MC1985A1 - AMINO ACID DERIVATIVES - Google Patents

Description

-1-

The present invention relates to amino acid derivatives. It relates particularly to amino acid derivatives of the general formula

I ? ï • H* ./ \ / \„/ \ / \ / a • n • •

i h (î)h • \_2 / \

K

' 1

10 in which R is hydrogen or methyl,

2

R, ethyl, propyl or imidazolyl-4,

3

R, isobutyl, cyclohexylmethyl or benzyl, R phenyl, furyl, vinyl, ethyl or 1,2-dihydroxyethyl, A is attached by the carboxy-15 group and represents dibenzosuberoacetic acid, 2-(2-pyridyl)benzoic acid, monoethyl ester of naphthylsuccinic acid, 2-indolylacetic acid or dihydrocinnamic acid or one of the groups

20

R*

W W

6

(has)

And

-y-z

(b)

25

-2-

in which the dashed line may represent an additional bond, R the phenyl, naphthyl, indolyl, pyrazolyl, β-imidazolyl or pyridylmethyl and R hydrogen, an alkyl, arylalkyl, 5-alkylcarboxylalkyl, alkylcarbonylalkyl, cycloalkylcarbonylalkyl, heterocycloalkylcarbonylalkyl, arylcarbonylalkyl, aminocarbonylalkyl, substituted aminocarbonylalkyl, aminocarbonyl, substituted aminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy, arylalkylcarbonyloxy, 10-alkylaminocarbonyloxy, alkoxycarbonylamino, cyano or 2,5-dimethylpyrrolyl-1, with the condition that R may not represent an alkoxycarbonylamino, when R represents phenyl or β-naphthyl, and Y is the N-terminal divalent remainder bonded to Z if applicable N- and/or 15-u-methyl of phenylglycine, cyclohexylglycine, phenylalanine, cyclohexylalanine, 4-fluorophenylalanine, 4-chlorophenylalanine, tyrosine, oi-naphthylalanine, homophenylalanine, ethyl ester of aspartic acid, β-butyl ester of glutamic acid or 20-benzyl ester of glutamic acid and Z hydrogen, an acyl group or one of the groups

25

30

35

• = •

/ n-ch,, çh-ch v J 2

• — •

2 / \

<. )-CH2-°"/

nh e;-cH2-°rsiH

(d)

(c)

and ch-Î~s\

)CR-

nc-n

(e)

q

-3-

in the form of optically pure diastereomers, mixtures of diastereomers, racemates of diastereomers, or mixtures of racemates of diastereomers, as well as pharmaceutically usable salts of these five compounds. These compounds are novel and distinguished by valuable pharmacodynamic properties.

The object of the present invention consists of compounds of formula I and their pharmaceutically usable salts as such and for application as therapeutic active ingredients, the preparation of these compounds, furthermore the drugs which contain them and the preparation of such drugs, as well as the use of compounds of formula I and their pharmaceutically usable salts for the fight against diseases or their prevention, or for the improvement of health, in particular in the prevention of high blood pressure and heart failure.

In this description, the term "alkyl" used—alone or in combination—means a branched hydrocarbon residue saturated with 1–8, preferably 1–4, carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, hexyl, and the like. The term "alkoxy" means an alkyl ether group, in which the term "alkyl" has the meaning above, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, t-butoxy, and the like. The term "cycloalkyl" refers to saturated, cyclic hydrocarbon residues with 3-8, preferably 3-6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and similar compounds. The term "heterocycloalkyl" similarly refers to saturated cyclic hydrocarbon residues of 3 to 8 members, preferably 5 or 6 members, in which one or two methylene groups are replaced.

by one or two oxygen, sulfur, or nitrogen atoms, possibly substituted by an alkyl, phenylalkyl, alkanoyl, or alkanoyloxy group, such as piperidinyl, pyrazinyl, N-benzylpyrazinyl, morpholinyl, N-methylpiperidinyl, 5-K-benzylmorpholinyl, and similar compounds. The term "aryl"

also designates a hydrocarbon residue of 6-14 mono- or bicyclic aromatic carbon atoms, substituted once or more, if necessary by an alkyl, alkoxy, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino, hydroxy, halogen, trifluoromethyl, or nitro group, such as phenyl, α- or β-naphthyl, indenyl, anthrylic, or phenanthryl group, and the like. The term "arylalkyl" designates linear or branched alkyl groups, in which one or more hydrogen atoms are replaced by aryl groups such as benzyl, diphenylmethyl, trityl, α- or naphthylmethyl, 2-phenylethyl, 3-phenyl-2-propyl, 4-phenyl-3-butyl, 2-(cu- or β-naphthyl)ethyl, 3-α-naphthyl-2-propyl, 4—a.-naphthyl-3-butyl and analogues, 20 the aromatic remainder being, each time, as indicated above, substituted once or more times. The expression "amino-substituted" represents an amino group mono- or disubstituted by an alkyl, arylalkyl, alkanoyl, alkoxycarbonyl or arylalkoxycarbonyl or an amino group disubstituted by a disubstituted alkylene group, optionally interrupted by an oxygen, sulfur, or nitrogen atom substituted by an alkyl, phenylalkyl, alkanoyl, or alkanoyloxy. The term "acyl" refers to the acyl group of a carboxylic acid, a carbonic acid hemister, an N-substituted carbamine or thiocarbamine acid, an N-substituted oxalamide, optionally an N-substituted sulfonic acid, or of an amidosulfonic acid, where appropriate N-substituted, in particular those of partial form Rb-CO-, Ra-O-CO-, (Rb)(Rb)N-CO-, (Rb)(Rb)N-CS-,

(Rb)(Rb)N-CO-CO-, Rb-SO2- or (Rb)(Rb)N-SOp-, in the-

3/

which R is a substituted or unsubstituted, saturated or unsubstituted, aliphatic, cycloaliphatic, cycloaliphatic-aliphatic hydrocarbon residue, which may have up to 18 carbon atoms, preferably 10, an unsubstituted or substituted aromatic, heteroaromatic, aliphatic aromatic or aliphatic heteroaromatic hydrocarbon residue, which may have up to 18 carbon atoms, preferably 10, or a heterocycle with 5 or 6 members,

saturated, substituted or not, and R is hydrogen or has the cl

meaning of R. The term "acyl" also refers to the mono-functional residue linked via the carboxyl group of an amino acid, dipeptide, or tri-

* 3, "b peptide. A hydrocarbon remainder R or R, substituted or unsubstituted, saturated or unsubstituted, aliphatic, cycloaliphatic, or aliphatic cycloaliphatic, is, for example, an alkyl, alkenyl, alkynyl, mono-, bi-, or tricycloalkyl, mono-cycloalkenyl, bicycloalkenyl, cycloalkylalkyl, cyclo-alkylalkenyl, or cycloalkenylalkyl, substituted or unsubstituted. "Substituted alkyl" represents an alkyl remainder in which one or more hydrogen atoms may be substituted by a hydroxy, alkoxy, aryloxy, alcanoyloxy, halogen, hydroxysulfonyloxy, carboxy, alkoxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, cyano, phosphono, phosphono esterified, amino, or oxo group, in which the substituents are only at position 1 of the alkyl remainder, when the latter is linked to the carbonyl group in the partial form R-CO-.

Examples of substituted alkyl groups are 2-hydroxyethyl, methoxymethyl, 2-methoxyethyl, phenoxymethyl, u- or p-naphtoxymethyl, acetoxymethyl, 2-acetoxyethyl, chloromethyl, bromomethyl, 2-chloro or 2-bromoethyl, hydroxysulfonyloxymethyl, 2-hydroxysulfonyloxyethyl, carboxymethyl, 2-carboxyethyl, methoxycarbonylmethyl, 2-methoxycarbonylethyl, ethoxycarbonyl-

-6-

methyl, 2-ethoxycarbonylethyl, carbamoylmethyl, 2-carbamoylethyl, methylcarbamoylmethyl, dimethylcarbamoylmethyl, cyanomethyl, 2-cyanoethyl, 2-oxopropyl, 2-oxobutyl, hydroxycarboxymethyl, 1-hydroxy-2-carbo-5-xyethyl, hydroxyethoxycarbonylethyl, hydroxymethoxycarbonylethyl, acetoxymethoxycarbonylmethyl, 1,2-dihydro-xy-2-carboxyethyl, 1,2-dihydroxy-2-ethoxycarbonylethyl, 1,2-dihydroxy-2-methoxycarbonylethyl, 1,2-diacetoxy-2-ethoxycarbonylethyl, 1,2-diacetoxy-2-methoxycarbonyl-10-ethyl, 1-o-naphtoxy-3-carboxypropyl, 1-u-naphtoxy-2-ethoxycarbonylethyl, 1-u-naphtoxy-3-t-butoxycarbonyl-propyl, 1-a-naphtoxy-2-benzyloxycarbonylethyl, 1-a-naphtoxy-3-carbamoylpropyl, a-naphtoxycyanomethyl, 1-tt-naphtoxy-3-cyanopropyl, 1-a-naphtoxy-4-dimethyl-15 aminobutyl or 1-a-naphtoxy-3-oxobutyl.

The term "alkenyl" refers to linear or branched, unsaturated hydrocarbon residues with 2-8, preferably 2-4, carbon atoms, in which the double bond can only be found at position 1, 20 when it is bonded to the carbonyl group in the partial form R-00. Examples of such alkenyl residues are vinyl, allyl, 2-butenyl, or 3-butenyl. Alkenyl residues can be substituted by the same substituents as alkyl residues. 25 The term "alkynyl" refers to hydro-

carbon compounds with 2-8, preferably 2-4, carbon atoms, which contain a triple bond such as ethynyl, 1-propynyl or 2-propynyl. The term "bicyclo-alkyl" refers to saturated bicyclic hydrocarbon residues with 5-10, preferably 6-9, carbon atoms, such as bicyclo[3-1.0]hexyl-1, bicyclo[3-1.0]hexyl-2, bicyclo[3-1.0]hexyl-3, bicyclo[4.1.0]heptyl-1, bicyclo-[4.1.0]heptyl-4, bicyclo[2.2.1]heptyl-2, bicyclo[3.2.1.]-octyl-2, bicyclo[3.3.0]octyl-3, bicyclo[3.3-1]nonyl-9, or -decahydronaphthyl and analogues.

H

-7-

The term "tricycloalkyl" refers to a tricyclic saturated hydrocarbon residue with 8-10 carbon atoms such as 1-adamantyl.

The term "cycloalkenyl" refers to an unsaturated cyclic hydrocarbon 5 residue with 3-8, preferably 3-6 carbon atoms, such as 1-cyclohexenyl, 1,4-cyclohexadienyl and analogue.

The term "bicycloalkenyl" refers to a bicyclic unsaturated hydrocarbon residue with 5-10, preferably 7-10 carbon atoms, such as 5-norbornenyl-2, bicyclo[2.2.2]octenyl-2, hexahydro-4,7-methanoindene-1-yl-6 and analogues.

Examples of cycloalkylalkyl groups include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, 15-cyclohexylmethyl, and analogs. Examples of cycloalkylalkenyl groups include cyclohexylvinyl and cyclohexylallyl and analogs. Examples of cycloalkenylalkyl groups include 1-cyclohexenylmethyl, 1,4-cyclohexadienylmethyl, and analogs.

20 Cycloaliphatic and cycloalipha- remnants

Aliphatic compounds can be substituted by the same substituents as alkyls.

An aromatic or aliphatic aromatic hydrocarbon residue, possibly substituted, is, for example, a substituted or unsubstituted 25-aryl, arylalkyl, or arylalkenyl. Examples of arylalkenyls include styryl, 3-phenylallyl, 2-(α-naphthyl)vinyl, 2-(β-naphthyl)vinyl, and analogs.

In a heteroaromatic hydrocarbon residue or 30 aliphatic heteroaromatic, the heterocycle is mono-,

bi- or tricyclic and contains one or two nitrogen atoms and/or one oxygen or sulfur atom and is linked by one of its ring carbon atoms to the group -CO-, -CO-, >N-CO-, /"N-CS-, "y N-CO-CO-, -SO2 or >N-SO2~. 36 Examples of such hydrocarbon residues are pyrro-

1

-8-

lyl, furyl, thienyl, imidazolyl, pyrazolyl, oxa-zolyl, thiazolyl, pyridyl, pyrazinyl, pyrimidinyl, indolyl, chinolyl, iso-chinolyl, chinoxalinyl, p-carbolinyl, or a benzo- or 5-cyclopenta-, cyclohexa-, or cyclohepta- condensed derivative of these residues. The heteroaromatic residue may be substituted at one nitrogen atom by an alkyl, phenyl, or phenylalkyl group, for example, benzyl, and/or at one or more carbon atoms by an alkyl, phenyl, phenylalkyl, halogen, 10-hydroxy, alkoxy, phenylalkoxy, or oxo- group, and may be partially saturated. Examples of such heteroaromatic residues are 2- or 3-pyrrolyl, phenylpyrrolyl, 4- or 5-phenyl-2-pyrrolyl, 2-furyl, 2-thienyl, 2-imidazolyl, 2-, 3- or 4-pyridyl, 2-, 3- or 5-indolyl, 15-substituted 2-indolyl, for example 1-methyl-, 5-methyl-, 5-methoxy-, 5-benzyloxy-, 5-chloro- or 4,5-dimethyl-2-indolyl,

1-benzyl-2-indolyl, 1-benzyl-3-indolyl, 4,5,6,7-tetrahydro-2-indolyl, cyclohepta[b]-5-pyrrolyl, 2-, 3- or 4-chinolyl, 4-hydroxy-2-chinolyl, 1-, 3- or 4-isochino-

èO lyl, 1-oxo-1,2-dihydro-3-isochinolyl, 2-chinoxalinyl,

2-benzofuranyl, 2-benzoxazolyl, 2-benzothiazolyl, benz[e]indolyl-2, -carboline and the like.

Examples of hetero-aromatic-aliphatic hydrocarbon residues are 2- or 3-pyrrolylmethyl, 2-, 3- or 4-pyridylmethyl, 2-(2-, 3- or 4-pyridyl)ethyl, 4-imidazolylmethyl, 2-(4-imidazolyl)ethyl, 2-indolyl-methyl, 3-indolylmethyl, 2-(3-indolyl)ethyl, 2-chinolylmethyl and analogues.

A saturated heterocycle with 5 or 6 members has at least one carbon atom, 1-3 nitrogen atoms, and possibly one oxygen or sulfur atom as a ring member, and is linked by one of its ring atoms to the group -CO- or -O-CO, >N-CO-, >N-CS-, >N-CO-CO, -SO2- or ^N-SO^. The heterocycle can be substituted at one of its carbon atoms or at one of its atoms

O

-9-

nitrogen of the ring by an alkyl, for example methyl or ethyl, phenyl or a phenylalkyl, for example benzyl, or on one of its carbon atoms by a hydroxy or an oxo and/or be condensed with benzene 5 on two contiguous carbon atoms. Examples of such heterocycles are pyrrolidinyl-3, 4-hydroxypyr-rolidinyl-2, 5-oxopyrrolidinyl-2, piperidinyl-2, piperidinyl-3, 1-methylpiperidinyl-2, 1-methylpiperidinyl-3, 1-methylpiperidinyl-4, morpholinyl-2, morpholinyl-3, 10-thiomorpholinyl-2, thiomorpholinyl-3, 1,4-dimethylpipera-zinyl-2, 2-indolinyl, 3-indolinyl, 1,2,3,4-tetrahydro-chinonyl-2, -3 or 4-, 1,2,3,4-tetrahydroisochinoyl-1, -3 or -4, 1-oxo-1,2,3,4-tetrahydroisochinolyl-3 and analogues.

The remaining amino acid linked by the 15-carboxyl group is taken into account, including natural L-configuration β-amino acids, homologs of these amino acids, for example those in which the amino acid chain has been lengthened or shortened by one or two methylene groups and/or a methyl group is replaced by hydrogen, substituted aromatic 20-amino acids, for example substituted phenylalanine or phenylglycine in which the substituent may be one or more times an alkyl, for example methyl, halogen, for example fluorine, chlorine, bromine or iodine, hydroxy, alkoxy, for example 25-methoxy, alkanoyloxy, for example acetoxy, amino, alkyl-amino, for example methylamino, dialkylamino, for example dimethylamino, alkanoylamino, for example acetylamino or pivaloylamino, alkoxycarbonylamino, for example β-butoxycarbonylamino, aryl methoxycarbonylamino, for example 30 benzyloxycarbonylamino and/or nitro of benzoate-concentrated phenylalanine or phenylglycine, such as α-naphthylalanine, or hydrated phenylalanine or phenylglycine, such as cyclohexylalanine or cyclohexylglycine, a 5- or 6-membered cyclic α-amino acid concentrated with benzoate, for example 35 indoline-2-carboxylic acid or 1,2,3,4-

-10-

tetrahydroisochinoline-3-carboxylic acid, a natural or homologous (i-amino) acid in which a carboxy group is in esterified or amidated form in the side chain, for example as an alkyl ester group, such as 5-methoxycarbonyl or t-butoxycarbonyl, or as carba-moyl-, alkylcarbamoyl, such as methylcarbamoyl, or as a dialkylcarbamoyl group, such as dimethylcarbamoyl; in which an amino group of the side chain is in acylated form, for example as an alkanoyl-10 amino, such as acetylamino or pivaloylamino, as an alkoxycarbonylamino, such as t-butoxycarbonylamino, or as an arylmethoxycarbonylamino group, such as benzyloxycarbonylamino; or in which a hydroxy group of the side chain is in etherified or esterified form, for example as an alkoxy group, such as methoxy, or as a group aryl-alkoxy, such as benzyloxy, or as a lower alkanoyloxy group, such as acetoxy, or epimers of such amino acids, i.e., with the non-natural D configuration. Examples of such amino acids are glycine, alanine, valine, norvaline, leucine, isoleucine, norleucine, serine, homoserine, threonine, methionine, cysteine, proline, trans-3- and trans-4-hydroxyproline, phenylalanine, tyrosine, 4-nitrophenylalanine, 4-amino-25-phenylalanine, 4-chlorophenylalanine, β-phenylserine, phenylglycine, u-naphthylalanine, cyclohexylalanine, cyclohexylglycine, tryptophan, indolin-2-carboxylic acid, 1,2,3,4-tetrahydroisochinolin-3-carboxylic acid, Aspartic acid, asparagine, aminomalonic acid, aminomalonic acid monoamide 30, glutamic acid,

mono-t-butyl ester of glutamic acid, glutamine, N-dimethylglutamine, histidine, arginine, lysine, N-t-butoxycarbonyllysine, β-hydroxylysine, ornithine, N-piva-loylornithine, α,γ-diaminobutyric acid or β-35-diaminopropionic acid and analogues.

20

-11-

The remainder linked by the carboxyl group of the amino acid can be N-terminally substituted by an alkyl, for example methyl or ethyl, to increase the stability of the compound of formula I with respect to enzymatic degradation.

The remainder linked via the carboxyl group of a di- or tripeptide consists of two or three of the amino acids mentioned above.

The term "pharmaceutically useful salts" includes salts with mineral and organic acids, such as hydrochloric acid, hydrobromidric acid, nitric acid, sulfuric acid, phosphoric acid, citric acid, fornic acid, maleic acid, acetic acid, succinic acid, α-15 tartaric acid, methanesulfonic acid, p-

toluenesulfonic acid and similar compounds. Such salts can be prepared by anyone skilled in the art without difficulty, taking into account the prior art and the nature of the compound to be transformed into a salt. Compounds of formula I possess at least three asymmetric carbon atoms, and therefore they occur as optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates. The present invention encompasses all such forms. Mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates can be separated by known methods, for example, column chromatography, thin-layer chromatography, HPLC, and similar techniques.

Compounds with formula I are preferred, in

2

in which R' is hydrogen. R preferably represents imidazolyl-4-. Furthermore, compounds of formula I in which R is cyclohexylmethyl are preferred. 35 Compounds of formula I are also preferred, in the-

1

-12-

Which R^ is vinyl or ethyl? Compounds of formula I, in which A is group (a) or (b), are also preferred. R^ preferably represents phenyl or naphthyl, phenyl being particularly preferred. The preferred meaning of R^ is alkylcarbo-nylalkyl, cycloalkylcarbonylalkyl, heterocycloalkyl-carbonylalkyl, aminocarbonylalkyl, or substituted aminocarbonylalkyl, preferably (alkyl-C^)-carbonyl-methyl, (cycloalkyl-C^-Cg)-carbonylmethyl, N-t-buto-10 μ-carbonylpyrrolidinylcarbonylmethyl, or aminocarbonylalkyl, which is disubstituted by an alkyl or an alkylene, optionally interrupted by an oxygen atom, or monosubstituted by a mono- or disubstituted amino-alkyl or alkoxycarbonyl. Furthermore, compounds of formula I are preferred, in which Y is the Z-linked N-terminal divalent residue of phenylalanine, cyclohexylalanine, 4-chlorophenylalanine, or α-naphthylalanine, with phenylalanine being particularly preferred. Z is preferably an acyl, especially preferred is a residue linked to Y by the terminal carbon of a natural N- and/or α-methylated amino acid of L configuration or of an epimer of such an amino acid with D configuration, or of a dipeptide obtained from two of the amino acids just described, where Z represents the group

Rb-CO- or Ra-CO-, wherein Ra is a hydrocarbon residue, optionally substituted, saturated or unsaturated, aliphatic, cycloaliphatic, cycloaliphatic-aliphatic with up to 18 carbon atoms, a hydrocarbo-5O residue, optionally substituted, aromatic, heteroaromatic, aromatic-aliphatic or aliphatic heteroaromatic with up to 18 carbon atoms, or a heterocycle, optionally substituted, saturated with 5 or 6 members

■u.

and R represents hydrogen or has the meaning of 35 Ra, Z represents in a particularly preferential way-

4

-13-

the remainder of proline, prolylproline or histi-dinylproline, whose C-terminus is linked to Y, the amino group being in each case substituted by t-butoxy-carbonyl, benzoxycarbonyl, isovaleryl or the 5 R^-CO group, in which R*5 is a hetero-aromatic-aliphatic hydrocarbon remainder with up to 10 carbon atoms or represents a saturated heterocycle with 5 or 6 members.

It follows from the above that the compounds of formula I that are particularly preferred are those

1 y 2

10 in which R is hydrogen, R is imidazolyl-4,

3 4-

R^ cyclohexylmethyl, R vinyl or ethyl and

R^ the phenyl and R^ an (alkyl C^-C^)carbonylmethyl, a (cycloalkyl C^-Cg)carbonylmethyl, the N-t-butoxy-carbonylpyrrolidinylcarbonylmethyl or an aminocarbonyl-15 alkyl, which is disubstituted by an alkyl or an alkylene

C4-C5, possibly interrupted by an oxygen atom, or mono-substituted by an aminoalkyl mono- or di-substituted by an alkoxycarbonylalkyl or alkoxycarbonyl, or Y is the divalent phenylalanine residue linked by the N-2C terminal to Z and Z is the residue linked to Y by the G-terminal of proline, prolylproline or histidinylproline, the amino group being in each case substituted by t-butoxy-

carbonyl, benzoxycarbonyl, isovaleryl or the group

"b "b

R -GO-, in which R is a hetero- hydrocarbon residue

25 aromatic-aliphatic with up to 10 carbon atoms or a saturated heterocycle with 5 or 6 members.

The compounds especially preferred for formula I are:

- t-butyl (R)-2-[ [(S)-a-[[(S)-1-[(1S,2S,4S)-1-(cyclo-30 hexylmethyl)-2-hydroxy-4~ isopropylhexyl]-2-imidazolyl-

4-éthyl]c a rbamoyl]phe ne thy1]c a rb amoy1]-1-pyrro1id in-carboxylate,

- (2S,3S,5S)-2-(Boc-D-Pro-Phe-His-NH)-1-cyc1ohexy1-5-isopropyl-6-heptenol-3,

has

-14-

- N-(S)-[(1S,2S,4S)-1-(cyclohexylmethyl-2-hydroxy-4-isopropyl-5-hexenyl] -a-[(S)-u-3-methylbutyramido] -imidazol-4-propionamide,

- t-butyl[(S)-a-[[(S)-1-[[(lS,2S,4S)-1-cyclohexyl-5 methyl-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]

-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamate;

- t-butyl[(S)-u-[[(S)-1-[[(1S,2S,4S)-1-cyclohexyl-methyl-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imida-zolyl-4-ethyl]carbamoyl]phenethyl]carbamate,

10 - t-butyl(S)-2-[[(R)-a-[[(S.)-1-[[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imida-zolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrrolidin-carboxylate;

- (S)-ïï-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-

15 isopropylhexyl] -«.-[[[[ (R)-u-2-hydroxy-1 -(hydroxy-

methyl)-1-methyl thyl]carbamoyl]methyl]hydro cinnamami-do]imidazol-4-propionamide,

- N-(S)-1-[[(1S,2S,4S)-1-(cyc1ohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]-r-

20 oxo-c<i-(1-naphthylmethyl)-4-morpholinbutyramide,

- t-butoxycai:bonyl[2-[[ (R and S)-3-[ [ (S)-1-[ [ (1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenyl-butyramido]ethyl]glycine t-butylester,

25 - (S) —IST—[ (1S,2S,4S)-1-(cyclohexylmethyl)-2-iiydroxy-4-

isopropyl-5-hexenyl] (R)-u-[ [ [ 2-hydroxy-1 -(hydroxy-methyl)-1-methylethyl]carbamoyl]methyl]hydroc innamido] -imidazol-4-propionamide,

- (S)-a-[(R)-2-benzyl-5,5-dimethyl-4-oxohexanamido]-

30 N-[(1S,2S,4S)-1-(cyclohexylmethyl-2-hydroxy-4-isopro-pyl-5-hexenyl]imidazol-4-propionamide,

- (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-ct-2-[N-(morpholinocarbamoyl)-3-phenyl-L-alanyl]amino]imidazol-4-propionamide,

-15-

- (S)-a-[ (R)-a-(carbamoylniethyl) hydro c innamamido] -N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4~ iso-propyl-5-hexenyl] imidazol-4-propionamide;

- (S ) -N-[ ..1S, 2S,4S) -1 -(cyclohexylmethyl)-2-hydroxy-4-5 isopropyl-5-hexenyl]-a.-[(R)-a-[(dimethylcarbamoyl)-

methyl]hydrocinnamamido]imidazol-4-propionamide,

- (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl] -u-[ (RS)-<*-[(cyclopentylcarbonyl)-methyl]hydrocinnamamido]imidazol-4-propionamide,

10 - (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-a-[(RS)-u-[(cyclopentylcarbonyl)-methyl]hydrocinnamamido]imidazol-4-propionamide and

- (2S,5S)-2-(Bo c-D-Pro-Pro-Ph e-Hi s-NH)-1-cyc1ohe xyl-5-isopropyl-6-heptenol-3.

15. Compounds of formula I in dia- form

optically pure stereoisomers, mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates, as well as their pharmaceutically usable salts, are prepared in that

20 a) A compound with the general formula is reacted

rale

/ ? ï3 . *4 H i I 4hY

\r2 0H / \

12 3 4 in which R, R, R and R have the meaning given above,

30 with an acylation agent giving the group

5 F6

• g

35 (a) (b)

C

-16-

5 6

in which R, S, I, Z and the dotted line have the meaning given above, or b) a compound of general formula is reacted

5 3

I.4

H,t/ V./ III

iU X

3 4

10 in which R and R have the meaning given above,

with a compound of general formula

15 A Aoh a • iv

V

12

in which R, R and A have the meanings given above,

20 or with one of its activated derivatives, or c) to prepare a compound of formula I, in which R is ethyl and the other symbols have the meaning given above, one hydrogenates a compound of formula I, in which R is vinyl and the au-

25 very symbols have the meaning given above, or d) to prepare a compound of formula I, in the-

/j.

which R is 1,2-dihydroxyethyl and the other symbols have the meaning given above we oxidize a

/ Zj. ,

compound of formula I in which R represents vinyl 30 and the other symbols have the meanings given above, or e) to prepare a compound of formula I, in which A contains a free amino group, the protecting group of N is removed from a corresponding compound

35 of formula I, in which A contains an amino group

C

-17-

N-protected, and/or f) if desired, a mixture of diastereomeric racemates is separated into the diastereomeric racemates or optically pure diastereomers, and/

5 or g) if desired, a diastereomer mixture is separated into optically pure diastereomers, and/or h) if desired, a compound 10 obtained is transformed into a pharmaceutically usable salt.

The acylation of a compound of formula II takes place according to processes known per se. Suitable acylation agents include activated acid derivatives such as esters, mixed esters, and acid anhydrides or mixed acid anhydrides. The reaction is carried out in an inert organic solvent under the reaction conditions, or in a mixture of solvents, at a temperature between approximately 0°C and room temperature. Suitable solvents include aromatic solvents such as benzene, toluene, or xylene; chlorinated hydrocarbons such as methylene chloride and chloroform; ethers such as diethyl ether, tetrahydrofuran, or dioxane; and similar solvents. In the case where the a-25 acylation agent is a peptide, the reaction takes place under the usual reaction conditions in peptide chemistry, that is preferably in the presence of a condensation agent such as HBTU (O-benzo-triazolyl-N,IT,N',N'-tetramethyluronium-hexafluorophos-30 phate), BOP (benzotriazolyl-1-oxy-bis-(dimethylamino)-phosphonium-hexafluorophosphate), HOBT (N-hydroxybenzo-triazol), DBIJ (1,8-diazabicyclo[5.4.0]undecene-7), DCC (dicyclohexylcarbodiimide), EDC (N-ethyl-N'-(3-di-methylaminopropyl)carbodiimide-hydrochloride), base 35 Hünig (ethyldiisopropylamine) and analogues.

-18-

The reaction is carried out appropriately in an inert organic solvent or mixture of solvents under reaction conditions at a temperature between approximately 0°C and 50°C, preferably near room temperature. Suitable solvents include, but are not limited to, dimethylformamide, methylene chloride, acetonitrile, tetrahydrofuran, and similar compounds.

The reaction of a compound of formula III with a compound of formula IV also occurs according to the processes known in peptide chemistry, that is, under the same conditions as those indicated above for the reaction of a compound of formula II with a peptide. Examples of activated derivatives of a compound of formula IV are acid halides, acid anhydrides, mixed anhydrides, esters, mixed esters, and analogs thereof.

The hydrogenation of a compound of formula I, in which R is vinyl, also takes place according to processes known per se in an inert organic solvent under the conditions of the reaction or a mixture of such solvents, at a temperature between approximately room temperature and 50°C, preferably at room temperature, in the presence of a noble metal catalyst such as platinum or palladium, preferably palladium. Suitable solvents include, in particular, alcohols such as methanol or ethanol, and similar compounds.

The oxidation of a compound of formula 1, 30, in which R^ is vinyl, takes place according to processes known per se in an inert organic solvent under the conditions of the reaction, or in a mixture of such solvents, at a temperature between approximately room temperature and the boiling point of the solvent 35 or the mixture of solvents, preferably at temperature

-19-

ambient. Osmium tetroxide is particularly suitable as an oxidizing agent. Pyridine and similar compounds are suitable solvents.

The removal of the protecting group of N according to process variant e) also takes place by methods known per se in an inert solvent under the reaction conditions or in a mixture of such solvents at a temperature between about 0°C and room temperature, with an acid such as hydrochloric acid, trifluoroacetic acid, and similar acids. Suitable solvents are ethers, such as tetrahydrofuran or dioxane, alcohols such as methanol, or chlorinated hydrocarbons, such as methylene chloride and similar acids.

The starting materials of formula II are novel and are also the subject of the present invention. These compounds can be prepared by reacting a compound of formula III with histidine, optionally N-methylated, norleucine, or norvaline. This reaction also takes place according to known methods of peptide chemistry, that is, under the reaction conditions described above for the reaction of a compound of formula II with a peptide.

The starting materials of formula III are also novel and are the subject of the present invention. They can, for example, be prepared by reacting an aldehyde of general formula in a Grignard reaction

F

b-hn7 xcho v

in which B is an amino protecting group, of

-20-

preferably t-butoxycarbonyl or benzyloxycarbonyl, and R^ has the meaning given above,

with a compound of general formula

5 41

/\ /R

X î VI

/ \

in which X represents chlorine, bromine or iodine, preferably bromine, and R^ is phenyl, furyl or vinyl.

This reaction also takes place according to methods known per se, for example in an inert solvent under the reaction conditions such as ether, at a temperature between 0° and 50°C, preferably at room temperature. Then the amino protecting group B of the resulting compound is removed according to known methods, for example with hydrochloric acid in dioxane at room temperature. If a compound of formula III is desired, in which R is ethyl or 1,2-dihydroxyethyl, the product of the Grignard reaction is hydrogenated under the reaction conditions of variant process c) or oxidized according to known methods, before the removal of the amino protecting group B. Suitable oxidizing agents are osmium tetroxide, potassium permanganate, ruthenium tetroxide, and similar compounds. The reaction is carried out in an inert organic solvent under the reaction conditions or a mixture of such solvents at a temperature approximately between room temperature and 50°C. Suitable solvents are pyridine, aromatic hydrocarbons such as benzene, and similar compounds.

Compounds of formula V are known or can be obtained by analogy with the preparation of

D

-21-

known compounds.

