AU622015B2 - Dipeptide derivatives having enzyme-inhibitory action - Google Patents

Description

Form COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952-69 COMPLETE SPECIFICATION (OR IGINAL) Class I t, Class Application Number: Lodged: Co~mplete Spccification Lodged: Accepted:- Published: 0 00 09000 Priority: 0 0 0 0 090 0000*0 Related Art: 9 0 00 0 00 a Name of Applicant 9 0a SAddress of Applicant Actual Inventor 0 Address for Service 0 a HOE CHST AKTI ENGESELLSCHAFT D-6230 Frankfurt am Main 80, Federal Republic of Germany, WOLF-ULRICH NICKEL, WOLFGANG LINZ, DIETER RUPPERT, HANSJORG URBAC- and ADALBERT WAGNER.

WATERMARK PATENT TRADEM,_ K ATTORNEYS.

LOCKED BAG NO. 5, H1AWTHORN, VICTORIA 3122, AUSTRALIA Complete Specification for the Invention entitled: DIPEPTIDE DERIVATIVES HAVING ENZYME-INHIBITORY ACTION The following staterrt Is a full description of this Invention, Including the best method of performing It known to

HOE

Des Di jIk la CHST AK(TIENGESELLSCHAFT HOE 89/F 128 1,)r.MY/gm cription eptide derivatives having enzyme-inhibitoxy action The invention relates to dipeptide derivatives which inhibit the action of the natural enzyme renin and to processes for their preparation and use and to pharmaceutical preparations containing them. In EP-A 0,255,082, dipeptide derivatives are described which carry N-terminal cycloalkylcarbor.,yl and cyc loalkyl-alkylcarbonyl substituents. In WO 838/05050, Priinodiol derivatives having renin-inhibitory action czr' described.

0 00 00 0 00.00 0 1 00 0 1 0 0 0 000 00 00 00 0 0 0 0 00 00 0 000 0 00*0 00 0000 a 0 00 020 000to 0 00 25 00 a Surprisingly, it has now been found that substituted cycloalkyl and cycloalkyl-alkyl derivatives contribute to the considerable improvement of the action in vivo and to the increased absorption of the compounkd.

The invention relates to compounds of the formula I R2OH

R

3 Ra~W -B NH-CH- CH- CH..R 4 which denotes (C 3 -cyc loalkyl, (CA-Cia) -bicyc loalkyl, -tricycloalkyl, -cycloalkyl- (C 1

-C

6 alkyll (CA-Cia) -bicycloalkyl- (Cl-Cr 8 -alkyl and

C,-C

12 -tricycloalkyl- (C 1

C

6 -alki, 1, in which in each case the cycloalkyl, bicycloalkyl and tricycloalkyl substituents can be substituted by one or two identical or different radicals from the series comprising F, Cl, Br, I, hydroxyl, (C,-C,)-alkoxy, -alkyl, carboxyl, (C 1 -alkoxycarbonyl, carbamoyl, carboxymethoxy,. amino, (C 1 -monoaljcylamino, (C 1-C6) -di alkyl amino, a~mino- -alkyl, (Cl-

C

8 -alkylamino- (C 1 -Cr 8 alkyl di- (CI-Cr,) -alkylamino-

(C

1 -C,)-alkyl, amidino, hydroxaiino, hydroximinor hydrazono, imino, guanidino, (Cl-C 8 -alkyloxy- -2sulf onyl, (C 1 -alkyloxysulf enyl, trif uorometly;-.

4 -alkoxyc arbonyl amino or (Cr,-C 12 -aryl- (Cl-C 4 alkoxycarbonylaxaino; W stands for -0-CO- or -SO 2

R

2 denotes hydrogen, (Cl-C 1 )-alkyl, (C 4

-C

7 )-cycloalkyl,

(C

4 -C7) -cycloalkyl- (CI-C 4 -alkyl, -aryl or (C6-C 14 -aryl- (Cl-CA) alkyl, denotes hydrogen, (C 1 -alkyl, (C 6

-C

14 -rl (Cs-C, 4 -aryl- Cl-C 4 -alkyl hydroxyl or amino, R4 denotes a radical of the formula II 00 (CH,)m-CHR 5 -D (I where R0 stands for hydrogen, (Cl-C 7 )-alkyl, (C,-C 5 0 0 0alkoxy, (C 1

-C

5 -alkylthio, (C 1

-C

5 -alkylamino, hydroxyl, azido, fluorine, chlorine, bromine or iodinpl. '4 0 0D deinotes a radical Het, where HIet denotes a 5- to 7-meinbered heterocyclic 0 0 0ring which can be f used to benze-ne, aromatic, 0 20 partially hydrogenated or completely hydro- 00 0 0genatedi which can contain one or two identical different radicals from the orour. N, 0, S, NO, So or SO 2 as heteroatoms and which can be 08 4 00'25 ferent radicals from the group comprising (CA-CO)-alkyl, (Cl-C 4 -alkoxy, hydroxyl, halogen, amino, mono- or di- (C 1

-C

4 )-al)ylamino or CF., and m denotes 0, 1, 2, 3 or 4; A and B independently of one anothe- denote a radical of an aptno ar(id linke~d at the nitrogen terminus to R"- -3 W or A and at the carbon terminus to B or NH-CHR 2 CHOH-CHR 3 -R 4 from the series comprising phenylalanine, histidine, tyrosine, tryptophan, methionine, leucine, isoleucine, asparagine, aspartic acid, p-2-thienylalanine, pi-3-thienylalanine, p-2-furylalanine, ,-3-furylalanine, lysine, ornithine, valine, alanine, 2,4-diaminobutyric acid, arginine, 4-c hiorophenylalanine, methionine sulfone, methionine sulfoxide, 2-pyridylalanine, 3-pyridylalanine, cyclohexylalanine, cyclohexyiglycine, immethyihistidine, 0-methyltyrosine, O-benzyltyrosine, 0-tert butyltyrosine, phenyiglycine, 1-naphthylalanine, 2 -naphthylalanine, 4 -nitrophenylalanine, norvaline, f-2-benzo[b]thienylalanine, p-3-benzo~bJthienylalanine, 2-f luorophenylalanine, 3-f luoro- 0phenylalanine, 4-pyridylalanine, 4-f luorophenylalanine, norleucine, cysteine, S-methylcysteine, 001,,3,-tetrahydroisoquinoline-3-carboxylic acid, 0 011 00 homophenylalanine, DOPA, 0-dimethyl-DOPA, N-methyl- 0 1: 20 histidine, 2 -amino- 4- 2-thienyl) -butyric acid, 2- 0a0 amino-4-(3-thienyl)-butyric ai,3(-hey) serine, (Z )-dehydrophenylalanine, -dehydrophenylalanine, 1, 3-dioxolan-2-ylalanine, N-pyrrolylalanine 0 and 3- or 4-pyrazolylalanine, 0 000a 0 :25 and their physiologically tolerable salts.

The chiral centers in the compounds of the formula I can have the S- or RS-configuration.

