of formula XIII wherein R ¹ represents

And n is 0.

Scheme C

1. acid

Base 2 &

3. ^ - (CR ^ VO-CO-Y (XII))

X- (CR ⁷ R ⁸ ) _m -Het 1 X 1 represents either hydrogen or a protecting group, for example

L t-butyldimethylsilyl $ Y is a group capable of succumbing to nucleophilic rearrangement; preference is given to succinimidooxy group

C, $.

s

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 treatment 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 treatment with sodium cyanoborohydride in the presence of ammonium acetate. Subsequent reaction 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 obtained 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, by reacting 1- (N-tert-butoxycarbonyl) -4-ketopiperidine with trimethylsulfonium iodide in the presence of a base,

4-Spiro-2'-oxyran. Treatment of this product with imidazole followed by elimination of the 4-hydroxy group gives 1- (N-tert-butoxycarbonyl) -4- (imidazol-1-yl) methyl-1,2,5,6-tetrahydropyridine from which is reduced by reduction to give the corresponding piperidine derivative.

The naturally occurring N-tert-butoxycarbonyl derivatives (Boc derivatives) of amino acids used in the synthesis of compounds of formula VII are commercially available, as are their hydroxysuccinimido esters. Corresponding intermediates derived from unnatural (i.e., non-natural) amino acids can be prepared by standard procedures (see, for example, M. J. O'Donell et al., J. Amer. Chem. Soc., 1989, 111, 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), 1921) by reaction with ethyl propionate (see AH Fray et al., J. Org. Chem., 1086, 51, 4828), followed by reduction to give

5-tert-butoxycarbonylamino-4-hydroxy-6-phenylhexanoate. The cyclization, which is carried out 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 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 procedures is therefore within the scope of this invention. The choice of a specific protecting group to a large extent depends on the availability of the necessary reagent, its effect on the solubility of the protected compounds, the ease of its cleavage, and the presence of other groups that could be affected by its use. For example, in the above procedure, certain amino groups need to be protected and then deprotected in order to carry out the next reaction on the regenerated amino groups. The choice of protecting group for a given amino group depends on the role of the amino group in the overall reaction sequence. Amino-protecting groups of varying levels can be used

- lability. Such protecting groups are known in the art and reference is made to Bodansky et al., Peptide Synthesis, 2nd Edition, John Wiley & Sons, New York (1976) in this regard; 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 Reactions of Organic Compounds, Pergamon Press, New York (1979), redig. E. Haslam, section 23.6, pp. 321-339.

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; diphenylmethylene; arylthio such as phenylthio, nitrophenylthio and trichlorophenylthio; phosphoryl derivatives such as dimethylphosphoryl; and

Ο, Ο-dibenzylfosforylskupina; tert-alkylsilyl such as trimethylsilyl; etc., as set forth in the US patent; no. A preferred amino-protecting group for use in the above reaction sequence is t-butoxy-carbonyl. Substitution procedures for the implementation of this

K groups into a given amino group are well known. Generally ; include acylating the corresponding amino compound corresponding to r

e:

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. If 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 ^r 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 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 that is 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 appropriately tailored to suit any of the compounds with various R ¹ to R ⁸ , X and Het meanings.

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).

R ¹ - (CR ⁵ R ⁶ ) _n

X- (CR ⁷ R ⁸ ) _m -Het (1X)

X'O

ABOUT

wherein X 'represents a selectively removable hydroxy protecting group, then isolating the compound of formula I and optionally converting it to a pharmaceutically acceptable salt thereof.

A preferred protecting group for X 'is t-butyldimethylsilyl and this 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, in addition to an inert carrier, flavorings. Tablets can be made by conventional compression or molding, the starting material for these processes being a powder mixture containing suitable additives, i. for example an antiviral agent in combination with a binder, diluent, lubricant 'i'

ΐ.

i:

And a surfactant. Rectal preparations may be p ', lace-like and vaginal preparations may be tampons, j | creams or foams. Parenteral formulations should be in sterile form.

Also in the form, for example, packaged in injection vials and containing aqueous or non-aqueous diluents, buffer solutions and antioxidants, | wherein the preparation must be isotonized with respect to blood.

A suitable dose of the anti-retroviral agents of formula (I) will range from 1 to 50, preferably 1 to 25, and ₄ mg / kg / day, and may be administered in up to six sub-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's responsiveness. Appropriate treatment must be determined by a physician.

í '· v í:

£ í p

i.

I '

The compounds of formula I can 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 pseudomonase toxin;

c) tat antagonists such as D-penicillamine;

d) other retroviral protease inhibitors such as Ro 31-8959; and

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

Compounds of the invention are evaluated for their antiviral activity by testing the test compound II; Dissolve in 50 μΐ DMSO, dilute the solution with RPMI 1640 and add β complex salt solution at pH 7.2 to a concentration of 1 mg / ml.

The assay is performed at concentrations of 0.001, 0.01, 0.1, 1 and even 10 µg / ml against HIV-1 (strain IIIB) in the human T-cell line (H9). At the same time, untreated control γ infections are also initiated.

«

Seven days after infection, the supernatant from each tissue culture is titrated for the presence of infectious virus on C8166 cells (human T-cell line). On days 3, 5, and 7, the cultures are also inspected to detect the presence or absence of syncytia. Typical viral cytopathic effects, such as syncytia formation and cell death, are evident in non-drug-treated control infections. The reported IC ₁₀₀ value is the lowest concentration of the test substance that provides 100% culture protection. Using this test method, the compounds of the invention have IC ₁₀₀ values ranging from 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 do not limit the scope of the invention in any way. The purity of the compounds is routinely monitored; thin layer chromatography using silica gel

First

Merck Kieselgel 60 F ₂₅₄ . The 1 H-nuclear magnetic resonance spectra are recorded either on a spectrometer f

| Nicolet QE-300 or Bruker AC-300 and in all cases are consistent with the proposed structure. Chemical values displacement are given in parts per million parts (ppm) in the direction

Even down from tetramethylsilane. For the labeling of the principal peaks

$ use conventional abbreviations, ie. s = singlet, d = doublet, * t = triplet, m = multiplet and b = wide. Optical rotation values are measured at 0.1% methanol concentration at 25 ° C unless otherwise specified. All temperatures are in degrees Celsius. IR = infrared spectrum.

Examples

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-inoate

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

- 25 ’C and a 1.6 M solution of n-butyllithium in hexane (24.4 ml) is added over 5 minutes while maintaining the temperature below - 20 C. 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 at -70 ° C for a further 20 minutes, and then a solution of N-tert-butoxycarbonyl-L-phenylalanalnal (6.5 g, see JR Luly et al., J. Org. Chem., 1987, 52, 1487) in dry tetrahydrofuran (15 mL), keeping the temperature again below -65 ° C. The clear yellow solution that formed was stirred at -70 ° C for 2 hours and then acetic acid (4 mL) was added. The cooling bath is removed, the mixture is allowed to warm to -30 ° C and when it reaches this value, water (100 ml) and ethyl acetate (100 ml) are 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 sodium chloride solution (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 silica gel chromatography using ethyl acetate / hexane (1: 4) as eluent. Product containing fractions were evaporated to give an oily residue which solidified overnight. This product was recrystallized from ether / hexane to give the title compounds as a mixture having a diastereomer ratio of 4S, 5S: 4R, 5S of about 2: 1 (4.3 g), mp 98-99 ° C.