4-1

The compound of formula VI, in which R is vinyl, is known. Compounds of formula VI, in 41

which R is phenyl or furyl, are novel 5 and are also the subject of the present invention.

Each diagram following I-III represents a process for preparing a compound of formula III, in which R represents a phenyl, furyl, or vinyl group. Diagrams II and III also allow us to deduce the pre-

10. Preparation of new compounds of formula VI, in the -41

Which R is phenyl or furyl? For the exact reaction conditions, please refer to the section with examples.

1

-22 -

Diagram I

-o-co-nh-cjih-cho • •

/ \ /

• •

II • •

\ /

Go

Br

/ \#/ ^ î

/ \ Via

/ \

n / \ /

8 î

-0/ VN/ N'./ ^

H

4b

IX

/ \ î î

/*\ /' • •

I

N./ ^ AH 7ix

II

-23 "

Diagram II

\ /

. . î

i3°oc y \ ^ ch3ooc/ n.

X

xiii xi

J

\ / \ / i i

H0CH2/ Y X|| + H00c/'Y'N|1

• V • • •

xii t

Br-CH

\ /

/*\ ^*\

2 î I* % /*

VIb

- |-o-co-nh-(j:h-cho

\ /

i

Vb

\ /

I H ?H î

"" î -0 N • • - •

i v,/ \ / \ / \ ^ \

g/- •

/ \

XIV

?H Y

h n • • y •

2 \ / \ / \ ^ \

/

/ \

K »

Illb

- 24 - Diagram III

y' = \ /COOC2h h3c0/ \/ ^ch3 \/ 'vcooc2h

16th century

y

X /

co2h

0

XVIII

I (rac) / \

1

y

V / \ /

0'

/

co2h (s)

/ \

/'~X °W'\

\/ >Ca /•

/ \

XVII

f \

\ X /C°2H

0 î (R) / \

S ■>

/ \

< i

/

XIXa

XLXb

XXb

V

/ \ / \

Oh

/ \

(R)

XXa

/ " .

V7 Y

/ \

VI c

-î-O-CO-NH-CH-CHO

1/

/ \

Vc

- 25 -

t

•y

/"■x. . ./! ?

V7 Y Y v vi

0 /i< ÔH « »

21st

\ /

I

S'~\ . /"

N/ Y ^6H NH2

/ V

iiic

-26-

The compounds of formula I and their pharmaceutically usable salts exhibit an inhibitory action on the natural enzyme renin. Renin passes from the kidneys into the blood and there causes the degradation of angiotensinogen, forming the decapeptide angiotensin I, which is then degraded in the lungs, kidneys, and other organs into the octapeptide angiotensin II. Angiotensin II increases blood pressure either directly by arterial constriction or indirectly by the release of the sodium-retaining hormone aldosterone from the adrenal glands, which is linked to an increase in extracellular fluid volume. This increase is due to the action of angiotensin II itself or of the heptapeptide angiotensin III, which is derived from it as a degradation product. Inhibitors of renin enzyme activity cause a decrease in the formation of angiotensin I and consequently a smaller amount of angiotensin II. The decrease in the concentration of this active peptide 20 hormone is the direct cause of the blood pressure-lowering action of renin inhibitors.

The efficacy of renin inhibitors can be demonstrated experimentally using the in-vitro test described below.

25 In-vitro test with human renin

The test is performed in Eppendorff tubes. The incubation mixture consists of (1) 100 µL of human renin in buffer A (0.1 M sodium phosphate solution, pH 7.4-, containing 0.1% bovine serum albumin 30, 0.1% sodium azide, and 1 mM ethylenediaminetetraacetic acid), sufficient for a renin activity of 2–3 ng angiotensin I/mL/hr; (2) 145 µL of buffer A; (3) 30 µL of human renin tetradecapeptide (hTD) substrate in 10 mL of hydrochloric acid; (4) 15 µL of dimethyl sulfoxide with or

-27-

without inhibitor and (5) 10 yul of a 0.03 molar solution of hydroxychinoline sulfate in water.

The samples were incubated for three hours at 37°C or at 4°C in triplicate. Two 100 mL samples per test tube were then used to measure angiotensin I production via RIA (standard radioimmunoassay; Clinical Assay Solid phase kit). The cross-reactivities of the antibodies used in the RIA are: angiotensin I 100%; angiotensin II 0.0013%; hTD (angiotensin I-Val-Ile-His-Ser-OH) 0.09%. Angiotensin I production is assessed by the difference between the assay at 37°C and that at 4°C.

The following witnesses were made in pa-

15 parallel:

(a) Incubation of renin-free and inhibitor-free hTD samples at 37°C and 4°C. The difference between these two values gives the baseline value of angiotensin I production.

20 (b) Incubation of hTD samples with renin,

However, without an inhibitor, at 37°C and 4°C. The difference between these values gives the maximum angiotensin I production value.

For each sample, the baseline value of angiotensin I production (25) is subtracted from the measured angiotensin I production. The difference between the maximum value and the baseline value gives the value of maximum substrate hydrolysis (= 100%) by renin. The results are presented as a value of 30, which gives the concentration of the inhibitor that inhibits 50% of the enzymatic activity. The IC50 values are determined from a regression line on a log-log plot.

The results obtained in this test are summarized in the following table:

-28-

Painting

Compound IC^q values enyuMol/lt

HAS

0.0035

B

0.0003

C

0.0090

D

0.0075

E

0.0079

F

0.0040

G

0.015a

H

0.0041

I

0.0460

K

0.0170

L

0.0070

M

0.0210

N

0.1400

0

0.0120

P

0.0460

Q

0.0680

R

0.0017

20 A = t-butyl-(R)-2-[[(S)-a-[[(S)-1-[(1S,2S,4S)-1-

cyclohexylmethyl-2-hydroxy-4-isopropylhexyl]-2-imida zolyl-4-ethyl]c arbamoyl]phene thyl]c arb amoy1]-

1-pyrrolidincarboxylate;

B = (2S,3S,5S)-2-(Boc-D-Pro-Phe-His-NH)-1-cyclohexyl-25 5-isopropyl-6-heptenol-3;

C = N-(S)-[(1S,2S,4S)-1-(cyclohexylmethyl-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-3-methylbutyramido]-imidazol-4-propionamide;

D = t-butyl[(S)-a--[[(S)-1-[[(1S,2S,4S)-1-cyclohexyl-30 methyl-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-

2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carba m ate; E = t-butyl[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-cyclohexyl-

methyl-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imida z oly1-4-ethyl]c a rb amoy1]phene thy1]c arbamate;

-29-

F = t-butyl (S)-2-[[(R)-u-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrrolidincarboxylate;

5 G = (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-a-[[[[(R)-a-2-hydroxy-1-(hydroxy-methyl)-1-methylethyl]carbamoyl]methyl]hydrocinnamamido] imidazol-4-propionamide;

H = N-(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-10 4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]-

i -oxo-a-(1-naphthylmethyl)-4-morpholinebutyramide;

I = t-butoxycarbonyl [2-[(R and S)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-nyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-15 phe nylbutyr amido]ethyl]glyc ine-t-butyl st er;

Z = (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(R)-a-[[[2-hydroxy-1-(hydro-xymé thy1)-1-methyl thyl]c arbamoy1]methyl]hydroc inna-mamido]imidazol-4-propionamide; 20 L = (S)-a-[(R)-2-benzyl-5,5-dimethyl-4-oxohexanamido]-N-[ ( 1 S , 2S ,4S ) -1 - ( cyclohexylm thy1-2-hydroxy-4-is o-propyl-5-hexenyl]imidazol-4-propionamide;

M = (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-2-[N-(morpholinocarbamoyl)-25 3-phenyl-L-alanyl]amino]imidazol-4-propionamide;

H = (S)-a-[(R)-a-(carbamoylmethyl)hydrocinnamamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-iso-propyl-5-hexenyl]imidazol-4-propionamide;

0 = (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-30 isopropyl-5-hexehyl] -a-[(R)-a-[(dimethylcarbamoyl)-

methyl]hydrocinnamamido]imidazol-4-propionamide;

P = (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(RS)-a-[(cyclopentylcarbonyl)methyl]hydrocinnamamido]imidazol-4-propionamide;

-30-

Q = (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropylhexyl] -a-[(RS)-a-[(cyclopentylcarbonyl)-methyl]hydrocinnamamido]imidazol-4—propionamide and

R = (2S,5S)-2-(Boc-D-Pro-Pro-Phe-His-NH)-1-cyclohexyl-5-5 isopropyl-6-heptenol-3-

Compounds of formula I, as well as their pharmaceutically usable salts, can find application as a drug, for example in the form of a pharmaceutical preparation. Pharmaceutical preparations can be administered enterally.

as oral, for example in the form of pills, coated pills, dragees, hard or soft gelatin capsules, solutions, emulsions or suspensions; nasal, for example in the form of nasal sprays; or rectal, for example in the form of suppositories. Administration can also be parenteral, such as intramuscularly or intravenously, for example in the form of injectable solutions.

For the preparation of pills, coated pills, dragees, and hard gelatin capsules, compounds of formula I and their usable pharmaceutical salts can be transformed with inert organic or mineral excipients. For example, excipients that can be used for hard gelatin pills, dragees, and capsules include lactose, corn starch or their derivatives, talc, stearic acid, or its salts.

Suitable excipients for soft gelatin capsules include, for example, vegetable oils, waxes, fats, and liquid and semi-solid polyols.

To prepare solutions and syrups, suitable excipients include, for example, water, polyols, sucrose, invert sugar, glucose, etc.

For injectable solutions, 35 are suitable as excipients, for example natural oils or

-31-

hardened waxes, greases, liquid or semi-solid polyols, etc.

In addition to also containing preservatives, solvents, gelling agents, wetting agents, emulsifiers, sweeteners, colorings, flavorings, salts to modify osmotic pressure, buffers, coating agents and antioxidants, they may also contain other valuable therapeutically active substances.

Formula I oils and their pharmaceutically usable salts for combating or preventing blood pressure and heart failure. 15 The dosage can vary within wide limits and must

The dosage should be adapted to individual characteristics. Generally, for oral administration, a daily dose of approximately 3 mg to approximately 3 g, preferably approximately 10 mg to approximately 1 g, should be adjusted. For example, approximately 300 mg per person, preferably divided into 1-3 unit doses, which, for example, may be identical, but the upper limit may also be exceeded if indicated. Children usually receive half the adult dose.

present invention, but they shall not limit it in any way. All temperatures are given in degrees Celsius. The following abbreviations are used for amino acids and amino acid derivatives

Pharmaceutical preparations can in

According to the invention, the components can be used

The following examples should explain the

H-His-OH H-Phe-OH H-Phe^-His-OH

L-histidine L-phenylalanine

N-[(S)-2-amino-3-phenylpropyl]-L-

histidine

L-proline

35 H-Pro-OH

-32-

H-Ser-OH H-Tyr-OH

L-serine L-tyrosine

Other abbreviations:

Boc 5 Bom t-but oxyc arb ony1

benzyloxymethyl

Isovaleryl benzyloxy methoxy piperidinyl

IVA OBz OME

Pip 10 Fmoc t-Bu

= 9-fluorenylmethoxycarbonyl = t-butyl

Example 1

A mixture of 313 mg (1.18 mmol) of t-butoxy-

carbonyl-L-phenylalanine, 460 mg (1.18 mmol) of (S)-α-15 amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, 0.15 ml (1.18 mmol) 4-ethylmorpholine, 320 mg (2.36 mmol) of HOBT and 292 mg (1.42 mmol) of DCC in 20 ml of dimethylformamide, is stirred overnight at room temperature. The precipitate formed is then filtered and

The solvent is evaporated under high vacuum. The remainder is dissolved in ethyl acetate and then washed successively with 2N sodium bicarbonate solution, saturated ammonium chloride solution, 25 with more 2N sodium bicarbonate solution, and again with saturated ammonium chloride solution. After drying the organic solution over sodium sulfate, the solvent is evaporated under reduced pressure, and the remaining 870 mg is chromatographed for purification on 30 g of silica gel using a 20:1:0.1 mixture of methylene chloride, methanol, and ammonia as the eluent. Crystallization of the crude product obtained in the methylene chloride/ether mixture gives 410 mg of t-butyl[(S)-a-[[(S)-1-[[(1S,2S, 35 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]

-33-

carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl] -carbamate, melting point 161°.

The (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imida z ol-4-5 propionamide used as a starting material was manufactured as follows:

315.3 g (1.28 mmol) of 2-t-butoxycarbonyl-L-phenylalanine methyl ester was hydrogenated according to the process described by J. Boger et al. in J. Med. Chem., 10 28 (1985) in the presence of 5% rhodium on aluminum oxide at room temperature, yielding 315.3 g of methyl ester of 2-t-butoxycarbonylamino-3-cyclohexylpropionic acid in oil form, which is used directly in the next step. 85.61 g (300 mmol) of methyl ester of

1.2 L of toluene is added dropwise at -75°C to 750 mL of a 20% diisobutyl hydride in hexane solution over 90 minutes. At the end of the addition, the reaction mixture is stirred for another 10 minutes at -75°C.

Then, within 25 minutes, at a temperature of -75° to -70°, first 70 ml of methanol and then 840 ml of a saturated solution of potassium sodium tartrate are added. The milky solution, slowly warmed to room temperature, is then extracted with ether, and the extracts are dried and evaporated, yielding 82 g of 2-t-butoxycarbonylamino-3-cyclohexylpropylaldehyde in oil form, which can be used in the next step without purification. To 24.65 g of magnesium shavings in 300 ml of ether under an argon atmosphere and at a temperature between 30° and the reflux temperature of the solvent, a solution of 179.7 g (1.01 mol) of 4-bromo-3-isopropyl-1-35-butene (which was prepared according to the process described by

-34»

R.G. Helmquist in Tetrahedron Letters, 2533 (1982)) in 900 ml of ether. At the end of the addition, the reaction mixture is heated for 3.5 hours under reflux. To the reaction mixture cooled to -60°C, a solution of 82 g of 2-t-butoxycarbonylamino-3-cyclohexylpropylalide in 900 ml of ether is added dropwise over 75 minutes, with the temperature maintained between -60°C and -70°C. After the addition, the cooling bath is removed and the reaction mixture is stirred for 21 hours at ambient temperature under argon. It is then cooled to 5°C and 225 ml of a saturated ammonium chloride solution is carefully added while stirring, raising the temperature to 20°C. The two phases are separated, and the organic phase is dried over sodium sulfate and evaporated, yielding 127.5 g of a yellow oil. Chromatographic separation of this oil on 2.5 kg of silica gel with methylene chloride containing 2.5% ether as the eluent gives 33.9 g of (aS,[*S)-β-t-butoxycarbonylamino-α-[(S)-2-isopropyl-3-20 butenylcyclohexanepropanol, 13.62 g of (aS,S)-β-t-butoxycarbonylamino-α-[(R)-2-isopropyl-3-butenyl]cyclohexanepropanol, and a mixture of the two diastereomers mentioned above as oils, MS: 354 (M+H)+.

25 1.0 g (2.83 mmol) of (<-«S, p> S)-£-t-butoxy-

Carbonylamino-α-[(S)-2-isopropyl-3-butenyl]cyclohexane-propanol is dissolved in 20 mL of 2,211 hydrochloric acid in dioxane and stirred for 2 hours at room temperature. The reaction mixture is then evaporated, and toluene is added twice successively, followed by evaporation to dryness. This yields (αS,ρ)-α-amino-α-[(S)-2-isopropyl-3-butenyl]cyclohexane-propanol as the hydrochloride, chromatographically pure in a thin layer, which is used directly in the next step.

-35-

To 1.7 g (2.83 mmol) of N-α-N-μ-Bis-με-L-histidine and 0.82 g (2.83 mmol) of (αS,μS)-P-amino-α-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol hydrochloride in 20 mL of dimethylformamide, 0.36 mL of ethylmorpholine and 0.77 g of HOBT are added, and the mixture is cooled in an ice bath. To the cooled solution, 0.7 g of DCC is added, and the mixture is stirred overnight under an argon atmosphere without removing the ice bath, allowing the reaction mixture to slowly return to room temperature. The precipitate then

The separated material is filtered, and the filtrate is evaporated under high vacuum. The remainder is poured onto ice and a 2N sodium bicarbonate solution and extracted twice, each time with 150 mL of ethyl acetate. The 15 organic extracts are washed once with ice and 70 mL of saturated ammonium chloride solution, once with ice and 70 mL of 2N sodium bicarbonate solution, and once with 70 mL of saturated sodium chloride solution, dried over magnesium sulfate, and evaporated. The crude product obtained in this manner (2.31 g) is stirred in 3 mL of piperidine and 75 mL of methylene chloride for 3.5 hours at room temperature. Next, it is evaporated at approximately 30°C, and the remainder is triturated with 60 ml of hexane, yielding 1.44 g of a solid, which is chromatographed on 120 g of silica gel with a 12:1:0.1 mixture of methylene chloride, methanol, and ammonia. Two recrystallizations in hexane give 580 mg of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethylethyl)-2-hydroxy-4-isopropyl 1,5-hexed-30-nyloid]imidazol-4-propionamide as crystals, chromatographically pure in thin layers, melting point 73°C.

-36-

Example 2

130 mg (0.24 mmol) of t-butyl[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-5-phenethyl]carbamate are dissolved in 30 mL of methanol and hydrogenated at room temperature in the presence of 65 mg on 5% activated carbon for 1.5 hours. At the end of hydrogen absorption, the catalyst is retained by filtration, and the filtrate is evaporated. Crystallization of the remainder in ether/hexane gives 110 mg of t-butyl [(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl~4-ethyl]carbamoyl]phenethyl]carbamate.

Example 3

15 A mixture of 83 mg (0.345 mmol) of di-acid

Benzylacetic acid, 90 mg (0.23 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, 0.045 mL (0.345 mmol) of 4-ethylmorpholine, 93 mg (0.69 mmol) of 20 HOBT, and 85 mg (0.41 mmol) of DCC in 15 mL of dimethylformamide are stirred overnight at room temperature. The separated precipitate is then filtered, and the filtrate is evaporated to dryness. The remaining precipitate is then dissolved in ethyl acetate, and the solution is washed.

•*

25 successively with 2N sodium bicarbonate solution, saturated ammonium chloride solution, again 2H sodium bicarbonate solution, and again saturated ammonium chloride solution, dried over sodium sulfate and evaporated. The remainder thus obtained 30 is chromatographed with a 14:1:0.1 mixture of methylene chloride, methanol, and ammonia on 30 g of silica gel. Crystallization of the remainder in methylene chloride/methanol/hexane gives 60 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-35-isopropyl-5-hexenyl]-α-(2,2-dibenzylacetamido)imidazol-4-

-37-

propionamide, melting point 98°.

Example 4-

A mixture of 880 mg (2.5 mmoles) of (S)-α-amino-N-[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-5 hexenyl]-imidazol-4-propionamide, 1.99 g (7.5 mmoles) of β-butoxycarbonyl-L-phenylalanine, 1.05 ml (7.5 mmoles) of triethylamine and 3.32 g (7.5 mmoles) of BOP in 100 ml of acetonitrile is shaken for 6 days at room temperature. The solvent is then evaporated, and the remaining 10% is dissolved in ethyl acetate and washed successively with 2N sodium bicarbonate solution, saturated ammonium chloride solution, 2N sodium bicarbonate solution again, and saturated ammonium chloride solution again. The organic solution is dried over sodium sulfate and evaporated, and the remainder is triturated twice with hexane. The remainder is chromatographed with a 98:2 mixture of chloroform and methanol on 100 g of silica gel. Recrystallization of the crude product thus obtained in methylene chloride and hexane gives 400 mg of t-butyl [(S)-a-[[(S)-1-[[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]c arb amoyl]-2-imida z olyl-4-ethyl]c arb amoyl]phen ethyl]-carbamate in the form of a white powder, melting point 25 82°.

The (S)-α-amino-N-[(1S,2S,4S)-2-hydroxy-1-iso-butyl-4-isopropyl-5-hexenyl]-imidazol-4-propionamide used as a starting material was prepared as follows:

30 The methyl ester of t-butoxycarbonyl-L-leu-

cine is reduced in a manner analogous to that of Example 1 with diisobutyl-aluminium hydride to give 2-t-butoxycarbonylamino-4-methyl-valeraldehyde which without further purification is directly used 35 in the next step.

-38-

To 16.8 g of magnesium shavings in 980 mL of ether, a solution of 14-2.3 g (691 mmol) of 4-bromo-3-isopropyl-1-butene 5 in 600 mL of ether is added dropwise under argon at room temperature over 75 minutes. After this addition, the reaction mixture is heated under reflux for 3.25 hours and then cooled to -60°C. Next, a solution of 50.1 g (203.8 mmol) of 2-t-butoxycarbonylamino-4-methylvaleraldehyde in 10,600 mL of ether is added dropwise over 75 minutes.

After the addition, the cooling bath is removed and the reaction mixture is stirred for 17 hours at room temperature. It is then cooled to 5°C and 150 mL of a saturated ammonium chloride solution is added dropwise while stirring, raising the temperature to 20°C. After a treatment similar to that described in Example 1, 88.4 g of a yellow oil is obtained, which is chromatographed on 2.5 kg of silica gel with a 98:2 mixture of methylene chloride and ether. After further chromatography of the middle fraction under the same conditions, a total of 30.2 g of chromatographically pure and homogeneous (4S)-t-butoxycarbonylamino-(5S)-hydroxy-(7S)-isopropyl-2-methyl-8-nonene, MS: 240 (M-t-Butoxy)+, is obtained. 25 In a manner analogous to that described in Example 1, the t-butoxycarbonyl group is finally removed by hydrochloric acid in the dioxane and the (4S)-amino(5S)-hydroxy-(7S)-isopropyl-2-methyl-8-nonene thus obtained is used directly in the following step 30 without further purification.

A mixture of 550 mg (1.75 mmol) of (4S)-amino-(5S)-hydroxy-(7S)-isopropyl-2-methyl-8-nonene, 1.547 mg (2.65 mmol) of N-amino-(5S)-bisamino-(7S)-isopropyl-2-methyl-8-nonene, 2.323 g of BOP, and 0.73 ml of triethylamine in 35 ml of acetonitrile is shaken for four days at room temperature.

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ambient temperature. Then the solvent is evaporated under reduced pressure and the remainder is treated in a manner analogous to that described in Example 1. Chromatography of the crude product on 500 g of silica gel with a 140:5:0.5 mixture of methylene chloride, methanol and ammonia gives 950 mg of fluorenyl-9-methyl [(S)-1-[[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]-carbamate in oil form which can be directly used in the next step. MS: 369 (M+H)+.

1.3 g of fluoranyl-9-methyl[(S)-1-[[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamate in 30 ml of methylene chloride and 2 ml of piperidine are stirred for 1.5 hours at room temperature. Then the solvent is evaporated.

and the remainder is triturated with hexane. The (S)-α-amino-N-[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]-imidazol-4-propionamide thus obtained is used without further purification in the next step. 20 Example 5

A mixture of 460 mg (1.18 mmol) of (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide, 313 mg (1.18 mmol) of t-butoxycarbojiyl-L-phenylalanine, 25 0.15 ml (1.18 mmol) of 4-ethylmorpholine, 320 mg

2.36 mmol of HOBT and 292 mg (1.42 mmol) of DCC in 20 ml of dimethylformamide is stirred for 18 hours at room temperature and then processed as usual. Chromatography of the remainder on 30 g of silica gel using a 20:1:0.1 mixture of methylene chloride, methanol, and ammonia as the eluent yields 410 mg of crystals with a melting point of 161°C.

2.5 g of the above compound are dissolved in 50 ml of 2.2N hydrochloric acid in a 35% solution and stirred for 6 hours at room temperature. Then the %

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The reaction mixture is evaporated under reduced pressure and the remainder is purified with a 190:10:1 mixture of methylene chloride, methanol and ammonia, giving N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-ct-(3-phenyl-L-alanyl)imidazol-4-propionamide as a homogeneous material, MS: 538 (M + H)+.

Example 6

Following a similar approach to the procedures described in Example 1, the following compounds are prepared:

- From (S)-a-amino-îT-[ (1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imida-zol-4-propionamide and α-benzyltetrahydro-oxo-4H-1,4-oxazinpropionic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(ES)-α-(morpho1inoc arb ony1)hydro c innamamid o]imid a z o1-4-propionamide as crystals with a melting point of 94° (in methylene chloride/ether/hexane);

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide and t-butoxycarbonyl-D-proline the

(2S,3S,5S)-2-(Boc-D-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 as crystals with a melting point of 132° (in methylene chloride/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-(3-phenyl-L-alanyl)-imidazol-4-propianamide and t-butylacetic acid the (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-et-(3,3-dimethylbutyramido)-hydrocinnamoyl]imidazol-4-propianamide in the form of crystals with a melting point of 105° (decomposed; in methylene chloride/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl] -<*-( 3-phenyl-L-alanyl)-

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imidazol-4-propionamide and isovaleric acid N-(S)-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-<*-[ (S)-α-3-methylbutyramido]-imidazol-4-propionamide in foam form. MS: 622 (M+H)+;

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide of (S)-α-[[(S)-2-benzylamino-3-phenylpropyl]amino]hydrocinnamic acid, benzyl [ce—[ [[et— [[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-

c arb amoyl]phen ethyl]amino]methyl]phen ethyl]c arb amat e in the form of crystals with a melting point of 89° (in methylene chloride/ether/hexane);

- From (S)-α-amino-1α-[(S)-1-[[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]hydrocinnamamide obtained from t-butyl[(S)-α-[[(S)-1-[[[(1S,2S,4S)-2-hydroxy-1-isobutyl-

4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]phenethyl]carbamate left for 18 hours at rest at room temperature in 3N methanolic hydrochloric acid, and cyclopentanecarboxylic acid (S)-a-[(RS)-cyclopentanecarboxamido]-N-[(S)-1-

[[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]hydrocinnamamide in the form of crystals with a melting point of 185° (Decomp.

in methanol/methylene chloride/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-(3-phenyl-L-alanyl)imidazol-4-propianamide and benzyl[(methyl-thio(mimidoyl)carbamate) of benzyl[1-[(S)-a-[[(S)-1-

[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-

5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-

GB Patent: 2,085,444

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phenethyl]amidino]carbamate in foam form, I-IS: 714 (M+H)+;

- From N-[(1S,2S,4S)-1-(cyclohexylmethylethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-

5-alanine)imidazol-4-propianamide and β-butoxycarbonylaminovaleric acid, β-butyl[4-[[(S)-α-[[(S)-1-[[(1S,2S,4-S)-1-(cyclohexylmethyl thy1)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]phenethyl]carbamoyl]butyl]carbamate in the form of

10 crystals with a melting point of: 110° (in methylene chloride/ether/hexane;

- From (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]imida-zol-4—propionamide and N-1-butoxycarbonyl-N-methyl-

15 phenylalanine the t-butyl [ (S)-ct-[[ (S)-1-(S)-[[ (1S ,2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4~ethyl]carbamoyl]-phenethyl]methylearbamate as crystals with a melting point of 93° (in ether/hexane);

20 - From (S)-a-amino-N-[(1S,2S,4R)-1-

(cyclohexylmethyl)-2-hydroxy-4— isopropyl-5-hexenyl]-imidazol-4—propionamide and t-butoxycarbonyl-L-phenylalanine the (2S,3S,5R)-2-(Boc-Phe-His-NH)-1-cyclo-hexyl-5-isopropyl-6-heptenol-3 in foam form, 25 MS : 638 (M+H)+;

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4—propionamide and dibenzosuberoacetic acid2 the N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-

30 hydroxy-4—isopropyl-5-hexenyl]-a-[2-(1O,11-dihydro-5H-dibenzo[a,d]cycloheptenyl-5)acetamido]imidazol-4— propionamide as crystals with a melting point of 135° (in methylene chloride/ether/hexane);

d EPA 0.056.616

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- From (S)-a-amino-N-[ (1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl] -imidazol-4—propionamide and 3-(3-indolyl)-propionic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl] -a-(3-indolyl-3-propionamido)imidazol-4—propionamide as a solid with a melting point of 122° (in ether/hexane);

- From (S)-ct-amino-N-[ (1S ,2S ,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-imidazol-4-propionamide and benzylmalonic acid monoamide (S)-a-[(ES)-a-carbamoylhydrocinnamamido ] -N-[ (1S, 2S ,4-S ) -1 - (cyclohexylmethyl ethyl ) -2-hydroxy-4—isopropyl-5-hexenyl]imidazol-4—propionamide in foam form, MS : 566 (M+H)+;

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and (S)-α-benzyl-2,5-dimethylpyrrol-1-acetic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl 1)-2-hydroxy-4-isopropyl-1-5-hexenyl 1]-α-[(S)-α-(2,5-dimethylpyrryl-1)hydrocinnamamido]-imidazol-4-propionamide as crystals with a melting point of 93° in ether/hexane;

- From (S)-a-amino-jN-[ (1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexeny1]-imidazol-4-propionamide and ot-[(2-hydroxyethyl)carbamoyl]hydrocinnamic acid (S)-N-[(1S,2S,4S)-1 - (cyclohexylmethyl thyl)-2-hydroxy-4~isopropyl-5-hexeny1]-a-[(RS)-a-[(2-hydroxyethyl)carbamoyl]hydrocinnamamido]-imidazol-4-propionamide in foam form, MS: 610 (M+H)+;

- From (S)-a-amino-N-[ (1S,2S ,4-S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and α-[(2-(dimethyl-amino)ethyl]carbamoyl]hydrocinnamic acid the (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-

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5-hexenyl-a-[(ES)-u-[[2-(dimethylamino)ethyl]carbamoyl]hydrocinnamamido]imidazol-4—propionamide as foam, NS: 637 (M+H)+;

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4—propionamide and dibenzylcyanoacetic acid (S)-a-(a-benzyl-a-cyanohydrocinnamamido)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4—propionamide as crystals with a melting point of 105° (in ether/hexane;

- From (S)-u-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-imidazol-4—propionamide and benzyl-cyano-acetic acid (RS)-a-cyano-N-[(S)-1-[[ (1S,2S ^S^I-^yclo-hexylmethyl^^-hydroxy^-isopropyl-^-hexenyl] carbamoyl] 2-imidazolyl-4-ethyl]hydrocinnamamide as crystals with a melting point of 97° (in methylene chloride/ether/hexane);

- From (S)-a-amino-lT-[ (1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4~isopropyl-5-hexenyl]-imidazol-4—propionamide and indole-2-carboxylic acid N-[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-2-benzimidazolcarboxamide as crystals with a melting point of 123° (in methylene chloride/ether/hexane);

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-imidazol-4—propionamide and isovaleric acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-(3-methylbutyramido)imidazol-4-propionamide in foam form, MS: 622 (M+H)+;

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-imidazole-4—propionamide and α-benzyl-P-oxo-

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1-pyrrolidin-propionic N-[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2.-hydroxy-4-isop^opyl-5-hexenyl] -a-[ (RS)-a-(l-pyrrolidinylcarbonyl)hydroc innamamido ] imidazo 1-4-propionamide in the form of crystals with a melting point of 89° (in methylene chloride/ether/hexane);

- From (S)-a-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-xmidazol-4-propionamide and a-[(benzyloxy)-carbamoyl]hydrocinnamic acid (S)-u-[(RS)-a-[(benzyloxy) carbamoyl]hydrocinnamamido]-N-[(1S,2S,4S)-1-cyclohexyl-

2-hydroxy-4-isopropyl-5-hexenylimidazol-4-propionamide in the form of crystals with a melting point of 104° (in methylene chloride/ether/hexane);

- From (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and 1-[a-(methoxycarbonyl)-hydrocinnamoyl]-4-piperidinecarboxylic methyl (RS) a-[[4-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]piperidino]carbonyl]-hydrocinnamate in the form of crystals with a melting point of 89° (in methylene chloride/ether/hexane).

The acids used as starting material are either known or have been prepared as follows:

α-Benzyltetrahydro-p-oxo-4H-1,4-oxazine propionic acid

The monomethyl ester of benzyl-maionic acid, which was prepared according to known processes, is amidated according to usual processes with morpholine and then hydrolyzed into α-benzyltetrahydro-β-oxo-4H-1,4-oxazinepropionic acid,

MS: 264 (M+H)+.

M. Goodman et al., J. Am. Chem. Soc., 77, 8675 (1969) and Int. J. Pept. Prof. Res., 21_, 84 (1W3)

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Acid (S) —cc—[[ (S)-2-benzylamino-3-phenylpropyl]amino] -

hydrocinnamate

In a manner analogous to the description in E-example 26 of the preparation of Boc-Phe-His(Bom)-OH, on 5 N-benzyl-Phe-Phe-OMe was treated by sulfation of the carbonyl amide group, desulfurization of the latter with Raney nickel and then hydrolysis of the ester obtained to give the (S)-a-[[(S)-2-benzylamino-3-pnényl-propyl]-amino]hydrocinnamic acid which is used 10 directly in the next step.

t-Butoxycarbonylaminovaleric acid

5-Aminovaleric acid was transformed under Schotten-Baumann conditions by di-t-butylcarbonate in the usual way into the corresponding N-protected acid, melting point 48-50° (in ether/hexane).