Alkyl can be straight-chain or branched. The same applies C to radicals derived therefrom, such as, for example, alkoxy, alkylthio, alkylamino, dialkyladino, alkanoyl and aralkyl. (C 3

-C

8 -Cycloalkyl is understood as meaning cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. (C 4 -C)-Bicycloalkyl or (C.-C 12 tricycloalkyl are understood as meaning an isocyclic kf 3 aliphatic, non-aromatic radical which can optionally contain unsymmetrically distributed double bonds, and4 i- -4which can optionally also be substituted with open-chain aliphatic side chains. The two or three rings as components of a radical of this type are condensed or linked in a spiro fashion and linked via a ring carbon atom or a side chain carbon atom. Examples of these radicals are bornyl, norbornyl, pinanyl, norpinanyl, caranyl, norcaranyl, thujanyl, adamantyl, bicyclo(3.3.0)octyl, bicyclo(1.1.0)butyl and spiro(3.3)heptyl substituents.

If the cyclic compounds mentioned carry more than one substituent, these can be both cis and trans to rne another.

on a o 00 0 a 0 0 o0 0 0 00o 0ao 0000 41

(C

6

-C

14 )-aryl is, for example, phenyl, naphthyl, biphenylyl or fluorenyl; phenyl is preferred. The same applies to radicals derived therefrom, such as, for example, 15 aryloxy, aroyl, aralkyl and aralkyloxy. Aralkyl is understood as meaning an unsubstituted or substituted

(C-C

4 )-aryl radical linked to (Cl-C)-alkyl, such as, for example, benzyl, a- and p-naphthylmethyl, halobenzyl and alkoxybenzyl, aralkyl, however, not being limited to the radicals mentioned.

A radical Het in the sense of the preceding definition is, for example, pyrrolyl, furyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, pyridyl, pyrazinyl, pyrimidinyl, indolyl, quinolyl, isoquinolyl, quinox- 25 alinyl, p-carbolinyl or a derivative of these radicals which is fused to benzene, cyclopentane, cyclohexcne or cycloheptane. This heterocycle may be substituted on a nitrogen atom by oxido, (C-C 6 s)-alkyl, for example methyl or ethyl, phenyl or phenyl-(C 1

-C

4 )-alkyl, for example benzyl, and/or on one or more carbon atoms by (C 1

-C

4 alkyl, for example methyl, phenyl, phenyl-(C,,-C 4 )-alkyl, for example benzyl, halogen, for example chlorine, hydroxyl, (C 1

-C

4 )-alkoxy, for example methoxy, phenyl-

(C

1

-C

4 )-alkoxy, for example benzyloxy, or oxo and can be partially saturated and is, for example, 2- or 3-pyrrolyl, phenylpyrrolyl, for example 4- or 5-phenyl-2pyrrolyl, 2-furyl, 2-thienyl, 4-imidazolyl, methylimidazolyl, for example 1-methyl-2-, 4- or 5-imidazolyl, 1,3thiazol-2-yl, 3- or 4-pyridyl, 1-oxido-2-, 3- or 4pyridino, 2-pyrazinyl, 4- or 5-pyrimidinyl, 3- or 5-indolyl, substituted 2-indolyl, for example 1-methyl-, 5-methoxy, 5-benzyloxy-, 5-chloro- or dimethyl-2-indolyl, 1-benzyl-2- or 3-indolyl, 4,5,6,7tetrahydro-2-indolyl, cyclohepta[b]-5-pyrrolyl, 3- or 4-quinolyl, 4-hydroxy-2-quinolyl, 3- or 4-isoquinolyl, 1-oxo-1,2-dihydro-3-isoquinolyl, 2-quinoxalinyl, 2benzofuranyl, 2-benzoxazolyl, 2-benzothiazolyl, benz[e)indol-2-yl or p-carbolin-3-yl.

Partially hydrogenated or completely hydrogenated heterocyclic rings are, for example, dihydropyridinyl, pyrolidinyl, for example 3- or 4-N-methylpyrrolidinyl, piperidinyl, piperazinyl, morpholino, thiomorpholino and tetrahydrothiophenyl.

SC Salts of compounds of the formula I are to be understood as meaning, in particular, pharmaceutically utilizable or non-toxic salts.

Such salts are formed, for example, from compounds of the formula I which contain acidic groups, for example t carboxyl, with alkali metals or alkaline earth metals, such as Na, K, Mg and Ca, and also with physiologically tolerable organic aminies, such as, for example, triethylamine and tri-(2-hydrox.yethyl)amine.

Compounds of the formula I which contain basic groups, for example an amino group or a guanidino group, form salts with inorganic acids, such as, for example, hydrochloric aid, sulfuric acid or phosphoric acid and with organic carboxylic or sulfonic acids, such as, for example, acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid and p-toluenesulfonic acid.

-6- Preferred sompounds of the formula I are those in which Ra and W are defined on page 1; R 2 denotes i~obutyl, benzyl or cyclohexylmethyl; R 3 denotes hydrogen, -alkyl,, (Cr-C 10 -aryl,

(C

6

-C

10 -aryl- (C 1

-C

4 -alkyl or hydroxyl; R 4 denotes a radical of the formula II, in which

R

5 standa for hydrogeA-,, (Cl-C 4 -alkyl, (Cl-C 4 aJkoxy, (Cl-C 4 -alkylthio, (CI-C 4 -alkylamiro, hydroxyl, azido, fluorine, chlorine,, bromine or iodine, D is defined as the radical Het on page 2, and in denotes 0, 1 or 2; and 0 00 00Q 0 0000 0 A and B independently of one another denote a bivalent radical from the series comprising 0 0 15 phenylalanine, histtdine, tyrsn, trpohn 000 methionine, leucine, isoleucine, asparagine, aspar- 00 tic acid, 86-2-thienylalanine, f-3-thienylalanine, pi- 2-furylalanine, p-3-furxylalanine, lysine, ornithine, valine, alanine, 2, 4-diaminobutyric acid, arginine, 4-chlorophenylalanine, inethionine sulfone, methi- 00 0 onine sulf oxide, 2-pyridylalanine, 4-pyridylalanine, 0040 0 3-pyridylalanine, cyclohexylalanine, cyclohexylglycine, im-melhylhistidine, 0-methyltyrosine, 0- 000 cine, 1-naphthylalanine, 2-:aaphthylalanine, 4nitrophenylalanine, .iorvaline, norleucine, cysteine, 0a 0 S-iethylcysteine, N-iethylhistidine, 1,2,3, 4-tetra- 0. 00 hydroisoqu ino line- 3-c arboxylic acid, homophenylalanine 2-amino-4-(2-thienyl) -butyric acid, 2-amino-4- (3-thienyl) -butyric acid, 3- (2-thienyl) -serine, dehydrophenylalanine, -dehydrophenylalanine, 1,3dioxolan-2-ylalani, N-pyrrolylalanine and 3or 4-pyrazolylalanine, and. their physiologically tolerable salts.