Analysis for ^C 19 ^H 25 ^NO 5 calculated :; C, 65.70; H, 7.20; N, 4.03% found: C, 65.62; H, 7.42; N, 4.33 NMR (CDC1 ₃₎ δ = 1. 30 to 1.39 (m, 3H); 1.43 (s, 9H); 2.90-3.11 (m, 2 H); 3.37-3.38 and 4.16-4.19 (2x brm, 1H); 3.93-4.04 (m, IH); 4.22-4.33 (m, 2 H); 4.51-4.56 (m, IH); 4.77-4.79 and 4.87-4.90 (2 * brm, 1H); 7.24-

7.35 (m, 5H).

b) 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), mp 125-126 ° C.

Analysis for ^C 19 ^H 29 ^NO 5: C, 64.95; H, 8.26; N, 3.98% found: c, 64.91; H, 8.40; N, 3.98 1 H NMR (CDCl ₃ ) δ = 1.24-1.33 (m, 3H); 1.39 and 1.42 (2X, 9H); 1.72-1.95 (m, 2 H); 2.38-2.62 (m, 2 H); 2.77 - 2.98 (m, 2H); 3.02-3.04 and 3.40-3.42 (2 × τη, 1H swap, f D ₂ 0); 3.59-3.91 (m, 2 H); 4.08-4.22 (m, 2 H); 4.58-4.61 _(a , 4.8 6-4.89 ₍ 2X m, 1H); 7.22-7.37 (m, 5H)).

c) (S) -5 - [(S) -1-tert-butoxycarbonylamino-2-phenylethyl] 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 chromatography on silica gel using diethyl ether / hexane (40:60) as eluent. The title compound (0.4 g) is obtained, m.p. 98-99 ° C.

G / analysis for ^C 17 ^H 23 ^NO 4 * calculated: c, 66.88 H, 7.54; N, 4.59%. Found: C, 66.77; H, 7.78; N, 4.38 m / e = 306 (MH +).

NMR (CDCl ₃ ) δ = 1.42 (s, 9H); 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 (m, 5H).

[a] ^{2 I} -22.6 ° (c = 1, MeOH).

D

IR (KBr) 1775, 1690, 1525 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 1.6M n-butyllithium in hexane (23 mL) over 3 minutes while maintaining the temperature below 0 ° C. . After an additional 5 minutes at 0 ° C, the solution is cooled to -70 ° C and a solution of (S) -5 - [(S) -1-tert-butoxycarbonylamino-2-phenylethyl] -gamabutyrolactone (5 g) in tetrahydrofuran (38 mL) while maintaining the temperature below -65 ° C. The resulting solution was stirred at -70 ° C for 15 minutes, then benzyl bromide (1.95 mL) in tetrahydrofuran (12.5 mL) was added over 1 minute, then the solution was stirred at -70 ° C for another 10 minutes. Acetic acid (6.5 ml) and cooling bath were added to the solution. shuts down. 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 over magnesium sulfate and the solvent was evaporated under reduced pressure. The crude product is obtained as an oil. The oil was purified by chromatography on silica gel using diethyl ether / hexane (50:50) as eluent. This afforded the title compound (3.52 g) as a clear oil.

Analysis for C ₂₄ H ₄ NO _{2 g} calculated? 'C 72.91; H, 7.34; N, 3.54% S ¹ * ¹ ®JC, 73.22; H, 7.50; N, 3.50 [α] _D ²⁵ -14 '(C = 0.1%, MeOH)

N.M.R. (CDCl 3) δ = 1.37 (s, 9H); 1.95-2.30 (m, 2H); 2,793.20 (m, 5H); 3.92-4.01 (m, IH); 4.21-4.25 (m, 1H); 4.521 4.56 (m, 1 H); 7.21-7.36 (m, 10 H).

- $ 27, f · x .1,

Si · tf> ·· 1 t; s

i.

K f · u i · r t.

b) (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 by addition of acetic acid. The mixture is allowed to stand for 30 minutes and the precipitate formed is filtered off and washed with water. This solid was washed with ethyl acetate, dried (MgSO 4) and evaporated at

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 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), brine (100 mL), dried (MgSO 4) and evaporated under reduced pressure 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}; (DMSO-d6) δ = 0.10 (s, 6H); 0.95 (s, 9H);

j1,30 (m, 10H); l, 35 (m, lH); l, 95 (m, lH); 2.40 (m, IH); ; _2f 72 (m, 2H); 2.85 (m, IH); 3.60 (m, IH); 3 _of 75 (m, 1H);

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

ú <‘ï í> í f | , s

A ξ

í t ’

I • V i

• y y *! ’T

V * ¾ t

Preparation

5-bromomethyl-isoquinoline

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) was added at rt

- 70 ’C. The reaction mixture was kept 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 successively 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 using 50:50 hexane / ethyl acetate as eluent. 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, then held at that 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 sulfate and the solvent was evaporated under reduced pressure. It will be obtained

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

Analysis for ^C 10 ^H 9 ^NO calculated: c, 75.45; H, 5.70; N, 8.80% found: c, 75.06; H, 5.72; N, 8.66.

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 basified with saturated aqueous bicarbonate solution.

solution. The phases are separated and the aqueous phase is extracted with methylene chloride. The combined organic phases were washed with a saturated aqueous solution of sodium bicarbonate, 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. (CDCl 3) δ = 4.90 (s, 2H); 7.50 (t, IH); 7.70 (d, IH); 7.90 (m, 2H); 8.65 (d, IH); 9.25 (s, IH).

Preparation 4

7-bromomethyl-isoquinoline

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

A solution of the above product hydrochloride (0.05 g) in thionyl bromide (0.5 ml) is heated to 60 ° C at

this temperature is maintained 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 pressure at room temperature. The residue was azeotroped with toluene to give the product as the hydrochloride as a colorless solid (0.04 g).

orig. 26

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).

d.

(IV) t;

"Prepare: Number." R ³ MH ⁺ C (theorists Analysis (%) of H value in N brackets) 5 54 6,1 66.06 (66.02 8.34 8.31 2.51 2.56) 6 ch ₂ - ^ 2 ^ - ^ci 562.2 7 F 5 64.6 and ^Xck2 C =) “ ^f and 1 8 654.1 54.9 (55.1 6.75 6.78 2.07 2:14) 9 596 62.2 (62.5 7.71 7.44 2.26 2.35) 10 ch ₂ ~ ^ ~ ^ —ocf ₃ 612.4 61,20 (60.90 7.30 7.30 2.30. 2.30) 11 ₂ CH = CH ^) 452 69.53 (69.40 8.59 8.55 2.52 2.53) 12 'ch ₂ 579 67.92 (68.26 7.92 8.02 4.72 4.82) ' 13 CH OO 579 67.77 (67.78 7.85 8.04 4.64 (4.79) ²

1. Ο, 1 mol Η ₂ 0 2. 0.33 mol Η ₂ Ο

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

a) Azetidine-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 using 50:50 ethyl acetate / hexane as eluent. 1- (N-tert-butoxycarbonyl) -3-hydroxyazetidine is obtained as a white solid (2.57 g), m.p. 51-53 ° C.

Analysis of the CGH ₁₅ NO _3: C, 55.47; H, S, 73; 8,0, 8.09; found: C, 55.29; H, 8.70; Ò, 7.98.

*

b) A solution of the above product (1.0 g) in methylene chloride (35 mL) was treated 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.

t.

(100 mL), dried (MgSO4) and evaporated under reduced pressure to give the title product as a colorless oil (1.4 g).

R '.