Benzylmalonic acid monoamide

The monomethyl ester of benzylmalonic acid is amided in the usual way with ammonia 20 and the methyl ester of 2-benzyl-2-formamidoacetic acid obtained is hydrolyzed with 1N sodium hydroxide solution in a 9:1 mixture of methanol and water in 30 minutes to give the monoamide of benzylmalonic acid, Rj> : 0.15 in a 90:10:1:0.5 mixture of chloroform, methanol, water and acetic acid.

(S)-α-Benzyl-2,5-Dimethylpyrrol-1-acetic acid

The hydrochloride of L-phenylalanyl methyl ester is transformed by stirring with a sodium bicarbonate solution in ether into the corresponding free base and is heated in glacial acetic acid with acetonylacetone for 2.5 hours at 100°C. The solvent is then evaporated, and the residue is filtered with toluene through silica gel, yielding (S)-α-benzyl-2,5-dimethylpyrrol-1-acetate, MS: 258 35 (M+H)+. The resulting ester is left to stand for

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1.5 hours at room temperature with sodium hydroxide solution in a 9:1 mixture of methanol and water. After extraction of the aqueous solution with ether and washing of the organic phases, (S)-u-5-benzyl-2,5-dimethylpyrrol-1-acetic acid, MS: 244(n+H)+, is obtained and is directly used in the next step. α-C (2-hydroxyethyl)carbamoyl-1-hydrocirmamic acid

In a manner analogous to what has been described above for α-benzyl-tetrahydro-P-oxo-4H-1,4-10 oxazinepropionic acid, the ethyl monoester of benzylmalonic acid is amidated with ethanolamine and the resulting compound is saponified to α-[(2-hydroxyethyl)carbamoyl]hydrocinnamic acid, MS: 251 (M)+, which is used directly in the next step.

15 α-[[2-(dimethylamino)ethyl(hydroxyhydroxychloroisocyanuric acid])

In a manner analogous to what has been described for α-benzyltetrahydro- α-oxo-4H-1,4-oxazinepropionic acid, the methyl monoester of benzylmalon-20 nic acid is amidated with N,N-dimethylaminoethylamine and is then saponified to α-[[2-(dimethylamino)-ethyl]carbamoyl]hydrocinnamic acid, [MS: 265 M+H+], which is directly used in the next step.

Dibenzylcyanoacetic acid 25 The ethyl ester of cyanoacetic acid is dibenzylated with an excess of benzyl bromide in ethanol to give the ethyl ester of dibenzylcyanoacetic acid, which is hydrolyzed with 1N sodium hydroxide solution in a 9:1 mixture of methanol and water to 30 give dibenzylcyanoacetic acid, melting point 194° (in methylene chloride/methanol/hexane). Benzylcyanoacetic acid

The ethyl ester of cyanoacetic acid is alkylated with benzyl bromide in the presence of sodium ethoxide 35 in ethanol to the ethyl ester of the acid

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Benzylcyanoacetic acid [MS: 203(M)+] is then hydrolyzed with 1N sodium hydroxide solution in a 9:1 mixture of methanol and water to give the corresponding acid. Recrystallization of the crude product in methylene chloride/hexane gives benzylcyanoacetic acid as crystals with a melting point of 97°C. α-Benzyl-1-oxo-pyrrolidinepropionic acid

In a manner analogous to that described for α-benzyltetrahydro- α-oxo-4-H-1,4-oxazinepropionic acid, 10 the monomethyl ester of benzylmalonic acid is amidated with pyrrolidine and the resulting product is hydrolyzed to α-benzyl-(3-oxo-1-pyrrolidine propionic acid, which is directly used in the next step, MS: 248 (M+H)+.

15 α-[(benzyloxy)carbamoylhydrocinnamic acid]

Similar to that described for α-benzyltetrahydro-β-oxo-4H-1,4-oxazinepropionic acid, the monomethyl ester of benzylmalonic acid is amidated with α-benzylhydroxylamine, and the resulting product [MS: 314 (M+H)+] is saponified with 1N sodium hydroxide solution in a 9:1 mixture of methanol and water to give the corresponding acid. Recrystallization of the crude product in methanol/methylene chloride/ether gives α-[(benzyloxy)carba-25 moyl]hydrocinnamic acid with a melting point of 147°.

1-[α-(methoxycarbonyl)hydrocinnamoyl-4-piperidine carboxyl acid]

In a manner analogous to the preparation of α-benzyltetrahydro-4-oxo-1,4-H-oxazinepropionic acid, the monomethyl ester of benzylmalonic acid is amided with the ethyl ester of piperidine-4-carboxylic acid, and the resulting compound [MS: 347(M)+] is saponified with 1N sodium hydroxide in a mixture of methanol and water for 2 hours at 0°C. The crude product obtained is purified by gel chromatography.

C

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silica with a 95:5:2 mixture of chloroform, ethanol and ethyl acetate and recrystallized in a mixture of methanol, methylene chloride and ether, which gives 1-[a-(methoxycarbonyl)hydrocinnamoyl]-4-piperi-5-dinecarboxylic acid, melting point 138°.

Example 7

Similar to that described in Example 1, N-[(1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-15-hexenyl]-α-(3-10-phenyl-L-alanyl)imidazol-4-propianamide is reacted with di-t-butoxycarbonylaminoethylglycine. Chromatographic purification and recrystallization of the crude product yields di-t-butyl N-[[[(S)-α-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-15-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-

phenethyl]carbamoyl]methyl]ethylenedicarbamate, melting point 109°.

150 mg N-[[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-20 carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]phenethyl]-carbamoyl] methyl ethylenedicarbamate are dissolved in 0.3N hydrochloric acid in methanol and left to stand overnight at room temperature. The solvent is then evaporated and the remainder is precipitated in methanol/ethyl acetate, yielding (S)-ct-

[(S)-a-[2-[(2-aminoethyl)amino]acetamido]hydrocinnamami-do]-N-[ (1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]imidazol-4—propionamide trichlorohydrate, MS: 638 (M+H)+. 30 The di-t-butoxycarbonylaminoethylglycine used as a starting material was prepared as follows:

N-(2-aminoethyl)glycine is prepared according to a process described by E.P. Heimer in Int. J. Pept. 35 Prot. Ees., 23, 203 (1984-) and is transformed from maQ

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known nière under Schotten-Baumann conditions with di-t-butylcarbonate in the corresponding N-protected acid, MS: 319 (M+H)+.

Example 8

5 60 mg of (2S,3S,5S)-2-(Boc-D-Pro-Phe-His-NH)-

1-Cyclohexyl-5-isopropyl-6-heptenol-3 is dissolved in 10 mL of 2.1N hydrochloric acid dissolved in dioxane and stirred for 1.5 hours at room temperature. The solvent is then evaporated, and two 10 mL portions of toluene are added successively to the residue, followed by drying. Crystallization of the resulting residue in methylene chloride/ether/hexane yields 40 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethylethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(S)-α-(D-prolylamino)hydrocinna-15mamido]imidazol-4-propionamid dihydrochloride as white crystals with a melting point of 159°C.

Example 9

N-Methyl-L-histidine is transformed into the corresponding benzyl ester with benzyl-20 alcohol in the presence of hydrochloric acid at room temperature and the latter is reacted according to the process described by D.H. Rich et al. in Proc. 9 American Pept. Sym. Toronto 1985, page 217, with β-butoxycarbonyl-L-alanine in the usual manner, then the N-[N-β-25 butoxycarbonyl]-3-phenyl-L-alanyl]-N-methyl-L-histidine ester obtained is hydrogenated for one hour in the presence of 5% palladium on activated carbon, giving N-[N-(β-butoxycarbonyl)-3-phenyl-L-alanyl]-N-methyl-L-histidine, R^: 0.2, in an 80:20:3:3 mixture of chloroform, 30 methanol, water, and acetic acid. This compound is reacted in the usual manner with (αS,βS)-α-amino-α-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol.

The crude product obtained is then chromatographed with a 20:1:0.1 mixture of methylene chloride, 35 methanol and ammonia on silica gel, which allows

-51-

separate the two epimeric compounds. Recrystallization of the crude product of the first fraction in ether/methylene chloride/hexane gives t-butyl[(S)-a_[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-5 4-isopropyl-5-hexenyl] carbamoyl] -2-imidazolyl-4-ethyl] -methylcarbamoyl]-phenethyl]carbamate as white crystals with a melting point of 95° •

The second fraction, after recrystallization in ether/methylene chloride/hexane, gives the epimeric compound t-butyl[(S)-u-[[(R)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl thy1)-2-hydroxy-4-isopropy1-5-hexeny1]-carbamoyl]-2-imidazolyl-4-ethyl]methylcarbamoyl]phenethyl]carbamate, in the form of white crystals with a melting point of 85°•

15. In a manner analogous to what is described above-

N-t-butoxycarbonyl-N-methyl-L-phenylalanine is reacted on top of this according to the process described by Rich et al.

with N-methyl-L-histidinebenzylester and the resulting ester is then hydrogenated to the corresponding acid, 20 and this is reacted in the usual manner with

(aS ,f>S)—p—amino-a—[ (S) -2-isopropyl-3-butenyl]cyclohexane-propanol. Purification by chromatography of the crude product on silica gel with a 140:1:0.1 mixture of methylene chloride, methanol and ammonia gives the pure 25 t-butyl [(S)-a-[[(S)-a-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]methylcarbamoyl]phenethyl]methylcarbamate in thin-layer chromatography as a foam: MS 666 (M+H)+.

Example 10

150 mg (0.20 mmol) of t-butyl-2-benzyl-2-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]-3-phenylpropyl]carbamate are dissolved in 35 10 ml of methanolic hydrochloric acid solution

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0.3N and left to stand overnight at room temperature. The solvent is then evaporated and the remainder is purified by chromatography with a 20:1:0.1 mixture of methylene chloride, methanol, and 5-ammonium chloride as the eluent on 30 g of silica gel. Recrystallization of the crude product thus obtained in methylene chloride/ether/hexane gives (S)-a-(3-amino-2,2-dibenzylpropionamido)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]imidazol-4,10 propionamide as white crystals with a melting point of 89°.

The starting material used, t-butyl-2-benzyl-2-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-15 4-ethyl]carbamoyl]-3-phenylpropyl]carbamate, was prepared as follows:

The methyl ester of dibenzylcyanoacetic acid was reduced with Nickel Raney in methanolic ammonia to the ethyl ester of di-20 benzylmethylaminoacetic acid (MS: 284 (M)+), the amino group of which was protected in the usual manner with di-t-butylcarbonate in triethylamine. The ester obtained in this way [MS: 398 (M+H)+] was saponified to the corresponding acid with 1N sodium hydroxide in a methanol/water mixture, which, after recrystallization in methanol/methylene chloride/hexane, was isolated as crystals with a melting point of 157°. Next, the acid is reacted in the usual way with (S)-a-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-30-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide to give t-butyl-2-benzyl-2-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-3-phenylpropyl] carbamate.

35 Recrystallization in the chloride mixture

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methylene/hexane/ether produces "white" crystals with a melting point of 97°C.

Example 11

150 mg of (S)-a-[ (RS)-β-[ (benzyloxy)carbamoyl]-5-hydrocinnamamido]-N-[(1S,2S,4S)-1-cyclohexyl-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide are hydrogenated in the presence of 5% palladium on activated carbon for 3 hours in methanol. At the end of hydrogen absorption, the catalyst is separated by 10 filtrations and the filtrate is evaporated. Recrystallization of the remainder in methanol/methylene chloride/ether gives (S)-11-[(1S,21,4S)-1-(cyclohexylmethyl)-4-ethyl-2-hydroxy-5-methylhexyl]-a-[(ES)-a-(hydroxy-carbamoy1)-hydroc innamamid o]imid a zo1-4-prop ionamide 15 as white crystals with a melting point of 123°.

Example 12

120 mg of t-butyl C(S)-a-[[(S)-1-(S)-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phen-20 thyl]methylcarbamate are dissolved in 0.3 N methanolic hydrochloric acid and stirred overnight at room temperature. The solvent is then evaporated under reduced pressure, and t-butylacetyl chloride and triethylamine in 25-tetrahydrofuran are added to the remaining solution, and the mixture is left to stand overnight at room temperature. After usual treatment and chromatographic purification on silica gel using a 20:1:0.1 mixture of methylene chloride, methanol and ammonia and recrystallization in ether/hexane on 30, (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-(N,3,3-trime-ethylbutyramido)hydrocinnamamido]imidazol-4-propionamide is obtained as white crystals with a melting point of 126°.

Example 13

35. In a manner similar to Example 1, we prepare

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the following compounds:

- From M-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-(3-phenyl-L-alanyl)-imidazol-4-propionamide and 3-(4-

5-hydroxyphenyl)propionic (S)-N-[(1S, 2S, 4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a.-[(S)-a-(p-hydroxyhydroc innamamido)hydroc innamamido]-imidazol-4-propionamide as white crystals, melting point 120° (in methylene chloride/

10 ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)-imidazol-4-propionamide and nicotinic acid, the (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-

15 hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-nicotinamido-hydrocinnamamido]imidazol-4-propionamide in solid form, m: 643 (M+H)+;

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-

20 L-alanyl)imidazol-4-propionamide and 3-(4-pyridyl)acrylic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-ct-[[(S)-u-L(E)-4-pyridinacrylamido]hydrocinnamido]imidazol-4-propionamide in the form of yellowish crystals, point

25 melting point 135° (Dec.; in methylene chloride/ ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 3-(3-

30 pyridyl)acrylic the (S)-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-(3-pyrid ine-acrylamido)hydroc innamamido]imida zo1-4-propionamide in the form of yellowish crystals, melting point 141° (in methylene chloride/ether);

1

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- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4—isopropyl-5-hexenyl]-^-(3-phenyl-L-alanyl)imidazol-4—propionamide and 3-(4-imidazolyl)acrylic acid (urocanic acid) the (S)-N-

[ (1S,2S,4-S )-1-(cyclohexylmethyl )-2-hydroxy-4— isopropyl-5-hex ny1]-a-[(S)-a-(imidazol-4—a c ry1amido)hydro c innamamido] imidazol-4—propionamide . in the form of "white" crystals, melting point 150-152° (in methylene/methanol/ether chloride);

- From N-[ (1S,2S,4-S)-1-(cyclohexyl-methyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-(3-phenyl-L-alanyl)imidazol-4—propionamide and D-(-)-China acid, (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-[ (s)-a-(1,3,4-, 5-tetrahydroxycyclohexanecarboxamido)hydrocinnamamido]-imidazo1-4—propionamide as a white solid, MS: 712 (M+H)+;

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-(3-phenyl-L-alanyl)imidazol-4—propionamide and chinaldic acid, N-[(S)-a-[[(S)-1—[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]-2-chinolinecarboxamide in the form of a white solid, MS: 693 (M+H)+;

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and levulic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(S)-u-(4-oxavaleramido)-hydro-cinnamamido]imidazol-4-propionamide in the form of white crystals, melting point 150° (Dec.; in methylene chloride/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4~isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4—propionamide and pyridyl-

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acetic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-(3-pyridine-acetamido)hydrocinnamamido]imidazol-4-propionamide as white crystals, melting point 195° (Dec.; in methanol/methylene chloride/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl)-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 4-chlorocinnamic acid, the (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-<*-[(S)-α-(4-chloro-

cinnamamido)hydrocinnamamido]imidazol-4-propionamide in the form of white crystals, melting point 139°

(in methylene chloride/methanol/ether);

- From the lT-[ (1S ,2S ,4S)-1 -(cyclohexylmethyl) -2-hydroxy-4-isopropyl-5-hexenyl}-a-( 3-phenyl-L-alanyl )imidazol-4-propionamide and trans-4-nitrocinnamic acid the (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl-ethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-(4-nitrocinnamamido)hydrocinnamamido]imidazol-4-propionamide in the form of yellowish crystals, melting point 120° (Dec.; .in methylene chloride/methanol/ ether);

- From H-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and the monoethyl ester of fumaric acid ethyl 3-[[(S)-ct-[[(S)-1-

[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyl]carbamoyl]acrylate in the form of white crystals, melting point 175° (Dec.; in methylene/methanol/hexane chloride);

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and diphenylcarbamoyl-L-phenylalanine, a mixture of (S)-N-[(1S,2S,4S)- epimers

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1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-[[N-(diphenylcarbamoyl)-3-phenyl-L-alanyl]amino]-imidazol-4—propionamide and (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4~isopropyl-5-hexenyl]-a-[[N-(diphenylcarbamoyl)-3-phenyl-D-alanyl]amino]imidazol-4—propionamide, the first epimer being predominant, melting point of the two epimers 106° (from methylene chloride/ether/hexane);

- From (S)-u-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexyl]-imidazol-4-propionamide and isobutoxycarbonyl-L-phenylalanine isobutyl [(S)-a-[[(S)-1-[[(1S,2S,4S)-

1-(cyclohexylmethyl)-2-hydroxy-4— isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]phenethyl]-carbamate in the form of white crystals, melting point 14-7° (from ether/methylene chloride/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and D-pyroglutamic acid, the (S)-N-[(1S,2S,4S)-1-[cyclohexylmethyl]-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(S)-α-[[5-oxo-D-

prolyl]amino]hydro c innamamido]imidazol-4-prop ionamid e in solid form, white, melting point 200°

(Dec., from methanol/ethyl acetate/hexane);

- Starting from N-[ (1S,2S,4-S)-1-(cyclohexyl-

methyl)-2-hydroxy-4-isopropionamide and L-pyroglutamic acid (S)-N-[(1S,2S,4S)-1-[cyclohexylmethyl)-

2-hydroxy-4—isopropyl-5-hexenyl]-a-[(S)-u-[[5-oxo-L-prolyl]amino]hydrocinnamamido]imidazol-4—propionamide as white crystals, melting point 148° (from methylene/methanol/ether chloride);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-α-(3-phenyl-

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L-alanyl)imidazol-4-propionamide and N-t-butoxy-carbonyl-a-methyl-L-alanine and t-butyl[l-[[(S)-a-[[(S)-1-[ [ ( 1S, 2S, 4S ) -1 - (cyclohexylmethyl)-2-hydroxy-4-iso-propyl-5-hexenyl]carbamoyl]-2-imidazolyl-4~ethyl]-carb amoyl]phenethyl]carb amoyl]-1-methylethyl]carb amate in the form of white crystals, melting point 112° (from chloride, methylene/tetrahydrofuran/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)-imidazol-4-propionamide and Fmoc-Ser(t-Bu)OH the 9H-fluorenyl-9-[(S)-2-t-butoxy-1-[[(S)-α-[[(s)-1-

[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyl]carbamoyl]ethyl]carbamate as a white solid, melting point 180° (Dec.; from methylene chloride/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-O-phenyl-L-alanyl)-imidazol-4-propionamide and dibenzyl-suberoacetic acid (S)-N-[(1S,2S,4S)]^-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[[N-(9H-fluorenyl-9-acetyl)-3-phenyl-L-alanyl]amino]imidazol-4-propionamide in the form of white crystals, melting point 119° (from methyl chloride/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)-imidazol-4-propionamide and adamantylacetic acid, the (S)-α-[[N-(1-adamantylacetyl)-3-phenyl-L-

a 1 any 1 ] amino ]-N-[(1S,2S,4S)-1-[cyc1ohexy lmé t hy 1 ) -2 -hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide in the form of white crystals, melting point 120° (from methylene chloride/ether/hexane);

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- From (S)-a-amino-N-[ (1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and phenyloxycarbonyl-L-phenylalanine benzyl [(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-c a rb amoyl]-2-imida zoly1-4-ethyl]c a rb amoyl]phen ethyl]-carbamate as white crystals, melting point 144°, (from methylene chloride/ether);

- From (S)-u-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and 2,2,2-trichloroethoxy carbonyl-L-phenylalanine and 2,2,2-trichloroethyl [(S)-1-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamate as white crystals, melting point 109° (from methylene/hexane chloride);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-[(3-phenyl-N-D-prolyl-L-alanyl)amino]imidazol-4-propionamide and 4-imidazolylpropionic acid, which was prepared by hydrogenation of urocanic acid, N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-

[(S)-a-[(B)-1-(3-imidazolyl-propionyl]-2-pyrrolidine-carboxamido]hydrociminamido]imidazo1-4-propionamide in the form of a foam, MS: 760 (M+H)+;

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and (R)-1-[(3-hydroxy-2-pyridyl)carbonyl]-2-pyrrolidinecarboxylic acid, (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-[(S)-α-[(r)_1_[(3_hy-droxy-2-pyridyl)carbonyl]-2-pyrrolidinecarboxamido]-hydrocinnamamido]imidazol-4-propionamide is a white solid with a melting point of 198°C (Dec., from

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methylene chloride/methanol/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl ethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-^-(3-phenyl-L-alanyl)-imidazol-4-propionamide and (R)-1- acid

5 [(dibenzylacetyl)-2-pyrrolidinecarboxylic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-tt-[(S)-a-[(R)-1-(dibenzylacetyl)-2-pyrroli-dinecarboxamido]hydrocinnamamido]imidazol-4-propionamide as white crystals, melting point 104° (from 10 from methylene chloride/ether/hexane);

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl )-2-hydroxy-4-isopropy 1-5-hexenyl]-and- (3-phenyl-L-alanyl)-imidazol-4-propionamide and 1-t-butoxycarbonylamino-cyclohexanecarboxylic acid, the t-butyl-

15 1-[[(S)-a-[[(S)—1 —[(1S,2S,4S)-1-(cyclohexylmethyl-2-hydroxy-4-isopropyl-5-hexenyl]-2-imidazolyl-4-ethyl]-carbamoyl]phenethyl]carbamoyl]cyclohexanecarbamate as a white solid, MS: 764 (M+H)+;

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-20 2-hydroxy-4-isopropylhexyl]-α-[(3-phenyl-N-D-prolyl-L-

alanyl)amino]imidazol-4-propionamide and isovaleric acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl] -a-[(S)-a-[(R)-l-isovaleryl-

2-pyrro1id ine c arb oxamido]hydro c inn amamid o]imid a z o1-4-23 propionamide in foam form, MS: 721- (M+H)+;

- From (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(S)-α-(D-prolylamino)hydrocinnamamido]imida z01-4-propionamid dihydrochloride and isovaleric acid, (S)-N-

30 [(1S,2S,4S')-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[[(S)-a-[(l-isovaleryl-L-propyl)carbonyl]-

3-phenyl-L-α1anyl]amino]imid a z o1-4-propionamide in foam form, MS: 7^9 (M+H)+.

The N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-35 4-isopropylhexyl]-a-[ (3-phenyl-N-D-prolyl-L-alanyl)-

L

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amino]imidazol-4-propionamide used as the starting material was prepared from t-butyl(R)-2-[[ (S)-a-[[ (S)-1-[ (1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl] -2-imidazolyl-4~ethyl] carba-5 moyl]phenethyl]carbamoyl]-1-pyrrolidinecarboxylate described in Example 16, by removing the t-butoxycarbonyl group in a manner analogous to that described in Example 8 with hydrochloric acid in dio-xanne.

10. The acids used as starting materials are either known or have been prepared as follows: Diphenylcarbamoyl-L-alanine

Treatment of L-phenylalanine with diphenylcarbamoyl chloride in dimethylforma-15, after recrystallization in methanol/methylene chloride/hexane, yields the desired acid in the form of white crystals. Isobutoxycarbonyl-L-phenylalanine

Treatment of L-phenylalanine with iso-20 butylchloroformate in a manner analogous to that described above, gives the desired acid in the form of an oil which crystallizes at rest [MS: 266 (M+H)+] which is directly used in the next step.

2,2,2-trichloroethoxycarbonyl-L-phenylalanine

25 L-Phenylalanine is reacted in a 2N sodium bicarbonate solution with 2,2,2-trichloroethylformate, which after usual treatment gives the desired acid in the form of crystals, melting point 126° (from methyl-30lene/hexane chloride).

(R)-1-f(3-hydroxy-2-pyridyl)carbonyl]-2-pyrroli-dinecarboxylic acid

3-Hydroxypicolic acid is condensed in the usual way with the benzyl ester of D-proline, and the resulting ester is transformed by hydrogenolysis into the a

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the desired cide, which is used directly in the next step.

(R)-1-(dibenzylacetyl)-2-pyrrolidinecarboxylic acid Dibenzylacetic acid is 5-condensed in the usual way with the benzyl ester of D-proline, and the resulting ester is transformed by hydrogenolysis into the desired acid, which is used directly in the next step.

1-t-Butoxycarbonylaminocyclohexanecarboxylic acid 10 1-Aminocyclohexanecarboxylic acid is transformed in the usual way with di-t-butyldicar-bonate into the corresponding N-protected acid, which is used directly in the next step.

Example 14

15. 40 mg of 9H-fluorenyl-9-[(S)-2-t-

butoxy-1-[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-^2-hydroxy-4—isopropyl-5-hexenyl]carbamoyl]-2-imida zolyl-4—ethyl]carbamoyl]phen ethyl1]carbamoyl]ethyl]-carbamate in 5 ml of methylene chloride and 0.4 ml 20 of piperidine and left for 2 hours to stand at room temperature and the remainder is chromatographed on silica gel with a 14:1:0.1 mixture of methylene chloride, methanol and ammonia. Recrystallization of the crude product thus obtained gives (S)-a-[[N-(3-t-2 5 but oxy-L-alanyl)-3-phenyl-L-alanyl]amino]( 1S, 4S ) -1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-propionamide as a white solid, melting point 103° (dec.: from methylene/ether/hexane chloride).

30 Example 15

t-butyl[4[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyl]carbamoyl]butyl1carbamate is dissolved in 0.3N methanolic hydrochloric acid and allowed to stand

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18 hours at room temperature. Then the reaction mixture is evaporated to dryness and the remainder is chromatographed on silica gel, giving (S)-a-(5-aminovaleramido)-N-[(S)-1-[[(1S,2S,4S)-1-(cyclohexyl-5 methyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imida azol ethyl]hydro c innamamide-dichlorohydrate as a foam, MS: 637 (M+H)+.

In a manner analogous to that described above, from t-butyl (R)-2-[[(S)-a-[[(S)-1-10 [(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-hexyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamoyl]-1-pyrrolidine-carboxylate, the preparation of which is described in Example 16, is obtained IT-[ (1S,2S,4S)-t-(cyclo-hexylmethyl)-2-hydroxy-4-isopropylhexyl]-&-[(3-phenyl-15 N-D-prolyl-L-alanyl)amino]imidazol-4-propionamide as a homogeneous white solid in thin-layer chromatography MS: 635 (M+H)+.

Example 16

Similar to what is described in Example 2, the following compounds are prepared by hydrogenation of the corresponding olefins:

- From ethyl-3-[[(S)-a-[[(S)-1-[ [(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl] -25 phenethyl]carbamoyl]ethyl acrylate [[(S)-a-[[(S)-1-

[ [ ( 1S, 2S, 4S ) -1 - ( c yc 1 oh exylmethyl)-2-hydroxy-4— is op r opy1-he xy1]c arb amoyl]-2-imid a zoly1-4—ethyl]c arb amoyl]phene-ethyl]carbamoyl]propionate as a homogeneous solid by thin-layer chromatography, MS: 668 (M+H)+; 30 - From (S)-a-[[N-(1-adamantylacetyl)-3-

phenyl-L-alanyl]amino]-N-[(1S,2S,4S)-1-[cyclohexyl1-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide (S)-a-[(S)-u-[2-(1-adamantyl)acetamido]-hydrocinnamamido]-N-[(1S,2S,4S)-1-cyclohexylmethyl)-2-35-hydroxy-4-isopropylhexyl]imidazol-4-propionamide

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white solid shape, MS: 717 (M+H)+;

- From (S)-N-[(1S,2S,4S)-1-(cyclohexyl-methyl}-2-hydroxy-4-i s opropy1-5-hexenyl]-a-[(S)-a-(3-pyridinacetamido)hydrocinnamamido]imidazol-4-propionami-

5 of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-[(S)-α-[2-(3-pyridyl)acetamido]-hydrocinnamamido]imidazol-4-propionamide as white crystals, melting point 211° (from methylene/methanol/ether chloride); 10 - A-From (S)-1T-[(1S,2S,4S)-1-[cyclohexyl-

methyl]-2-hydroxy-4-isopropyl-5-hexenyl]-u[(S)-a-[[5-oxo-L-proly1]amino]hydr1c innamamid o]imid a z o1-4-p rop i o-namide le (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-a-[(S)-u-[(S)-5-oxo-2-pyrro-15 lidinecarboxamido]hydrocinnamamido]imidazol-4-propiona-mide as a homogeneous solid by thin layer chromatography, MS: 651 (M+H)+;

- From the mixture of (S)-N- epimers

[(1S,2S,4S)-1-[cyclohexylmethyl]-2-hydroxy-4-isopropyl-20 5-hexenyl] -a-[(S)-a-[[5-oxo-D-prolyl]amino]hydrocinnama-mido]imidazol-4-propionamide and (S)-N-[ (1S,2S,4S)j-1 -[cyclohexylmethyl]-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(s)-a-[[5 -oxo-L-prolyl]amino]hydrocinnamamido]imida-zol-4-propionamide (S)-N-[(1S,2S,4S)-1-(pyclohexyl-25) methyl)-2-hydroxy-4-isopropylhexyl]-u-[(S)-ct-[(RS)-5-oxo-2-pyrrolidinecarboxamido]hydrocinnamamido]imidazol-4-propionamide as a white solid, MS: 651 (M+H)+;

- From (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[[N-(9H-fluo-

30 renyl-9-acetyl-3-phenyl-L-alanylaminoimidazol-4-

propionamide (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-a-[(S)-a-[2-(10,11-dihydro-5H-dibenzo[a,d]cycloheptenyl-5)acetamido]hydrocinnamami-do]imidazol-4-propionamide as a homogeneous white solid, 35% homogeneous in thin-layer chromatography, MS: 775

C

-65-

(M+H)+;

- From (S)-N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[[N-(di-phenylcarbamoyl)-3-phenyl-D-alanyl] amino] imidazol-4-

5 propionamide ethyl [(S)-u-[[(S)-1-[[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hyd r oxy-4-is opropylhexyl]c a rb amoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamate in the form of white crystals, melting point 196° (from methanol/methylene chloride/ether);

10 - From benzyl[(S)-a-[[(S)-i-[[(1S,2S,

4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-nyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl] carbamate phenyl[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyllcarbamoyl]-2-

15 imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamate in the form of white crystals, melting point 157° (from methylene/methanol/ether chloride);

- From (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-is opropy1-5-hexenyl]-a-[ (S) -a- (4-

20 nitrocinnamamido)hydrocinnamamido]imidazol-4-propionamide (S)-a-[ (S)-a-(4-aminohydrocinnamamido)hydrocinnamamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]imidazol-4-propionamide as white crystals, Melting point 174° (from the chloride-

25 methylene/methanol/ether ride);

- From (2S,3S,5S)-2-(Boc-D-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 the t-butyl(E)-2-[[(S)-a-[[(S)-1-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-2-imidazolyl-4-ethyl]carba-

30 moyl]phenethyl]carbamoyl]-1-pyrrolidinecarboxylate in the form of white crystals, melting point 173° (from methylene/methanol/hexane chloride);

- From (S)-N-[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-a-(4-oxa-

35 valeramido)hydrocinnamamido]imidazol-4-propionamide

C

-66-

(S)-N-[(1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4— isopropylhexyl] -a-[(S)-a-(4—oxovaleramido^hydroxycinnama-midojimidazol^-propionamide in foam form, MS: 638 (M+H)+;

3 -From (S)-N-[ (1S,2S,4-S)-1-(cyclohexyl-

methyl)-2-hydroxy-4~ isopropyl-5-hexenyl]-u-[ (S)-a-(4— hydroxyhydrocinnamamido)hydrocinnamamido]imidazo1-4— propionamide le (S)-N-[ (1S ,2S, 4-S )-1-( cyclohexylmethyl )-2-hydroxy-4— isopropylhexyl]-a-[ (S)-a-(4—hydroxyhydro-10 c innamamido)hydro c innamamido]imidazo ol-4—propionamide as a white solid, melting point 120° (Dec., from methylene chloride/ether/hexane);

- From (S)-[ (1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4~isopropyl-5-hexenyl]-a-[(S)-a-

15 (3-pyridine-acrylamido)hydrocinnamamido]imidazol-4-

propionamide le (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl] -a-[ (S)-a-[3-(3-pyridyl)-propionamido]hydrocinnamamido]imid a zo1-4-propionamide in the form of white crystals, melting point 195° 20 (from methanol/ether/hexane);

- From (S)-N-[ (1S,2S,4-S)-1-(cyclohexyl-methyl)-2-hydroxy-4—isopropyl-5-hexenyl] -a-[(S)-u-nic o-t inamidohydro c innamamido]imida zol-4—prop ionamide the N-[ (S)-a-[[ (S)-1-[[ (1S,2S,4-S)-1-(cyclohexylmethyl)-2-25 4-hydroxy—isopropylhexyl]carbamoyl]-2-imidazolyl-4—

ethyl]carbamoyl]phenethyl]nicotinamide, as a white solid, melting point 112° (Dec., from methylene chloride/ether/hexane);

- From (S)-N-[(1S,2S,4S)-1-(cyclohexyl-30 methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(S)-α-(imidazole-4-acrylamido)hydrocinnamamido]imidazol-4—propionamide, N-[(1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylα-[(S)-α-imidazol-4-propionamidohydrocinnamamido]imidazol-4—propionamide in crystal form

35 whites, melting point 118° (from methanol/

17

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ether/hexane);

- From t-butyl[l-[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-r-imidazolyl-4-ethyl]carbamoyl]-5-phenethyl]carbamoyl]-1-methylethyl]carbamate, t-butyl-[l-[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]hydrocinnamoyl]carbamoyl]-1-methylethyl]carbamate in foam form, MS: 725 (M+H)+. 10 Example 17

A mixture of 95 mg (0.135 mmole) of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-[(3-phenyl-N-D-prolyl-L-alanyl)amino]imidazol-4-propion-† amide, 0.025 ml (0.199 mmole) of piva-15 loyl chloride and 0.185 ml (1.35 mmole) of triethylamine in 10 ml of tetrahydrofuran is shaken overnight at room temperature. After usual processing and chromatographic purification on silica gel with a 20:1:0.1 mixture of methylene chloride, methanol, and 20% ammonia, (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-[(S)-α-[(E)-1-pivaloyl-2-piperidinecarboxamido]hydrocinnamamido]imida-zol-4-propionamide is obtained as a homogeneous solid by thin-layer chromatography, MS: 722 (M+H)+. Example 18

20 mg of (S)-a-[[N-(3-t-butoxy-L-alanyl)-3-

phenyl-L-alanyl]amino]-N-[(1S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-propionamide are dissolved in 1 ml of aqueous tri-30 fluoroacetic acid solution (55%) and stirred for 2.5 hours at room temperature. Evaporation of the reaction mixture and purification by silica gel chromatography with an 8:1:0.1 mixture of methylene chloride, methanol, and ammonia gives (S)-N-[(1S,2S,4S)-1-(cyclohexyl-35 methyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[[N-(L-

h

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serylamino)-3-phenyl-L-alanyl]amino]imidazol-4—propionamide , MS : 625 (M+H)+, homogeneous in thin layer chromatography.