-7- Particularly preferred compounds of the formula I are those in which R a denotes (C 4 'C)-cycloalkyl and (C 4 -C)-'cyc1oalkyl-

(CI-C

2 -alkyl, in which in each case the cycloalkyl substituent can be substituted by one or two identical or different radicals from the series comprising hydroxyl, (Cl-C 4 -alkoxy, (Cl-C 2 -alkyl, carboxyl, (Cl-C 2 -alkoxycarbonyl, carbamoyl, carboxymethoxy, amn, (CI-C 2 -xonoalkylamino, (Cl-CA)-dialkylamino, amino- (C 1

-C

2 -alkyl, (f* 1

-C

2 '-alkylamino- (C 1

-C

2 alkyl, di- (Cl-C 2 -alkylanino- (Cl-C 2 -alkyl, aniidino, trifluoromethyl, (C -C 4 -a 1koxyc arbonyl amino, such as tert .butoxycarbonylamirio,

(C

6

-C

12 -aryl- (Cl-C 2 alkoxycarbonylamino such as benzyloxycarbonylamino and methanesulfonylamino or trifluoromethane- ,~IIsulf onylamino; stands for -CO0-;

*R

2 denotes isobutyl, benzyl or cyclohexylmethyl; R 3 denotes hydrogen or hydroxyl;

R

4 denotes a radical of the formula II, in which too*R staftds for hydrogen or fluorine, tad ora2- -or 4-pyridine radical, a 4- or 5-imidazole radical or a 2-oxazoline radicial, where the heterocycles mentioned can in each case be substituted by one or two radicals Zrom the group comprising methyl, ethyl, propyl, allyl, fluorine, chlorine,bromine, CF 3 and methoxy, and m denotes 0, 1 or 2; and A and B independently of one another denote a bivalent radical from the series, comprising phenylalanine, histidine, tytosine, tryptophan, methionine, leucine, isoieucine, ae ,aragine, aspartic acid, p-2-thienylalanine, p-3-thienylalanine, pig ~imYY~ 1.

8 0 0015 0 0 0 000000 o 0 0 0 0 o020 00 0 0000 o0 0 0000 00 2-furylalanine, lysine, ornithine, valine, alanine, 2,4-diaminobutyric acid, arginine, 4-chlorophenylalanine, methionine sulfone, methionine sulfoxide, 2-pyridylalanine, 3-pyridylalanine, 4-pyridylalanine, cyclohexylalanine, cyclohexylglycine, immethylhistidine, 0-methyltyrosine, O-benzyltyrosine, O-terL butyltyrosine, phenylglycine, l-naphthylalanine, 2-naphthylalanine, 4-nitrophenylalanine, norvaline, norleuci.e, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, homophenylalanine, 2-amino- 4-(2-thienyl)-butyric acid and 3- and 4-pyrazolylalanine, and their physiologically tolerable salts.

The invention furthermore relates to a process for the preparation of compounds of the formula I which comprises coupling a fragment having a terminal carboxyl group or its reactive derivative with a corresponding fragment having a free amino group, optionally removing (a) protective group(s) temporarily introduced for the protection of other functional groups and optionally converting the compound thus obtained into its physiologically tolerable salt.

Fragments of a compound of the formula I having a terminal carboxyl group possess the formulae IIIa, IIIb or IITc below: R -W-A-OH Ra-W-A-B-OH

IE

.4r O 4 E IIIa IIIb SI Ic Fragments of a compound of the formula I having a terminal amino group possess the formulae IVa, IVb or IVc below: 1 -9- ~2 H R

H

2 N-A-B-MNL 12H- CH- CH- R IVa 22 HN- CH- CH- CH- R 4 Methoden which are suitable for the preparation of an bond are~ described, for example, in Houben-Weyl, Metode dj oraniche Chmie(Methods of Organic Chemistry), volume 15/2; Bodanszky et al., Peptide syntheis,2nd ed. (Wiley Sons, New York 1976) or Gross, Meierihofer, The Peptid~es. 2Analysiso synthesis, biology *1O (Academic Press, New York 1979). The following met~hods are preferably usedz active ester method using N-hydroxysuccinimide or 1hydrw~ybenzotriazole as the ester component, coupling with a carbodiimide such as dicyclohaxylcarbodiimide or with propanephosiphonic anhydride and the mixed anhydride method with pivaloyl chloride.

The preparation of the optically active amines used as starting comporuiftd of the formula IVc R2OH

R

out st Rtin active- IV; h in which R 2

R

3 and R 4 are as defined above, is carried asymetrc cnte ofwhih i reaind. or hispurpoise, an N-protected amino acid aldehyde is prepared in a known manner, coupled in an aldol-analogous addition to a corresponding heteroarylalkyl building block and, after removing the N-protective group, gives amino alcohols of 10 the formula IVc. Diastereomeric mixtures relative to the OH-carrying center are obtained which can be separated in a manner known per se, for example !by fractional crystallization or by chromatography. The checking of the diastereomer'ic purity is carried out by means of HPLC, and the enantiomeric purity can be checked in a known manner by converting into Mosher derivatives Mosher et al., J. Org. Chem. 34, 2543 (1969)).

The preparation of N-protected amino acid aldehydes is carried out according to B. Castro et al. (Synthesis 1983, 676).

The aldol-analogous addition to N-protected amino acid aldehydes (preferably N-tert.-butoxycarbonyl and benzyloxycarbonyl protective groups) is carried out in a solvent inert towards bases, such as ether, THF, toluene, DMF, DMSO or dimethoxyethane.

Bases which can be used for the deprotonation of the Sheteroarylalkyl component are alkali metal alcoholates, such as potassium O-tert.-butylate, sodium methylate, alkali metal hydrides, such as sodium hydride or potassium hydride, organometallic bases, such as n-butyl- S lithium, s-butyllithium, methyllithium or phenyllithium, Ssodium amide and alkali metal salts of organic nitrsg@? bases, such as lithium diisopropylamide.

The preceding and subsequent operations necessary for the preparation of compounds of the formula I cuch as intro- S, duction and removal of protective groups are known from 4" the literature and described, for example, in T.W.

Greene, "Protective Groups in Organic Synthesis". Salts of compounds of the formula I with salt-forming groups are prepared in a manner known per se, for example by reacting a compound of the formula I having a basic group with a stoichiometric amount of a suitable acid.

Mixtures of se omer, in particular mixtures of Mixtures of stereoisomers, in particular mixtures of J case the cycloalkyl, bicycloalkyl and tricycloalkyl substituents can be substituted by one or two identical or different radicals from the series comprising F, Cl, Br, I, hydroxyl, (CI-C,)-alkoxy, (C,-Ce)-alkyl, carboxyl, (C-Cs) -alkoxycarbonyl, /2 .4: 11 diastereomers, which are obtained using racemic amino acids A cr B, can be separated by fractional crystallization or by chromatography in a manner known per se.