I Analysis for C 8 H ₁₇ NO ₅ S | Calcd .: Jc, 43.03; H, 6.77; N, 5.58%.

found: C, 43.00; H, 6.80; N, 5.50 m / e 269 (MNHJ *)

NMR (DMSO-d ₆ ) δ = 1.37 (s, H); 3.24 (s. 3H); 3,883,96 (m, 2H); 4,17-4 _; 28 (m, 2H); 5.25 (m, IH).

Preparation 15

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

The title compound was prepared from 1- (N-benzyloxycarbonyl) - (R) -3-hydroxypyrrolidine (J. Med. Chem., 1992, 35, 1764) using the procedure described in Preparation 14, except that pyridine using triethylamine as the base. The product is obtained as an oil.

Calcd for ^C 13 ^H 17 ^NO 5 ^S : c, 51.63; H, 5.78; N, 4.63% Found: C, 51.60; H, 5.80; n, 4.30 m / e MH ⁺ 300.

N.M.R. (CDCl 3) δ = 2.15 (m, 1H); 2.55 (m, IH); 3.0 (s, 3H);

3.45-3.80 (m, 4H); 5.10 (s, 2H); 5.25 (m, IH); 7.30 (m, 5 H).

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) triphenylphosphine (13.58 g) was added under nitrogen.

The resulting solution was cooled to -30 ° C and methyl tosylate (10.06 g) was added over 0.5 h, and diethylazo was added. dicarboxylate (10.41 g). After a further one hour, the reaction is continued

The mixture was allowed to warm to room temperature

I ·. is maintained for 65 hours. The reaction mixture is evaporated under reduced pressure, the residue is dissolved in dichloromethane, washed with water, dried

magnesium sulfate and evaporated under reduced pressure to an oil.

The residue was purified by silica gel chromatography eluting with methylene chloride / methanol (98: 2 to 96: 4). A second chromatography (80:20 to 50:50 hexane / ethyl acetate) was then performed. Obtained as a gold-colored oil (10.22 g).

Analysis for ^C 19 ^H 21 ^NO 5 ^S: C, 60.78; H, 5.64; N, 3.73% found:. ^c , 60.59; H, 5.68; N, 3.67

4-9 ° (c = 0.1% MeOH)

D 'N.M.R. (CDCl 3) δ = 1.85-2.25 (m, 2H); 2.45 (s, 3H); 3.40-3.65 (m, 4H); 4.95-5.15 (m, 3H); 7.20-7.35 (m, 7H);

7.75 (d, 2H).

Preparation 17

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

The title compound was prepared from 1- (N-tert-butoxycarbonyl) -4-hydroxypiperidine as described in Preparation 14. The product obtained had a melting point of 85-86 ° C.

Calcd for ^C 11 ^H 21 ^NO 5 ^S :, c, 47.3; H, 7.58; N, 5.02% found: Jc, 47.2; H, 7.66; N, 4.91. @ ₁ H NMR (CDCl3) .delta. = 1.5 (s, 9H); l, 85 (m, 2H); 2.0 (m, 2H); | 3.08 (s, 3H); 3.35 (m, 2H); 3.75 (m, 2H); 4.9 (m, IH).

Preparation 18 1- (N-tert-butoxycarbonyl) -3- (imidazol-1-yl) and zetidine

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- (methanesulfonyloxyazetidine (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 224 (MH) &lt; ^{+ &gt};

• .N.M.R. (DMSO-d6) = 1.40 (s, 4.26-4.35 (m. 2H); 5.12 (m, 1H); 7.79 (s, 1 H). 9H); 3.95-4.06 (m, 2 H); 1H); 6.96 (s, 1 H); 7.43 (s, 4 Preparations 19 to 2 5

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

Preparing the number PX- (CR ⁷ R ⁸ ) _m -Het (MH) &lt; ^{+ &gt;} Analysis (%) ‘N brackets) C (theoretical h H odnoty v 19 nh. 239.3 55.1 (55.4 7.50 7.67 22, 9 23.5) 1 Boc-N ^/ \ -N'Z ^! * N \ = J 20 Boc-N NMR: 1 (m, 2H); 5.1 (m, 18.8, 10 o (s, 33 (s, 9H) 4.05 4.2 4.2 (m, 2H); H); 7.99 (s, 1H); 1H) - 21 ' / X (S) of X Cbz-N y-N N \ = J 272 6 5.57 (65.60 6,3 / 6, 25 15.04 15.25) ¹ 22 Cbz-N J = J 272 6 5.69 (65.60 6.33 6.25 15.25 15.25) ¹ 23 Boc-N ^ N ^ N 252 24 BOC-N Y-N ^ N 253 25 CH, r = Boc-N4 _/ XNN 265.9 61.0 (61.3 8.92 8.82 15, 6 15.3) ²

j: 1. 0.05 moles of methylene chloride 2. hemihydrate

Preparation 26

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

Reaction of 4-methylimidazole with 1- (N-t-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};;

NMR (DMSO-d ₆ ) δ = 1.43 (s, 9H); 1.7 (dq, 2 H); 1.93 (bd, 2H); 2.07 (s. 3H); 2.83 (m, 2 H); 4.05 (m. 3H);

6.95 (s, 1 H); 7.55 (s, 1 H).

b) 1- (N-tert-butoxycarbonyl) -4- (5-methylimidazpl-1-yl) piperidine

Minor isomer, Rf 0.52; m / e 265.9 (MH) &lt; ^{+ &gt;.}

NMR (DMSO-d ₆ ) δ = 1.43 (s, 9H); 1.7 (dq, 2 H); 1.9 (bd, 2H); 2.05 (s. 3H); 2.88 (m, 2 H); 4.05 (m. 3H); 6.6 ' (s, 1H); 7.65 (s, IH).

Preparation 27

1- (N-t-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. To the mixture was added n-butyllithium (25 mL, 1.6 N in hexane) at such a rate that the reaction temperature remained below -35 ° C. Then 1- (N-tert-butoxycarbonyl) -4-ketopiperidine (2.65 g) in dry tetrahydrofuran (10 ml) was added dropwise over 10 minutes, the temperature was kept below -40 ° C and the resulting mixture was stirred for 2 hours. at - 40 ° C. The reaction mixture was further stirred with hydrochloric acid (50 mL, 0.1 N) minutes, then ethyl acetate (50 mL) was added and the resulting mixture was stirred 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) and then with 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) ⁺

b) This product (267 mg) was mixed with diisopropylamine (350 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 hours. The resulting mixture was diluted with water (4 mL), adjusted to pH 9 with 1 M sodium hydroxide solution and extracted with ethyl acetate (3 x 10 mL). The combined organic extracts were dried (MgSO 4) and evaporated to the resin (260 mg). The residue was purified by silica gel chromatography, eluting with dichloromethane: methanol: concentrated aqueous ammonia (95: 5: 1). The title compound was obtained as a yellow resin (144 mg), m / e 250.1 (MH) ⁺ [nTm.R. (CDCl 3) δ = 1.46 (s, 9H); 2.6 (bs, 2 H); 3.53 (t, 12H); 4.0 (bs, 2 H); 6.3 (m, IH); 7.0 (s. 2H); 9.4 (bs,

1H).

y Preparation 28

C.

? l- (N-t-butoxycarbonyl) -4- (imidazol-2-yl) piperidine

The product of Preparation 27 (0.55 g) was dissolved in ethanol (30 ml) and hydrogenated at 0.2 MPa in use; 10% palladium on carbon catalyst. The catalyst was

It is filtered from the reaction mixture and the solvent is evaporated to give the product as a foam (0.55 g), m / e 252.1 (MH) ^{@ +} @ 1 H NMR (CDCl3) .delta. 9H); 1.68 (qd, 2 H); 1.95 (bd, 2 H); 2.75 (tt, 2H); 2.92 (bt, 1 H); 4.1 (bd, 2 H); 6.9 (s. 2H); 8.77 (bs, 1 H).