Example 19

195 mg of N-[ (R)-2-benzyl-5,5-<ii^lethyl-4-oxo-hexanoyl]-l!i-methyl-L-histidinebenzylester are hydrogenated for 2 hours in the presence of 25 mg of palladium on activated charcoal (5%) and the N-[(R)-2-benzyl-5,5-dimethyl-4-oxohexanoyl]-N-methyl-L-histidine obtained is reacted in the usual way with (aS,pS)-^-amino-a-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol. Chromatographic purification of the crude product obtained, on silica gel with a 140:10:0.1 mixture of methylene chloride, methanol and ammonia gives N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(R)-2-benzyl-N,5,5-trimethyl-4-oxohexanami-do]imidazol-4-propionamide as a homogeneous substance in thin-layer chromatography, MS: 635 (M+H)+.

As a by-product, we obtain the less polar epimer N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(R)-2-benzyl-N,5,5-trimethyl-

4-oxohexanamidoj-imidazol-4-propionamide, MS: 635 (M+H)+.

The N-[(R)-2-benzyl-5,5-dimethyl-4-oxohexanoyl]-N-methyl-L-histidinebenzylester used as a starting material was prepared by condensation of N-(a-methyl)-L-histidinebenzylester with 2-(R)-benzyl-4-oxo-5,5,5-trimethylvaleric acid and used directly in the next step.

Example 20

100 mg (0.164 mmol) ethyl (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-

5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyrate are stirred in a 30% hydrazine hydrate solution in methanol for 3 hours

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at room temperature. The reaction mixture is then evaporated, and the remainder is chromatographed with a 14:1:0.1 mixture of methylene chloride, methanol, and ammonia on silica gel. Crystallization of the crude product thus obtained from methanol/methylene chloride/ether gives (H)-5-[[(S)-1-[[(1S,2S,4S)-

1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl-carbamoyl-2-imidazolyl-4-ethyl-carbamoyl-4-phenyl-butyrohydrazide in the form of white crystals, dot

10 of fusion 109°. (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-

2-imida zolyl-4-ethyl]c arb amoyl]-4-phenylbutyrate e ut ili-sé as a starting point was prepared as described in Example 40.

15 Example 21

0.083 ml (0.54 mmole) of ethyl ester of 4-piperidine carboxylic acid is reacted with 210 mg (0.36 mmole) of ethyl 1-[(R)-3-[[(S)-1-[[(1S,2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-20 hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyry-l]carboxylate in 20 ml of acetonitrile in the presence of bis-(2-oxo-3-oxazolidinyl)phosphine chloride at room temperature. After usual processing and silica gel chromatography, ethyl-1-[(R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyryl]-4-piperidinecarboxylate is obtained as a foam, MS: 720 (M+H)+.

30. In a manner analogous to what has been described above-

The following compounds were prepared on top of this:

- From (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbuty-35 ric acid and L-prolinol, (S)-N-[(1S,2S,4S)-1-(cyclo-

H

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hexylmethyl ) -2 -hydroxy-4~is op ropy1-5-hexé nyl ] -a -[ (R)-a-[[(2-hydroxymethyl)-1-pyrrolidinyl]carbonyl]methyl]-hydrocinnamamido]imidazol-4—propionamide as a solid "white, melting point 103° (Dec.; from 5 methylene chloride/ether/hexane);

- From (R)-3-[[ (S)-1-[[ (1S,2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]-4-phenylbutyric acid and N-methylethanolamine (S)-N-[(1S,

10 2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(R)-a-[[(2-hydroxyethyl)methylcarbamoyl]-methyl]hydrocinnamamido]imidazol-4-propionamide as a white solid, melting point 82° (Dec.; from methylene chloride/ether/hexane); 15 - From (R)-3-[[(S)-1-[[(1S,2S,

4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexe-nyl]carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]-4—phenyl-butyric and N,N-dimethylethylenediamine the (S)-N-[(1S ,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-20 5-hexenyl]-a-[(R)-a-[[2 -(dimethylamino)ethyl]carbamoyl]-methyl]hydrocinnamamido]imidazol-4—propionamide as a white solid, melting point 98° (from methylene/hexane chloride);

- From the acid (R)-3-[[(S)-1-[[(1S,2S,

23 4S) 1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl] carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]-4—phenylbutyric and N,N-dimethylpropylenediamine the (S)-N-[(1S ,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-[ (R)-cc-[[ [3-(dimethylamino)propyl]carba-30 moyl]methyl]hydrocinnamamido]imidazol-4-propionamide as white crystals, melting point 98° (from methylene chloride/ether).

'The acid (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclo-

hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-35 2-imidazolyl-4—ethyl]carbamoyl]-4—phenylbutyric has been

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prepared in the following manner:

To 100 mg (0.16 mmol) of ethyl (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyrate (see Example 40) was added,

In 3 ml of ethanol, 0.66 ml of 0.5N sodium hydroxide (0.33 mmol) was mixed and the mixture was stirred for 2.5 hours at room temperature. Then, it was neutralized with 0.33 ml (0.33 mmol) of 1N hydrochloric acid, ethyl acetate was added, and the mixture was washed with saturated sodium chloride solution.

Next the organic phase is dried over sodium sulfate, and the solvent is removed under reduced pressure, and 91 mg (55%) of (R)-3-[[(S)-1-[[(1S, 2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-15-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyric acid is obtained as an amorphous powder, MS: 581 (M+H)+.

Example 22

70 mg (0.1 mmol) ethyl 1-[(R)-3-[[(S)-1-20 [[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopro-

[pyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyryl]-4-piperidinecarboxylate are suspended in 1N sodium hydroxide in a mixture of methanol and water and stirred for 3 hours at room temperature. Then the solvent is evaporated and the remainder is recrystallized in methanol/methylene chloride/hexane, yielding 1-[(R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenyl-30 butyryl]-4-piperidinecarboxylic acid as white crystals, melting point 169°.

Example 23

The reaction of (S)-N-[(1S,2S,4S)-1-(cyclohexyl-methyl1)-2-hydroxy-4-isopropy1-5-hexony1]-a-[(S)-a-(D-35 prolylamino)hydrocinnamamido]imidazol-4-propionamide di-

-72-

hydrochloride (see Example 8) with Fmoc-Ser(t-Bu)-OH in a manner analogous to that described in Example 1, treatment of the protected Fmoc compound with piperidine and then chromatography of the crude product on silica gel with a mixture of methylene chloride, methanol and ammonia gives the pure (S)-a-[[N-[l-(3-butoxy-L-alanyl)-D-propyl]-3-phenyl-D-alanyl]amino]-N-[(2S,S,4S)-3-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexenyl]-imidazol-4-propionamide by 10-thin-layer foam chromatography, MS: 779 (M+H)+.

Example 24

A mixture of 100 mg (0.17 mmol) of 2-t-butyl-[(S)-u-[[(S)-1-[[(1S,2S,4S)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-15 carbamoyl]phenethyl]carbamate and 65 mg (0.25 mmol) of osmium tetroxide is stirred in 20 mL of pyridine overnight at room temperature. Then, 4 mL of a 33% sodium bisulfite solution is added, and the mixture is stirred for another hour. The chromography of the crude product obtained after usual treatment of the crude product on 20 g of silica gel with a 9:1:0.1 mixture of chloroform, methanol and ammonia gives 65 mg of (2RS,3S,5S,6S)-6-(Boc-Phe-His-NH)-3-isopropyl-8-methyl-1,2,5-nonanetriol, MS: 632 (M+H)+, 25 homogeneous to thin-layer chromatography.

Example 25

A mixture of 411 mg (0.82 mmol) of (RS)-α-[[(S)-β-(Boc-Pro-Nα)phenethyl]carbamoyl]imidazol-4-propionic acid, 200 mg (0.82 mmol) of (αS,βS)-β-amino-α-30[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol, 0.1 mL (0.82 mmol) of 4-ethylmorpholine, 222 mg (1.64 mmol) of HOBT, and 203 mg (0.99 mmol) of DCC is dissolved in 20 mL of dimethylformamide and stirred for 2 days at room temperature. The separated precipitate is filtered, and the solvent is evaporated under reduced pressure. After

THAT

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Usual treatment 011 yields 590 mg of a crude product which is chromatographed on 30 g of silica gel with a 20:1:0.1 mixture of methylene chloride, methanol, and ammonia. Recrystallization of the homogeneous product 5 by thin-layer chromatography in methylene chloride/ether/hexane gives 260 mg of t-butyl(S)-2-[[(S)-a-[(BS)-a-[[(1S,2S,4S)-1-(cyclohexylmethyl-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]imidazol-3-propionamido]phenethyl]carbamoyl]-1-pyrrolidinecarboxy-10 late in the form of crystals, melting point 112°.

The (ES)-o.-[[ (S)-a-(Boc-Pro-NH) phenethyl]-carbamoyl]imidazol-4-propionic acid used as a starting material was prepared as follows:

10.8 g (30 mmols) of Boc-Pro-Phe-OH and 3.0 g (30 mmols) of N-methylmorpholine were dissolved in 120 mL of tetrahydrofuran and cooled to -20°C. To this solution, 4.1 g of isobutyl ester of chloroformic acid in 15 mL of tetrahydrofuran was added dropwise, and after 5 minutes, a solution of 3.9 g (60 mmols) of sodium azide in 30 mL of water was added dropwise at -20°C. At the end of the addition, the mixture was stirred for 30 minutes at 0°C, diluted with ice-cold ethyl acetate to double the volume, and the reaction mixture was treated successively with cold saturated sodium bicarbonate solution and then with cold saturated sodium chloride solution. The organic phase is separated, dried over magnesium sulfate, and evaporated under reduced pressure at a temperature of 20–25°C. The remainder is dissolved under a nitrogen atmosphere in 300 mL of toluene and heated for 30 minutes at 80°C. Then, 4.9 g (45 mmol, 1.5 molar equivalents) of benzyl alcohol is added, and the reaction mixture is heated for 6 hours under reflux. After cooling and evaporation of the reaction solution, the remaining 35% is mixed with a 1:1 mixture of ether and he-

c

-74-

xane, and the separated precipitate is filtered. After drying the filtrate under reduced pressure, 11 g (79%) of [(R)-1-(Boc-Pro-NH)-1-(benzylamino)ethyl]benzene is obtained.

melting point 155-157°•

5.47 g (10 mmol) of the above compound are hydrogenated in 300 ml of tetrahydrofuran in the presence of 1.0 g of palladium on 10% activated carbon for 3 hours at 343.25 kPa. At the end of hydrogen absorption, the catalyst is filtered and the filtrate is evaporated under reduced pressure. The remainder is dissolved in 60 ml of dimethylformamide and 1.98 g (10 mmol) of monomethyl ester of (RS)-imid a zo lyl-4-methyl ) malonic acid is added.

The resulting solution is then cooled to 0°15 and 3.0 g (20 mmol) of HOBT and 2.2 g are added

(12 mmol) of EDO. The reaction mixture is then allowed to warm to room temperature and stirred for 48 hours at this temperature. After evaporation of the reaction mixture under high vacuum, the remaining 20% is dissolved in ethyl acetate, washed with water, and then extracted with 1N citric acid. The citric acid extract is neutralized with sodium bicarbonate and extracted with ethyl acetate. The organic extract is then dried over sodium sulfate and evaporated under reduced pressure.

Silica gel chromatography of the remainder and recrystallization of the crude product obtained in ethyl acetate/ether gives 2.5 g (49%) methyl ester of (RS)-a-[[(S)-u-(Boc-Pro-NH)-phenethyl]-30 carbamoyl]imidazol-4-propionic acid, melting point 142-143°.

M. Goodmann et al., Int. J. Pept. Prot. Res. 17, 72-88 (1981)

Of

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1.54 g (5 mmol) of the above ester are dissolved in 6 mL of ethanol, and 3.15 mL (3.15 mmol, 1.05 mol equivalent) of 1N sodium hydroxide are added. After 3 liters of stirring, 3.15 mL (3.15 mmol) of 1N sodium hydroxide is added under ice-cooled conditions, and the reaction mixture is evaporated under reduced pressure. The remainder is then stirred in 300 mL of ethanol, and the insoluble sodium sulfate is filtered out. The concentration of the ethanolic solution yields 1.47 g × 10⁻¹⁰ (98% concentration) of (RS)-α-[[(S)-α-(Boc-Pro-NH)phenethyl]-carbamoyl]imidazol-4-propionic acid, melting point 120°C (Dec.).

Example 26

A mixture of 1.05 g (2.06 mmol) of Boc-Phe-15 His(Bom)OH, 0.5 g (2.06 mmol) of (aS,pS)-p>-amino-a-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol, 0.26 mL (2.06 mmol) of 4-ethylmorpholine, 0.56 g (4.12 mmol) of HOBT, and 0.51 g (2.47 mmol) of DCC in 40 mL of dimethylformamide is stirred at room temperature overnight. The precipitated urea is then retained by filtration, and the solvent is evaporated under reduced pressure. The usual treatment of the remainder gives 1.69 g of t-butyl [(S)-1-benzyl-2-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-25 isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-amino]ethyl]carbamate, which is used directly in the next step.

1.69 g (approximately 2.06 mmol) of t-butyl[(S)-1-benzyl-2-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-30 2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]amino]ethyl]carbamate is hydrogenated overnight at room temperature in 50 ml of a 4:1 mixture of acetic acid and water and 0.8 g of N,N'-dimethylethylenediamine in the presence of 0.5 g of palladium on 35% activated carbon, at the end of the hydroq absorption

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The catalyst is filtered, and the filtrate is evaporated. Toluene is added twice successively, and then the mixture is evaporated again to dryness. Chromatography of the remainder on 180 g of silica gel with a 5 20:1:0.1 mixture of methylene chloride, methanol, and ammonia as the eluent gives 1.13 g of crude product, which is recrystallized in methylene chloride/ether/hexane, yielding 730 mg of t-butyl[(S)-1-benzyl-2-[[(S)-1-[[(1S,2S,4S)-1~(cyclohexylmethyl)-2-hydroxy-4-10 isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]amino]-ethyl]carbamate in crystal form, melting point 144°.

Iî y

The Boc-Phe-His(Bom)-OH used as a starting material was prepared as follows:

15 58 g of Boc-Phe-His(Bom)OMe which has been prepared

according to the usual processes by condensation of Boc-

Phe-OH with His(Bom)OMe, 30.35 g of Lawesson's reagent, and 700 mL of benzene were heated under reflux overnight. The reaction mixture was then cooled.

20 and it evaporates completely. The remainder is then filtered through

1.5 g of silica gel, first with methylene chloride and then with methylene chloride, gradually adding ethanol (up to 7%).

This yields 35.56 g [(S)-2-(Boc-NH)-3-phenylthio-

25 propanyl]His(Bom)OMe in the form of a foam, MS: 533

(M+H)+.

0.5 g of the above thio compound and 5 g of Raney nickel are stirred in 30 ml of ethanol for 2 hours under a hydrogen atmosphere. Then the catalyst is filtered and the filtrate is evaporated. Chromatography of the crude product on 30 g of silica gel with chloroform and a 95:5 mixture of chloroform and ethanol as the eluent gives 0.25 g of [(S)-2-(Boc-NH)-3-phenylpropyl]His(Bom)OMe as a foam, MS: 523 (M+H)+.

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170 mg of the above ester is stirred in 2 mL of 1N sodium hydroxide in a 1:1 mixture of methanol and water overnight at room temperature. Then 1 mL of concentrated acetic acid is added and the reaction mixture is evaporated under reduced pressure. The remainder is reheated three times in 150 mL of methanol and decanted. The methanolic phase is evaporated, and the remainder is chromatographed on 30 g of silica gel with a 4:1:1 mixture of butanol, acetic acid, and 10 mL of water, giving 100 mg of [(S)-2-(Boc-NH)-3-phenyl-propyl]-His'(Bom)OH as a foam, MS: 509 (M+H)+.

Example 27

0.17 g (0.56 mmol) of 2-(RS)-benzyl-15 4-(4-chlorophenyl)-4-oxo-butyric acid are dissolved in

10 ml of dimethylformamide is mixed with 0.20 g (0.51 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, 0.057 g (0.56 mmol) of triethylamine, and 0.21 g (0.56 mmol) of HBTU. The mixture is shaken overnight at room temperature and then evaporated under high vacuum. The remaining product is dissolved in ethyl acetate and subsequently washed with saturated sodium bicarbonate and sodium chloride solutions. After drying the organic phase over sodium sulfate, the solvent is evaporated under reduced pressure, and the remaining product is purified by silica gel chromatography using a [missing information]. mixture of methylene chloride and methanol which contains 0.5% ammonium hydroxide. From 30 in this way 270 mg (78%) of (S)-a-[(RS)-ct-(4-chlorophenylacetyl)hydrocinnamamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide is obtained in the form of a yellowish foam MS: 675 (M+).

35 2-(RS)-benzyl-4-(4-chlorophenyl)-4-oxo- acid

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butyric acid used as a starting material was prepared as follows:

To a suspension of 0.87 g of sodium hydride dispersion (55% in oil) in 20 mL of 5-dimethylformamide, 5.0 g (20 mmol) of benzylmalonic acid diethyl ester is added dropwise at room temperature. The reaction mixture is then stirred for 20 minutes at room temperature, and 4.67 g (20 mmol) of omega-10 bromo-4-chloroacetophenone in 25 mL of dimethylformamide is added dropwise. After stirring, the dark brown reaction mixture is stirred overnight at room temperature and then evaporated under high vacuum. The remaining residue is dissolved in methylene chloride, washed with 15% water, and dried over sodium sulfate. Next the solvent is evaporated under reduced pressure and the remainder is purified by silica gel chromatography with a 6:1 mixture of hexane and ether on silica gel, giving 5.17 g (64%) of 2-(RS)-"benzyl-2-carbethoxy-20 4-(4-chlorophenyl)-4-oxobutyric acid as a yellowish solid, MS: 351 (M-OC^)"1", 311 (M-benzyl) + .

To a solution of 1.64 g (4.1 mmol) of the ethyl ester mentioned above in 10 mL of ethanol and 9 mL of water, 6.15 mL (3 molar equivalents) of 2N sodium hydroxide are added, and the reaction mixture is stirred for 16 hours at room temperature. The pH is then adjusted to 3 with 1N hydrochloric acid, and the mixture is extracted with methylene chloride. The organic phase is dried over sodium sulfate and evaporated under reduced pressure. The crystalline residue is heated to 150–160°C until complete CO2 evolution. After cooling, the residue is dissolved in ether and crystallized by the addition of hexane. This yields 970 mg (79%) of 2-(RS)-benzyl-4-(4-chlorophenyl)-4-oxo-butyric acid as yellowish crystals, with a melting point of

-79-

137-138°.

Example 28

To a solution of 1.4-5 S (4.4-3 mmol) of 2-(ES)-(1-naphthylmethyl)-3-morpho1inocarbonyl-5-propionic acid in 60 mL of dimethylformamide, 1.57 S (4.03 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-μclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide, 0.45 g (4.43 mmol) of triethylamine, and 1.68 g (4.43 mmol) of HBTU are added. The resulting yellowish reaction solution is stirred for 15 hours at

The solution was heated to room temperature and then evaporated under high vacuum. The remainder was dissolved in 150 mL of methylene chloride. The organic phase was washed twice with 30 mL of saturated sodium bicarbonate solution and 30 mL of water, dried over sodium sulfate, and evaporated under reduced pressure. The remaining solution was chromatographed for purification and separation of the two epimers with a 95:5 mixture of methylene chloride and methanol, containing 0.5% ammonium hydroxide, on a 20% silica gel. This yielded 855 mg (30%) of an isomer with a B value of 0.45 and 726 mg (26%) of an isomer with a B value of 0.33. Both isomers exhibited the same mass spectrum, I_S: 700 (M+H)₊.

2-(ES)-(1-naphthylmethyl)-3-morpholino-25 carbonylpropionic acid was prepared as follows:

To a solution of 3.63 g (12.6 mmols) of ethoxycarbonyl-4-(1-naphthyl)butyric acid in 50 ml of dimethylformamide, 1.10 g (12.6 mmols) of morpholine, 2.42 g (12.6 mmols) of EDC, and 3.43 g (25.2 mmols) of HOBT are added. The reaction solution is stirred for 15 hours at room temperature and then evaporated under vacuum. The remainder is dissolved in ethyl acetate, and the organic phase is washed with water.

EPA 0 200 406 6 EPA 0 181 110

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Saturated sodium bicarbonate solution and saturated sodium chloride solution were dried over sodium sulfate and evaporated under reduced pressure. The remainder was chromatographed with a 98:2 mixture of methylene chloride and ethanol on silica gel, yielding 3.78 g (84%) of the ethyl ester of 2-(RS)-(1-naphthylmethyl)-3-morpholine carbonylpropionic acid as a yellowish oil, which began to crystallize after prolonged standing. MS: 355(M+). 1.7 g (4.78 mmols) of the aforementioned ethyl ester were dissolved in 6 mL of ethanol, and after the addition of 7.2 mL (7.2 mmols) of 1N sodium hydroxide, the mixture was stirred for 3 hours at 30°C. After cooling, 7.5 mL (7.5 mmol) of 1N hydrochloric acid is added to the dark yellow reaction solution and then evaporated under reduced pressure. The remainder is dissolved in 100 mL of methylene chloride, and the organic phase is washed twice with 20 mL of water, dried over sodium sulfate, and evaporated under reduced pressure, yielding 1.45 g (92%) of 2-(RS)-(1-naphthylmethyl)-3-morpholinocarbonyl-propionic acid as a yellow foam, which is used directly in the next step. MS: 327 (M+).

Example 29

109 mg (0.38 mmole) of ethyl ester of α-25 3-(RS)-(1-naphthylmethyl)succinic acid are dissolved in 6 ml of acetonitrile and 1 ml of dimethylformamide, and after the addition of 13.5 mg (0.35 mmole) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropopyl-5-hexenyl]imidazol-4-propionamide, 39 mg (0.38 30 mmole) of triethylamine and 144 mg (0.38 mmole) of HBTU, the mixture is shaken at room temperature for 6 hours. Next, the reaction solution is evaporated under high vacuum, and the remainder is dissolved in 50 mL of ethyl acetate and washed with three 10 mL portions of saturated sodium bicarbonate solution. The organic phase is i

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dried on sodium sulfate and evaporated under reduced pressure, and the remainder is chromatographed on silica gel with a 95% mixture of methylene chloride and methanol, which contains 0.1% ammonium hydroxide, giving the ethyl ester of the acid (45%)

(RS)-Ê [[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-1-naphthylbutyric acid as a yellowish foam, MS: 594 (M-ethanol-water)+. 10 The 1-ethyl ester of 3-(RS)-(1-

naphthylmethyl succinic acid was prepared as follows:

820 mg (3.44 mmol) of 2-(1-naphthylmethylene)succinic acid anhydride were heated in 5 mL of ethanol for 4 hours under reflux. The reaction mixture was then evaporated under reduced pressure, and the remainder was resuspended in 10 mL of saturated sodium bicarbonate solution and extracted with ether. The aqueous phase was acidified with 1N hydrochloric acid and extracted with ether. After drying the ether extracts over sodium sulfate and evaporating under reduced pressure, 10 mg (73%) of 3-(RS)-(1-naphthylmethylene)succinic acid 1-ethyl ester remained.

These 710 mg (2.5 mmol) of the ethyl ester are dissolved in 5 ml of ethanol and hydrogenated in the presence of 100 mg of palladium on activated carbon (10%) for 15 hours. The catalyst is then filtered, the filtrate is evaporated under reduced pressure, and the remainder is silica gel chromatographed with a 98:2 mixture of methylene chloride and methanol, yielding 30,190 mg (27%) of 1-ethyl ester of 3-(RS)-(1-naphthylmethyl1)succinic acid. MS: 286 (M)+.

6 EPA 0 181 110

(

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Example 50

Similar to what is described in Example 27, (aRS,S)-α-benzyl-1-t-butoxycarbonyl-f-oxo-2-pyrroli-5-dinebuturic acid is prepared by condensation with (S)*α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and then separation of the epimers by chromatography to produce t-butyl(S)-2-[[(S)-α-[[(S)-

1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydro xy-4-is o-

10 propyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrrolidinecarboxylate (least polar isomer) and t-butyl(S)-2-[[(R)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-15 phenethyl]acetyl]-1-pyrrolidinecarboxylate. MS: 734 (M+H)+.

The acid (u.RS,S)-a-benzyl-1-t-butoxycarbonyl-

-V-oxo-2-pyrrolidinebutyric acid used as a starting material was obtained in analogy with the preparation

20 of 2-(RS)-benzyl-4-(4-chlorophenyl)-4-oxobuty- acid

rique (see Example 27) from N-t-butoxycarbonyl-

7

prolyl-bromomethane and diethyl ester of benzylmalonic acid. MS: 362 (M+H)+.

Example 31

25. In a manner analogous to that described in Example 2 by catalytic hydrogenation of t-butyl-(S)-2-[ [ (S)-c<.-[ [ (S)-1-[ [ (1S ,2S, 4S)-1-( cycl ohexylmethyl thyl) -

2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrro1idinecarbo-

30 xylate and t-butyl (S)-2-[[(R)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]-acetyl]-1-pyrrolidinecarboxylate, t-butyl-

7 EPA 0 029 163

-85-

(S)-2-[[(S)-a-[[(S)-1 —[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrrolidinecarboxylate (least polar isomer) and t-butyl(S)-2-3 [[(E) -u-[[(S)-1-[[( '1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl] carbamoyl] -2-imidazolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrrolidinecarboxylate. MS: 736 (M+H)+.

Example 52

10 In a manner similar to that described in E

Example 1: 1-[1-[(ES)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyryl]-D-prolyl]-L-prolinebenzylester is prepared as a mixture of 15 2 epimers by condensation of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-imidazol-4—propionamide with (αRS,2R)-α-benzyl-2-[[(S)-2-[(benzyloxy)carbonyl]-1-pyrrolidinyl]carbonyl]-X -oxo-1-pyrrolidinebutyric acid, MS: 867 (M+H)+. 20 The acid used as a starting material was

prepared in the following manner:

Starting with 2-benzyl-3-methoxycarbo-

0

nylpropionic is prepared according to the process described by "Widmer in Synthesis 1983, page 135 the t-buty-25 lic ester of 2-benzyl-3-methoxycarbonylpropionic acid, MS: 222 (M-C/|Hg)+.

4-15 mg of the above t-butyl ester are dissolved in t-butyl alcohol and stirred in the presence of 1.5 mL of 1N sodium hydroxide for 3 hours at room temperature. Then the reaction solution is

evaporated under reduced pressure, and the remainder is dissolved in water and washed with ether. The aqueous phase is in-

8 L.D. Byers and R. Volfenden, J. Biol. Chem. 247, 606 (1972)

o

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The suite was neutralized with 1N hydrochloric acid and extracted with methylene chloride. The organic extracts were dried over sodium sulfate and evaporated under reduced pressure, giving the t-5 butyl ester of 2-benzyl-3-carboxypropionic acid as a colorless oil, MS: 208 (M-CZjHg)+.

370 mg (1.4 mmol) of the hemiester mentioned above, 409 mg (1.2 mmol) of H-D-Pro-Pro-OBz, and 270 mg/2 molar equivalents of triethylamine are dissolved in 8 mL of dimethylformamide. To this solution, 530 mg (1.4 mmol) of HBT11 is added, and the reaction mixture is stirred for 15 hours at room temperature and then evaporated to dryness under high vacuum. The remainder is dissolved in ethyl acetate and washed with saturated sodium bicarbonate solution. The organic phase is dried over sodium sulfate and evaporated under reduced pressure. Silica gel chromatography of the remainder with a 98:2 mixture of methylene chloride and methanol gives the t-butyl ester of 20(aRS,2R)-a-benzyl-2-[[(S)-2-[(benzyloxy)carbonyl]-1-pyrrolidinyl]carbonyl]-8-oxo-1-pyrrolidinebutyric acid as a colorless foam, MS: 549 (M+H)+.

505 mg (0.9 mmol) of the above t-butyl ester are stirred in 10 mL of hydrochloric acid/25 2N acetic acid for 3 hours at room temperature. The reaction mixture is then evaporated under reduced pressure, giving (aRS,2R)-u-benzyl-2-[[(S)-2-[(benzyloxy)carbonyl]-1-pyrrolidinyl]-carbonyl]-V-oxo-1-pyrrolidinebutyric acid as a colorless foam. MS: 493

The (S)-α-amino-β-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]imidazol-4-propionamide used as the starting material was prepared as follows:

35 50 mg of (S)-a-amino-N-[(1S,2S,4S)-1-(cyclo-

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α-amino-N-[(1-,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]imidazol-4-propionamide was dissolved in 3 mL of methanol and hydrogenated in the presence of 7 mg of palladium on activated carbon under normal pressure for 3 hours at room temperature. The catalyst was then filtered and washed with methanol. The alcoholic solution was then evaporated under reduced pressure, yielding α-amino-N-[(1-,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]imidazol-4-propionamide as a colorless foam. MS: 393 (M+H)+.

Example 33

In a manner similar to that described in Example 1, the following compounds are prepared:

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and (aRS,2R)-a-benzyl-2-[[(S)-2-[(benzyloxy)carbonyl]-1-pyrrolidinyl]-carbonyl]-y -oxo-1-pyrrolidine butyric acid and 1-[1-[(RS)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl] -carbamoyl] -2-imidazolyl-4-ethyl]carbamoyl]-4-phenyl-butyryl]-D-prolyl]-L-pro-linebenzylester as a 1:1 epimer mixture, MS: 865 (M+H)+;

- From (S)-u-amino-N-[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and N-(t-butoxycarbonyl)-N-[2-[(RS)-3-

c arb oxy-4-phenylbutyramido]é thyl]glyc ine-1-buty1ester the two epimeric compounds t-butoxycarbonyl [2-[(R and S)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carb amoy1]-4-phenylbutyramido]é thyl]glyc ine-1-butylester, MS : 837 (M+H)+;

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmida-zol-4-propionamide and (R)-α-[[[2-hydroxy-1-

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(hydroxymethyl)-1-methyl]carbamoyl]carbamoyl]carbamoyl]hydro-5 cinnamamido]imidazol-4-propionamide, MS: 668 (M+H)+;

- From (S)-a-amino-K-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and t-butyl ester of 5-[(RS)-3-carboxy-4-phenylbutyramido]valeric acid 10 t-butyl 5-[(RS)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl] -2-imidazolyl-4-ethyl]carbamoyl]-4-phenylbutyrami-do]valerate, MS: 736 (M+H)+;

- From (S)-α-amino-α-[(1S,2S,4S)-1-15 (cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

imidazol-4-propionamide and 3-carboxy-4-phenylbutyryl-L-proline-t-butylester 1-[(RS)-3-[[(S)-1-[[(1S,2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-nyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenyl-20 butyryl]-L-proline-t-butylester, MS: 734 (M+H)+;

- From (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and the monomethyl ester of (RS)-2-benzylmalonic acid^ the methyl (RS)-u-

25 [[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]hydrocinnamate as a 1:1 mixture of the two epimers, MS: 580 (M+);

- From (S)-u-amino-N-[(1S,2S,4S)-1-30 (cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

imidazol-4-propionamide and 3-(RS)-ethoxy-

carbonyl-4-(1-naphthyl)butyric^ ethyl (RS)-a--

F: Texier et al., Tetrahedron, 1974, 3185, 5 EPA 0.181.11Q

-87-

[[[ (S)-1-[[ (1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]methyl]-1-naphthalenepropionate as a 1:1 mixture of the two epimers, MS: 659 (M+H)+;

5 -From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and (ES)-[ (4-benzyl-4-piperidinyl)carbamoyl]imidazol-4-propionic acid (ES)-a-(1-benzyl-1-4-piperidinyl)-N-[ (S )-1-[ [ ( 1S, 2S, 4S )-1 -10 (cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

carbamoyl]-2-imidazolyl-4-ethyl]imidazol-4-propionamide as a 1:1 mixture of the two epimers, MS: 729 (M+H)+;

- From (S)-α-amino-N-[(1S,2S,4S)-1-15 (cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

imidazol-4-propionamide and 2-(RS)-benzyl-3-morpholinocarbonylpropionic acid and after chromatographic separation of the two epimers, the less polar (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-20-pyl-5-hexenyl]-α-[(S)-α-['morpholinocarbonyl)methyl]-hydrocinnamamido]imidazol-4-propionamide, MS: 650 (M+H)+, and the polar (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(E)-α-[(morpholinocarbonyl)methyl]-hydrocinnamamido imidazol-25-4-propionamide, MS: 650 (M+H)+.