The compounds of the formula I according to the invention exhibit enzyme-inhibiting properties; in particular they inhibit the action of the natural enzyme renin. Renin is a proteolytic enzyme of the aspartylprotease class whj h, as a result of various stimuli (volume depletion, Aium deficiency, p-receptor stimulation), is secreted by the juxtaglomerular cells of the kidney into the blood circulation. In the latter it cleaves the decapeptide angiotensin I from the angiotensinogen liberated from the liver. This is converted into angiotensin II by the "angiotensin converting enzyme" (ACE). Angiotensin II plays an essential role in the regulation of blood pressure, since it directly increases the blood pressure by meani of vascular contraction. Additionally, it o .stimulates the secretion of aldoste7one from the adrinal S gland and in this manner increases the extracellular fluid volume via the inhibition of sodium excretion, which for its part contributes to an increase in blood pressure. 1i .ibitors of the enzy-'atic activity of renin cause a decreased formation of angiotensin I, which has a decreased formation of angiotensin II as a consequence.

The lowering of the concentration of this active peptide hormone is thu direct cause of the hypotensive action of renin inhibitors.

The activity of renin inhibitors can be checked by in vitro tests. In this connection, the decrease in the formation of angiotensin I is Reasured in various systems (human plasma, purified human renin).

1. Test principle Human plasma, for example, which contains both renin and angiotensinogen, is incubated at 37*C with the compound 12 to be tested. In the course of this, angiotensin I is released from angiotensinogen under the action of renin and can subsequently be measured using a commercial radioimmunoassay. This a1agiotensin release is inhibited by renin inhibitors.

2. Obtaining the plasma The blood is obtained from volunteer subjects (about 1 per person; Bluko sampling device from ASID Bonz und Sohn, U.nterschleiBheim) and collected in partially evacuated bottles with ice cooling. The clotting is prevented by adding EDTA (final concentration 10 mM), 9 After centrifuging (HS 4 rotor (Sorvall), 3,500 rpm, 0- 4°C, 15 min; repeat if necessary), the plasma is care- 0 0 fully pipetted off and frozen in suitable portions at 0°0 15 -30 0 C. Only plasmas having a sufficiently high renin activity are used for the test. Plasmas having low renin o 0 o 0o activity are activated by cold treatment (-4 0 C, 3 days) (prorenin renin).

3. Carrying out the test 0' 0 20 Angiotensin I is determined using the renin Maia@ kit 0 0 0 (Serono Diagnostics Coinsins, Switzerl.and). The O, incubation of the plasma is carried out according to the oo instructions given theret Incubation batch: 1000 Al of plasma (thawed at 0-4°C) 100 pl of phosphate buffer (pH 7.4) I (addition of 10-4 M ramiprilate) p0 of PMSF solution Al of 0.1 Genapol PFIC 12 Al of DMSO or test preparation The test preparations are in general dissolved at 10 2

M

in 100 dimethyl sulfoxide (DMSO) and diluted correspondingly with DY1O; the inuubation batch contains a maximum 13 13 of 1 DMSO.

The batches are mixed in ice and placed in a water bath (37"C) for 1 hour for incubation. A total of 6 samples (in each case 100 1p) are taken without further incubation from an additional batch without inhibitor for tha determination of the starting angiotensin I content of the plasma used.

The concentration of the test preparations are chosen so that approximately the range from 10-90 enzyme inhibition is covered (at least five concentrations). At the end of the incubation time, three 100 pl samples from each batch are frozen on dry ice in precooled Eppendorf o co vessels and stored at about -25°C for the angiotensin I determination (mean value of three individual samples).

0 o 004 Angiotensin I radioimmunoassay (RIA) o I o. The instructions for use of the RIA kit (renin Maia® kit, Serono Diagnostics Coinsins, Switzerland) are followed exactly.

o0oo 0 The calibration curve includes the region from 0.2 to 00 20 25.0 ng of anqiotensin I per ml. The base angiotensin I content of the plasma is taken away from all measured S. values, The plasma ren, activity (PRA) is given as ng of Ang I/ml x hour. PRA values in the presence of the test substances are based on a batch without inhibitor t 100 and given as residual activity. The IC 50 value is read off on the plot of residual activity against the concentration of the test preparation (logarithmic scale).

The compounds of the general formula I described in the present invention show inhibitory actions at concentrations of about 10 5 to 10"0 mol/1 in the in vitro test.

Renin inhibitors cause a lowering of bloo pressure in 14 *salt-depleted animals. Since human renin differs from the renin of other species, primates (marmosets, rhesus monkeys) are used for the in vivo test of renin inhibitors. Primates' renin and human renin are substantially homologous in their sequence. An endogenous effusion of renin is stimulated by i.v. injection of furosemide. The test compounds are then administered and their action on blood pressure and heart rate is measured. The compounds of the present invention are in this case active in a dose range from about 0.1 5 mg/kg and on intraduodenal administration by gastroscope in the dose range from about 1-50 mg/kg. The compounds of the general formula I described in the present invention can be used Sas antihypertensives and also for the treatment of 0oo.o 15 cardiac insufficiency.

c ,aoou 0 3 r° The HIV protease is excised autocatalytically from the 0 0 0 oo GAG-POL polypeptide and then cleaves the precursor o o peptide p5i in the core antigens p17, p 2 4 and p14. It is o° thus an essential enzyme, whose inhibition interrupts the life cycle of the virus and suppresses its reprocation.

0oe In biological tests, it has been shown that the compounds oooo according to the invention have enzyme-inhibitory action °oo and also inhibit viral enzymes such as HIV protease. The ooo HIV protease-inhibiting action has a particular impor- 0" 0 25 tance, which qualifies the compounds according to the invention in particular for the therapy and prophylaxis Sof disorders caused by infection with HIV. The compounds o 0 of the general formula I according to the invention show S0 inhibitory actions at concentrations of about 10-4 to 10 8 mol/l in the in vitro tests used.

The invention furthermore relates to the use of compounds of the formula I for the preparation of medicaments for the therapy of high blood pressure and the treatment of congestive cardiac insufficiency and for the therapy and prophylaxis of viral disorders, in particular disorders which are caused by HIV, and also to the medicaments 15 mentioned.

Pharmaceutical preparations contain an effective amount of the active compound of the formula I together with an inorganic or organic phariaceutically utilizable excipient. Administration can be carried out intranasally, intravenously, subcutaneously or perorally. The dosage of the active compounds depends on the warm-blooded animal species, the body weight, the age and the manner of administration.

The pharmaceutical preparations of the present invention are produced in dissolving, mixing, granulating or tablet-coating processes known per se.