Preparation

1- (N-t-butoxycarbonyl) -4- (imidazol-l-yl) methyl

1,2,5,6-tetrahydropyridine

(a) 60% dispersion of sodium hydride in oil (4 g) in. dry dimethylsulfoxide (100 mL) was washed with hexane to remove oil and then heated to 70 ° C for 1 hour with stirring. Dry tetrahydrofuran (100 mL) was then added and the resulting mixture cooled to -20 ° C. To this mixture was added a solution of trimethylsulfonium iodide (20.4 g) in dimethylsulfoxide; (80 mL) followed by a solution of 1- (N-t-butoxycarbonyl) -4-keto piperidine (19.9 g) in dry tetrahydrofuran (100 mL) and stirred at room temperature for 0.5 h. - 10 ºC and then 1 hour at room temperature. Next, water (500 mL) was added and the mixture was extracted with ethyl acetate

IN

I? (3 x 250 mL). The combined extracts were washed with sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure.

Pressure. The residue was purified by silica gel chromatography eluting with cyclohexane: diethyl ether: isopropanol (60: 40: 1). 1- (N-tert.).

s £

f: 'f

butoxycarbonyl) piperidine-4-spiro-2'-oxirane as a colorless solid (19.6 g), m.p. 65-66 ° C.

Analysis for ^C 11 ^H 19 ^NO 3 calculated: C, 61.97; H, 8.92; N, 6.57% found: 'C' θ ² · ¹¹ * H, 9.06; N, 6.55.

.9 b) To a solution of imidazole (1.91 g) in dry acetonitrile (30 mL) under stirring under a nitrogen atmosphere is added a * 80% oil dispersion of sodium hydride ( 0.84 g) and the resulting mixture is heated to 60 ° C until a solution is 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 (0.888 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 for ^C 13 ^H 23 ^N 3 ° 3. 1/10 CH ₂ Cl ₂ calculated: c, 58, 43; H, 8.07; N, 1.4.50¾ found: ^{C / 58} ' ³⁵ ' * 8.30; n, 14.53 n / e 282 (MH) &lt; ^{+ &gt};.

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 with stirring a solution of methanesulfonyl chloride (2.20 ml) in; dry methylene chloride (10 mL) and the resulting mixture was allowed to warm to room temperature and maintained at this temperature for 1414 hours. The reaction mixture was washed with water, dried

Μ. '

f.

ΐ i

Magnesium sulphate and evaporated under reduced pressure. The residue is purified by chromatography on silica gel, eluting with of methylene chloride, methanol and concentrated (with a density of | 0,88) aqueous ammonia in a ratio of 96: 4: 0 to 95: 4: 1 as eluent | agents. Obtained as a gold-colored oil

I (1.36 g).

í · a í ’ l · ' Analysis for ^C 13 ^H 21 ^N 3 ° 2 ’ 1/4 CH ₂ C1 ₂ t. calculated: c. 60, 14 ; h. 7.61; N, 14.77 % * found: C. 59.93 ; h. 7.60; N, 14.79 ? '· m / e (MH) &lt; ^{+ &gt;.} 264 ť N.M.R. (CDCl) 8 = 1.45 (ε 9H); 1.97 (M, 2H); 3.50 (t, ? - 2H); 3.90 (with, 2H); 4.50 (s, 2H); 5.50 (with, 1H); 6.90 (s, 1H); 7.10 (with, 1H); 7.55 (s, 1 H).

x

Preparation 30 1- (N-tert-butoxycarbonyl) -4- (imidazol-1-yl) methyl | piperidine

A solution of the product of Preparation 29 (1.33 g) in absolute i-ethanol (25 mL) was hydrogenated with stirring in the presence of 10% palladium on activated carbon (0.3 g) as a catalyst 4) at room temperature and pressure for 4 hours. 0.35 MPa. Reaction mixture? The mixture is filtered and evaporated under reduced pressure and the residue is taken up in vacuo

azeotropically evaporated 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 ° 2 ' ^CH 2 ^CX 2 | Calcd .: 1 C, 63.36; h, 8.74; N, 15.84% found: J ^{c '63} , 08; h, 8.60; n, 15.50 | Im / e (MH) &lt; ^{+ &gt;} 266

N.M.R. (CDCl) δ = 1.15 (M, 2H); 1.45 (s, 9H); 1.58 (m, 2H); 1.85 (m, 1H); 2.65 (T, 2H); 3.80 (d, 2H); 4.10 (m, 2H); 6.90 (s, 1H); 7.10 (with, 1H); 7.45 (s, 1 H). P r í p r a v a 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 added dropwise after 2 hours. a solution of 1- (N-tert-butoxycarbonyl) -4-keto-piperidine (6.0 g) in dry methylene chloride (50 mL). 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). The combined extracts were 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, 9H); 2.55 (s, 2 H); 3.70 (t, 2 H); 4.05 (s, 2 H); 5.80 (s, IH); 7.10 (s, 2 H); 7.65 (s, 1 H).

Preparation 32 f 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 i. room 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 sulphate 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). It will be obtained

4-Amino-1- (N-benzyloxycarbonyl) piperidine as a yellow oil. A solution of this product (1.5 g) in toluene (20 ml) was treated 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 is purified by chromatography on silica gel using a mixture of methylene chloride, methanol and concentrated aqueous solution.

93: 7: 1 ammonia as eluent. The title compound is obtained as a colorless oil, m / e (MH ⁺ ) 287; NMR (DMSO-d ₆ ) δ = 1.82 (m, 2H); 2.01 (m, 2 H); 2.833.09 (m, 2H); 4.12 (m. 2H); 4.40 (m, IH); 5.11 (s, 2H);

7.28-7.45 (m, 5H); 8.65 (s, 2 H).

t t:

£

I

f. ? · $

Preparation 33

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

A solution of 1- (N-tert-butoxycarbonyl) -3- (imidazol-1-yl) azetidine from Preparation 18) (0.72 g) in methylene chloride (30 mL) was saturated with hydrogen chloride at 0 ° C and maintained at this temperature. another hour. The solvent was evaporated under reduced pressure and the amine hydrochloride obtained was dissolved in N, N-dimethylformamide (25 ml). To this was added N-tert-butoxycarbonyl- (S) -valine ester and N-hydroxysuccinimide (1.0 g) and N, N-diisopropylethylamine (1.7 mL) at room temperature. The reaction mixture was stirred at room temperature for 18 hours and then the solvent was evaporated 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) ⁺ ;

NMR (DMSO-d ₆ ) δ = 0.89 (m, 6H); 1.38 (s, 9H); 1.90 (m, 1H); 3.61-3.78 (m, 1H); 3.92-4.10 (m, IH); 4,24-

4.80 (m, 3H); 5.19 (m, IH); 6.98 (s, IH); 7.06 (dd, IH); 7.38 (s, 1 H); 7.77 (s, 1H).

Preparations 34 to 43

The following compounds of formula VII, wherein R ⁴ 7 f 1 is (S) -isopropyl and R and R 0 are each hydrogen, are prepared as described in Preparation 33 using the appropriate intermediates from Preparations 18 to 32, which are condensed with an ester N-tert-butoxycarbonyl - (S) -valine and N-hydroxysuccinimide.