The acids used as starting materials were prepared as follows:

M-(t-butoxycarbonyl)-N-[2-(ES)-5-carboxy-4-phenylbutyr-

amidoethyl]glycine-t-butylester

30 0.84 g (2.4 mmol) of N-(t-butoxycarbonyl)-N-

10

(2-benzyloxycarbonylaminoethyl)glycine is suspended in 5 ml of toluene and heated to 80°.

10 Patent ÏÏS 4 145 337

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To the clear solution obtained in this way, 1.96 g of N,N-dimethylformamide-di-t-butylacetal is added dropwise over a period of 20 minutes. After the addition, the reaction mixture is stirred for 30 minutes at 80°C, then cooled and washed successively with water, saturated sodium bicarbonate solution, and saturated sodium chloride solution. The organic phase is washed over sodium sulfate and evaporated under reduced pressure, yielding N-(t-10 butoxycarbonyl)-N-(2-benzyloxycarbonylaminoethyl)glycine-t-butyl ester as a colorless oil, MS: 4-09 (M+H)+.

0.78 g (1.9 mmol) of N-(t-butoxycarbonyl)-N-(2-benzyloxycarbonylaminoethyl)glycine-t-butyl ester is dissolved in 20 mL of methanol and hydrogenated in the presence of 0.1 g of palladium on activated carbon at pH 4.5. The pH is kept constant using a hydrochloric acid solution in methanol. At the end of hydrogen absorption, the catalyst is filtered, the filtrate is evaporated, and the remainder is dissolved in methylene chloride and washed with a saturated sodium bicarbonate solution. After drying the organic phase over sodium sulfate and evaporating under reduced pressure, N-(t-butoxycarbo-25 nyl)-2-(2-aminoethyl)glycine-t-butyl ester is obtained as an oil.

HMN (250 MHZ, TMS, CDCl^: ô 1.44-, 1.4-7 (2xs, 18H) 2.83 (s, 2H) 3.36 (S, 2H), 3.79, 3.87 (2xs, 2H).

The condensation of 3-ethoxycarbonyl-30 4-phenylbutyric acid with N-(t-butoxycarbonyl)-2-(2-aminoethyl)glycine-t-butyl ester gives N-(t-butoxy-carbonyl)-N-[2-[(ES)-3-(ethoxycarbonyl)-4—phenylbutyramido]ethyl]glycine-t-butyl ester (MS: 492 (M)+], which in turn is hydrolyzed with 1N sodium hydroxide in 35% ethanol at 50°C to give the desired acid.

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carbonyl)-N-[2-[(RS)-3-carboxy-4-phenylbutyramido]-ethyl]glycine-t-butylester, MS: 408 (M-C^Hg)+.

Acid (R)-a-[[[2-hydroxy-1-(hydroxymethyl)-1-methyl]-

carbamoyl

5

and 2-amino-2-methyl-1,3-propanediol are condensed in the presence of EDO and HOBT in dimethylformamide at room temperature, giving the ethyl ester of the acid (RS)-o.-[[[2-hydroxy-1-(hydroxymethyl)-1-

10 methyl] carbamoyl"1 -methyl] hydrocinnamic, MS: 324 (M+H)+.

ethyl ester of (ES)-α-[[[2-hydroxy-1-(hydroxymethyl)-1-methyl]c arb amoyl]methyl]hydro c innamic acid 15 in 13 ml of water, 50 mg of α-chymotrypsin is added,

The pH was maintained at 7.1 by the addition of 0.1 N sodium hydroxide (temperature 25°C). After 18 hours, 9 mL of 0.113" sodium hydroxide were added, and the reaction mixture was extracted with ether. The aqueous phase was placed at pH 4 with 20 1 N hydrochloric acid and then evaporated under reduced pressure. The remainder was digested with ethyl acetate. The organic phase was evaporated under reduced pressure, and the remainder was flash-chromatographed with a 9:1 mixture of methylene chloride and methanol on 25 silica gel, giving (R)-a-[[[2-hydroxy-1-(hydroxymethyl thy1)-1-methyl]carbamoyl]methyl]hydrocinna-mic acid as a colorless foam, MS: 264 (M-CE^OH)4". 5-[(RS)-3-carboxy-4-phenylbutyramido[butyl valerate] 3-Ethoxycarbonyl-4-phenylbutyric acid and 30 5-aminovalerate of t-butyl are condensed in the presence of EDC and HOBT in dimethylformamide to give 5-[(RS)-3-(ethoxycarbonyl)-4-phenylbutyramido[butyl]

A suspension of 0.54 g (1.67 mmol)

11

W.G. Kofron and L.G. Wideman, H. Org. Chem. 37, 555 (1972) —

-90-

t-butyl valerate, MS: 392 (M+H)+. Saponification of this ester with 1N sodium hydroxide in 50° ethanol gives 5-[(ES)-3-carboxy-4-phenylbutyramido]t-butyl valerate, MS: 307 (M-CZ{_Hg)+.

5 3-carboxy-4-phenylbutyryl-L-proline-t-butylester

The synthesis of 3-carboxy-4-phenylbutyryl-L-proline-t-butylester [MS: 361 (M)+] occurs by condensation of 3-ethoxycarbonyl-4-phenylbutyric acid and L-proline-t-butylester to give 3-ethoxycarbo-10-nyl-4-phenylbutyryl-L-proline-t-butylester [MS: 389 (M)+], the subsequent saponification of which takes place in a manner analogous to the description of the preparation of 2-(ES)-benzyl-3-morpholinocarbonylpropionic acid. (BS)-[(4-benzyl-4-piperidinyl)carbamoyl]imidazol-4—15-propionic acid

2 g (10 mmol) of monomethyl (ES)-(imidazolyl-4-methyl)-malonate (prepared according to the general process known from dimethyl malonate by alkylation with chloromethylimidazole and successive saponification) and 1.91 g (10 mmol) of 4-aminobenzyl-

Piperidine is stirred with 1.91 g (10 mmol) of EDC in 100 mL of dimethylformamide for 72 hours at room temperature. The reaction solution is then evaporated under reduced pressure, and the remainder is dissolved in 100 mL of methylene chloride and washed with 30 mL of water. The organic phase is then dried over sodium sulfate and evaporated to dryness. The remainder is digested with ether and then filtered, giving (ES)-[(4-benzyl-4-piperidinyl)carbamoyl]imidazol-4-30 methyl propionate as a colorless powder, MS: 370 (M)+.

371 mg (1 mmol) of the above methyl ester are dissolved in 10 mL of a 1:1 mixture of methanol and water and stirred with 1 mL of 1N potassium hydroxide in methanol for 24 hours at temperature

-91-

ambient temperature. The reaction solution is then evaporated under reduced pressure, the remainder is dissolved in water and washed with ether. The aqueous phase is then acidified with one molar equivalent of α-5 hydrochloric acid and evaporated under reduced pressure, yielding (RS)-[(4-benzyl-4-piperidinyl)-carbamoyl]imidazol-4-propionic acid as a colorless oil, MS: 312 (M-CO2)+-

2-(ES)-benzyl-3-morpholinocarbonylpropionic acid 10 1.0 g (4.2 mmols) of 3-ethoxycarbonyl-

4-Phenylbutyric acid is dissolved in 25 mL of dimethylformamide, and 0.37 g (4.2 mmol) of morpholine, 0.81 g (4.2 mmol) of EDC, and 1.3 g (8.4 mmol) of HOBT are added. The mixture is stirred for 48 hours at room temperature. The reaction solution is then evaporated under vacuum, and the remaining solution is dissolved in ethyl acetate. It is then washed with saturated sodium bicarbonate solution and saturated sodium chloride solution. Next the organic phase is dried over 2C sodium sulfate and evaporated under reduced pressure, and the remainder is chromatographed on silica gel using a mixture of methylene chloride and ethanol, giving ethyl 2-(ES)-1-benzyl-3-morpholino-carbonylpropionate as a colorless oil 25 MS: 305 (M)+.

0.42 g (1.4 mmol) of the above ester is dissolved in 2 mL of ethanol, and 2.1 mL (1.5 mol equivalents) of 1N sodium hydroxide are added. The reaction solution is then stirred for 4 hours at 50°C, and the remaining 30 mL is dissolved in water and washed with ether. The aqueous phase is acidified with 2.3 mL of 1N hydrochloric acid and extracted with methylene chloride. The organic extracts are dried over sodium sulfate and evaporated under reduced pressure, yielding 2-(ES)-benzyl-3-morpholinocarbonylpropionic acid.

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in the form of colorless oil, which is implemented directly in the next step, MS: 277 (M)+.

Example $4-

100 mg (0.13 mmol) of t-butoxycarbonyl[2-[ (BS)-3-[[ (S) —1 —[[ (1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4~isopropyl-5-kexenyl] carbamoyl] -2-imidazolyl-2-ethyl] carbamoyl] -4—phenylbutyr amido] ethyl] glycine-t-butyl ester are dissolved in 2 ml of acetic acid and stirred for 3 hours with 4 ml of hydrochloric acid/

1N acetic acid at room temperature. Then the reaction mixture is evaporated under reduced pressure, and the remainder is digested with ether and filtered, giving N-[2-[(RS)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4~isopropyl-5-hexenyl] carbamoyl]-2-imidazolyl-4~ethyl]carbamoyl]-4—phenyl-butyramido]ethyl]glycine-dihydrochloride in powder form, MS 681 (M+H)+.

Example 35

54- mg of t-butyl-5-[ (RS)-3-[ [ (S ) —1 —[ [ (1S ,2S, 4S )-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexe-nyl]carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]-4—phenyl-butyramidolvalerate are dissolved in 1 ml of acetic acid and 2 ml of hydrochloric acid/acetic acid (1.5 N) are added at room temperature and the mixture is stirred for 2.5 hours at this temperature. Next the reaction mixture is evaporated under reduced pressure and the remainder is digested in ether and filtered, giving the hydrochloride of the acid 5-[(RS)-3-[[(S)-1-[ [ ( 1S , 2S, 4-S ) -1 - ( cy c 1 ohe xylmethyl thyl ) -2-hyd r o xy -4— isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4—ethyllcarbamoyl]-4~ phenylbutyramido]valeric, MS : 680 (M+H)+.

Example 36

In a manner similar to that described in Example 27, the following compounds are prepared:

-93-

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4—propionamide and (RS)-2-benzyl-6,6-dimethyl-4~oxoheptanoic acid (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl thyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-[(RS)-a-(4,4—dimethyl-2-oxopentyl)hydrocinnamamido]-imidazol-4—propionamide, MS: 635 (M+H)+;

- Starting from (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and (RS)-α-benzyl-Y-oxo-cyclohexanebutyric acid and successive separation of the two epimers, the less polar (S)-α-[(S)-α-

[(cyclohexylcarbonyl)-methyl]hydrocinnamamido]-N-[(1S,2S, 4-S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl] imidazol-4-propionamide [MS: 64-7 (M+H)+]

and the polar, (S)-a-[ (R)-a-[ (cyclohexyl carbonyl )-methyl] hydroc innamamido] -N-[ (1S,2S ,4-S)-1 -(-cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, MS : 64-7 (M+H)+;

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4—propionamide and (RS)-α-benzyl-V-oxo-cyclopentanebutyric acid the (S)-N-[(1S,2S,4S)-1-

(cyc lohexylmethyl)-2-hyd ro xy-4—isopropylT-5-hexenyl] -a-[(RS-a-[(cyclopentylcarbonyl)methyl]hydrocinnamamido]-imidazol-4—propionamide, MS: 633 (M+H)+;

- From (S)-a-amino-N-(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazo-1-4-propionamide and (RS)-2-benzyl-5,5-dimethyl-4-oxohexanoic acid the (S)-a-[ (RS)-2-benzyl-5,5-dimethyl-4-oxohexananamido]-N-[ (15,2S ,4-S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazo-1-4-propionamide, MS : 621 (M+H)+;

- From (S)-a-amino-H-[ (1S ,2S,4-S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

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imidazol-4-propionamide and (R)-2-benzyl-5,5-dimethyl-4-oxohexanoic acid (S)-a-[(R)-2-benzyl-5,5-dimethyl-4-oxohexane-amido]-N[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl1imidazol-5 4-propionamide, MS: 621 (M+H)+;

- From (S)-a-amino-N-[(1S,2S,4S)-1-

(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and (aRS,S)-a-benzyl-l-(benzyloxycarbonyl)-Y -oxo-2-pyrrolidinebutyric acid 10 benzyl(S)-2-[[(RS)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]acetyl]-1-pyrrolidinecarboxylate, MS: 768 (M+H)+.

The acids used as starting material 15 were prepared by analogy with the preparation of 2-(RS)-benzyl-4-(4-chlorophenyl)-4-oxobutyric acid (see Example 27);

- From 1-chloro-4,4-dimethyl-2-pentanone, which itself is prepared from chlor-

20 rure of 3,3-dimethylbutyric acid by reaction with diazomethane and hydrochloric acid and of diethyl benzylmalonate (RS)-2-benzyl-6,6-dimethyl 4-oxoheptanoic acid;

13

- From bromacetylcyclohexane and diethyl 25-benzylmalonate, (RS)-α-benzyl-Y-

oxocyclohexanebutyric acid;

' 1-5

- From bromacetylcyclopentane and diethyl benzylmalonate, (RS)-α-benzyl-Y-

oxocyclopentanebutyric acid;

30 - From N-benzyloxycarbonyl-prolyl-

bromomethane which was prepared in analogy to N-t-

14

butoxycarbonyl-prolyl-bromomethane and benzylmalo-

13 M. Gaudry and A. Marquet, Tetrahedron, 1970, 5611

14 EPA 0.129.163

-95-

diethyl nate (aRS,S)-a-benzyl-1-(benzyl-oxycarbonyl)-1f-oxo-2-pyrrolidinebutyric acid.

The acids (RS)-2-benzyl-5,5-dimethyl-4-oxo-hexanoic and the acid enantiomers (R)-2-benzyl-5-5-dimethyl-4-oxo-hexanoic, also used as starting materials, are known from European Patent 0 184 550.

Example 57

In a manner analogous to what was described in Example 2, the following compounds were prepared by catalytic hydrogenation of the corresponding olefins.

- (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-4-ethyl-2-hydroxy-5-methylhexyl]-a-[(R)-a-[(morpholino-carb onyl)-methyl]hydroc innamamido]imida zol-4—prop ionamide, MS : 652 (M+ïï)+;

- (S)-a-[(R)-a-(3,3-dimethyl-2-oxobutyl)-hydroc innamamido] -N-[ (1S,2S, 4S)-1-(cyc lohexylmethyl) -2-hydroxy 4—isopropylhexyl]imidazol-4—propionamide, MS: 625 (M+H)+;

- (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropylhexyl]-a-[[[[(R)-a-2-hydroxy-1-(hydr oxym ethyl ).-1 -methyl ethyl] c arb amoyl] methyl] hydr o c innamamido] imidazol-4—propionamide, MS: 620 (M+H)+;

- N-[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl] -Y"-oxo-ot-(1-naphthylmethyl)-4-morpholinobutyramide, MS: 702 (M+H)+.

Example 38

To 90 mg (0.23 mmol) of (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-prop±onamide and 0.047 mL of Hunig's base in 2 mL of acetonitrile, a solution of 60.7 mg of (S)-ct-[(3,3-dimethylbutyryl)oxy]hydrocinnamic acid and 101 mg of BOP in 5 mL of acetonitrile is added dropwise at room temperature. The mixture is stirred for 4 hours at room temperature, then drained.

-96-

in a 2N sodium bicarbonate solution and extracted with methylene chloride. The organic phase is washed with ammonium chloride solution and dried over sodium sulfate. The solvent is removed under reduced pressure and the remainder is chromatographed on silica gel, and after elution, 50 mg (54%) of (S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-10 2-imidazolyl-4-ethyl]carbamoyl]phenethyl-3,3-dimethyl-butyrate is obtained as a yellow oil.

MS: 637 (M+H)+.

(S)-u-[(3,3-dimethylbutyryl)oxy]hydro- acid

Cinnamic acid was prepared as follows:

15 A 2.1 g (8.2 mmol) of (S)-2-hydroxy-3-

1 5

Benzyl phenylpropionate and 1.24 mL of triethylamine are mixed with 100 mL of methylene chloride. Over 2 hours, 1.26 mL of t-butylacetyl chloride is added dropwise at room temperature. The reaction mixture is then stirred for 6 hours at 40°C. It is then poured over water and extracted with methylene chloride. After drying over sodium sulfate, the solvent is removed under reduced pressure, and the crude product is purified by flash chromatography on silica gel with a 1:3 mixture of ether and petroleum ether, yielding 1.4 g (48%) of (S)-u-[(2,3-dimethylbutyryl)oxy]benzyl hydrocinnamate. PIS: 238 [M-(GE3)5-CCH2COOH]+.

The 1.4 g (3.95 mmol) of benzyl ester 30 are dissolved in 80 ml of ethanol and hydrogenated for 2 hours at room temperature in the presence of 0.4 g of palladium on activated carbon. After filtration of the catalyst-

15 Isv. Acad. Navk SSE, Ser. Khim. 1966(3), 5^9

-97-

lysing and solvent removal under reduced pressure yields 1 g (96%) of (S)-a-[(3,3(dimethylbutyl)-oxy]hydrocinnamic acid in the form of a colorless oil which is used directly in the next step.

5 Example 39

To 100 mg (0.394 mmol) of (aS, fcS)-p-amino-α-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol in a mixture of 3.5 mL of dimethylformamide and 7 mL of acetonitrile, 0.079 mL of Hünig base is added. Then, 174 mg of BOP and 150 mg (0.394 mmol) of (S)-α-(dibenzylacetoxy)imidazol-4-propionic acid are added, and the solution is stirred for 6 hours at room temperature. It is then diluted with 60 mL of ethyl acetate and washed with 1 N hydrochloric acid and sodium bicarbonate solution. The organic phase is then dried over sodium sulfate, and the solvent is removed under reduced pressure. The remainder is chromatography with a 10:1 mixture of methylene chloride and methanol on silica gel, yielding 179 mg of (S)-1-20 [[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-iso-

propyl-5-hexenyl]carbamoyl]-2-(imidazolyl-4)ethyldiben-zylacetate in amorphous powder form, MS: 596 (M-0H)+.

The (S)-u-(dibenzylacetoxy)imidazul-4-25 propionic acid used as a starting material was prepared as follows:

Dry hydrochloric gas is bubbled at 0°C for 3 minutes through a suspension of 2.5 g of α-hydroxyimidazolpropionic acid in 80 ml of 30 benzyl alcohol and the resulting solution is then left to stand for 12-20 hours at room temperature.

16 C.E. Baker and A. Merster, JACS 73, 1336

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Next, the excess benzyl alcohol is distilled under vacuum, and the remainder is dissolved in methylene chloride and washed with sodium bicarbonate solution. The organic phase is dried over sodium sulfate and evaporated under reduced pressure. The remainder is purified by flash chromatography with a 10:1 mixture of methylene chloride and methanol on silica gel, yielding 3.5 g (89%) of benzyl (S)-u-hydroxyimidazolpropionate as a yellow oil. MS: 246 (M)+.

To 10.53 g (43.8 mmol) of dibenzylacetic acid in 50 mL of pyridine, 6.19 mL (48.3 mmol) of benzene-tallow onyl chloride is added dropwise at -5°C, and the reaction solution is stirred for 30 minutes at room temperature. Next, a 6 g (24.4 mmol) solution of benzyl (S)-α-hydroxyimidazeppropionate is added dropwise at -5°C, and the solution is stirred first for 2 hours at 0°C and then for another 2 hours at room temperature. The reaction mixture is then poured onto ice and 3N hydrochloric acid and extracted with ethyl acetate. The organic extracts are then washed with 2N sodium bicarbonate over sodium sulfate and evaporated under reduced pressure. The remaining 25 is flash-chromatographed on silica gel using a 15:1 mixture of methylene chloride and ethyl acetate, giving 6.35 g (47%) of a 1:1 mixture of (S)-a-dibenzylacetoxy-N-(phenylsulfonyl)imidazol-4-propionate of benzyl and (S)-u,N-Bis(dibenzyl-30 acetoxy)imidazol-4-propionate of benzyl as a yellow oil, which can be used without further purification in the next step.

To 5.74 g (9.4 mmol) of the above mixture in 25 ml of methanol, add 95 ml of a 0.3 N hydrochloric acid solution in methanol and stir.

-99-

2 hours at room temperature. Then poured onto ice and sodium bicarbonate solution and extracted with ethyl acetate, dried over sodium sulfate, and solvent removed under reduced pressure. The remainder was purified by flash chromatography on silica gel, yielding 3.2 g (70%) of (S)-α-(dibenzylacetoxy)imidazol-4-benzyl propionate.

MS: 468 (M)+.

10 3.2 g (6.83 mmol) of (S)-a-(dibenzyl-

Benzyl acetoxyimidazol-4-propionate in 150 mL of methanol is hydrogenated for 2 hours at room temperature in the presence of 660 mg of palladium on activated carbon. The catalyst is then filtered, and the residue on the filter is washed several times with a 1:1 mixture of methylene chloride and methanol. The solution is then concentrated under reduced pressure until crystallization occurs. Recrystallization of the crystallized material in the methylene chloride/methanol mixture yields 2.5 g (96%).

of (S)-a-(dibenzylacetoxy)imidazol-4-propionic acid, melting point 210°.

Example 4-0 A 236.2 mg (1 mmol) of R-(+)-u-benzyl-25 monoethyl succinate and 390.6 mg (1 mmol) of

(S)-amino-N-[(1S,2S,4S)-1-(cyclohexylmethylethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide is added to acetonitrile. 4,42 mg (1 mmol) of BOP and 0.204 mL of Hünig's base are then added, and the resulting solution is stirred for 20 hours at room temperature. The reaction mixture is then poured onto ice/1N hydrochloric acid and extracted with acetate.

17 S.G. Cohen, A. Milovanovic, JACS 90, 3495 (1968)

-100-

of ethyl. The organic extracts are washed with sodium bicarbonate solution, dried over sodium sulfate and evaporated. The remainder is chromatographed with a 10:1 mixture of methylene chloride and 5-methanol as eluent on silica gel, giving 465 mg (76%) of ethyl (R)-3-[C(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl1)-2-hydroxy-4-isotropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-4-phenyl-butyrate as an amorphous powder.

10 MS : 609 (M+H)+.

Example 41

At 66.5 mg (0.404 mmol) of (S)-(+)-u- acid

- 18

methylhydrocinnamic acid and 157 mg (0.404 mmol) of (S)-

α-amino-N-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-15-4-isopropyl-5-hexenyl]imidazol]-4-propionamide is dissolved in 10 mL of acetonitrile. 181 mg (0.404 mmol) of BOP and 0.083 mL of Hiinig base are then added and the mixture is stirred for 16 hours at room temperature. The reaction solution is then poured over ice/1N hydrochloric acid and extracted with ethyl acetate. The organic extracts are washed with sodium bicarbonate solution and dried over sodium sulfate. Next the solvent is removed under reduced pressure, and the remainder is chromatographed on silica gel with 25 a 10:1 mixture of methylene chloride and methanol which gives 136 mg (63%) of N-[(1S,2S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-u-methylhydrocinnamamido]imidazol-4-propionamide as an amorphous solid.

30 MS : 537 (M+H)+.

Example 42

A 73 mg of u-methylcinnamic acid and 117 mg of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-

18 A.W. Schrecker, J. Org. Chem. 22, 33 (1957)-

-101-

In 5 mL of acetonitrile, 135 mg of BOP and 0.062 mL of H1N1 base are added to hydroxy-4-isopropyl-5-hexenyl imidazol-4-propionamide, and the mixture is stirred for 20 hours at room temperature. The reaction solution is then poured over ice and 3N hydrochloric acid and extracted with ethyl acetate. The organic phase is washed with 2T sodium carbonate solution and dried over sodium sulfate. Next the solvent is removed under reduced pressure and the remainder 10 is chromatographed with a 10:1 mixture of methylene chloride and methanol on silica gel, giving 100 mg of N-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-iliexenyl] -a-[a-methylcinnamoyl] -amino]-imidazol-4—propionamide as an amorphous solid 15.

MS: 535 (M+H)+.

Example 43

To 106 mg (0.404 mmol) of t-butoxycarbonyl-α,p>-dehydrophenylalanine and 157 mg of (S)-α-amino-N-20[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide in 10 mL of acetonitrile, 181 mg of BOP and 0.083 mL of Hünig's base are added, and the mixture is stirred for 16 hours at room temperature. The resulting solution is then poured onto ice and 1N hydrochloric acid, extracted with ethyl acetate, washed with 2U sodium carbonate solution, dried over sodium sulfate, and the solvent removed under reduced pressure. The remainder was chromatographed on silica gel with a 30 20:1 mixture of methylene chloride and methanol, yielding 144 mg (54%) of (S)-u-[u-(t-butoxycarbonylamino)-cinnamoyl]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-

19 H. Poisel, Chem. Ber. 110, 948 (1977)

-102-

4-isopropyl-5-bexenyl] imidazol-4-propionamide in amorphous solid form.

MS: 636 (M+H)+.

Example 44

5 To 100 mg (0.256 mmol) of (S)-a-amino-N-

[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopro-

pyl-5-hexenyl]imidazol-4-propionamide and 0.052 ml of

"Hiinig base in 2 ml of acetonitrile, 92 mg of (S)-α-(diphenylacetoxy)hydrocinnamic acid are added and

10,113 mg BOP in 2 ml of acetonitrile and shake

12 hours at room temperature. Then the reaction solution is poured onto ice and 1N hydrochloric acid, and extracted with ethyl acetate.

wash the organic phase with a carbo- solution

15% sodium nate, dried over sodium sulfate, and finally the solvent is removed under reduced pressure. The remainder is chromatographed on silica gel with a 10:1 mixture

of methylene chloride and methanol, which gives

130 mg of (S)-a-[[(S)-1-[[(1S,2S)-1-(cyclohexylmethyl)-

20 2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-

4-ethyl]carbamoyl]phenethyldibenzylacetate in the form of an amorphous solid.

MS: 733 (M+H)+.

(S)-u-(diphenylacetoxy)hydrocinnamic acid

25 was prepared as follows:

To 1.06 g (5 mmol) of diphenylacetic acid in 5 ml of pyridine, add, drop by drop, at -5°C

0.833 ml (6.5 mmol) of benzene-suifonyl chloride is stirred for 30 minutes at temperature

30 ambient temperature. Then we add 1.28 g (5 mmol) of (S)-2-

20

benzyl 3-hydroxyphenylpropionate in 5 ml of

20 Isv. Acad. Navk SSR, Ser. Khim. 1966(3), 519

-103-

Pyridine and O11 are stirred for 2 hours at 0-10°C and then for another 2 hours at room temperature. Water is added to the reaction solution and extracted with ethyl acetate. The organic phases are washed with 1N hydrochloric acid and sodium bicarbonate solution and dried over sodium sulfate. The solvent is evaporated under reduced pressure and the remainder is flash-chromatographed on silica gel with a 4:1 petroleum ether/ether mixture, yielding 1.8 g (80%) of benzyl(S)-α-(diphenylacetoxy)hydrocinnamate as a yellow oil. MS: 359 (M-Benzyl)+.

The 1.8 g of benzyl ester mentioned above are dissolved in 180 mL of ethyl acetate. 300 mg of palladium is added to activated carbon, and hydrogen is added until the theoretical amount of hydrogen at room temperature is absorbed (approximately 2 hours). The catalyst is then filtered, and the solvent is removed under reduced pressure, initiating the precipitation of the desired acid. Filtration of the precipitated crystals yields 1.3 g (90%) of (S)-α-(diphenylacetoxy)cinnamic acid.

MS: no molar peak.

212 (diphenylacetic acid)+

167 (diphenylmethyl)+

Example 45

To 100 mg of (aS,f>S)-j5-amino-Cfc-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol in 20 mL of acetonitrile, add 0.080 mL of Hunig's base, 174 mg of BOP, and 140 mg of (S)-3-carbamoyl-2-(dibenzylacetoxy)propionic acid and stir for 12 hours at room temperature. Then, pour the reaction solution onto ice and 1N hydrochloric acid, extract with ethyl acetate, wash the extracts with sodium carbonate solution, and finally dry them on

-104-

sodium sulfate. Then the solvent is removed under reduced pressure, and the remainder is chromatographed with a 20:1 mixture of methylene chloride and methanol on silica gel, giving 142 mg (60%) of (S)-2-carbamoyl-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]ethyl-dibenzylacetate in the form of a resin.

MS: 591 (M+H)+

(S)-3-carbamoyl-2-(dibenzylacetoxy)propionic acid was prepared as follows:

To 480 mg (2 mmol) of dibenzylacetic acid in 3 mL of pyridine, 0.34 mL of benzenesulfonyl chloride is added at -5°C, and the mixture is stirred for 30 minutes at room temperature. Next, 447 mg (2 mmol) of benzyl(S)-α-malamidate is added, and the reaction solution is stirred first for 2 hours at 0°C and then for 2 hours at room temperature. The reaction solution is then diluted with 100 mL of ethyl acetate, dilute hydrochloric acid is added, and the two resulting phases are separated. The organic phase is washed with sodium carbonate solution, dried over sodium sulfate, and evaporated under reduced pressure.

Silica gel chromatography of the remainder using a 4:1 mixture of methylene chloride and ethyl acetate as eluent gives 539 mg (59%) of benzyl-(S)-3-carbamoyl-2-(dibenzylacetoxy)propionate as a yellow oil.

MS: 354 (M-benzyl)+

520 mg of the benzyl ester described above in 5 ml of methanol are hydrogenated in the presence of 100 mg of palladium on activated carbon at room temperature for two hours. The catalyst is then filtered and the solvent removed under reduced pressure, yielding 370 mg (90%) of (S)-3-carbamoyl-2-

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(Dibenzylacetoxy)propionic acid as a white solid. MS: 338 (M-NH3)+

Example 46

100 mg (0.256 mmol) of (S)-tt-amino-N-5 [(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 0.052 mL of Hunig's base are mixed in 2 mL of acetonitrile at room temperature. 100 mg of N-(dibenzylcarbamoyl)-3-phenyl-L-alanine and 113 mg of BOP are added, and the mixture is stirred for 12 hours at room temperature. The reaction solution is then poured onto ice and 1N hydrochloric acid, extracted with ethyl acetate, dried over sodium sulfate, and the solvent removed under reduced pressure. Silica gel chromatography of remainder 15 using a 10:1 mixture of methylene chloride and methanol as eluent gives 130 mg of 1,1-dibenzyl-3-[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropoxy1-5-hexeny1]-carbamoyl]-2-imidazoyl-4-ethyl1]carbamoyl1]phenethyl1]-20 urea as an amorphous solid.

MS: 761 (M+H)+

The N-(dibenzylcarbamoyl)-3-phenyl-L-alanine used as a starting material was prepared as follows:

25 A 1 g (5.06 mmol) of dibenzylamine and

1.73 mL (10.1 mmol) of Hiinig base in 50 mL of methylene chloride was mixed dropwise with 2.6 mL of phosgene in toluene (20%, 5.06 mmol) under ice-cooled conditions. The resulting solution was stirred for 3 hours at 0°C. Next, 1.1 g of L-phenylalanine methyl ester was added, and the mixture was heated for 12 hours at 40°C. The reaction mixture was then poured onto water, extracted with methylene chloride, and dried over sodium sulfate. The solvent was then removed under reduced pressure, and the remaining

C

-106-

is chromatographed on silica gel with a 1:1 mixture of methylene chloride and ether as the eluent, giving 900 mg (45%) of N-(dibenzylcarbamoyl)-3-phenyl-L-alanine methyl ester as a white solid 5 MS:402 (M)+

To 900 mg (2.23 mmoles) of the above methyl ester in 20 ml of ethanol, add 9 ml of 0.5 N sodium hydroxide (4.46 mmoles) and heat for 1 hour at 40°.

Next, the reaction solution is acidified, and the product is extracted with methylene chloride. The organic phase is dried over sodium sulfate, and the solvent is removed under reduced pressure, yielding 100 mg (92%) of N-(dibenzylcarbamoyl)-3-phenyl-L-alanine as an amorphous solid.