00 00 0 0OO o°oo0 For a form for oral administration, the active compounds o oare mixed with the additives customary therefor such as Q 000 o B 15 excipients, stabilizers or inert diluents and, by custo- 0 0 0 mery methods, brought into suitable forms for oo administration, such as tablets, coated tablets, hard gelatin capsules, aqueous, alcoholic or oily suspensions or aqueous, alcoholic or oily solutions. Inert excipients 0o, 20 which can be used are, for example, gum arabic, magnesia, magnesium carbonate, potassium phosphate, lactose, o 00 o0 glucose, magnesium stearylfumarate or starch, in parti- 000 cular maize starch. The preparation can be carried out both as dry and moist granules here. Suitable oily excipients or solvents are, for example, vegetable or animal oils, such as sunflower oil and cod liver oil.

oo For subcutaneous or intravenous administration, the active compounds or their physiologically tolerable salts are brought into solutions, suspensions or emulsions, if desired with the substances customary therefor such as solubilizers, emulsifiers or other auxiliaries. Possible solvents are, for example: water, physiological saline solutions or alcohols, for example ethanol, propanediol or glycerol, and in addition also sugar solutions such as glucose or mannitol solutions, or else a mixture of the

I

i ~4U

I

16 different so.vents mentioned.

List of the abb.;; _dtiofl5 used: o 0 0 0 0 000 0 4 00 00 4

ACC

Boc BuLi

DCC

DCI

DNP

DME

DMF

EA

El

FAB

h HOBt

MS

MTB

NTE

N-PrpP

THF

4 -aminocyclohexylcarbonyl tert. -butox~ycarbonyl n-butyll1ithiuam dicyc loherylc arbodiimide desorption chemical ionization 2, 4-dinitrqphenyl dimethoxyethane dimethyl formamide ethyl acetate electron impact fast atom bombardment hour 1 -hydroxybenzotriazole molecular peak mass spectrum methyl tert.butyl etler N-ethylmorpholime n-propylphosphonic anhydride tetrahydro furan The other abb7 v.-iat ions u'~ed for amino acids correspond 0 to the three letter code customary in peptide chemistry as is described, for example, in Eur. J. Biochem. 138~, 9- 37 (1984). If not expressly stated otherwise, the amino acids are of the L-configuration.

The eluents used are abbreviated as follows: Ell E12 E13 E14 E16

CH

2 Cl 2

/CH

3 QH 20:1

CH

2 Cl 2

/CH

3 OH 9:1

CH

2 Cl2/CH 3 0H 12: 1

CH

2 Cl 2

/CH

3 OH/conc. NH 3 10:1:0* cyclohexane/ethyl acetate 9:1 n-hexane/ethyl acetate 2:1 The examples below serve to illustrate the present invention without limiting it thereto.

Example 1 N- (N-Tert-butoxycarbonyl-ciB-4-aminocycloh--xylcarbonyl) L-phenylalanine-L-norvaline- (l-S-cyc ioheiqylmethyl-2-S- -pyridyl) )pentylamide 98 mg of HOBt, 130 mg of DCC and 0.08 ml of NEX were added to 235 mg of N-(N-tert-butoxycarbonyl-cis-4-amino-/ cyclohexylcarbonyl)-L-phenylalanine dissolved in 3 ml of DMF. 230 mg of L-norva line-( 1-S-cyc lohexylmethyl- 2-Shydroxy-5-(2-pyridyl))-pentylamide, dissolved in 3 ml of 0 00 DMF, were added to this solution and the mixture was stirred overnight at RT. The DMIF was distilled off in a high vacuum, the solution remaining was taken up in ethyl acetate, precipitated urea was filtered off and the 0 0 2 solution, and the organic phase was dried over IMgSO 4 and concentrated in vacuo. 315 mg were isolated by column chromatography (silica gel, Ell).

Melting point 100-16~3 0

C

MS (FAB) 748 (M+l) 04 a) N- (N-Tert -butoxyc arbonyl -cis -4 -aminocyc lohexyloarbonyl) -L-phenylalanine 5.5 g of N- (N-tert-butoxycarbonyl-cis-4-aminocyclohexyl-] carbonyl)-L-phenylalanine benzyl ester were hydrogenated 0 in 230 ml of ethanol over 1 g of Pd/carbon under normal.

pressure. After completion of the reaction, the catalyst was filtered off and 'the solvent was removed by distillation. 4. 1 g of colorless product were obtained af ter xecrystallization from n-heptane/ethyl acetate.

Melting point 160-161*C (decomposition) MS (DCI) 391 (M+1) -4 H b) N- (N-Tert -butoxycarbonyl-cis-4-aminocyclohexylcarbonyl) -L-phenylalanine benzyl ester g of N-tert-butoxycarbonyl-cis-1,4-aminocyclohexanecarboxylic acid and 6.3 g of L-phenylalanine benzyl ester were dissolved in 75 ml of DMF and mixed with 24 ml of nprop PA and 15.7 ml of NEM at 0 0 C and the mixture was made to react overnight at RT. The solution was diluted with CHC1 2 and washed in each case with semisaturated NaHCO 3 solution, 10 strength citric acid and water, dried over MgSO 4 and concentrated in vacuo. Flash chromatography gave 5.7 g of pure product.

-14.6 (c 1.1 in CH 3

OH)

o 00 00 0 040004 00 00 Example 2 0 5 N- (cis-4-Aminocyclohexylcarbonyl) L-phenylalanine-L- 15 norvaline-(1-S-cyclohexylmethyl-2-S-hydroxy-5-(2-pyri- 0 dyl) -pentylamide 00 0 0404 77.7 mg of the compound described in Example 1 were dissolved in 2 ml of CH 2 Cl 2 and 2 ml of CFaCOOH were added :0 at OOC under N 2 The solution was concentrated after A 20 30 min, the residue was taken up in ethyl acetate and washed twice each time with satd. NaHCO 3 and NaC1 solution.

Column chromatography (silica gel, Ell) gave 24.7 mg of product.

MS (FAB) 648 (M+1) -19- Example 3 N- (N-Tert-butoxycarbonyl-trans-4-aminocyclohexy1carbonyl) -L-phenylalanine-L-norvaline- 1-S-cyclohexylmethyl- 2-S-hydroxy-5-(2-pyridyl) )pentylamide Analogously to the process described in Example 1, the title compound was prepared from N-(N-tert-butoxycarbonyl-trans-4-aminocyclohexylcarbonyl) -L-phenylalanine V and L-norvaline-( 1-S-cyclohexylmethyl-2-S-hydroxy-5-(2pyridyl))pentylaiide. The yield 'was 270 mg.

Melting point: 219-220*C.

MS (FAB) 748 (M+X) a) N- (N-Tert-butoxycarbonyl -trans -4 -aminocyc lohexylc arbonyl) -L-phenylalanine 444.45 g of N-(N-tert-butoxycarbonyl-trans--4-aminocyclohexylcarbonyl)-L-phenylalanine benzyl ester were catalytically hydrogenated under normal pressure over 900 mg of palladiun-cairbon in 200 ml of ethanol at After completion of the reaction, the catalyst was filtered off and the solvent was removed by distillation in vacuo.

2.67 g of pure product were obtained after 4444 recrystallization.