(VII)

To prepare: number -X- (CH ₂ ) _m Het ΜΉ * Analysis (%) C H N (theoretical values in brackets) 34 nh ₂ —N ^Z \ -N ^/ ^ N \ = J 338 35 I —N ^z \ -n ' ^z ^ n CF _: 391 52.09 6.73 14.03 (52.32 6.46 14.36) 36 ^. (S · a N ^ N ^ yl-N 337 58.76 8.51 16.63 (59.11 8.46 16.22) ¹ 37 and ZJR '. alpha -n ^ - ^ nn \ ___ / \ =. ' 337 59.46 8.54 15.96 (59.54 8.24 lo, 24) ^: 38 —F / N ^ N 351 • 39 CH ₃ -R ( ^N ) ^-N 3 c 5 61.1 9.23 14.9 (61.1 8.91 15.0) ' 40 —N SN ^ N ch ₃ 364.9 61.9 9.15 14.9 (61.9 8.9 15.2) ³ 41 351, 1 42 -O ^ch ^ 3 6 5 59.29 8.15 14.29 (59.44 8.43 14.37) ⁴ 43 _ _n ^ and_ _n ^ _n \ = j 349 61.51 8.11 15.68 (61.46 8.03 15.89) ⁵

1 ^ Hemihydrate T1 0.25 mol H ₂ O 5 0.05 mol CH ₂ Cl ₂

2. 0.1 M CHjC1 ₂ 4 0.3 mol of CH ₂ C1 ₂

Preparations 44 to 48 /

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.

(VII)

Preparation number -X-Het MH ⁺ C (theorists Analysis (%) of the H value in N brackets) 44 365 62,01 (61,99 9.28 8 _f 87 14.94 15.22) ' 45 —N ^? In on 366.3 46 ch ₃ / - \ A —N> ~ NN \ __ / \ = J 379 61.7 (62.0 9.22 9.10 14.5 14.5) ² 47 —H ν'Ά ch ₃ 379.9 62.2 (62.0 8.98 9.10 14.33 14.5) ² 48 1 H 2 -N 3 N ₃ CH ₃ 379.9

1. 0.02 mol H ₂ O

2. 0.5 mol H, O

Preparation

1- (N-t-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 instead of 3- (imidazol-1-yl) azetidine hydrochloride and (S) -isoleucine ester with N-hydroxysuccinimide. m / e 313 (MH) ⁺ [a] ²⁵ -16 (c = 0.34%, MeOH)

D

N.M.R. (CDCl3) < 5 = 0.9 (m, 6H); 1.2 (m, IH); 1.4 (s, 9H); 1.6 (m, IH); 1.75 (m, IH); 2.5 (m, 4 H); 3.7 (m, J 2H); 4.15 (m. 2H); 4.55 (m, IH); 5.2 (d, 2 H).

Preparation

N - ((R) -2-Benzyl- (S) -5-terc.butoxykarbonylamino- (S) -4- (tert-butyldimethylsilyloxy) -6-phenylhexanoyl) - (S) -valine

The title compound was prepared as described in

S. J. de Solms et al., J. Med. Chem., 1991, 34, 2852.

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

The title compound was prepared from p-toluenesulfonylmethyl isocyanide and isobutyraldehyde by the method described by Van Leusen, Schaart and Van Leusen Recueil, 98, No. 5, p. 5, 258 (1979). IR (nujol) 2100 cm &lt; ^{-1 &gt;.}

Δη / θ 2 67 '- (M + N + H +) 1 NMR (CDCl 3) δ = 1.15 (d, 6H); 2.5 (s, 3H); 2.84 (m,> 1H); 6.88 ( d, 1H), 7.4 (d, 2H), 7.85 (d, 2H).

Preparation

3-oxetanyloxykarbonyloxysukcínimid

Oxethan-3-ol (2.0 g) and N, N-diisopropylethylamine (7.76 g) were dissolved in methylene chloride (50 mL) and added dropwise to a solution of bis (trichloromethyl) carbonate over 15 minutes under a nitrogen atmosphere. (2.69 g) in methylene chloride (100 mL) kept at -20 ° C. The solution was stirred at 20 ° C for a further 15 minutes and N-hydroxysuccinimide (3.45 g) was added in one portion. 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. 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- (imidazol-l -ylJazetidín

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

1- [N-tert-Butyloxycarbonyl- (S) -valyl) -

Of 3- (imidazol-1-yl) azetidine (from Preparation 33, 0.79 g) in methylene chloride (50 mL) was saturated with hydrogen chloride at

’C and maintain 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-butoxycarbonylamino- (S) -4- (tert-butyldimethylsilyloxy) - 6-phenylhexanoic acid (from Preparation 2, 1.32 g), 1-hydroxybenzotriazole (0.36 g), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.58 g) and N, N-diisopropylethylamine ( 1.75 mL) in dimethylformamide (75 mL) (20 min). 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 is obtained as a colorless oil. The oil was purified by chromatography on silica gel 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 41 ^H 58 ^N 5 ° 5 ^sX . H ₂ O Calcd: C, 66.39; H, 8.36; N, 9.44% found: C, 66.04; 11,8,26; N, 9.61 m / e 733 (MH) ⁺ [α] ^{21 D} -6 ° (c = 0.29%, MeOH)

D

NMR (DMSO-d ₆ ) δ = 0.12 (m, 6H); 0.90 (m, 15H); 1.25 and 1.29 (2 xs, 9H); l, 96 (m, lH); 2.32-2.77 (m, 4H);

j 2.89 (m, 1H); 3.48-3.74 (m, 2 H); 3.88-4.06 (m, 3H); 4.18 and 4.66 (m, 4H); 5.12 (m, IH); 6.77 (t, IH); 6.98 (s, IH); 7,077,32 (m, 10 H); 7.40-7.82 (2.times.s, 1H); 7.76-7.96 (2.times.s, 1H);

8.0-8.20 (2X d, 1H).

b) 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -3- ( imidazol-l-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 is a colorless solid (0.52 g), m.p. 124-126 ° C.

^C 35 ^H 47 ^N 5 ^O 5 requires C, 68.05; H, 7.67; N, 11.34% found: C, 67.99; H, 8.01; N, 11.06 m / e 618 (MH) ⁺ [α] ²⁵ -3 ° (c = 0.14%, MeOH)

D

NMR (DMSO-d ₆ ) δ = 0.80 (m, 6H); 1.21 to 1.47 (m, 1H); ; 1.27 (s, 9H); 1.58-1.71 (m, 1H); 1.81-1.98 (m, 1H); 2,44j2,62 (m, 2H); 2.68 to 2.92 (m, 3H); 3.37 to 3.61 (m, 2H); 3,854,02 (m, 2H); 4.13-4.36 (m, 2 H); 4.41-4.56 (m, 2 H);

| 5.18 (m, IH); 6.43 (d, IH); 6.98 (s, IH); 7. 05-7.30 (m, 10H); 7.26-7.77 (2 × S, 1H); 7.38-7.82 (2.times.s, 1H); 7.88-7.98 (2x: d, 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), an appropriately protected intermediate carboxylic acid of formula IV and a suitable amine of formula VIII, and then removing the tert-butyldimethylsilyl protecting group as described in step b). .