15 MS: 388 (M)+

Example 47

To 100 mg (0.52 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 0.052 mL of 20 Hiinig base in 2 mL of acetonitrile, 99 mg of N-(morpholinocarbamoyl)-3-phenyl-L-alanine and 113 mg of BOP are added at room temperature to 2 mL of acetonitrile, and the mixture is stirred for 12 hours at room temperature. The reaction solution is then poured onto a mixture of ice-cold 25 and 1N hydrochloric acid, extracted with ethyl acetate, and the organic extracts dried over sodium sulfate. The solvent is then removed under reduced pressure. Next, the remainder is chromatographed with a 10:1 mixture of methylene chloride and methanol on silica gel 30, which gives 130 mg (67%) of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-nyl]-Cfc-2-[1T-(morpholinocarbamoyl)-3-phenyl-L-alanyl]-amino]imidazol-4-propionamide as an amorphous solid.

35 MS: 651 (M+H)+

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The N-(morpholinocarbamoyl)-3-phenyl-L-alanine used as a starting material was prepared as follows:

To 0.6 mL (6.95 mmol) of morpholine and 2.4 mL (13.9 mmol) of Hiinig base in 20 mL of methylene chloride, 3.6 mL of phosgene in toluene (20%, 6.95 mmol) is added dropwise while cooling in ice water, and the solution is stirred for 3 hours at 0°C. Then, 1.5 mL (6.95 mmol) of phenylalanine methyl ester hydrochloride is added, and the reaction solution is stirred for 12 hours at 40°C. The reaction solution is then poured onto ice and extracted with methylene chloride. The organic phase is dried over sodium sulfate and evaporated under reduced pressure. 15 Silica gel chromatography of the remainder using a 2:1 mixture of methylene chloride and ether as eluent gives 400 mg (20%) of N-(morpholinocarba-moyl)-3-phenyl-L-alaninemethyl ester as a resin.

20 MS: 292 (M)+.

To 400 mg (1.37 mmol) of the methyl ester mentioned above in 10 mL of ethanol, 5.5 mL (2.75 mmol) of 0.5 N sodium hydroxide solution is added. The reaction mixture is then stirred for 1 hour at 40°C, poured onto ice and 3 N hydrochloric acid, extracted with methylene chloride, dried over sodium sulfate, and finally removed from the solvent under reduced pressure. This yields 210 mg (55%) of N-(morpholinocarbamoyl)-3-phenyl-L-30 alanine as an amorphous solid.

MS: 278 (M)+

Example 48

A solution of 100 mg (0.256 mmol) of (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-35 hydroxy-4-isopropyl-5-hexenylmimidazol-4-propionamide is shaken,

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0.052 ml of Hiinig base, 113 mg of BOP and 81 mg of

21

N-(t-butoxycarbonyl)-p-pyrazolyl-1-L-alanine in acetonitrile, 12 hours at room temperature. Then the reaction mixture is poured onto ice and 1N hydrochloric acid, extracted with ethyl acetate, dried over sodium sulfate, and the solvent removed under reduced pressure. Silica gel chromatography of the remainder with a mixture of methylene chloride and methanol gives 10 80 mg (50%) of white t-butyl[(S)-1-

[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl-2-pyra z oly1-1-é thyl]c arbamate e.

MS: 628 (M+H)+

15 Example 49

To 100 mg (0.256 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4—propionamide, 0.052 ml of Hünig's base in 5 ml of acetonitrile, a solution of 103 mg of N-[[2-(4—biphenyloxy)ethyl]carbamoyl]-

3-Phenyl-L-alanine and 113 mg of BOP are mixed in acetonitrile and stirred for 12 hours at room temperature. The reaction mixture is then poured onto ice and 1N hydrochloric acid and extracted with acetate.

25 of ethyl, dry the extracts over sodium sulfate and evaporate under reduced pressure to dry. By silica gel chromatography with a 10:1 mixture of methylene chloride and methanol, the crude product is purified, yielding 90 mg (45%) of 1-[2-(4-biphenyloxy)ethyl]-30 3-[(S)-[-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-

4-ethyl]carbamoyl]phenethyl]urea in amorphous powder form. MS: 777 (M+H)+.

21 L.D. Arnold, T.H. Kalantar, J.C. Vederas, JACS 107, 7105 (1985)

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N-[[2-(4-biphenyloxyethyl]carbamoyl]-3-phenyl-L-alanirie was prepared as follows:

A 3 S (14.06 mmol) of 2-(4-biphenyloxy)-22

ethylamine in 100 ml of toluene is added at 0°C, along with 4.8 ml of Hining base and 7 ml of a 20% phosgene solution in toluene, and the reaction mixture is then stirred for one hour at 0°C. Next, 6 g of L-phenylalaninebenzyl ester is added, and the solution is stirred for 12 hours at 90°C. The reaction mixture is then processed in the usual manner, and the crude product is purified by silica gel chromatography with a 10:1 mixture of methylene chloride and methanol on silica gel, giving 4.5 g (67%) of N-[[2-(4-biphenyloxy)ethyl]carbamoyl]-3-phenyl-L-alaninebenzyl-15 ester.

4.5 g (9.4 mmol) of the benzyl ester mentioned above is hydrogenated in the presence of 0.5 g of palladium on activated carbon until the theoretical amount of hydrogen is absorbed at room temperature. 20 Then the catalyst is filtered, the solvent is removed under reduced pressure and the remainder is recrystallized in methylene ether/chloride/methanol, giving 2.1 g (55%) of N-[[2-(4-biphenyloxy)ethylcarbamoyl]]-3-phenyl-L-alanine, melting point 175° 25 C^Q9 = +8.4° (c = 1%, methanol).

Example 50

To 100 mg (0.256 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 0.052 ml of Hiinig base 30 in 5 ml of acetonitrile, a solution of 100 mg of (S)-ct-[(dibenzylcarbamoyl)oxy]-hydrocinnamic acid and 113 mg of BOP in 2 ml of aceto-

22 DOS 2.500.692

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nitrile and then shake for 12 hours at room temperature. The treatment takes place in the usual way. This yields 150 mg (77%) of (S)-a-[[(S)-1-[[(1S,2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-5 hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyldibenzylcarbamate.

MS: 762 (M+H)+

(S)-α-(dibenzylcarbamoyloxy)hydrocinnamic acid was prepared as follows: 10 A 1.0 g (3.9 mmol) of (S)-2-hydroxy-

3-Phenylpropionic acid and 1.68 mL (3.9 mmol) of Hinig base are added to 20 mL of tetrahydrofuran. 2.23 mL (4.3 mmol) of a 20% phosgene in toluene solution is then added dropwise while cooling on ice, and the mixture is stirred for 1.3 hours at 0-10°C. Next, 0.824 mL (4.3 mmol) of dibenzylamine is added dropwise to 20 mL of tetrahydrofuran, and the reaction solution is stirred for another 12 hours at room temperature. Finally, it is poured onto ice, neutralized with potassium bicarbonate, and extracted with ethyl acetate. The crude product obtained is flash chromatographed for purification on silica gel using a 3:1 mixture of petroleum ether and ether, giving 825 mg (42%) of (S)-a-25 [(benzyloxy)carbonyl]phenethyldibenzylcarbamate as resin.

MS: 388 (M-benzyl)+

0.8 g of the benzyl ester described above is hydrogenated in 30 mL of a 4:1 mixture of methanol 30 and ethyl acetate in the presence of 80 mg of palladium on activated carbon for 2.5 hours at room temperature. The catalyst is then filtered and the solvent evaporated under reduced pressure. This yields 461 mg (73%) of (S)-u-[(dibenzylcarbamoyl)oxy]-35 hydrocinnamic acid in the form of a resin.

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MS: 389 (M)+.

Example 5!

A solution of 100 mg of ethyl (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-5 4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]-4-phenylbutyrate is saturated in 30 ml of methanol for 1-2 minutes with ammonia and then shaken for 12 hours at room temperature. The solvent is then removed under reduced pressure, and the remainder is chromatographed with 10% methanol in methylene chloride as the eluent on silica gel, giving 63 mg (66%) of (S)-u-[(R)-a-(carbamoylmethyl)-hydrocinnamamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide as a resin. MS: 580 (M+H)+.

Example 52

A solution of 84.62 mg (0.3 mmol) of t-butoxycarbonyl-α-methyl-D,L-phenylalanine, 135 mg of BOP, 118 mg of (S)-cu-amino-N-[(1S,2S,4S)-1-(cyclohexyl-20 methyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-

Propionamide and 0.062 mL of Hiinig base are stirred for 12 hours at room temperature. The reaction mixture is then processed as described in Example 48, yielding 125 mg (64%) of β-butyl[(R or S)-α-25[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-carbamoyl]-α-methylphenethyl]carbamate as an epimeric mixture. By silica gel chromatography with 10% methanol in methyl-30-lene chloride, the two epimers are separated. The polar epimer has an R_p value of 0.38, while the less polar one has an R_p value of 0.46. MS: 652(M+H)_p.

The t-butoxycarbonyl-α-methyl-D-L-phenylalanine 35 used as a starting material was prepared as

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follows:

To 2.0 g (11.16 mmol) of u-methyl-D-phenylalanine in 11.16 mL of 1N sodium hydroxide and 10 mL of β-butyl alcohol, 2.44 g (11.16 mmol) of di-β-butyl dicarbonate in 5 mL of β-butanol is added dropwise at room temperature, and the reaction mixture is then stirred overnight at room temperature. Water is then added, the mixture is washed with pentane, the aqueous phase is adjusted to pH 2 by adding potassium bisulfite solution, and it is extracted with ethyl acetate, which precipitates 1.56 g (50%) of β-butoxycarbonyl-β-methyl-D,L-phenylalanine as a white powder, which is directly used in the next step.

15 Example 55

A mixture of 100 mg (0.45 mmol) of N-benzyl-3-carbamoyl-D-alanine, 187 mg (0.45 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, 20 0.093 mL of Hunig's base, and 202 mg of BOP is stirred in 10 mL of a 2:1 mixture of acetonitrile and dimethylformamide for 18 hours at room temperature. The reaction mixture is then poured into aqueous sodium chloride solution, extracted with α-25-ethyl acetate, and washed with sodium carbonate solution. It is dried over sodium sulfate and evaporated to dryness under reduced pressure. Silica gel chromatography of the remainder with 10% methanol in methylene chloride as the eluent gives 87 mg (32%) of (S)-α-[(R)-2-(benzylamino)-3-carbamoylpropionamido]-

N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmimidazol-4-propionamide as a resin. MS: 595 (M+H)+.

The N-benzyl-3-carbamoyl-D-alanine used as a starting material was prepared as follows:

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To 1.5 g (10 mmols) of D-asparagine-hydrate in 5 ml of sodium hydroxide 21T, 1.01 ml (10 mmols) of benzaldehyde was added, and the reaction mixture was stirred homogeneously for 20 minutes at room temperature.

5. Next, 114 mg (3 mmol) of sodium borohydride is added per portion, the mixture is stirred for 30 minutes at room temperature, another 114 mg of sodium borohydride is added, and the mixture is stirred for a further 30 minutes at room temperature. The aqueous phase is then washed with methylene chloride, and the pH is adjusted to 6-7 with 1N hydrochloric acid, which initiates crystallization. The precipitated crystals are filtered, washed with water and ether, and finally dried under high vacuum, yielding 0.6 g (27%) of N-13-benzyl-3-carbamoyl-D-alanine as a white powder. MS: 223 (M+H)+

Example 34-

80 mg (0.13 mmole) of ethyl (R)-3-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4~ isopropyl-20 5-hexenyl]carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]-4-phenylbuty.rate are taken up in 2 ml of a 5.6 molar alcoholic solution of dimethylamine and heated for 3 hours under reflux. The solution is then evaporated, and the remainder is chromatographed with a 20:1 mixture of methylene chloride and methanol on silica gel, yielding 33 mg (20%) of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-α-[(R)-α-[(dimethylcarbamoyl)methyl]hydrocinnamamido]-imidaazol-4—propionamide as an amorphous powder. 30 MS: 608 (M+H)+

Example 55 In a manner analogous to that described in Example 40, the following compounds are prepared:

- From 117 mg (0.3 mmol) of (S)-a-35 amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-

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isopropyl-5-hexenyl] imidazol-4-propionamide and 75 mg (0.3 mmol) of N-benzyl-3-ethoxycarbonyl-D-alanine, 85 mg (45%) of ethyl(R)-3-(benzylamino)-3-[[(S=-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-5 isopropyl-5-hexyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]propionate as an amorphous powder. MS: 624 (M+H)+;

- From 117 mg (0.3 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-

10 isopropyl-5-hexenyl]-imidazol-4-propionamide and

88 mg (0.3 mmol) N-benzyloxycarbonyl-L-asparagine, 136 mg (71%) benzyl [(S)-1-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-2-carbamoyl-15 ethyl]carbamate in amorphous form, MS: 639 (M+H)+.

- From 157 mg (0.4 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide and 136 mg (0.4 mmol) of N-benzoyl-Y"-benzyl-D-glutamate

20 115 mg (40%) benziyl (S)-4-benzamido-4-[[ (3)-1_

[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-butyrate, MS: 714- (M+H)+;

- From 100 mg of (S)-α-amino-N-[(1S,2S,25,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-

nyl]imidazol-4-propionamide and 64 mg of 1 (RS)-a-[[(2-hydroxyethyl)carbamoyl]methyl]hydrocinnamic acid, 50 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydro-xy-4-isopropyl-5-hexenyl]-ct-[(RS)-a-[[(2-hydroxyethyl)-30 carbamoyl]methyl]hydrocinnamido]imidazol-4-propionamide, as a 1:1 mixture of epimers, MS: 624 (M+H)+;

- From 100 mg of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide and 80 mg of <*- acid

35 [(phenethylcarbamoyl)-methyl]cinnamic, 65 mg (37%)

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of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-[a-[(phenethylcarbamoyl)methyl]-c innamamido] imidazol-4—propionamide , MS : 682 (I+H)+;

- From 214 mg (0.55 mmol) of (S)-u-5 amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-

isopropyl-5-hexenyl]imidazol-4—propionamide and 160 mg (0.55 mmole) of (RS)-ct-[2-(morpholinocarbonyl)ethyl]hydrocinnamic acid, 210 mg (60%) of (S)-N-[ (1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-10 5-hexenyl]-a-[(ES)-a-[2-(morpholinocarbonyl)ethyl]-hydrocinnamyl]imidazol-4—propionamide as a 1:1 mixture of epimers, MS: 664- (M+H)+;

- From 100 mg (0.25 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—

15 isopropyl-5-hexenyl]imidazol-4-propionamide and

100 mg (0.28 mmole) of N-[(4—biphenylmethyl)carbamoyl]-3-phenyl-L-alanine, 150 mg (77%) of 1-(4-biphenyl-methyl)-3-[(S)-1-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl]-2-hydroxy-4~isopropyl-5-hexenyl]carbamoyl]-2-20 imidazolyl-4—ethyl]carbamoyl]phenethyl]urea, MS: 748 (M+H)+.

The acids used as a starting material are prepared as follows: N-benzyl-3-ethoxycarbonyl-D-alanine 25,574 mg (2.9 mmol) of ethyl-D-aspar-

The tic, which was prepared according to the Pivitti process described in Gazetta, 18, 4-80 (1888) for the preparation of racemic compounds, was dissolved in 2.9 ml of sodium hydroxide and 0.3 ml of benzaldehyde was added. The mixture was then vigorously stirred for one hour at room temperature. Next, 33 mg of sodium borohydride was stirred, and the mixture was stirred for another hour at room temperature. Then another 33 mg of sodium borohydride was added, and the mixture was stirred for another hour at room temperature. Finally, the mixture was extracted with ether and the phase adjusted.

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aqueous solution at pH 6-7, which induces crystallization. The precipitated crystals are filtered and dried under high vacuum, yielding 81 mg (10%) of N-benzyl-3-ethoxycarbonyl-D-alanine, which is used directly in the next step. MS 206 (M-COOH) +

N-benzoyl-f-benzyl-D-glutamate

Y-Benzyl-D-glutamate is benzoylated according to the process described in Arch. Biochem. Biophys. 1952, 35, 176 by Miller and Waelsch for the benzoylation of V-ethyl-L-glutamate, which gives the desired product with a yield of 75%, MS: 341 (M)+.

(RS)-α-[[(2-hydroxy7/-ethyl)carbamoyl]methyl]hydrocinnamic acid

To 150 mg (0.8 mmol) of benzalsuccinic acid anhydride [S.G. Cohn et al., JACS, 90, 3495 (1968)] in 10 ml of methylene chloride, 0.048 ml of ethanolamine is added and the mixture is stirred for 2 hours at room temperature. The precipitated crystals are filtered and dried, yielding 155 mg (78%) of α-[[(2-(hydroxyethyl)-carbamoyl]methyl]cinnamic acid in the form of colorless crystals. MS: 249 (M)+.

180 mg of the acid mentioned above are hydrogenated in 5 mL of ethanol in the presence of 20 mL of palladium on activated carbon for 2 hours at room temperature. The catalyst is then filtered and the solvent is removed under reduced pressure. The remainder is dissolved in ethyl acetate and washed with 0.1% hydrochloric acid. The organic phase is dried over sodium sulfate and the solvent is evaporated under reduced pressure, yielding 125 mg (70%) of (SS)-u-[[(2-hydroxyethyl)carbamoyl]methyl]hydrocinnamic acid, which is used directly in the next step. tt-[(phenethylcarbamoyl)methyl]cinnamic acid

To 226 mg (1.2 mmol) of benzalsuccinic anhydride in 20 ml of methylene chloride is added

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0.15 ml of phenethylamine is added and then the mixture is stirred for 2 hours at room temperature. The solvent is removed under reduced pressure and the remainder is chromatographed on silica gel, giving 260 mg (70%) of tt-5 [(phenethylcarbamoyl)methyl]cinnamic acid.

MS: 309 (M)+.

(RS)-α[2-(morpholinocarbonyl)ethyl]hydrocinnamic acid

To 2.5 g (10 mmol) of di-10 ethyl benzylmalonate and 1.45 mL (10 mmol) of t-butyl acrylate in 5 mL of acetonitrile, 1.49 mL of DBU is added and the mixture is stirred for 6 hours at room temperature. It is then poured onto dilute hydrochloric acid solution (pH 2), extracted with ether, the organic phase is washed with a little water, dried over sodium sulfate, and the solvent is removed under reduced pressure. This yields 3.7 g (97%) of 4-t-butyl-2,2-diethyl-1-phenyl-2,2,4-butanetricarboxylate as a light yellow oil, which is used directly in the following step 20.

To 1.9 g (5.4 mmol) of 4-t-butyl-2,2-diethyl-1-phenyl-2,2,4-butanetricarboxylate in 20 mL of ethanol, 27 mL of 2N sodium hydroxide (54 mmol) is added, and the mixture is stirred for 6 hours at 50°C. The reaction mixture is then extracted with ether, dried over sodium sulfate, and the solvent evaporated under reduced pressure. The remainder is chromatographed using a 10:1 mixture of methylene chloride and methanol as the eluent, yielding 0.8 g (50%) of 3-ethyl-2,4-dihydrogen-1-phenyl-2,2,4-butanetricarboxylate as an oil, MS: 250 (MC^)*.

800 mg of 3-ethyl-2,4-dihydrogen-1-phenyl-2,2,4-butanetricarboxylate are heated for 2 hours at 170°C under high vacuum and then chromatographed on 35 silica gel with a 10:1 mixture of chloride of

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methylene and methanol which gives 600 mg (88%k of (RS)-4-(ethoxycarbonyl)-5-phenylvaleric acid in oil form, which is directly used in the next step, MS: 250 (M+).

To 600 mg (2.4 mmols) of (RS)-4-ethoxy-carbonyl-5-phenylvaleric acid in 20 ml of acetonitrile, 1.06 g (2.4 mmols) of BOP, 310 mg of Hunig's base, and 0.21 ml of morpholine are added, and the mixture is stirred first for 1 hour at room temperature and then for 2 hours at 50°C. After usual processing and silica gel chromatography using a 10:1 mixture of methanol and chloroform as the eluent, 400 mg (52%) of ethyl (HS)-α-[2-(morpholinocarbonyl)ethyl]hydrocinnamate is obtained, which is used directly in the next step, MS: 319 (M+).

To 400 mg (1.25 mmol) of the ether mentioned above in 10 ml of ethanol, 5 ml of 0.5 N sodium hydroxide are added and then the mixture is stirred for 2 hours at 50°C. After usual processing and silica gel chromatography using a 10:1 mixture of methylene chloride and methanol, 170 mg (47%) of (RS)-α-[2-(morpholinocarbonyl)ethyl]hydrocinnamic acid is obtained in the form of an oil which is used directly in the next step, MS: 291 (M+).

N-[(biphenylmethyl)carbamoyl-3-phenyl-L-alanine]

To 3.9 g (21.3 mmol) of biphenylmethylamine and 7.2 mL of Hunig's base in 200 mL of toluene, 10.5 mL of phosgene in toluene (20%) is added while cooling in an ice bath, and the solution is stirred for 1 hour at 0°C. Then, 9 g of L-phenylalanine-benzyl ester-4-toluene sulfonate is added, and the solution is stirred for 12 hours at 80°C. Finally, the chloride is distributed

23 A. Zamil et al., Czech. 163, 819 (1976)

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of methylene and water, dries the organic phase on sodium sulfate and removes the solvent under reduced pressure and isolates the product by silica gel chromatography using a 10:1 mixture of 5 methylene chloride and methanol, giving 7.3 g (74%) of N-[(4-biphenylmethyl)carbamoyl]-3-pheny 1-L-benzyl ester.

3.2 g (6.7 mmol) of the benzyl ester mentioned above in 100 ml of ethanol are hydrogenated in the presence of 320 mg of palladium on activated carbon at

room temperature until the theoretical amount of hydrogen is absorbed. Then the catalyst is filtered, the filtrate is evaporated and the remainder is crystallized in the ethanol/methylene chloride/15 ether mixture, giving 2.1 g (65%) of N-[(4-biphenylmethyl]carbamoyl]-3-L-alanine in the form of white crystals, melting point 169-170°•

Example 56

57 mg of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-20 2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)-imidazol-4-propionamide, 21 mg of t-butoxycarbonyl-L-azetidine-2-carboxylic acid which was prepared by t-butoxycarbonylation from azetidine carboxylic acid according to the process described by O. Keller et al. 25 in Org. Synth. 63, 160, 1985, 4-7 mg of BOP and 0.021 ml of Hiinig base in 5 ml of acetonitrile were shaken for 8 hours at room temperature and then processed in the usual way. The crude product is purified by silica gel chromatography with a 10:1 mixture of methanol and chloroform, yielding 69 mg t-

butyl [(S)-2-[(S)-a-[(S)-1 —[[(1S,2S,4S)-1-(cyclohexyl-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imida z olyl-4-ethyl]carbamoyl]phen ethyl]carbamoyl]-1-azetidinecarboxylate in the form of white crystals. 35 MS : 721 (M+H)+ .

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-Example 57

56 mg of t-butyl [(S)-2-[(S)-a-[(S)-1-[[(1S, 2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropy1-5-hexenyl]carbamoyl]-2-imidazolyl—4-ethyl]carbamoyl]-5 phenethyl]carbamoyl]-1-azetidinecarboxylate in 5 ml of ethanol are hydrogenated in the presence of 10 mg of palladium on activated carbon for 2 hours at room temperature. Next the catalyst is filtered and the solvent removed under reduced pressure, giving 47 mg 10 of t-butyl (S)-2-[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclo-hexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imid a z o ly 1-4-ethy1]carbamoyl1]phenethyl]carbamoyl1]-1-azetidinecarboxylate in the form of white crystals. MS: 723 (M+H)+, similarly, by hydrogenation of 90 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-[(RS)-α-[(cyclopentylcarbonyl)methyl]hydrocinnamamido]imida-zol-4-propionamide, 78 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]-α-[(RS)-20 u-[(cyclopentylcarbonyl)methyl]hydrocinnamamido]imida-zol-4-propionamide in the form of an amorphous solid as a 1:1 mixture of epimers. , MS: 635 (M+H)+.

Example 58 In a manner analogous to that described in Example 56, the following compounds are prepared:

- From 75 mg of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 30 mg of (3R,8aR)-hexahydro-8α-methyl-5-oxo-5H-thiazolo-30[3,2-α]pyridine-3-carboxylic acid2Zl", 64 mg (64%) of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(RS)-α-[(3R,8aR)-hexahydro-8α-methyl-

24 J.E. Baldwin et al., JACS 100, 4597

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5-oxo-5H-thiazolo-[3,2-a]pyridine-3-carboxamido]-hydrocinnamamido]-imidazol-4-propionamide in the form of a white solid, MS: 735 (M+H)+:

- From 75 mg of N-[(1S,2S,4S)-1-(cyclo-5-hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-(3-

phenyl-L-alanyl)imidazol-4-propionamide and 30 mg of (3R,7aR)-tetrahydro-7a-methylpyrrolo[2,1-b]-thiazol-3-carboxylic acid^Zl" 86 mg (3R,7aR)-N-[(S)-a-[[(S)-

1-[ [(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-iso-10 propyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-

carbamoyl]phenethyl]hexahydro-5-oxo-pyrrolo[2,1-b]-thiazol-3-carboxamide as an amorphous solid, MS: 721 (M+H)+;

- From 75 mg of N-[(1S,2S,4S)-1-(cyclo-15 hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-(3-

phenyl-L-alanyl)imidazol-4-propionamide and 34 mg of N-t-butoxycarbonyl-L-methionine, 90 mg of t-butyl [(S)-1-[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-

2-hydroxy-4-isopropyl-5-hexethyl]carbamoyl]-2-imidazolyl-20 4-ethyl]carbamoyl]phenethyl]carbamoyl]-2-(methylthio)-

ethyl]carbamate in the form of an amorphous solid, MS: 769 (M+H)+;

- From 75 mg of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-*x-(3-

25 phenyl-L-alanyl)imidazol-4-propionamide and, 32 mg of N-isovaleryl-L-methionine, which was prepared by reaction of L-methionine methyl ester with isovaleric acid chloride and then saponification of the methyl ester, 61 mg of (S)-N-[(1S,2S,4S)-30 1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

a-[(S)-a-[(S)-4-(methylthio)-2-isoval ramidobutyramido]-hydrocinnamamido]-imidazol-4-propionamide, MS: 753 (M+H)+;

24 J.E. Baldwin et al., JACS 100, 4597

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Example 59

A mixture of 95 mg of (RS)-a-[(2-morpholinoethyl)carbamoylmethyl]-1-naphthalenepropionic acid, 100 mg of (S)-cu-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl-5-ethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 97 mg of HBTU in acetonitrile is stirred for 3.5 hours at room temperature and then processed in the usual way. The remainder is chromatographed on silica gel with a 4:1 mixture of 10 methylene chloride and methanol as the eluent, giving 60 mg of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[3-[(2-morpholinoethyl)carbamoyl]-2-(1-naphthylmethyl)propionamido]imidazol-4-propionamide in foam form,

15 MS : 7^3 (M+H)+.

The (RS)-α-[(2-morpholinoethyl)carbamoyl-methyl]-1-naphthalenepropionic acid used as a starting material was prepared as follows:

we add to 1.0 g (3.5 mmol) of acid (ES)-

25

20 3-ethoxycarbonyl)-4-(1-naphthyl)butyric acid is dissolved in 50 mL of methylene chloride. 0.72 g of DOC is then stirred for 2 hours at room temperature. Next, 0.46 mL (3.5 mmol) of 4-(2-aminoethyl)morpholine is added, and the mixture is stirred for another 18 hours at room temperature. After standard processing, the remainder is flash-chromatographed on silica gel with a 20:1 mixture of methylene chloride and methanol as the eluent, yielding 0.5 g of (RS)-α-[(2-morpholinoethyl)carbamoylmethyl]-1-naphthalene ethyl propionate as a light yellow resin.

MS: 398 (M)+

To 0.5 g (1.25 mmol) of the ethyl ester mentioned above in ethanol, 2.5 ml of 1N sodium hydroxide is added.

25 EPA 0.181.110

c

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The mixture is stirred for 6 hours at 80°C. The reaction mixture is then adjusted to pH 6 and the solvent is removed under reduced pressure. The remaining material is mixed with a 1:1 solution of methylene chloride and methanol, the insoluble salts are filtered out, the filtrate is dried over sodium sulfate, and the solvent is evaporated under reduced pressure. This yields 500 mg of ethyl (RS)-α-[(2-morpholinoethyl)carbamoylmethyl]-1-naphthalenepropionate as a light yellow resin.

MS: 398 (M)+

To 0.5 g (1.25 mmol) of the above ethyl ester in ethanol, 2.5 mL of 1N sodium hydroxide solution is added and the mixture is stirred for 6 hours at 80°C. The reaction mixture is then adjusted to pH 6 and the solvent is removed under reduced pressure. The remainder is mashed in a 1:1 mixture of methylene chloride and methanol, the insoluble salts are filtered out, the filtrate is dried over sodium sulfate, and the solvent is evaporated under reduced pressure. The following is obtained in this manner:

500 mg of (ES)-<£-[ (2-morpholinoethyl)carbamoyl-methyl]-1-naphthalenepropionic acid in the form of a yellow foam, which is used directly in the next step.

Example 60

25 A mixture of 153 mg (0.4 mmol) of (S)-a-

amino-N-[(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, 98 mg of (E)-α-(pivaloylmethyl)hydrocinnamic acid, 175 mg of BOP, and 0.14 mL of Hunig's base in 10 mL of acetonitrile was stirred for 30 minutes at room temperature and then processed in the usual manner. Silica gel chromatography of the remainder using a 10:1 mixture of methylene chloride and methanol as the eluent gave 120 mg of N-[(1S,2S,4S)-1-benzyl-2-hydroxy-4-35-isopropyl-5-hexenyl]-α-(α-pivaloylhydrocinnamamido)-

C

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4-propionamide in the form of white crystals, MS: 615 (M+H)+.

The (S)-u-amino-N-[(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexenylmimidazol-4-propionamide 5 used as a starting material was prepared as follows:

Similar to the preparation described in Example 1 of (aS,f?>S)-£ -t-butoxycarbonylamino-u-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol from 10 of 56.6 g (320 mmols) of (RS)-2-isopropyl-3-butenyl-

bromide, 7.7 g of magnesium and 24.2 g of N-t-butoxy-

✓ 26 ✓ ✓

15.6 g (49%) of carbonyl-L-phenylalanal was prepared

t-butyl [(1S,2S,4ES)-1-benzyl-2-hydroxy-4-isopropyl-

5-hexenylcarbamate as a mixture of epimers. 15 The desired (1S,2S,4S) epimer was isolated by flash-

silica gel chromatography using a mixture of methylene chloride and ether as the eluent as the least polar isomer, MS: 256 (M-ben-zyl)+.

20 2.2 g of t-butyl [(1S,2S,4S)-1-benzyl-2-hydro-

[xy-4-isopropyl-5-hexenyl]carbamate was stirred in 70 mL of 1.8 N hydrochloric acid in dioxane for 2 hours at room temperature. The reaction mixture was then poured into a sodium bicarbonate solution and extracted at pH 7 with ether. Drying and evaporation of the ether extracts and chromatography of the remainder on silica gel with a 5:1 mixture of methylene chloride and methanol gave 900 mg (60%) of (2S,3S,5S)-2-amino-5-isopropyl-1-phen-30-nyl-6-heptenol-3 as an oil, MS: 248 (M)+.

A mixture of 1.22 g (2.83 mmoles) of N-a-N-im-Di-t-butoxycarbonyl-L-histidine, 700 mg (2.83 mmoles) of (2S,3S,5S)-2-amino-5-isopropyl-1-phenyl-6-heptenol-3.

26 Evans et al., J. Org. Chem., 50, 4615 (1985)

c

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1.25 g of BOP and 0.58 ml of Hunig's base in 20 ml of acetonitrile were stirred for 12 hours at room temperature and then processed in the usual manner. Silica gel chromatography of the remainder using a 2:1 mixture of methylene chloride and ether gave 1.0 g (10%) of t-butyl 4-[(S)-2-[[(1S,2S,4S)-

1-benzyl-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-(l-t-butoxyformamido)ethyl]imidazol-1-carboxylate in foam form, MS: 585 (M+H)+

10 1g (1.7 mmol) of t-butyl 4-[(S)-2-[[(1S,2S,

4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-

2-(1-t-butoxyformamido)ethyl]imidazol-1-carboxylate was dissolved in 7 ml of 5N hydrochloric acid in dioxane and then stirred for 2 hours at temperature

15 ambient. Then the reaction mixture is poured into sodium carbonate solution and extracted with ethyl acetate. The organic extract is dried and evaporated and the remainder is chromatographed on silica gel, yielding 340 mg (52%) of (S)-a-amino-N-20 [(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexenyl]-imidazol-4-propionamide as a foam, MS: 385 (M+H)+.