Melting point: decomposition from 160 0

C

MS (DCI): 391 b) N- (N-Tert-butoxycarbonyl-trans-4-aminocyclohexylcarbonyl) -L-phenylalanine benzyl ester V3 ?ofi: acid and 3.2 g rof l-phenyl4aain eylestner 3abxyi gci ofd 3-trtbutgyof hnyla4aninoceylheaner weedissolved in 40 ml of DMF and 7.85 ml of NEM and the 12mlof n-propPA were added at 0 0 C. The mixture was mdtoreact overnight at RT. After diluting with 200 ml of ehylacetate, the solution was washed twice each with H20110 %strength citric acid, satd. NaHCO 3 and satd.

Nalsolution. The remaining organic phase was dried uigMgSO 4 and freed from the solvent in vacuo. The crude product was triturated with diethyl ether, and the precipitated crystals were filtered off with suction and dried. 4.45 g of colorless crystals 'ere obtained.

Melting point: 176-177 0

C

MS (DCI) 481 (M+1) Example 4 N-(N-trans-4-Aminocyclohexylcarbonyl)-L-phenylalanine-Lnorvaline-(l-S-cyclohexylmethyl-2-S-hydroxy-5-(2-pyridyl))-pentylamide The title compound was prepared by the process described in Example 2 from the compound described in Example 3.

.0 Yield after chromatography (silica gel, E12) 27 mg.

,00n MS (FAB) 648 (M+1) RF 0,17 (CH 2 Cl 2

/CH

3 0H 8:2) F2 2 0 Example N-(N-Tert-butoxycarbonyl-trans-4-aminocycloheylcarbonyl)-L-phnylalanine-N(im)-trityl-L-histidine-( 1-Scyclohexylmethyl-2-S-hydroxy-5-(2-pyridyl))pentylamide The title compound was prepared from the phenylalanine derivative described in Example 3a) and from FMOC-trityl- His-(l-S-cyclohexylmethyl-2-S-hydroxy-5-(2-pyridyl))pentylamide by the process described in Example 1.

Yield after chromatography (silica gel, Ell) 0.4 g MS (FAB) 1028 1034 R 0,5 (toluene/ethanol 8:2) F .i_ 21 Example 6 N-(N-Tert-butoxycarbonyl-trans-4-aminocyclohexylcarbonyl)-L-phenylalanine-L-histidine-(1-S-cyclohexylmethyl- 2-S-hydroxy-5-(2-pyridyl))pentylamide 40 mg of the substance obtained in Example 5 were dissolved i- 5 ml of 90 strength acetic acid aiid the mixture was heated to 60"C for two hours. The solution was adiuted to pH 8 using Na 2

CO

3 and extracted three times using ethyl acetate. The organic phase was washed twice each with satd. NaHCO 3 solution and with satd. NaC1 solution, then dried over MgSO 4 and concentrated in vacuo.

Chromatography (silica gel, Ell) gave 16 mg of pure S*product. 155'C decomposition o MS (FAB) 786 (M+1) Example 7 "N-(trans-4-Aminocyclohexylcarbonyl)-L-phenylalanine-Lhistidine-(l-S-cyclohexylmethyl-2-S-hydroxy-5-(2-pyridyl))pentylamide 300 mg of the substance described in Example 5 were 0 20 stirred for 15 min at 0 0 C and for 2 h at RT with 5 ml of

CF

3 COOH and 0.25 ml of H 2 0. The solution was concentrated 4 in vacuo, and the residue was -endered alkaline with 44 NaHCO 3 solution and extracted using ethyl acetate. The organic phase was concentrated in vacuo and the residue was chromatographed (silica gel, E12). MeXting point 114-1166C.

MS (FAB) 686 (M+1) Example 8 N-(N-Tert-butoxycarbonyl-cis-4-aminocyclohexylcarbonyl)- L-phenylalanine-N (im) -trityl L-histidine- 1-S-cyclohexylmethyl-2-S-hydroxy-5-(2-pyridyl))pentylamide The title compouna was prepared from the phenylalanine

I,

~4 U

I

22 derivative described in Example la) and from Fmoc-trityl- His-(l-S-cyclohexylmethyl-2-S-hydIroxy-5-(2-pyridyl)pentylamide by the process described ifl Example 1. Yield after chromatography (silica gel, Ell) 0.5 g.

MS (FAB) 1,028 1034 R F0,26 (El 1) Example 9 N- (N-Tert-butoxycarbonyl-cis-4-aminocyclohexylcarbonyl) L-phenylalanine-L-histidine- -S-cyclohexylmethyl-2-S- (2-pyridyl) )pentylamide The title compound was prepared by the proceso described in Examnp1- 6 from the compound described i~n Example 8.

900 MS (FAB) 786 (M+1) 00V Iyield: 61 mg. Melting poinit 127-130C. RF 0,21 (El 2) Example ri ao; N-(cis-4-Aminocyclohexylcarbonyl)-L-phenylalanine-L~.histidine-( l-S-cyclohexylmethyl-2-S-hydroxy-5-(2-pyri.

dyl))pentylamide The title compound was prepared by the process described 0 in Example 7 from the compound described in Example 8.

4 Yield 180 mg. Melting point 1770C(decomp.) MS (FAB) 686 (M+1) Example 11 N- (N-Tert-butoxycarbonyl-trans-4-aminocyclohexylcarbonyl methyl-L-tyrosine-N- (im) -trityl-L-hJistidine- (1- S -cyc lohexylmethyl4=2 -S-hydroxy-5 -pyridyl) )pentylamide The title compound was prepared from N-(N-tert-butoxycarbonyl-trans -4 -aminocv.r lohexylcarbonyl) -methyl-L-tyrosime and trityl-L-His- -S-cyelohexylmethyl-2-S-hydroxy-5- (2pyridyl))pentylamide by th~s process described in Example 1R 0,54 (toluene/etha7.ol 8:2) X; (fAB) 1058 (M+1) -23- Example 12 N- (N-Tert-butoxycarbonyl-trans-4-amiflocyclohexylcarbonyl )-O-methyl-L-tyrosine-L-hist-dile (l-$-cyclohexy1- (2-pyridyl) )pentylamiA'de The title compound was prepared by the process described in Example 6 from the compound described in Example 11.

MS (FAB) 816 180 0 C decomposition; R F 0,2 (toluene/ ethanol 8:2) Exa!Tple 13 N- (trans -4 -Aminocycl1ohr,,yl carbonyl) -O-methyl tyroP ine- L-histidine (l-S-cyclohexylmethyl-2-S-hydroxy-5-(2pyridyl) )pentylamide The title compound was prepared by the process described in Example 7 from the compound described in Example 11.

MS (FAB) 716

G

0xmle1 o 0Tr-uoxcroy-i-4aioylheycroy) 14tepoes eciedi xml 0. RVI7 E MS(FB)158(Ml IJi 24 Example N- (N-Tert-butoxycarbonyl-cis"4-aminocyclohfxylcarbonyl) 0-methyl-L-tyrosine-L-histidine (l-S-cyclohexylmethY-l- 2- S-hydroxy-5-(2-pyridyl))pentylamide The title compound was prepared by the process described in Examuple 6 from the compound described in Exaymple 14.