33 Q in co and VD O Ch z λ; σχ oo · σχ o m » rh R-4 * · * » ABOUT o o «H R * co cn Rľ. <P f — i »“ H : . - i N <· R- m r * vo σχ m o VO VO within within σν co N X G w • w «* «. '> <Ό »- <o r- r- KD kD in in <0 Ä 5 '0) Λί U m n CO o • H -P o o τ * within σχ r * 0) M σχ cn σχ σχ r * r * o o \ o o m m this Φ * - · -L> *** s o 0 1 I about 53 T 1 about CJ fX ' about F and • R * t-1 4> 1 o 1 CO -P 17 • e r— ( r> LL Het about 7 CM X z— <] tl 1 I «C. 1 n LL LL r ⁱ > i ω ω f 0 about about in 1 1 f » T x à ll about eye Q L 0 11 r CM at X x x from O / U \ C 1 73 «5 0 WHAT σχ o rH • u fi> υ

Example 11

1 '[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, then tert. the butyldimethylsilyl protecting group is removed as described in step 1b). The title compound is obtained, m.p. 111-112 ° C.

Calcd for ^C 37 ^H 48 ^F 3 ^N 5 ^O 6: J, 62.08; H, 6.76; N, 9.78% found: _c , 61.90; H, 6.85; N, 9.61 m / e (MH) ⁺ 716 '[α] ²⁵ + 2 ° (c = 0.1%, MeOH) [α] ²⁵ 4-14 ° (c = 0.1%, MeOH)

D 365

N.M.R. (DMSO-d 6) δ = 0.65-0.90 (m, 6H); 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, IH); 6.40 (d, IH); 6.90-6.95 (2x s, 1H); 7.10-7.30 (m, 10H); 7.70-7.80 (2x s, 1H); 7.90-8.0 (2x d, 1H).

Example 12

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

r.

A

V | ·. The procedure described in Example 1 was repeated using 1- (N-tert-butoxycarbonyl- (S) -valyl) -3- (R) - (imidazol-1-yl) pyrrolidine to give the title product, m.p. C.

£ i - '.

Analysis for ^C 37 ^H 48 ^F 3 ^N 5 ° 6 ^{'2/5 H} 2 ° calculated: C, 61.47; H, 6.80 ; N, 9.69% found: C, 61.44; H, 7.05 ; N, 9.71 ^m / e (MH) ⁺ 716 1; [α] ²⁵ -23 ° (c = 0.1%, MeOH). D N.M.R. (DM SO-d6) δ = 0.70-0 85 (m. 6H) ; 1.15-1.40 (m, Í10H); 1.60 (m, 1 H); 1.85 (m, 1H); 2.00 -2.90 (m, 7H); ΐ 3.25 - 3.70 (m, 5H); 3.95 (m, 1H); 4.20 (m, 1 H); 4,45-

4.65 (m, IH); 4.75-4.95 (2.times m, 1H); δ, 40 (m, 1H); 6.90 (s, 1H); 7.05-7.25 (m, 10H); 7.70-7.75 (2.times.s, 1H);

7.90-8.05 (2x d, 1H).

Example 13

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-l-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).

Calcd for ^C 43 ^H 65 ^N 5 ° 5 ^Si * ^{1/4 H} 2 °: No, 67.54; H, 8.56; N, 9, 5% ^found:? C, 67.32; H, 8.44; N, 9.22 m / e 760 ([delta]) ⁺

NMR (DMSO-d ₆ ) δ = 0.12 (m, 6H); 0.91 (m, 15H); 1.13 l, 30 (m, 2H); l, 26 (s, 9H); 1.47-1.76 (m, 2H); 1,892,08 (m, 4H); 2.36 to 2.97 (m, 6H); 3.50-3.75 (m, 2H); 3,994,61 (m, 4H); 6.79 (m, IH); 6.90 (d, IH); 7.07-7.29 (m, 11H); 7.68-7.73 (2X, 1H); 7.93-7.99 (2 × d, 1H).

b) 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -4 (imidazole -l-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 for ^C 37 ^H 51 ^N 5 ° 5 - ^{1/4 H} 2 ° calculated: C, 68.30; H, 7.92; N, 10.77% ¹ found:: C, 68.38; H, 7.96; N, 10.59 m / e 646 (MH) ⁺ [α] ²¹ -10 ° (c = 0.1, MeOH)

NMR (DMSO-d ₆ ) δ = 0.77 (m, 6H); 1.19-1.37 (m, 1H);

l, 28 (s, 9H); 1.48-1.72 (m, 3H); 1.82 to 2.04 (m, 3H); 2.42, 2.94 (m, 7H); 3.38-3.60 (m, 2H)); 3.96-4.15 (m, IH);

4.28 (m, IH); 4.37-4.58 (m, 3H); 6.43 (d, IH); 6.88 (s, IH); , 7.04 to 7.29 (m, 11 H); 7.65-7.69 (2 × S, 1H); 7.84-7.88 (2 x id, 1H).

Examples 14 to 30

The following compounds were prepared as described in Example 13, but using the appropriately 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 ).

Het à ₍ ·, í '·, ·

É · C y x t ·· ι: λ

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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. the amine product is condensed with N - ((R) -2-benzyl- (S) -5-tert-butoxycarbonylamino- (S) -4-tert-butyldimethylsilyloxy) -6-phenylhexanoyl) - (S) -valine as described in Example 1 of step a) to give 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 calculated for ^C 36 ^H 50 ^N 6 ° 5 · ^H 2 ° _g : C, 65 _from 06; H, 7.83; N, 12.65% found: C, 64.90; H, 7.93; N, 12.48 [α] ²⁵ -9 ° (c = 0.12 MeOH)

D

NMR (DMSO-d ₆ ) δ = 0.70-0.92 (m, 6H); 1.15-1.41 (m, 10H);

1.46-1.76 (m, 3H); 1.81-2.16 (m, 3H); 2.53 to 2.96 (m, 7 H);

P, 3.38-3.64 (m, 2H); 3.95-4.15 (m, IH); 4.27-4.61 (m, 4H);

r 6.36-6.44 (m, IH); 7.01-7.12 (m, 10H); 7.78-7.91 (m, 1H);

δ 8/54 (s, 1H); 8.60 (s, 1H).

i ¹ f

I k

Ϊ ¹ · irj

í · í {ľ

Example 32

1- [N - ((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) -4-tert-butyldimethylsilyloxy) - (R) -2- (4-trifluoromethoxybenzyl) -6-phenylhexanoic acid with an amine prepared by deprotection of 1- (N- (tert. butoxycarbonyl) -4-imidazol-2-yl (1,2,5,6-tetrahydropyridine) by the method described in Example 1a) gives 1- (N- (S) -5-tert-butyloxycarbonylamino) - (S) -

4-tert-Butyldimethylsilyloxy) -6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S) -valyl] -4- (imidazol-1-yl) -

1,2,5,6-tetrahydropyridine.

Analysis for C ₃₃ H ₆₂ ^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, 62.19; H, 7.39; N, 8.38. m / e 842 (M) &lt; ⁺ &^gt; [[alpha]] -10 [deg.] (c = 0.1%, MeOH) D

N. M.R. (CDCl 3) δ = 0.10 (m, OH); 0.80 (d, 6H);

0.90 (s, 9H); l, 35 (s, 9H); 1.20 to 1.95 (m, 5H); 2,352,55 (m, 3H); 2.70 (m, 2H); 3.41-4.10 (m, OH); 4.55 (m, IH); 4.65 (d, IH); 5.60-5.80 (2 x s, 1H); O 25 (d, lH); 6,907,30 (m, llH); 7.65 (s, IH).

b) The title compound as the free base was 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. There was obtained 1-tartaric acid salt as a colorless solid, m.p. 92-152 ° C (0.61 g).