Example 61

120 mg of N-[(1S,2S,4S)-1-benzyl-2-hydroxy-4-25 i s op ropyl-5-hexenyl] -o.- (a -p ival oy lhydr 0 c innamamido) -

Imidazol-4-propionamide in 10 ml of ethanol is hydrogenated in the presence of 20 mg of palladium on activated carbon for 2 hours at room temperature. Then the catalyst is filtered and the solvent is evaporated under 30 reduced pressure, giving (S)-N-[(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropylhexyl]-a-[(R)_a_(3 y 3-dimethyl-2-oxobutyl)hydrocinnamamido]imidazol-4-propionamide as a foam, MS: 617 (M+H)+.

Example 62

35 A mixture of 256 mg (1.03 mmol) of (2S,3S,

°t,

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5S)-2-amino-5-isopropyl-1-phenyl-6-heptenol-3, 414 mg of Boc-Phe-His-OH, 0.354 ml of Hunig's base and 455 mg of BOP in 20 ml of acetonitrile is stirred for 12 hours at room temperature and then processed in the usual way. Flash chromatography of the remainder on silica gel using a 7d.e mixture of methylene chloride and methanol yields 385 mg (61%) of t-butyl [(S)-u-[[(RS)-1-[[(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexenyl]-carbamoyl]-2-imida zolyl-4-ethyl]carbamoyl]phenethyl]-carbamate as a 4:1 epimer mixture. MS: 632 (M+H)+.

Example 63

334 mg (0.53 mmol) of t-butyl[(S)-α-[[(RS)-1-[[(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexe-nyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamate are dissolved in 3 mL of 5.2N hydrochloric acid in dioxane and stirred for 2 hours at room temperature. After the usual preparation, 100 mg of the crude product obtained is dissolved in 10 mL of acetonitrile, 40.4 mg (0.188 mmol) of t-butoxycarbonyl-D-proline, 0.064 mL of Hiinig base, and 83 mg of BOP are added, and the reaction mixture is stirred for 12 hours at room temperature. After usual processing, the remainder was chromatographed on silica gel using a 10:1 mixture of methylene chloride and methanol, yielding 78 mg (56%) of t-butyl (R)-2-[[(S)-u-[(RS)-1-[[[(1S,2S,4S)-1-benzyl-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyl]carbamoyl]-1-pyrrolidinecarboxylate as white crystals as a 4:1 epimer mixture. MS: 729 (M+H)+.

Example 64

0.65 g of (S)-a-amino-N-[(1S,2S,4RS)-2-hydroxy-1-i s obuty1-4-is opropy1-5-h exeny1]imid a zo1-4-propionamide

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In 20 ml of acetonitrile, 0.36 ml of ethyldiisopropylamine is added to a solution of 0.4-3 g of 2-benzyl-3-phenylpropionic acid (prepared according to the method described in J. Am. Chem. Soc. 71, 5 1863 (194-9)) and 0.8 g of BOP in 30 ml of acetonitrile and stirred for 20 hours at room temperature. The reaction mixture is then evaporated under reduced pressure, the remainder is dissolved in 50 ml of ethyl acetate, and the organic solution is washed twice, each time with 100 ml of water, dried over calcium sulfate, and evaporated. The remainder is separated by flash chromatography on 100 g of silica gel with a 250:15 mixture of methylene chloride and methanol as the eluent, giving 0.17 g of (S)-a-15 (2,2-diεβαβαμεναλαμον)-N-[(1S,2S,4-S)-2-hydroxy-1-

isobutyl-4—isopropyl-5-hexenyljimidazol-4-propionamide as a white powder, melting point 120°. This compound corresponds to the least polar isomer and has a value of 0.14-. 20 The (S)-α-amino-N-[(1S,2S,4RS)-2-hydroxy-1-

isobutyl-4-isopropyl-5-hexenyl-imidazol-4-propionamide used as a starting material was prepared as follows:

0.72 g of (4S)-amino-(5S)-hydroxy-7-isopropyl-25 2-methyl-8-nonene in 20 mL of dimethylformamide is added with 0.7 mL of ethyldiisopropylamine to a solution of 2 g of α-α-N-μ-Bis-Emo-L-histidine and 1.4–7 g of BOP in 20 mL of dimethylformamide and stirred for 20 hours at room temperature. The reaction mixture is then evaporated under reduced pressure, the remainder is dissolved in 60 mL of ethyl acetate, and the organic phase is washed twice with 200 mL of water each time, dried over sodium sulfate, and evaporated. The crude product is purified by flash chromatography on 100 g of silica gel with a 250:20 mixture of chlor-

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methylene and methanol mixture as eluent giving 1.25 g of fluorenyl-9-methyl [(S)-1-[[(1S,2S,4RS)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexed nyl] carbamoyl]-2-imidazolyl-4-ethyl]carbamate as a white powder, MS: 573 (M+H)+.

1.25 g of fluorenyl-9-methyl [(S)-1-[[(1S,2S,4RS)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamate are dissolved in 20 mL of piperidine and stirred for 3 hours at room temperature. The reaction mixture is then poured onto 200 mL of ice and water and filtered. Evaporation of the filtrate yields 0.65 g of (S)-a-amino-N-[(1S,2S,4RS)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]imidazol-4-propionamide as a white powder, MS: 269

15 (m-C4H5N2)+.

Example 65

0.3 g of (S)-2-[[N-(t-butoxycarbonyl)-3-phenyl-L-alanyl]oxy]succinamidine acid is dissolved with 0.35 g of BOP in 20 mL of acetonitrile. To this, 2 mL of dimethylformamide is added, followed by a solution of 0.17 g of (4S)-amino-(5S)-hydroxy-7-isopropyl-2-methyl-8-nonene and 0.15 mL of ethyldiisopropylamine in 20 mL of acetonitrile. The mixture is stirred for 20 hours at room temperature. The reaction mixture is then evaporated for 25 minutes under reduced pressure, and the remainder is dissolved in

50 mL of methylene chloride. The organic solution is washed successively with 50 mL of 3N hydrochloric acid, 50 mL of sodium bicarbonate solution, and 50 mL of saturated sodium chloride solution, dried over sodium sulfate, and evaporated. The remainder is purified by flash chromatography on 50 g of silica gel using a 10:1 mixture of methylene chloride and methanol as the eluent, and then crystallized in the ether/methylene chloride mixture. This yields 0.3 g of t-butyl[(S)-a-

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[[(S)-2-carbamoyl-1-[[(1S ,2S,4RS)-2-hydroxy-1-isobutyl-4-isopropyl-5-hexenyl]carbamoyl]ethoxy]carbonyl]phenethyl]carbamate in the form of a white powder, melting point 75°.

5 The acid (S)-2-[[N-(t-butoxycarbonyl)-3-phenyl-

L-Alanyl oxy succinamide, used as the starting material, was prepared as follows:

0.8 g of t-butoxycarbonyl-L-phenylalanine were dissolved in 5 ml of pyridine, 10 0.3 ml of oxalyl chloride were added at -5° and the mixture was stirred for 10 minutes at 0-5°. After adding 0.7 g of benzyl-(S)-3-carbamoyl-2-hydroxypropionic acid (prepared according to the instructions in Agr. Biol. Chem. 40, 1651 (1976) and Int. J. Peptide Protein Res. 20, 35 (1982)) to 15 ml of pyridine, the mixture was stirred for 2 hours at 0-5°C and then for 1 hour at room temperature. The reaction mixture was poured onto ice and extracted twice with 50 ml of ethyl acetate each time. The combined organic extracts were washed with 20-50 ml of 3N hydrochloric acid and 50 ml of sodium bicarbonate solution, dried over sodium sulfate, and evaporated. The remainder was purified by flash chromatography on 50 g of silica gel using a 10:1 mixture of methylene chloride. and methanol 25 as the eluent, yielding 0.9 g of benzyl (S)-2-[,[N-(t-butoxy-carbonyl)-3-phenyl-L-alanyl]oxy]succinamidate as a yellow powder, MS: 397 (M-

0.9 g of the above benzyl ester were dissolved in 40 ml of ethanol and hydrogenated in the presence of 0.1 g of palladium on 10% activated carbon under normal pressure. At the end of hydrogen absorption, the catalyst was filtered and the filtrate was evaporated. Crystallization of the remainder in methylene chloride gave 0.52 g of (S)-2-[[N-(t-butoxycarbonyl)-3-phenyl-L-35 alanyl]oxy"]succinamide as a white powder.

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MS: 381 (M+H)+.

Example 66

A 200 mg solution of (2S,3S,5S)-2-[Boc-His-(3-Bom)-Pro-His-NH]-1-cyclohexyl-5-isopropyl-6-heptenol-5,3 is thoroughly hydrogenated in a 4:1 mixture of glacial acetic acid and water in the presence of 20 g of palladium on 10% activated carbon. After the hydrogen absorption period (4 hours), the catalyst is filtered, and the filtrate is evaporated to dryness. The crystals obtained after purification on silica gel are recrystallized in ethyl acetate-hexane, yielding (2S,3S,5S)-2-(Boc-His-Pro-His-NH)-1-cyclohexyl-5-isopropyl-3-heptanol in 69% yield, with a melting point of 135–136°C.

15 The preparation of (2S,3S,5S)-2-[Boc-His(3-Bom)-

Pro-His-NH]-1-cyclohexyl-5-isopropyl-6-heptenol-3 used as a starting material is described in Example 68.

Example 67

To a solution of 100 mg of (S)-α-[[N-(t-20 butoxycarbonyl)-P-phenyl-L-alanyl]oxy]imidazol-4-propionic acid, 78 mg of (ctS,^S)-p»-amino-α-[(S)-2-isopropyl-3-butenyl]cyclohexanepropanol, and 55 mg of N-methylmorpholine in 3 mL of dimethylformamide, 102 mg of HBTU is added under stirring and argon bubbling. After resting for 25 minutes, the reaction solution is poured into 5 mL of concentrated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic extracts are dried and evaporated under reduced pressure and the crude product obtained is purified by silica gel chromatography with a 95:5 mixture of methylene chloride and methanol as eluent, giving t-butyl[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethoxy]carbonyl]phenethyl]carbamate with a yield of 22%, melting point 72-75° (Dec.).

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The (S)-a-[[N-(t-butoxycarbonyl)-3-phenyl-L-alanyl]oxy]imidazol-4-propionic acid used as a starting material was prepared as follows:

1.99 g of N-t-butoxycarbonyl-L-phenylalanine and 1.58 g of benzyl(S)-u-hydroxyimidazolpropionate were suspended in 25 mL of methylene chloride and stirred. After 15 minutes of stirring at room temperature, 305 mg of 4-dimethylaminopyridine was added and the reaction mixture was cooled to -15°C. After the dropwise addition of 2 mL of N-methylmorpholine and a solution of 6.6 g of propane-phosphoric anhydride in 5 mL of methylene chloride, the reaction mixture was stirred for another 2 hours at -15°C and then left to stand for 6 days at -18°C. After adding a few drops of water, the reaction mixture is evaporated under reduced pressure, and the remaining oil is stirred for 2 hours with 100 ml of ethyl acetate and 25 ml of water. The organic phase is washed successively with aqueous sodium bicarbonate solution (5%) and 1.5 M citric acid. The colorless oil obtained after

evaporation of the solvent under reduced pressure is chromatographed on silica gel with a 95:5 mixture of methylene chloride and methanol as eluent, giving (S)-a-[[N-(t-butoxycarbonyl)-3-phenyl-L-25 alanyl]oxy]imidazol-4—benzyl propionate as a yellow oil with a yield of 13%, MS: 4-94- (M+H)+.

A solution of 960 mg of the benzyl ester described above in 20 mL of absolute ethanol is hydrogenated in the presence of 500 mg of palladium on 10% activated carbon until the theoretical amount of hydrogen is absorbed. The catalyst is then filtered and the filtrate is evaporated under reduced pressure, giving an oily residue, which crystallizes after the addition of methanol/ethyl acetate. In this way, the acid (S)-a-[[N-(t-butoxycarbonyl)-3-phenyl-L-

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alanyl]oxy]imidazol-4-propionic, with a yield of 18%, melting point 130-131°»

Example 68

In a manner analogous to that described in example 67, the following compounds are prepared:

- From (S)-tt-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide and N-(t-butoxycarbonyl)-5-(1-naph-tyl)-L-alanine in a 50% yield, t-butyl[(S)-1-

10 [[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl] carbamoyl] -2-imidazolyl-4—ethyl] -carbamoyl]-2-(1-naphthyl)ethyl]carbamate, melting point 14-1-14-3° (from ethyl/hexane acetate);

- From (S)-a-amino-N-[(1S,2S,4S)-1-(cy-15 clohexylmethyl)-2-hydr0xy-4—isopropyl-5-hexenyl]imidazol-

4-propionamide and N-(t-butoxycarbonyl)-4-chlorophenylalanine in 41% yield t-butyl [(S)-1 — [[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]-20 carbamoyl]-2-(4-chlorophenyl)ethyl]carbamate, melting point 220-221° (from ethanol/ethyl acetate);

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide and N-t-butoxycarbonyl-L-phenylglycine

25 in a yield of 39% the t-butyl[(S)-a-[[(S)-1-[[(1S,2S,

4S)-1-(cyclohexylmethyl)-2-hydroxy-4-is-opropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]benzyl]carbamate, melting point 125-126° (from ethyl acetate/hexane);

30 - From (S)-cc-amino-N-[(1S,2S,4S)-1-(cy-

clohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and N-t-butoxycarbonyl-L-cyclohexyl-glycine in a yield of 54% t-butyl[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-35 5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-

<

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cyclohexylmethylcarbamate, melting point 139-140° (from ethyl acetate/hexane).

- From (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmida-

5 zol-4-propionamide of t-butoxycarbonyl-D-phenylalanine in 48% yield t-butyl[(H)-u-[[(S)-1-[ [ ( 1S, 2S, 4S ) -1 - (cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyl]carbamate, melting point 195° (from 10 ethyl acetate);

- Starting from (S)-u-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenylmidazol-4-propionamide and t-butoxycarbonyl-L-cyclohexylalanine in a 22% yield, t-butyl[(S)-2-cyclohexyl-

15 1-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-(2-imidazolyl-4-ethyl]-carbamoyl]ethyl]carbamate, melting point 116° (from ethyl acetate/diisopropyl ether).

- From N-[(1S,2S,4S)-1-(cyclohexylmethyl)-20 2-hydroxy-4-isopropyl-5-hexenyl]-a-(3-phenyl-L-alanyl)-

imidazol-4-propionamide and N-[5-(1-t-butoxyformami-do)valeryl]-^-alanine in 40% yield t-butyl[4-[[—2—[[(S)-a-[[(S)-1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]-2-imidazolyl-4-25 ethyl]carbamoyl]phenethyl]carbamoyl]ethyl ]-carbamoyl]butyl]-carbamate, melting point 114-115° (from ethyl acetate);

- From (S)-a-amino-IT-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-

30 propionamide and Boc-His(3-Bom)-Pro-OH in 51% yield of (2S,3S,5S)-2-[Boc-His(3-Bom)-Pro-His-NH]-1-cyclo-hexyl-5-isopropyl-6-heptenol-3, melting point 121-122° (from ethyl acetate/hexane).

The acids used as part-products are either known and commercially available or have been pre-existing.

-1 Separated as follows:

N-[ 5-(1-t-butoxyformanido)valeryl"l-fr-alanine

To a suspension of 7.68 g of α-alanine ethyl ester hydrochloride in 50 mL of 5-methylene chloride, 5.56 g of triethylamine, 10.86 g of N-t-butoxycarbonyl-5-aminovaleric acid dissolved in 50 mL of methylene chloride, and 11.35 g of DCC are added in each portion, with stirring. After the addition, the reaction mixture is stirred at room temperature overnight, and the precipitated urea is then retained by filtration. The filtrate is washed successively with 250 mL of 5% aqueous sodium bicarbonate solution, 250 mL of 0.1 M citric acid, and finally with saturated sodium chloride solution. The solvent is then evaporated under reduced pressure, and the remaining oil is purified by silica gel column chromatography, yielding... N-[5-(1-t-butoxyformamido)valeryl]--alanine-ethyl ester with a yield of 59%, melting point 28° (from hexane).

3.94 g of the ester mentioned above is dissolved in 20 mL of methanol. Under stirring at 0°C, 1 mol equivalent of sodium hydroxide is added first, followed after 30 minutes by another 0.2 mol equivalent of 1 N sodium hydroxide. The mixture is then stirred for one hour at room temperature. 1.2 mol equivalents of 1L hydrochloric acid (pH 4) are added dropwise, the solvent is evaporated under reduced pressure, and the aqueous phase is extracted several times with methylene chloride. Drying and evaporation of the extracts yields an oily residue that crystallizes upon the addition of hexane. Recrystallization in hexane gives α-[5-(1-t-butoxyformamido)valeryl]-alanine in 87% yield, with a melting point of 70°C.

Example 69

35 A 1.06 g (3 mmol) of t-butyl [(1S,2S,4S)-4-

(

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(2-furyl)-2-hydroxy-1-isobutyl-5-methylhexyl]carbamate is dissolved in 8 mL of ethyl acetate. 5 mL of a 4-mol solution of hydrochloric acid in dioxane is then added. After 30 minutes, the solvent is evaporated under reduced pressure. The remaining solvent is digested with toluene and evaporated again. This residue is dissolved in 10 mL of absolute dimethylformamide. In this solution, 1.8 g (3 mmol) of N-u-N-im-Bis-Fmoc-L-histidine is dissolved in 10 mL of tetrahydrofuran, along with 0.35 mL of 10 N-methylmorpholine and 0.8 g (5.6 mmol) of N-hydroben-zotriazol. This solution is cooled to -10°C and 5 mL of a 1 M solution of DCC in tetrahydrofuran is added dropwise. The reaction mixture is stirred overnight and then filtered. To the solution, 5 mL of a 50% piperidine in dimethylformamide solution is added and the mixture is evaporated under high vacuum after 30 minutes. The remaining solution is dissolved in ether, and the organic phase is washed successively with water, a sodium carbonate solution, and then with water. The organic phase is then dried over sodium sulfate and evaporated under reduced pressure. Next, the remainder is dissolved in 10 mL of dimethylformamide, and 0.8 g (3 mmol) of N-t-butoxycarbonylphenylalanine is added to 10 mL of tetrahydrofuran, along with 0.35 mL of N-25-methylmorpholine, 0.8 g (5.6 mmol) of 11-hydroxybenzotriazole, and 5 mL of a 1 M solution of DCC in tetrahydrofuran. The reaction mixture is stirred overnight and then filtered. The filtrate is evaporated under high vacuum, and the remainder is dissolved in ether. The organic solution is washed with water, a sodium carbonate solution, and again with water.

dried over sodium sulfate and evaporated under reduced pressure. The remainder is chromatographed for purification with a 19:3 mixture of methylene chloride and ethyl acetate as the eluent, yielding 0.65 g of

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(S,S,S)-1-(Boc-Phe-His-NH)-methylbutyl- y" -isopropyl-2-furanepropanol as a viscous oil, MS: 638 (M+H)+.

The t-butyl-[(1S,2S,4S)-4-(2-furyl)-2-hydroxy-1-5-isobutyl-5-methylhexyl]carbamate used as a starting material was prepared as follows:

To a solution of 217 g (1.67 moles) of 2,5-dimethoxy-2,5-dihydrofuran, 1500 ml (10 moles) of diethyl malonate, 167 ml of water and 330 ml of glacial acetic acid, 340 g (2.5 moles) of zinc chloride are added at once, raising the temperature to 35°• The dark solution is stirred overnight at room temperature, and then poured onto ice, extracted three times with diethyl ether, washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution and then dried. After evaporation of excess diethyl malonate (70°/13Pa), the remaining oil is distilled on a Vigreux column, yielding 20,109.2 g (29°) of diethyl 2-furylmalonate (purity 94%) boiling point 100-110°/39 Pa.

24 g (1 mole) of sodium hydride are suspended in 100 ml of absolute tetrahydrofuran.

For 30 minutes, a solution of 218.4 g (0.97 mole) of diethyl 2-furylmalsate is added dropwise to 200 ml of absolute tetrahydrofuran, which raises the temperature to 30° (blue suspension).

After 30 minutes of stirring at 30°C, 340 g (200 mL; 2 mol) of isopropyl iodide is added over 30 minutes. The reaction mixture is heated under reflux overnight, then evaporated under reduced pressure and dissolved in ether. After evaporation of the solvent, the remaining oily residue is distilled under high vacuum, yielding 240.6 g (92.5%) of diethyl 2-furyl-2-35-isopropylmalonate (93.6% purity) in the form

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of a yellowish oil, boiling point 85-88°/13 Pa.

240.6 g (0.9 mole) of diethyl 2-furyl-2-isopropyl-malonate are dissolved in 500 ml of dimethyl sulfoxide, 2000 ml of 2N sodium hydroxide is added and the mixture is heated under reflux for 2 hours.

After cooling, the reaction mixture is poured into dilute sulfuric acid and extracted with ether until exhaustion. The organic extracts are then dried and the solvent is evaporated under reduced pressure. In this way, 148.7 g (98%) of (-)-2-(2-furyl)-3-methylbutyric acid is obtained in the form of a thick, yellowish oil.

161.0 g (0.96 mol) of (-)-2-(2-furyl)-3-methylbutyric acid are dissolved in 1000 ml of ether, and 109 g (0.9 mol) of S-(-)-α-phenylethylamine in 100 ml of ether are added. The precipitated crystals are filtered and recrystallized three times in 600 ml of ethyl acetate. This yields 80 g of white crystals, melting point 135-136°, α = -14.1° (c = 2%).

20 methanol).

These crystals are suspended in ethyl acetate and acidified by ice-cooling with a 3N hydrochloric acid solution. The organic solution is washed with water, dried, and evaporated under reduced pressure, yielding 46.7 g of (+)-2-(2-furyl)-3-methylbutyric acid as a viscous oil (98.6% purity), at +29.9°C (c = 1%, ethyl acetate).

thyle).

The mother liquors from the above 30 crystallizations are collected, and the free acids are isolated as described above, dissolved in 1000 ml of ether, and the corresponding amount of R-(+)-α-phenylethylamine is added. After three recrystallizations in ethyl acetate, 80.3 g of white crystals with a melting point of 135-136°, [α]^ = +15° (c = 1%, methanol) are obtained.

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The transformation of this salt according to the process described above gives 45 g of (-)-2-(2-furyl)-3-methylbutyric acid in the form of a viscous oil with a purity of 99.5%, [a]^° = -31.1° (c = 1%, ethyl acetate).

5. 10.5 g (0.277 mol) of lithium aluminum hydride are suspended in 100 mL of absolute ether, and 100 mL of absolute tetrahydrofuran is added with stirring over 10 minutes. To this stirred suspension, 10. 46.7 S (0.277 mol) of (+)-2-(2-furyl)-3-methylbutyric acid in 100 mL of absolute tetrahydrofuran is added dropwise under reflux over 6–8 hours. The reaction mixture is then cooled, and a 3N hydrochloric acid solution is slowly added. The organic phase is separated, washed with water, dried, and evaporated under reduced pressure. The remaining oil is vacuum-distilled, yielding 32 g (75%) of (R)-(+)-2-(2-furyl)-3-methyl-1-butanol as a colorless liquid with a purity of 96%.

boiling point 58°/39 Pa, [a]^ = 2.5° (c = 1%, 20 ethanol).

To a solution of 17.8 g (0.1154 mol) of (R)-(+)-2-(2-furyl)-3-methyl-1-butanol in 150 mL of carbon tetrachloride, 121.7 g (0.464 mol) of triphenylphosphine is added in portions over a period of 8 hours and 25 minutes. After 24 hours of stirring the reaction mixture, 500 mL of pentane is added and the precipitate formed is filtered. After evaporation of the solvent, the remaining precipitate is dissolved in pentane and purified by silica gel chromatography, yielding 16.5 S (92%) of 2-[(S)-(-)-30 1-(chloromethyl)-2-methylpropylfuran, which is used directly in the next step.

A solution of 14.6 g (0.084 mol) of 2-[(S)-(-)-1-(chloromethyl)-2-methylpropylfuran in 46 mL of ether is slowly added to a suspension of 3.22 g of magnesium in 12 mL of ether. The reaction mixture

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is vigorously shaken overnight at room temperature and then cooled to -70°C. Next, 8.3 g (0.038 mol) of t-butoxycarbonyl-L-leucinal is added dropwise to 40 mL of ether. After the addition is complete, the reaction mixture is left to stand for another hour at 0°C. Then, 100 mL of a saturated ammonium chloride solution is added, the organic phase is dried over magnesium sulfate, and the solvent is evaporated under reduced pressure. The remaining 16 g is chromatographed on silica gel with a 1:4 mixture of ether and hexane on silica gel, yielding 4.8 g of t-butyl[(1S,2S,4S)-

4-(2-furyl)-2-hydroxy-1-isobutyl-5-methylhexyl]carbamate which is directly implemented in the next step.

Example 70

15 In a manner similar to that described in E

Example 69, from t-butyl [(1S,2S,4S)-2-hydroxy-1-isobutyl-5-methyl-4-phenylhexyl]carbamate, we obtain [4S,5S,7S)-4-(Boc-Phe-His-£!H)-2,8-dimethyl-7-phenyl-

5-nonanol in the form of a viscous oil. MS: 648 20 (M+H)+.

The t-butyl[(1S,2S,4S)-2-hydroxy-1-isobutyl-5-methyl-4-phenylhexyl]carbamate used as a starting material was prepared as follows:

To 1500 ml of liquid ammonia, 25-23 g of small pieces of sodium are added at -35°C over 90 minutes. At the end of the addition, 150 mg of ferric (III) nitrate 9% is added. To this suspension, 150 g (1 mole) of methyl phenylacetate in 500 ml of absolute tetrahydrofuran is added dropwise under vigorous stirring, and 30 half hours later, 100 ml (1 mole) of iso- iodide is added.

Propyl. At the end of the addition, the ammonia is distilled overnight. Water is slowly added to the remaining liquid, and the mixture is extracted with ether. The organic extracts are washed with water, dried over sodium sulfate, and evaporated under reduced pressure, yielding

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165.8 g (87%) of methyl 2-phenyl-3-methylbutyrate in the form of yellow oil, which is directly implemented in the next step.

A solution of 165.8 g (0.87 mol) of methyl 2,5-phenyl-3-methylbutyrate, 52.0 g (1.30 mol) of sodium hydroxide, 1200 mL of methanol, and 600 mL of water is heated under reflux for one hour. After evaporation of the methanol under reduced pressure, dilution with water, and acidification with concentrated hydrochloric acid, the aqueous solution is extracted with ether. The ether extracts are washed with water, dried over sodium sulfate, and evaporated under reduced pressure, yielding 146.3 g (95%) of 2-phenyl-3-methylbutyric acid as a yellow oil, which is used directly in the next step:

To a solution of 146.3 g (0.82 mol) of rac-2-phenyl-3-methylbutyric acid in 820 ml of acetonitrile, 78.5 ml (0.615 mol) of S-(-)-α-phenylethylamine is added. The crystals thus formed are filtered and washed with acetonitrile. After three recrystallizations in acetonitrile, 88.5 g of acid is obtained.

(+)-2-phenyl-3-methylbutyric acid in the form of a salt of α-

on phenylethylammonium, melting point 198-200°, [and]^ +43° (c = 1% methanol). This salt is suspended in 1000 ml of ethyl acetate and 3N hydrochloric acid is added while cooling in an ice bath. The acetic extracts are washed with water, dried over sodium sulfate, and evaporated under reduced pressure, yielding 54.5 g of (S)-(+)-2-phenyl-3-methylbutyric acid

20

30 in the form of a slightly yellowish oil, [a]^ = 61.3° (c = 1%, chloroform).

A suspension of 19.5 g (0.51 mol) of lithium aluminum hydride is heated under reflux in 750 ml of absolute ether. Within 4 hours, a solution of 54.5 g (0.307 mol) of (S)- acid is added.

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(+)-2-phenyl-3-methylbutyric acid is mixed in 250 mL of absolute ether. After another 2 hours of heating under reflux, 1 U of hydrochloric acid is carefully added to the suspension, and the etheric phase is washed with water, then with 5 mL of dilute sodium bicarbonate solution, and finally with water again. After drying the organic phase over sodium sulfate and evaporating under reduced pressure, 53.1 g of (S)-(+)-2-phenyl-3-methylbuta-nol is obtained as a viscous oil, which is used directly in the next step.

To a solution of 49.3 g (0.3 mol) of (S)-(+)-2-phenyl-3-methylbutanol, 80 g (0.3 mol) of trimethylphosphine is added. Then, with vigorous stirring, 53.4 g (0.3 mol) of N-bromosuccinimide is added in 15 portions. After 16 hours, the reaction mixture is filtered, and the filtrate is evaporated under reduced pressure. The remainder is dissolved in hexane and purified by silica gel chromatography, yielding 34.9 g (5%) of (S)-(-)-2-phenyl-3-methylbromobutane as hui-20 volatile, [a]^ = -19.6° (c = 1%, chloroform).

A solution of 40.9 g (0.18 mol) of (S)-(-)-2-phenyl-3-methylbromobutane in 110 mL of tetrahydrofuran and 0.5 mL of 1,2-dibromoethane is slowly added dropwise to a suspension of 6.6 g of magnesium in 25 mL of ether, raising the temperature to 50°C. The reaction mixture is stirred for another 2 hours at 50°C and then cooled to -50°C. Next, a solution of 13 g (0.06 mol) of N-t-butoxy-carbonyl-L-leucinal in 40 mL of tetrahydrofuran is slowly added. After 16 hours at room temperature, 300 mL of a 20% ammonium chloride solution is added to the reaction mixture. The organic phase is dried over magnesium sulfate, and the solvent is evaporated under reduced pressure. The remainder (24.3 g) is chromatographed with a 4:1 mixture of hexane and ether on silica gel.

C

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which gives 9.3 g of t-butyl[ (1S,2S,4S)-2-hydroxy-1-iso-butyl-5-methyl-4—phenyl-hexyl] carbamate, which has a melting point of 95-96° after crystallization in hexane.

Example 7^

Similar to what was described in Example 1, the following compounds are prepared:

- From 78 mg (0.20 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 57 mg (0.24 mmol) of ethoxycarbonyl-L-phenylalanine 57 mg of (2S,3S,5S)-1-cyclohexyl-2-[(ethoxycarbonyl)-Phe-His-NH]-

5-isopropyl-6-heptenol-3 in solid form, US: 610 (H+H)+;

- From 78 mg (0.20 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]imidazol-4—propionamide and 70.8 mg (0.25 mmol) of phenylacetyl-L-phenylalanine

97 mg of (2S,3S,5S)-1-cyclohexyl-2-[(phenylacetyl)-Phe-His-NH]-5-isopropyl-6-heptenol-3 as a solid MS: 656 (M+H)+;

- From 78 mg (0.20 mmol) of (S)-α-amino-N-[(1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 68 mg (0.25 mmol) of 2-pyridylcarbonyl-L-phenylalanine 88 mg of 1-cyclohexyl-5-isopropyl-2-[(2-pyridylcarbonyl)-Phe-His-NH]-

6-heptenol-3 in solid form, MS: 64-3 (M+H)+;

- From 78 mg (0.20 mmole) of (S)-a-amino-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4— isopropyl-5-hexenyl]imidazol-4—propionamide and 75 mg (0.25nmole) of 2-phenylacetyl-L-tyrosine 110 mg of (2S,3S,5S)-1-cyclohexyl-5-isopropyl-2-[(phenylacetyl)-Tyr-His-NH)-6-heptenol-3 in solid form, MS: 672 (M+H)+;

- From 78 mg (0.20 mmol) of (S)-α-amino-N-[(1S,2S,4S)-1-(cyclohexylmethylethyl)-2-hydroxy-4—

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isopropyl-5-hexenyljimidazol-4-propionamide and 72 mg (0.25 mmole) of 2-pyridylcarbonyl-L-tyrosine 96 mg of (2S,3S,5S)-1-cyclohexyl-5-isopropyl-2-[(2-pyridylcarbo-nyl)-Tyr-His-NH]-6-heptenol-3 in solid form, 5 ÎIS : 659 (M+H)+;

- From 175 mg (0.45 mmol) of (S)-α-amino-N-[(1S,2S,4-E)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 145 mg (0.5 mmol) of 2-pyridylcarbonyl-L-tyrosine

10 150 mg of (S)-N-[(1S,2S,4E)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-[(S)-p-hydroxy-a-picolinamidohydrocinnamido]imidazol-4-propionamide in solid form, MS: 659 (M+H)+;

- From 175 mg (0.45 mmol) of (S)-a-

15 amino-IT-[ (1S,2S,4E)-1-(cyclohexylmethyl)-2-hydroxy-4—

isopropyl-5-hexenyl]imidazol-4-propionamide and 150 mg (0.5 mmol) of phenylacetyl-L-tyrosine 150 mg of (S)-N-[ (1S,2S,4-E)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-[(S)-p-hydroxy-^-(2-phenylacetamido)hydro-

20 cinnamamido]imidazol-4-propionamide in solid form, MS: 672 (M+H)+;

- From 175 mg (0.4-5 mmol) of (S)-α-amino-N-[(1S,2S,4-E)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]imidazol-4—propionamide and 137 mg

25 (0.5 mmole) of 2-pyridylcarbonyl-L-phenylalanine 150 mg of (S)-N-[ (1S,2S, 4-R)-1-( cyc lohexylmethyl thyl) -2-hydroxy-4— isopropyl-5-hexenyl] -a -[(S)-a-pic o1inamido-hydro c innamamido] imidazol-4—propionamide as solid, MS : 64-3 (M)+;

30 - From 175 mg (0.4-5 mmol) of (S)-a-

amino-N-[(1S,2S,4-E)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]imidazol-4—propionamide and 141 mg (0.50 mmol) of phenylacetyl-L-phenylalanine 150 mg of (S)-K-[(1S,2S,4E)-1-(cyclohexylmethyl)-2-hydroxy-4—iso-

35 propyl-5-hexenyl]-a-[(S)-a-(2-phenylacetamido)hydro-

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cinnamamido)imidazol-4-propionamide in solid form, PIS: 656 (H+H) +;

- From 107 mg (0.20 mmol) of R-[(1S, 2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-

5 hexenyl]-a-(3-phe nyl-L-alanyl)imida zol-4-prop ionamide and 66 mg of 1-(imidazolyl)-3-propionyl-D-proline 126 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl] -a-[(s)-a-[(R)-1_[3_(1H-imidazolyl-1)propionyl]-2-pyrrolidinecarboxamido]-10 hydrocinnamamido]imidazol—4-propionamide as solid, PIS: 757 (M+H)+;

- From 111 mg (0.20 mmol) of (S)-u-

[p-fluoro-L-alanyl)amino]-N-[(1S,2S,4S)-1-(cyclohexylmethyl) -2-hydroxy-4-is opr opy 1-5-hexenyl] imidazol-4- • 15 propionamide and 47.8 mg (0.24 mmol) of 2-(2-pyri-dyl)benzoic acid 120 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl) -2-hydroxy-4-isopropyl-5-hexenyl] -a-[(S)-u-[2-(2-pyridyl)benzamido]-p-fluorohydrocinnamamido]imidazol-4-propionamide as solid, MS: 737 (M+H)+; 20 - From 107 mg (0.20 mmol of ÏÏ-[(1S,2S,

4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-u-(3-phenyl-L-alanyl)imidazol-4-propionamide and 75 mg (0.24 mmole) of Boc-D-Pro-Pro-OH 122 mg (2S,5S)-2-(Boc-D-Pro-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-hept-25 nol-3 as solid, MS: 832 (M+H)-+.