MS (FAB) 916 RF 0,80 (El 4) Example 16 N- (cis-4-Aminocyclohexylcarbonyl) -0-methyl-L-tyrosine-Lhistidine (1-S-cyclohexylmethyl-2-S-hydroxy-5-(2pyridyl))pentylamide The title compound was prepared by the process described 0 in Example 7 from the compound described in Example 14.

MS (FAB) 716 RF 0,50 (El 4)

~F

0 0 0 0 Example 17 N-(14-(N-(cis-4-Tert,butxycabonylamino-ylheycr hexyl-3S,4Rdihydroxy-n-hdxyl)-pyridine mg of the compound from Example 17 a) are stirred with 40 v~g of thiophenol in 2 wl of' acetonitrile for: 2 h.

After co~ncentra~ing, the rosidtue is ohromt~ographed (silica gal, 214) and 45 mg are obtaind as an Amorphous powdr.

MS (FAB) 802 000 0 Human plasma, for example, which contains both renin and angiotensinogen, is incubated at 37*C with the compound a) flOC-CiS-CC-Phe-DNP-His-2- (5-amkino-6 -cyclohexy.",'- 3S, 4R-dihydroxy-n-hexyl )pyridine mg of BOC-DNP-.His-(3S,4R,5S)-2A-(5-amtino-6-cyclohexyl- 3,4-dihydroxyhexyl~pyridine are htirred in 5 ml of DME/HCl for 2 hi. After concentrating, the crude product is dissolved in 3 ml of DMF together with 50 mg of N-(Nte-rt .butoxycarboniyl-cis-4-aminovrclohexyicarbonyl phenylalanine, 120 iag of DCC and 80 mg of I"OBt. The solution is adju~sted to pH 5) using NEM and stirred for 10 h. The bolutio 11,s filtered, diluted with ethyl -cetate and wzashed on-e each with 3 strength NaHCO 3 solution., H 2 0 and sacd. NeiCl solution, dried using MgSO41 concentrated and chromati,-graphe1 on silica gel (E13).

mg of the title compound are obtained as a yellow MS(FA3) 968 (14+1) b) BOC-DNP-His. ,3S,4R,5S)-2-(5-amino-6-cyctohexyl-3,4dihydroxyhexyl )pyridine 0. 5 xmol of the (3S, 4Rl,5S) is'umer f rom Example 17 c are stirred with 5 ml oil HCl in DME (saturated) for 2 h.

After concentrating in vacuo, the residue is dissolved in 3ml of absolute D1M'. 0.5 mmol each of BOC-DNt--His-OH, dicyclohexyicarbodiimide and HOBt are added. The solution ,s adjusted to pH 9 using NEI4 and stirred for 24 h. After filtration, it is diluted with EA and washed with each 3 strength XaHCO, solution, dried usj* 3 MgSO 4 and concentrated. Chromatography on silica gel (Eli) gives the title compound as a yellow resin.

MS (FAB) 696 (M+1) c) 2-((3S,4R,5S)-5-tert.butoxcycarbonylamino-3,4-dihr--roxy- 6-cyc lohexyl-n-hexyl )pyridine 1.4 ml (1 mzrtol) of n-BuLi are added at -78*C to 93 mg (1 mmol) of 2-picioline in 10 ml of THF. After warming to RT, the mixture is stcirred for 30 min, then cooled to 26 0 C. 1 mmol of (2RS,3R,4S)-3-tert.-butyldilethylsilylox-y-4-tert.-butoxycarbonylamino-5-cyclohexyl-1,2-oxopentane (known from EP-A 189,203, Example 6) are added (dissolved in 5 ml of THF). After 10 hours at RT, the mixture is diluted with water and extracted using MTBE.

The crude product (0.4 g) is dissolved in THF and stirred at OOC with 5 ml of a 1M solution of tatrabuty.am" noni um fluoride in THF for 1 h After diluting with water and extracting with EA, 0.15 g of the (3S,4R,5S)-isomer (MS (FAB): 391 and 0.±2 g of the (3S,4S,5S)-isomer (MS (FAB): 391 are obtained.

Example 18 cis-4-Aminocyclohexylcarbonyl-L-Phe-L-His-(l-S-cyclohexylmethyl *2R, 3S-dihydroxy-5-(2 -pyridyl)) -n-pentylamide tristrifluoroacetate 0 25 mg of the compound described in Example 17 are stirred 00 00 o i 1 ml of TFA/1 ml of CHzC1 2 for 2 h. After concentrating, the residue is taken up in H2O and lyophilized. 20 mg of product are obtained as a colorless powder.

MS (FAB) 702 (14+l) 0 0 00 Example 19 BOC-trans-ACC-L-Phe-L-His- (1S-cyclohexylmethyl-2R,3dihydroxy-5-(2-pyr! 3yl) )-n-pentylamide The title compoiund was prepared by the process described in Example 17 from the compound described in Example 19a.

MS (FAID) 802 Yield: 79,5 mg. RF 0,19 (El 2) a) BOC-trans-ACC-L-Ph--L-DNP-Hi 1S-cycJIohexylmthyl- 2Rg3S-dihydro,"yi- 5-(2-pyridyl))-n-pentylamide The title compound was prepared by the process described -f -27in Example 17 a) f rom. the compounds described in Examples 3 a) and 19 R F 0,40 (El 2) MS (FAB) 968 (M+1) b) DNP-H-His-(3S,4R,5S)-2-(5-amino-6-cyclohexylN3,4dihydroxy-n-hexyl) -pyridine hydrochloride 400 mg of the compound from Example 17 b) are stirred in ml of DME/HC7. (satd.) for 2 h. After concentrating, the residue is taken up in toluene twice and in each case concent-ated again. The title compound is obtained as a yellow foam.

MS (FAB) 596 (M+l) Example 0 04 0 a 4 trans -ACC-L-Phe-L-His -(1S-cyclohexylmethyl-2R, 3S-dihydroxy- 5-(2 -pyridyl) -n-pentylamide o 00 a 0 15 The title compound was preparedX by the pro.-ess described 0 in Example 18 from the compound described in 4 Example 19.

MS (FAB) 702 Yield: 49 xng Example 21 0 BOC-trans-ACC--methyl-L-Tyr-L-His- iS -cycltohexylmeothyl- 2 R, 3 S -dihydro xy- 5-(2 -p7 -d dyl) n-pentyl1amide The title compound was prepared by the procesz. described in Example 17 from the com~pound described in Example 21 J a).

MS (FAB) 832 (M+1) a) BOC-trans-ACC-O'-methyl-L-Tyr-DNP-L-His- IS-cyclohexylmethyl-2R, 3S-dihydroxy-5- 2-pyridyl) n-pentylamide The title compound was ptcepared by the process described in Example 17 a) f rom thO compound, ~scribed in Examples -28- 21 b) and 19 b).