Analysis for ^C 38 ^H 38 ^F 3 ^N 5 ° 6 ° C • ^CH 16 ° 6 * ^{1/3 H} 2 ° calculated: C, 57.11; H, 6.23; N, 7.93% found: 57.40; H, 6.64; N, 7.51 m / e (MH) &lt; ^{+ &gt;.} 728

[α] ²⁵ + 6.7 '(c = 0.1%, MeOH)

D [α] ^{2 S} - 6.7 ° (c = 0 _f %, MeOH)

65

N.M.R. (DMSO-d 6) δ = 0.75 (m, 6H); 1.25 (s. 9H); 0.951.40 (m, IH); 1.0 (m, 1H); 1.90 (m, IH); 2.35-2, 95 (m, 7H); 3.0-3.85 (m, 4H); 4.0 (m, IH); 4.10 (m, IH); 4.25 (s, 2 H); 4.40-4.60 (m, 2 H); 6.0 (m, IH); 6.40 (d, IH)

7.0 (s, 1 H); 7.10-7.30 (m, 9H); 7.50 (s, 1 H); 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-isopropylimidazol-l-yl) piperidine

a) Deprotection of 1- (N-tert-butoxycarbonyl- (S) isoleucyl) -4-ketopiperidine (Preparation 49) and subsequent reaction with (S) -5-tert-butoxycarbonylamino- (S) -4- (tert-butyldimethylsilyloxy) -6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoic acid (Preparation 10) gave

1- (N - ((S) -5- (tert-butyloxycarbonylamino) - (S) -4- (tert-butyldimethylsilyloxy) -6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S) isoleucyl] -4-ketopiperidine.

Calcd for ^C 42 ^H 62 ^F 33 ° 7 ^S : C, 62.60; H, 7.80; N, 5.20 Found: C, 62.50; H, 8.00; N, 5.20: m / e 806.5 (MH) +. [α] ²⁵ -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 1 hour at 0.2 MPa in the presence of palladium on carbon ⁴ (5100 mg). The catalyst was filtered off and the solvent was evaporated. The resulting colorless foam (2.0 g) was purified by chromatography on silica gel, eluting with methylene chloride: methanol: concentrated aqueous ammonia (90: 10: 1). Concentration of the appropriate fractions gives 1- (N - ((S) -5-tert-butyloxycarbonylamino- (S) -4-tert-butyldimethylsilyloxy-6-phenyl- (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 mixed 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 eluting with methylene chloride: methanol: concentrated aqueous ammonia (98: 2: 0.4). 1- (N - ((S) -5- (tert-butyloxycarbonylamino) - (S) -4-tert-butyldimethylsilyloxy-6-phenyl (R) -2- (4-trifluoromethoxybenzyl) hexanoyl- (S)) was obtained. 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 tetrabutylammonium fluoride as described in Example 1b). This gives the title compound.

Analysis for ^C 42 ^H 58 ^F 3 ^N 5 ° 6 · 1 ^H / ^{2 H} 2 ° calculated: C, δ 3.5; H, 7.48; N, 8.81% found:; ^c , 63.5; H, 7.70; N, 8.70 im / e 785.9 (MH) ⁺

NMR (DMSO-d ₆ ) δ = 0.8 (m, 6H); 1.05 (m, IH); 1.2 (m,

6H); 1.3 (s. 9H); 1.3-2.0 (m, 7H); 2.5-3.0 (m, 6.5H);

3.2 (br. 0.5H); 3.3 (m, IH); 3.58 (m, IH); 4.13 (m,

3H); 4.4-4.65 (m 1, 4H); 6.43 (m, IH); 6.63 (s, 1H);

7.3 (m. 9H); 7.47, 7.6 (s, s, 1H); 7.95 (bt, IH).

Example 34

1- [N - ((S) -4-Hydroxy- (S) -5- (oxetan-3-yloxycarbonylamino) 6-phenyl- (R) -2- (4-trifluórmetoxybenzy1) hexanoyl- (S) valyl] - 4- (imidazol-l-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 (MgSO 4), 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 well as

eluent. 1- [N- (S) -5-amino- (S) -4-hydroxy-6-phenyl- (R) -2- (4-trifluoromethoxybenzyl) hexanoyl (S) -valyl] -4- (imidazole-1) was obtained. -yl) piperidine as a white solid (2.98 g), m / e 630 (MH) ⁺ ;

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, 8H); 3.05-3.29 (m, 3H); 4,054,56 (m, 5H); 6.88 (s, IH); 7.10-7.27 (m, 10H); 7.67 (d, IH); 7.95 (m, IH);

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), then the resulting mixture was stirred at room temperature for 1 hour. The solution was washed with 0.5 M 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 and so. to give the title compound as a white solid, mp 201-203 ° C.

For ^C 37 ^H 46 ^F 3 ^N 5 ° 7 * ^-1 / ^{2 H} 2 ° calculated: C, 60.15; H, 6.41; N, 9.48% found: [c, 60.32; H, 6.22; n, 9.15 m / e 730 (MH) ⁺ [α] ²⁵ + 3 ° (c = 0.1%, MeOH), [α] ²⁵ + 32 ° (c = 0.1, MeOH)

D 365

NMR (DMSO-d ₆ ) δ = 0.77 (m, 6H); 1.22-2.05 (m, 7 H); 2.50; 2.93 (m, 8H); 3.42-3.60 (m, 2H); 4.08 (d, IH); 4.22 and 4.73 (m, 7H); 5.12 (m, IH); 6.89 (s, IH); 7.11-7.28 (m, 11 H);

7.67 (d, 1H); 7.86 (d, IH).

in.

i:

Example 35

1- [N- (S) -4-hydroxy- (S) -5- (oxethan-3-yloxycarbonylamino) -6-phenyl- (R) -2- (3-phenylprop-2-enyl) hexanoyl- (S) ) isoleucyl] -4- (imidazol-l-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.

= ⁴ The reaction mixture was concentrated under reduced pressure and the residual resin was azeotroped 3 times with toluene and dried. A solution of the crude amine product and diisopropylethylamine (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 chromatography on silica gel using methylene chloride: methanol: concentrated aqueous ammonia = 95: 4: 1 as eluent _w. Obtained as a colorless foam (0.26 g), m.p. 157-160 ° C.

For ^C 38 ^H 49 ^N 5 ° 6 * ^{3/4 H} 2 ° calcd: c, 66.60; H, 7.43; N, 10.22% found: c, 66.45; H, 7.49; N, 10.22 [α] ²⁵ +32 ° (c = 0.1%, MeOH)

NMR (DMSO-d ₆ ) δ = 0.75 (m, 6H); 1.25-2.95 (m, 12H);

3.05-3.65 (m, 4H); 3.95-4.75 (m, 10H); 5.05 (m, IH);

δ 6.10 (m, 1H); 6.30 _(d , _IH ); 6.80-6.85 (2.times.s, _1H );

« ° r ^{9: 3} ~ ^{7 »35} (m, 11 H); 7.55-7.65 (2 xs, 1H); 7.95 (d

1H).

f I ý

í í.

í '

Example 36

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

The title compound was prepared as described in Example 35, except that isopropyl chloroformate was used to react with the intermediate amine. The crude product was purified by silica gel chromatography 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.