- From 269 mg (0.6 mmol) of N-[(1S,2S,

4R )-1-(cyclohe xylmethyl)-2-hydro xy-4-is opropyl-5-h exenyl]-a-(3-phenyl-L-alanyl)imidazol-4-propionamide and 187 mg (0.6 mmol) of Boc-D-Pro-Pro-OH 400 mg of t-butyl (R)-2-30 [[(S)-2-[[(S)-a-[[(S)—1—[[(1S,2S,4R)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]carbamoyl]- 2-imidazolyl-4-ethyl]carbamoyl]phenethyl]carbamoyl]-1-pyrrolidinyl]-carbonyl]-1-pyrrolidinecarboxylate in solid form, MS: 832 (M+H)+.

35 - From 269 mg (0.5 mmol) of N-[(1S,2S,

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4R)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexe-nyl]-a-(3-phenyl-L-alanyl]imidazol-4—propionamide and 260 mg (0.6 mmole) of t-butoxycarbonyl-D-phenylglycine 110 mg of t-butyl[[(R)-a-[[(S)-u-[[(S)-1-[[(1S,2S,4-R)-1-5 (cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-carbamoyl]-2-imidazolyl-4—ethyl]carbamoyl]phenethyl]-carbamoyl]benzyl]carbamate in solid form, MS: 771 (M+H)+;

- From 269 mg (0.5 mmol) of N-[(1S,2S,104R)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexe-

nyl]-u-(3-phenyl-L-alanyl)imidazol-4—propionamide and 119 mg (0.6 mmole) of 2-(2-pyridyl)benzoic acid 290 mg of (S)-N-[(1S,2S,4R)-1-(cyclohexylmethyl)-2-hydroxy-4—is opropyl-5-hexenyl]-u-[(S)-a-[2-(2-pyridyl)-15 benzamido]hydrocinnamamido]imidazol-4—propionamide as solid, MS: 719 (M+H)+.

- From 107 mg (0.20 mmol) of N-[(1S,2S, 4S)-1-(cyc1oh exylmethyl)-2-hyd r oxy-4—is op ropyl-5-hexe-nyl]-a-(3-phenyl-L-alanyl)imidazol-4—propionamide and

20 75 mg (0.24 mmol) of Boc-Pro-Pro-OH 137 mg of (2S,3S,

5S)-2-(Boc-Pro-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 in solid form, MS: 832 (M+H)+;

- From 107 mg (0.20 mmol) of N-[(1S,2S,4R)-1-(cyclohexylmethyl)-2-hydroxy-4~isopropyl-5-hexenyl]-

25 ct-(3-phenyl-L-alanyl)imidazol-4—propionamide and 7^ mg (0.24 mmole) of IYA-D-Pro-L-Pro-OH 1J0 mg of (S)-N-[(1S,2S,4R)-1-(cyclohexylmethyl)-2-hydroxy-4— isopropyl-5-hexenyl]-a-[[N-[1-[(1-isovaleryl-D-prolyl)-L-prolyl]-3-phenyl-L-alanyl]amino]imidazol-4—propionamide in 30 solid form, MS: 816 (M+H)+;

- From 107 mg (0.20 mmol) of N-[(1S,2S,4S)-1-(cyclohexylmethyl1)-2-hydroxy-4-isopropy1-5-hexenyl]-&-(3-phenyl-L-alanyl)imidazol-4-propionamide and 67 mg (0.24 mmol) of N-t-butoxycarbonyl-N-methyl-L-

35 phenyl-alanine 100 mg t-butyl [(S)-u-[[(S)-a-[[(S)-

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1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-iso-propyl-5-hexenyl]carbamoyl]-2-imidazolethyl]carbamoyl] phenethyl]carbamoyl]phenethyl]methylcarbamate in solid form, MS: 799 (M+H)+;

5 -From 107 mg (0.20 mmol) of N-[(1S,

2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-&-(3-phenyl-L-alanyl)imidazol-4-propionamide and 71 mg (0.24 mmole) of IVA-D-Pro-Pro-OH 141 mg of (2S,3S,5S)-1-cyclohexyl-2-[(3-methylbutyryl)-D-Pro-Pro-10 Phe-IIis-NH]-5-isopropyl-6-heptenol-3 in solid form, MS: 816 (M+H)+;

- From 107 mg (0.20 mmol) of R-[(1S, 2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide

15 and 74 mg (0.24 mmole) of Boc-(£3-deshydro)-Pro-Pro-OH 125 mg (2S,3S,5S)-2-[[£(S)-1-Boc-3-pyrrolinyl-2]carbonyl] -Pro-Phe-His-NH]-1-cyclohexyl-5-isopropyl-6-hexenol-3 in solid form, MS: 830 (M+H)+;

- From 107 mg (0.20 mmol) of N-[(1S,2S,20 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-

a-(3-phenyl-L-alanyl)imidazol-4-propionamide and 75 mg (0.24 mmol) of Boc-Pro-D-Pro-OH 135 mg (2S,3S,5S)-2-(Boc-Pro-D-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 in solid form, MS: 832 (M+H)+; 25 - From 107 mg (0.20 mmol) of N-[(1S,2S,

4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-a-(3-phenyl-L-alanyl)imidazol-4-propionamide and 75 mg (0.24 mmole) of Boc-D-Pro-D-Pro-OH 133 mg (2S,3S,5S)-2-(Boc-D-Pro-D-Pro-Phe-His-IïH)-1-cyclohexyl-5-isopropyl-6-30 heptenol-3 in solid form, MS: 832 (M+H)+;

- From 110 mg (0.20 mmol) of (S)-a-(Phe-methylamino)-N-α(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide and 75 mg (0.24 nmol) of Boc-D-Pro-Pro-OH 87 mg of (S)-a-[Boc-D-

35 Pro-Pro-Phe-H(OH^)] -N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-

c

-147-

hydroxy-4-isopropyl-5-hexenylimidazol-4-propionamide in solid form, MS: 846 (M+H)+;

- From 107 mg (0.20 mmol) of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropy1-5-hexe-

5 nyl]-a-(3-phenyl-L-alanyl)imidazol-4-propionamide and

55 mg (0.24 mmol) of (S)-N-t-butoxycarbonyl-piperidine-2-carboxylic acid 127 mg (2S,3S,5S)-1-[[[(S)-

1-Boc-2-piperidinyl]carbonyl]-Phe-His-NH]-1-cyclohexyl-5-isopropyl-6-heptenol-3 in solid form, MS: 749 10 (M+H)+;

- From 54 mg (0.10 mmol) of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-nyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 49 mg (0.12 mmol) of Boc-D-Pro-D-Pro-Pro-OH 57 mg of

15 (2S,5S,5S)-2-(Bo c-D-Pro-D-Pro-Pro-Phe-His-NH)-1-cyc1o-hexyl-5-isopropyl-6-heptenol-3 in solid form, MS: 927 (M+H)+;

- From 107 mg (0.20 mmol) of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexe-

20 nyl]-a-(3-phenyl-L-alanyl)imidazol-4-propionamide and

56 mg (0.24 mmol) of (R)-N-t-butoxycarbonyl-thiazolidine-2-carboxylic acid 126 mg of (2S,3S,5S)-2-

[[[(R)-3-Bo c-4-1h ia z o1idiny1]c arbony1-Phe-His-NH]-1-cyclohexyl-5-isopropyl-6-heptenol-3 in solid form, 25 MS : 753 (M+H)+;

- From 80 mg (0.123 mmol) of (2S,3S,5S)-^-[[[(S)-2-piperidinyl]carbonyl]-Phe-His-NH]-1-cyclo-hexyl-5-isopropyl-6-heptenol-3 and 64 mg (0.30 mmol) of Boc-D-Pro-OH 106 mg of (2S,3S,5S)-2-(Boc-D-Pro-Pip-

30 Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 in solid form, MS: 846 (M+);

- From 107 mg (0.20 mmol) of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 79 mg

35 (0.24 mmol) of (R)-N-t-butoxycarbonyl-thiazolidinyl-l-

1

-148-

carbonyl-proline 163 mg of (2S,3S,5S)-2-[[[(R)-3-Boc-

4-thiazolidinyl]carbonyl]-Pro-Phe-His-NH]-1-cyclohexyl-

5-isopropyl-6-heptenol-3 in solid form, MS:850 (M+H)+;

- From 107 mg (0.20 mmol) of N-[(1S,2S,4-S)-1-(cyclohexylmethylethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 71 mg (0.24 mmol) of 2-(2-pyridyl)benzoic acid, 140 mg of

(2S,3S,5S)-1-cyclohexyl-5-isopropyl-2-[[2-(2-pyridyl)-benzoyl]-Pro-Phe-His-NH]-6-heptenol-3 as dihydrochloride, MS: 816 (M+H)+;

- From 107 mg (0.20 mmole) of (N-[(1S,2S,4-S )-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexenyl]-a-(3-phenyl-L-alanyl)imidazol-4—propionamide and 57 mg (0.24- mmole) of 3-dimethyl-3-(1-imidazolyl)-propionyl-Pro-Pro-OH 160 mg of (2S,3S,5S)-1-cyclohexyl-2-[(3,3-dimethyl-3-imidazolyl-1-propionyl)-Pro-Pro-Phe--His-NH]-5-isopropyl-6-heptenol-3, MS: 882 (M+H)+;

- From 107 mg (0.20 mmol) of (N-[(1S, 2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-

hexenyl]-and-(3-phenyl-L-alanyl)imidazol-4-propionamide and 77 mg (0.24 mmol) of N-(1-imidazolyl)acetyl-D-Pro-Pro-OH 160 mg (2S,3S,5S)-1-cyclohexyl-2-[(imidazol-1-

ylacetyl)-D-Pro-Pro-Phe-His-NH-5-isopropyl-6-heptenol-3

in the form of solid dihydrochloride, MS: 840 (M+H)+;

- From 107 mg (0.20 mmol) of (N-[(1S,2S,4-R)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide and 77 mg (0.24 mmol) of N-(1-imidazolyl)acetyl-D-Pro-Pro-OH 148 mg (2S,3S,5R)-1-cyclohexyl-2-[(imidazol-1-ylacetyl)-D-Pro-Pro-Phe-His-NH]-5-isopropyl-6-heptenol-3 as dihydrochloride, in solid form,

MS: 840 (M+H)+;

- From 107 mg (0.20 mmol) of (N-[(1S, 2S, 4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isotropyl-5-

-149-

hexenyl] -a-(3-phenyl-L-alanyl)imidazol-4-propionamide 83 mg (0.24 mmol) benzyloxycarbonyl-Pro-Pro-OH 155 mg (2S,3S,5S)-2-(benzyloxycarbonyl-Pro-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 in solid form, MS: 866 (M+H)+.

The (S)-a-(phemethylamino)-N-[(1S,2S,4S)-1-(cyclohexylmethyl thyl)-2-hydroxy-4-isopropyl-3-hexenyl]-imidazol-4-propionamide used as a starting material was prepared as follows:

10 A mixture of 1.50 g (4 mmol) of N-a-N-im-

Di-t-butoxycarbonyl-N-α-methylhistidine, 1.0 g (3.4 mmol) of (S,S,S)-6-amino-6-cyclohexylmethyl-4-isopropyl-1-hexenol-5-hydrochloride, 1.33 g (3.5 mmol) of HBTU, 7.8 mL of N-methylmorpholine, and 20 mL of dimethyl-15-formamide are stirred for 4 hours at room temperature. The solvent is then removed under reduced pressure, and the remaining oil is dissolved in ether. The organic solution is then washed with saturated sodium chloride solution and, after drying over sodium sulfate, evaporated under reduced pressure, yielding

2.1 g of (S)-3-(t-butoxycarbonyl)-α-[l-(t-butoxycarbonyl)-N-methylformamido]-N-[(1S,2S,4S)-1-(cyclohexylmethylethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide which is directly used in the next step: 25 2.1 g of (S)-3-(t-butoxycarbonyl)-α-[l-(t-buto-

(xycarbonyl)-N-methylformamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide are dissolved in 30 mL of hydrochloric acid in dioxane and left to stand for 5 hours at room temperature. Then 500 mL of ether is added and the precipitate formed is filtered. The precipitate is then washed with ether and dissolved in water. Solid sodium bicarbonate is added to the aqueous solution and the solution is extracted several times with ethyl acetate, yielding 0.85 g of (S)-a-(N-methylformamido)-N-[(1S,2S,4S)-

t

-150-

1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl-imidazol-4-propionamide, which is directly used in the next step:

A mixture of 0.85 S (2.1 mmol) of (S)-a-(N-5-methylformamido)-N-[(1S, 2S, 4S)-1-(cyclohexylmethyl)-

2-hydroxy-4-isopropyl-5-hexenyl] i midazol-4-propionamide, 0.67 g (2.5 mmol) of N-t-butoxycarbonylphenylalanine, 0.84 g (2.2 mmol) of HBTU, 0.3 ml of N-methylmorpholine and 20 ml of dimethylformamide is left to stand

10 is left at room temperature for 60 hours and then poured onto a saturated sodium bicarbonate solution. The resulting precipitate is filtered and dissolved in ethyl acetate. The organic solution is dried over magnesium sulfate and evaporated under reduced pressure. The remaining 15 is ground in hexane and filtered, yielding 1.35 g of (S)-a,-(Boc-Phe-methylamino)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4—propionamide as a powder, which is used directly in the next step.

20 1.3 g of (S)-u-(Boc-Phe-methylamino)-N-[(1S,2S,

4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide is dissolved in 30 mL of 3N hydrochloric acid in dioxane and left to stand for 1 hour at room temperature. Then, 200 mL of ether is added and the mixture is stirred for 16 hours at room temperature. The resulting precipitate is filtered, washed with ether, and dissolved in water. Solid sodium bicarbonate is then added to the aqueous solution, and the mixture is extracted several times with ethyl acetate, yielding 0.97 S of (S)-u-(Phe-methylamino)-N-[(1S,2S,4-S)-1-(cyclo-30hexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, which is used directly in the next step.

The (2S,3S,5S)-1-[[[(S)-2 -piperidinyl]carbonyl]-Phe-His-NH]-1-cyclohexyl-5-isopropyl-6-heptenol-3 used as a starting material was obtained by elimination

H

-151-

of the t-butoxycarbonyl group of (2S,3S,5S)-1-[[[(S)-1-Boc-2-piperidinyl]carbonyl-Phe-His-NH]-1-cyclohexyl-5-isopropyl-6-heptenol-3 by means of hydrochloric acid in dioxane and directly used in the following step 5.

Example 72

A mixture of 31 mg (0.21 mmol) of N-cyano-dimethyldithiocarbamate, 107 mg (0.20 mmol) of N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-10 5-hexenyl]-α-(3-phenyl-L-alanyl)imidazol-4-propionamide, and 1 mL of methanol is left to stand for 24 hours at room temperature. The solvent is then evaporated under reduced pressure, and the remaining mixture is digested in a 1:1 mixture of hexane and ether, yielding 112 mg of 15(S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-

isopropyl-5-hexenyl]-a-[[N-[ (cyano imino)(methylthio)-methyl]-3-phenyl-L-alanyl]amino]imidazol-4-propionamide in solid form, MS: 636 (M+H)+.

Example 73

20. 100 mg of (2S,5S)-2-(Boc-D-Pro-Pro-

Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3 in 3.2 mL of 3.2N hydrochloric acid in dioxane, the solution is allowed to stand for 1 hour and then the desired product is precipitated by the addition of ether. The precipitate is filtered and dried under reduced pressure, giving 100 mg of [2S,3S,5S]-1-cyclohexyl-2-(D-Pro-Pro-Phe-His-NH)-5-isopropyl-6-heptenol-3 as the dihydrochloride solid, MS: 732 (M+H)+.

Example 74

30 41 mg (0.05 mmol) of (2S,3S,5S)-1-cyclohexyl-

2-[(3-methylbutyryl)-D-Pro-Pro-Phe-His-NH]-5-isopropyl-6-heptenol-3 are dissolved in 5 ml of methanol and hydrogenated in the presence of 20 mg of palladium on activated carbon (5%). At the end of hydrogen absorption, the ca-35 analyzer is filtered and the solvent is evaporated under pressure.

-152-

reduced ion. Grinding the remainder with hexane and drying gives 34 mg of (2S,3S,5S)-1-cyclohexyl-5-iso-propyl-2-(isovaleryl-D-Pro-Pro-Phe-His-NH)-3-heptanol as a solid, MS: 818 (M+H)+.

The hydrogenation can also be directed in such a way that the isomeric compound is obtained when the hydrogenation is carried out under the following conditions:

200 mg of starting material are hydrogenated in the presence of 50 mg of palladium on activated carbon (at 5%) in 15 ml of methanol and treated in the usual way, giving (2S,3S,5R)-1-cyclohexyl-5-iso-propyl-2-(isovaleryl-D-Pro-Pro-Phe-His-NH)-3-heptanol in solid form, MS: 818 (M+H)+.

Example 75

80 mg (0.25 mmol) of 3-(1-imidazolyl)propionyl-Pro-Pro-OH, 110 mg (0.20 mmol) of (S)-α-(Phe-methyl-amino)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide, 84 mg (0.22 mmol) of HBT11, 0.03 mL of N-methylmorpholine, and 2 mL of dimethylformamide were left to stand for 16 hours at room temperature. The reaction mixture was then poured onto 50 mL of a 5% sodium carbonate solution, and the milky suspension was extracted several times with ethyl acetate. The organic extracts were dried over magnesium sulfate and evaporated under reduced pressure. The organic extracts were dried over magnesium sulfate and evaporated under reduced pressure. The remainder was dissolved in 10 mL of ethyl acetate, and 1.5 mL of a 3.0 Nm hydrochloric acid in dioxane solution was added to the solution. The resulting precipitate was filtered and dried, yielding 143 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[[3-(imidazo-lyl-1)propionyl]-D-Pro-Pro-Phe-N-(methyl)]imidazol-4-propionamide hydrochloride as a solid, MS: 868

-153-

(M+H)+.

Similar to what has just been described, 155 mg of (S)-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide is prepared from 77 mg (0.24 mmol) of 2-(1-imidazolyl)acetyl-Pro-Pro-OH and 110 mg of (S)-5 α-(Phe-methylamino)-N-[(1S,2S,4-S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[(imidazolyl-1-acetyl)-D-Pro-Pro-Phe-N-(methyl)]imidazol-4-propionamide di-10 hydrochloride in solid form, MS: 84-0 (M+H)+.

Example 76

105 mg of (S)-N-[(1S,2S,4S)^-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-5-hexenyl]-α-[[3-(imidazo-lyl-1)propionyl-D-Pro-Pro-Phe-N-(methyl)]imidazol-4—15 propionamide hydrochloride are dissolved in 15 mL of methanol and hydrogenated in the presence of 50 mg of palladium on activated carbon. At the end of hydrogen absorption, the catalyst is filtered and the filtrate is evaporated to dryness. The remainder is ground with ether and filtered, giving 65 mg of (2S,3S,5S)-1-cyclohexyl-2-[(3-imidazolyl-1-propionyl)-D-Pro-Pro-Phe-His-NH]-5-isopropyl-3-heptanol-dihydrochloride in solid form, MS: 856 (M+H)+.

Example 77

25 A sterile, filtered aqueous solution of (S)-a-

[(R)-2-benzyl-5,5-dimethyl-4~oxohexanamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4—isopropyl-5-hexe-nyl]imidazol-4—propionamide is mixed by heating under aseptic conditions with a sterile gelatin solution containing phenol as a preservative, so that 1.0 ml of solution has the following composition:

(S)-a-[(R)-2-benzyl-5,5-dimethyl-4~oxohexanamido]-N-[(1S,2S,4S)-1-(cyclo-35 hexylmethyl)-2-hydroxy-4—isopropyl-5-

-154-

hexenylimidazol-4-propionamide 3.0 mg, gelatin 150 mg, phenol 4.7 mg, distilled water up to 1.0 ml

5 The mixture is placed, under aseptic conditions, in ampoules, up to 1 ml.

Example 78

5 mg of (S)-u-[(R)-2-benzyl-5,5-dimethyl-4-oxohexanamido]-N-[(1S,2S,4S)-1-(cyclohexyl-10-methyl)-2-hydroxy-4-isopropyl-5-hexenyl]imidazol-4-propionamide is dissolved in aqueous solution with 20 mg of mannitol. The solution is filtered to sterility and filled into a 2 mL ampoule under aseptic conditions, then frozen and lyophilized. Before use, the lyophilized powder is dissolved in 1 mL of distilled water or 1 mL of physiological saline. The solution is administered intramuscularly or intravenously. This formulation can also be packaged in dual-chamber injectable ampoules. Example 79

In a mixture of 3.5 ml of Myglyol 812 and 0.08 g of benzyl alcohol, a finely ground (5.0 µm) (S)-[(R)-2-benzyl-5,5-dimethyl-4-oxohexanamido]-N-[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropyl-25-5-hexenyl]imidazol-4-propionamide is suspended. This suspension is packaged in a container with a dispensing valve. 5.0 g of Freon 12 is introduced under pressure through the valve. By shaking, the Freon dissolves in the Myglyol-benzyl alcohol mixture. This aerosol can contains approximately 100 unit doses that can be administered individually.

c

Claims (1)

-155- - DEMANDS - 1 - Process for preparing amino acid derivatives of general formula S' ? f ■ _/l\ / V/ \ / \ T A • N • • J- i H ] \r2 / \ 1 v 2 10 in which R is hydrogen or methyl, R is e- ethyl, propyl or imidazolyl-4, Br isobutyl, cyclohexylmethyl or benzyl, R phenyl, furyl, vinyl, ethyl or 1,2-dihydroxyethyl, A is attached by the carboxyl group and represents the acid 15 dibenzosuberoacetic acid, 2-(2-pyridyl)benzoic acid, naphthylsuccinic acid monoethyl ester, 2-Indolylacetic acid or dihydrocinnamic acid or one of the groups 20 r6 5 T ■R -i -Y-Z -y • G and (a) (b) 25 in which the dotted line may represent an additional bond, R^ phenyl, naphthyl, indolyl, pyrazolyl, imidazolyl or pyridyl-methyl and R^ hydrogen, an alkyl, arylalkyl, alkoxy-30 carbonylalkyl, alkylcarbonylalkyl, cycloalkylcarbonyl-alkyl, heterocycloalkylcarbonylalkyl, arylcarbonylal-kyl, aminocarbonylalkyl, substituted aminocarbonylalkyl, aminocarbonyl, substituted aminocarbonyl, alkoxycarbonyl, alkylcarbonyloxy, arylalkylearbonyloxy, alkylami-35 nocarbonyloxy, alkoxycarbonylamino, cyano or 2,5-dime- -156- thylpyrrolyl-1, with the condition that R cannot 3 represent an alkoxycarbonylamino, where R represents phenyl or α-naphthyl, and Y is the N-terminal divalent remainder bonded to Z, if presently N- and/or β-methylated, of phenylglycine, cyclohexylglycine, phenylalanine, cyclohexylalanine, 4-fluorophenylalanine, 4-chlorophenylalanine, tyrosine, α-naphthylalanine, homophenylalanine, ethyl ester of aspartic acid, β-butyl ester of glutamic acid, or benzyl ester of glutamic acid, and Z is hydrogen, an acyl group, or one of the groups 15 • S • .( y • -CH_- •Ç_'.>-ca*-or/;ÇH-CH2-NH;/;»;V;\;y;-CH--OÇ-N-Ç-;mi;(d);(c);20;et;CH3-S\;NC-N;CH-;(e);25;in the form of optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates or mixtures of diastereomeric racemates as well as pharmaceutically usable salts of these compounds characterized in that a) a compound of general formula;-157-;5;fi??' is reacted 4;VS 6h A;12 3 4 in which R, E, R and R have the meanings given above, with an acylation agent giving the group;10 5 ?6;R N A„/ "Y"Z;• g or;(a) (b);5 S;15 in which E, E, I, Z and the dotted line have the meanings given above, or b) a compound of general formula r3;20 f 4;2 À 111;3 4; in which E^ and E have the meanings given above with a compound of general formula;À A0H;30 A î Iv;•v;1 2; in which E, E and A have the meanings given above,;35 or with one of its activated derivatives, or;-158-;c) to prepare a compound of formula I, in which R^ is the ethyl and the other symbols have the meanings given above, to hydrogen a compound of formula I, in which R is vinyl and the other symbols have the meaning given above, or d) to prepare a compound of formula I, in which R is 1,2-dihydroxyethyl and the other symbols have the meaning given above, one oxidizes a compound of formula I in which R represents vinyl and the other symbols have the meaning given above, or e) to prepare a compound of formula I, in which A contains a free amino group, one removes the protecting N group from a corresponding compound of formula I, in which A contains an N-protected amino group, and/or f) if desired, one separates a mixture of diastereomeric racemates into the diastereomeric racemates or the optically pure diastereomers, and/or g) if desired, one separates a diastereomeric mixture into the optically pure diastereomers, and/or h) if desired, a compound obtained is transformed into a pharmaceutically usable salt ion.;25 2 - Process according to claim 1, in which R represents hydrogen, R is imidazolyl-4-, R is cyclohexylmethyl and R is vinyl or ethyl.;3 - Process according to claim 1 or 2, in which A represents group (a) or (b), in which R is phenyl or naphthyl, R is an alkylcarbonyl-alkyl, cycloalkylcarbonylalkyl, heterocycloalkylcarbonylalkyl, aminocarbonylalkyl or substituted aminocarbonylalkyl, Y is phenylalanine, cyclohexylalanine, 4-chlorophenylalanine or α-naphthylalanine and Z is a 35 acyl.;t;;-159-;4 - Process according to claim 3, in which R is phenyl, R is an alkylcarbonylmethyl;C^-C^, cycloalkylcarbonylmethyl C^-Cg, N-t-butoxycarbo-;nylpyrrolidinylcarbonylmethyl or aminocarbonylalkyl;5 which is disubstituted by an alkyl or an alkylene;C.-Cc, optionally interrupted, by an oxygen atom, or monosubstituted by a mono-aminoalkyl or disubstituted by an alkoxycarbonylalkyl or an alkoxycarbonyl, and Y is phenylalanine, and Z is the remainder linked to Y by the C-terminal of a natural N- and/or α-methylated amino acid, optionally substituted by the R-CO- or R-CO- groups, of L configuration or of a epimer of such an amino acid with the D configuration, or of a dipeptide of two of these amino acids, or Z re-15 presents the R^-CO- or Ra-0-CO- group in which Ra is a hydrocarbon remainder, optionally substituted, saturated or unsaturated, aliphatic, cycloaliphatic, cycloaliphatic-aliphatic with up to 18 carbon atoms, a hydrocarbon remainder, optionally substituted, aromatotic, heteroaromatic, aromatic aliphatic or heteroaromatic-aliphatic with up to 18 carbon atoms or a heterocycle, optionally substituted, saturated with 5 or 6 members and R represents hydrogen or has the meaning of Ra, preferably Z is the remainder linked by the C-25 terminal to Y of proline, prolylproline or histidinyl-proline, the amino group being in each case substituted by the t-butoxycarbonyl, benzoxycarbonyl, isovaleryl;•j. v or the R-C0 group, wherein R is a heteroaromatic-aliphatic hydrocarbon residue with up to 30 carbon atoms or represents a saturated heterocycle with 5 or 6 members; 5 - A process according to any one of claims 1-4, 1-2, wherein R is hydrogen, R is 4-imidazolyl, 3-4, R is cyclohexylmethyl, R is vinyl or ethyl, and 3-5, R is phenyl, and R is an alkylcarbonylmethyl C-C-160, cycloalkylcarbonylmethyl C-C-160, N-butoxycarbonylpyrrolidinylcarbonylmethyl, or aminocarbonylalkyl that is disubstituted by an alkyl C-C4 or an alkylene, optionally interrupted by an oxygen atom, or monosubstituted by an aminoalkyl mono- or disubstituted by an alkoxycarbonylalkyl or alkoxycarbonyl, where Y is the divalent phenylalanine residue linked to Z by the N-terminus and Z is the proline, prolylproline, or histidinylproline residue linked to Y by the C-terminus, the amino group being in each case substituted by t-butoxycarbonyl, benzoxycarbonyl, iso-valeryl, or the R-CO- group, wherein R is an aliphatic heteroaromatic hydrocarbon residue having up to 10 carbon atoms or a saturated heterocycle with 5 or 6 members. A process according to claim 1, characterized in that t-butyl(R)-2-[[(S)-a-[[(S)-1-[,2S,4S)-1-(cyclohexylmethyl-2-hydroxy-4-isopropyl-hexyl]-2-imida z olyl-4-e thyl]c arb amoyl]phen thyl]c arba-;20 moyl]-1-pyrrolidine carboxylate.;7 -- Process according to claim 1, characterized in that the (2S,3S,5S)-2-(Boc-D-Pro-Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3- is prepared;8 - Process according to claim 1, characterized in that the t-butyl(S)-2-[[(R)-a-[[(S)-;1-[[(1S,2S,4S)-1-(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-2-imidazolyl-4-ethyl]carbamoyl]-phenethyl]acetyl]-1-pyrrolidinecarboxylate is prepared.;9 - Process according to claim 1, 30. A process characterized in that it prepares N-(S)-1-[[(1S,2S,4S)-1-;(cyclohexylmethyl)-2-hydroxy-4-isopropylhexyl]carbamoyl]-;2-imidazoly 1-4-ethyl} - V-oxo-o. - (1-naphthylmethyl)-4-mor-pholinebutyramide.; 10. A process according to claim 1, characterized in that it prepares (2S,5S)-2-(Boc-D-Pro-Pro-;C;-161-;Phe-His-NH)-1-cyclohexyl-5-isopropyl-6-heptenol-3.; 11. A process for preparing medicinal products, particularly for use in the control or prevention of hypertension or insufficiency. cardiac;5 characterized in that an amino acid derivative of formula I indicated in claim 1 is administered in the form of an optically pure diastereomer, a mixture of diastereomers, a racemate of diastereomers or a mixture of racemates;10 diastereomers, or one of their pharmaceutically usable salts.;12 - Use of an amino acid derivative of formula I indicated in claim 1 in the form of an optically pure diastereomer, a mixture of diastereomers, a racemate of diastereomers or a mixture of racemates or a pharmaceutically usable salt for preparing drugs against hypertension and/or heart failure.;20 13 - Compounds of general formulas;25;t f;T»/ \ /'\/\ /"\ /;H ! I;R;V;AH /JN;II;H_N;r;<5H >';R';III;St /^\ /^~0H;30 A N|/ IV;•'V;1 2;in which R is hydrogen or methyl, R is ethyl, propyl, or imidazolyl-4, R is isobutyl, cyclohexylmethyl, or benzyl, R is phenyl, furyl, vinyl, ethyl, or 1,2-dihydroxyethyl, X is chlorine, bromine, or iodine, and R is phenyl or furyl.;q f* l G i N A L in S>89®S l References cooten<,n . " . . crossed-out word, no added word — José CURAU Industrial Property Consultancy 26b,l< Princess Charlotte Blvd. uiauc^ .has Base. , S f\ -