Analysis for ^C 38 ^H 51 ^N 5 ° ^5'H 2 ° calculated: C, 61.75; H, 7.16; N, 8.57 found: C, 61.60; H, 6.87; N, 8.38 m / e 658 (MH) &lt; ^{+ &gt;} [α] ²⁵ +2 6 ° (c = 0.1%, MeOH). D

0.75 (m,

NMR (DMSO-d ₆ )? = 14H);

6.65 (d,

6H); 1.0 (m, 6H); 1.253.65 (m, 14H); 3.95-4.70 (m, 10H); 6.10 (m, IH); 6.30 (d, IH); 6.65 (d, IH); 6.85-6.90 (2.times.s, 1H); 7.05-7.35 (m, 11H); 7.65-7.75 (2x s, 1H); 7.90 (d, 1H).

Example 37

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

a) Dissolve N-benzyloxycarbonyl-L-valine (1.03 g) and dicyclohexylcarbodiimide (0.51 g) in dichloromethane (25 mL) and stir for 3 hours. The precipitated dicyclohexylurea is filtered off and the filtrate is evaporated to a white foam. This white foam was treated 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 over magnesium sulfate 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] -4- (imidazol-yl) piperidine as a white foam.

Analysis for ^C 51 ^H 65 ^F 3 ^N 6 ° ^9'H 2 ° calculated: C, 62.43; H, 6.88; N, 8.56% found: C, 6 2.63; H, 6.73; N, 8.37. b) The product of step (a) (0.95 g) is dissolved in

absolute ethanol (50 mL) and 10% palladium on charcoal (0.1 g) was added followed by hydrogenation at 40 psi for 4 hours 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 is obtained in the form of a white glass (0.48 g), m.p. 122 ° C.

Analysis for calculated: found:

^C 43 ^H 59 ^F 3 ^N 6 ° 7 ^C 4 ^H 6 ° 6 ^H 2 °; C, 56.61; 1c, 566, 74;

NKR _(DMSO-d6) delta =; 1.35-1.77-3.51 (m, FH-466 (m, 12H);

(M,

1H);

2H)

H, Ί / * i (m,

6.77;

6.78;

0.8-0.95 c-2.15 (

1H); 4.1

N,

N, > (m (m, R 4.81 (m, 1H);

1H); 7.73-8 (s, 11H);

(m, 8H);

(m, 1 H);

Claims (24)

Peptides of the general formula I X- (CR ⁷ R ⁸ ) _m -Het (I) wherein R ¹ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl, heterocyclyl or CONR 4 R 10; alkyl, R ² C 1 -C 6 alkyl, (C až-C v-C až) or Cyk-C alebo-heterocyclylalkyl; Cycloalkylalkyl having a cycloalkyl moiety and 1 alkyl moiety, arylalkyl having 1 to 4 carbon atoms in the alkyl moiety having 1 to 4 atoms. R ³ 1 to 6 carbon atoms, up to 8 to 4 carbon atoms; carbon atoms of up to 4 atoms are alkyl of cycloalkyl of 3 to cycloalkylalkyl of cycloalkyl and alkyl, of arylalkyl of 1 carbon of alkyl, of arylalkenyl of 2 to 6 carbon atoms; 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 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-membered group up to eight 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, said heterocyclic group being saturated or partially unsaturated and excluding - (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 ORR ^xx; and 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 ; R ⁹ and R ¹⁰ are each independently hydrogen, C 1 -C 6 alkyl or cycloalkyl, C 3 -C 8 carbon atoms or R 8 and R 8 are taken together with the nitrogen to which they are attached, 74 represent a nitrogen-containing 4 to 8 membered heterocyclic group; s I 11 R @ ¹ represents a hydrogen atom, an alkyl group having from 1 to 6 @ 1 carbon atoms or a cycloalkyl group having from 3 to 8 carbon atoms; i. 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, and pharmaceutically acceptable salts and bio-precursors thereof. Peptides according to claim 1, having stereochemistry corresponding to the general formula Ia X- (CR ⁷ R ⁸ ) _m -Het (Ia) wherein R ¹ to R ⁸ , 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. Peptides according to any one of claims 3 to 5, wherein R is benzyl optionally substituted in the phenyl ring by methyl, fluorine, chlorine, iodine, trifluoromethyl or trifluoromethoxy or wherein R is 3-phenylpropyl or 3-phenylprop-2- enyl. Peptides according to any one of claims 3 to 6, wherein each of R ⁵ , R ⁶ , R ⁷ and R ⁸ represents a hydrogen atom. 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- yl) azetidine, 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) valyl] -4- (imidazol 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-trifluórmetoxybenzy1) hexanoy1) - (S) valyl] -4- (imidazol-l-yl) -piperidine, l ^- [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-l-yl) piperidine. A process for the production of peptides of the formula I according to claim 1, characterized in that the protecting groups of the compound of the formula IX are cleaved off. X- (CR ^{7 R)} _m-Het (IX) wherein X is a selectively removable protective group Ί p "hydroxy groups and R and R, X and Het are as defined in Claim 1, isolating the compound of formula I, and optionally it turns into its pharmaceutically acceptable salt. 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. Use of the peptides of formula I or Ia or pharmaceutically acceptable salts thereof according to any one of claims 1 to 9 for the manufacture of a medicament for use in the treatment or prophylaxis of human retroviral infections. 14. A method of treating human retroviral infections, comprising administering to a patient an effective amount of a peptide of formula I or Ia according to any one of claims 1 to 9. A process for producing peptides of formula I wherein R ¹ is C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl, aryl, heterocyclyl or ο n n of CONR 1 R ^X ; r ² is (C 1 -C 6) alkyl, (C 3 -C 8) cycloalkylalkyl (C 1 -C 4) alkyl, (C 1 -C 4) arylalkyl or C 1 -C 4 heterocyclylalkyl (C 1 -C 4) an 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 and R ⁸ are taken together to form a three-membered group up to eight 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, said heterocyclic group being saturated or partially unsaturated and excluding - (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 with C 1 -C 6 alkyl, C 3 -C 8 cycloalkyl or NR ⁹ R 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 and R ^{10 are} linked and together with the nitrogen atom to which they are attached represent a 4- to 8-membered heterocyclic a nitrogen-containing group; R ( ¹¹⁾ is hydrogen, (C1-C6) alkyl or (C3-C8) 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, or a pharmaceutically acceptable salt thereof, characterized in that the protecting group is removed from the compound of formula IX t and X- (CR ⁷ R ⁸ ) _m -Het (ix) wherein X 'represents a selectively cleavable hydroxy protecting group and R ¹ to R ⁸ , X and Het are as defined in claim 1, the compound of formula I is isolated and optionally converted to a pharmaceutically acceptable salt thereof. 16. The process of claim 15 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. 17. The method of claim 15, wherein the compound of Formula I has the stereochemistry of the Formula Ib wherein R ^{@ 8} and R ,, n, m, X and Het are as defined in Claim 15 °. The process of claim 15, 16 or 17, wherein R ¹ is tert-butyl, isopropyl or oxetanyl and n is 0. The method of claim 18, wherein R ² is benzyl. A process according to claim 18 or 19, wherein R ⁴ is isopropyl or sec-butyl. The method of any one of claims 18 to 20, 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. 22. wherein each of hydrogen. The method according to any one of claims R ⁵ , R ⁶ , R ⁷ and R ⁸ represents 18 to 21, atom The method according to wherein m is 0 23rd any of the claims; or. 18 to 22, The process of formula I is claim 15, wherein the compound 1- [N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -3 (imidazole- yl) azetidine, 1 '[N - ((R) -2-benzyl- (S) -5- (tert-butoxycarbonylamino) - (S) -4-hydroxy-6-phenylhexanoyl) - (S) -valyl] -4 (imidazole- 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.