MS (FAB) 998 (M+1) b) BOC -trans -ACC -Tyr (OCH 3

-OH

0.05 mol of BOC -trans -ACC -Tyr (OCHO)- OCH 3 are dissolved in 250 ml of DM.E and the mixture igs stirred with an equimolar amount of 1N NaOH solution for 5 h. The solution is then concentrated and the residue is taken up with water, acidified to pH 4 using KHS0 4 and extracted with EA. The crude product is recrystallized from n-heptane/EA and the title compound is obtained as ai white powder.

MS (DCI) 421 .Melting point: 1810C(decomp.) R F 0,20 (toluene/ethanol 8:2) c) BOC trans -ACC -Tyr (OCH 3

-OCH

3 0 0. 1 mol of BOC -trans -ACC -OH and 0. 1 mol of L-tyrosine- 0 (OCH 3 methyl ester are dissolved in 500 ml of DMF and o all propPA (50 strength in C'1 2 Cl 2 and 0.5 niol of trielthyl- 000 0amine. The solution is made to react overnight. After completion of the reaction (TLC checking), the solvent is removed by distillation in a high vacuum and the residue is taken up in The organic phase is washed twice each 0000 with 10 strength citric acid, water,, semisaturated 6)060 U NaHCO 3 solution and satd. NaCl solution, dried over MgSO4 00'4 and concentrated in vacuo, and the residue is chromatographed (silica gel, Melting point: 150-155 *C.

MS (DCI) 435 (M+1) 00 00 Example 22 trans-AC'7-O-methyl-L-Tyr-L-His-( iS-cyclohexylmethyl- 2R, 3S-dihydroxy-5- (2-pyridyl) )-n-1p ,antylamide The title compound was prepared by the process described in Example 18 from the compound described in Example 21.

MS (FAB) 732 (MI) -29 Example 23 BOC-cis -ACC-O-methyl-li-Tyr-L-His- 1S-cy( ,lohexylmet.,-'l- 2Rf 3S-dihydroxy-5- (2-pyridyl) )-n-pentylamide The title compound was prepared by the process described in Example 17 from the compound described in Example 23 RF 0,21 (El 2) MS (FAB) 832 (M+1) a) BOC-cis-ACC-O-methyl-L-Tyr-DNP-L-His- S-cyclo-_ hexylmethyl-2R, 3S-dihydroxy-5- 2-pyridyl) -n-pentylamide The title compound was prepared by the process described *off#in E'xample 17 a) from the compounds described in Examples 23 b) and 19 b).

of MS (FAB) 998 (M+1) 04 15 b) BOC-cis-ACC-Tyr(OCH 3

)-O

t4 The title compound is obtained analogously to the process Example 21 z425- 18,6 0 CH OH) MS (DCI) 421 (M+1) c) BOC-cis-ACC-Tyr(0CH 3 methyl ester The title compound was prepared analogously to the process in Example 21 c).

MS (DCI) 435 (M+1) Example 24 cis-ACC-O-methyl-L-Tyr-L-His- S-cyclohexylmethyl-2R, 3Sdihydroxy-S-(2-pyridyl) )-n-pentylamide The title compound was prepared by the process described in Example 18 from the compound described in Example 23.

30 MS (FAB) 732 RF 0,56 (El 4) Example BOC-cis-ACC-Phe-His- (lS-cyclohexylmethyl-2S-hydroxy-5- (Npropyl-2-imidazolyl))pentylamide The title compound was prepared by the process described in Example 17 from the compound described in Example MS (FAB) 817 (M+1) a) BOC-cis-ACC-Phe-DNP-His-(lS-cyclohexylmethyl-2S- (N-propyl-2-imidazolyl))pentylamide 80 mg of the compound from Example b are stirred with 4 ml of HCl in DME (satd.) for 2 h. After concentrating ^a in vacuo, the reaidue is dissolved in 3 ml of absolute DMF. 44 mg of BOC-cis-ACC-Phe-OH and 0.11 mmol each of oo DCC and HOBt are added. The solution is adjusted to pH 9 using NEM and stirred for 24 h. After filtering, it is diluted with EA and washed twice each with 3 strength NaHCO 3 solution, H20 and satd. NaCI solution, dried using MgSO 4 and concentrated. Chromatography on silica gel gives the title compound as a yellow resin.

MS (FAB) 983 (M+1) 4 4 b) BOC-DNP-His- propyl-2-imidazolyl) pentylamide 100 mg of the compound from Example c are stirred with ml of HCl in DME (satd.) for 2 h. After concentrating in vacuo, the residue is dissolved in 3 ml of absolute DMF. 105 mg of BOC-DNP-His-OH and 0.3 mmol each of DCC and HOBt are added. The solution is adjusted to pH 9 using NEM and stirred for 24 h. After filtration, it is diluted with EA and washed twice each with 3 strength NaHCO 3 solution, H20 and satd. NaCl solution, dried using MgSO 4 and concentrated. Chromatography on silica gel gives the title compound as a yellow resin.

31 MS (FAB) 711 (M+1) c) 3-BOC-4S-cyclohexylmethyl-2,2-dimethyl-5-(3-(2-Npropylimidazolyl)-propyl)oxazolidine 500 mg of the compound from Example d are dissolved in 5 ml of absolute THF. Under an argon atmosphere, 1.9 ml of a 1.5 molar solution of n-butyllithium in n-hexane are added at -60 0 C. After 15 min, 500 mg of the compound from Example e dissolved in 5 ml of absolute THF are added.

After 1 h at -60 0 C, 10 ml of a satd. NaHCO 3 solution are added. After warming to RT, the mixture is extracted three times with EA, dried using NazSO 4 and concentrated.

The title compound is obtained by chromatography on silica gel (El EA).

R(f) 0.3 MS (FAB) M+1) d) 2-Methyl-N-n-propylimidazole g of 2-methylimidazole are heated to boiling in 40 ml of n-propylbromide for 2 h. The solution is then poured into 150 ml of a 5 strength NaHSO0 4 solution, and the mixture is extracted three times using EA, dried using NaS0 4 and concentrated. The residue is taken up in ether and separated from insoluble startinq material by filtration. In this manner, the title compound is obtained as a colorless oil.

R(f) 0.2 (EA/CH 3 OH 10:1) e) 3-Boc-cyclohexylmethyl-2,2-dimethyl-5-(2-bromoethyl)oxazolidine 1.6 ml of diethyl azodicarboxylate are added dropwise at under argon to 690 mg of 3-BOC-1S-cyclohexylmethyl- 2,2-dimethyl-5-(2-hydroxyethyl)oxazolidine, 2.6 g of triphenylphosphine and 1.6 g of pyridinium bromide in ml of CH 2 Cl 2 After 16 h at RT, water is added and the mixture is diluted using 100 ml of CH 2 C1 2 The organic phase is washed twice with satd. NaHCO, solution and once '4 -32with NaCI solution. The organic phase dried with Na 2 SO is concentrated, and the residue is taken up in a little EA and filtered to separate PPh 3 Purification on silica gel yields the title compound (E16).

R(f) 0.3 (E16); MS 404 f) 3-BOC-4S-cyclohexylmethyl-2,2-dimethyl-5-(2-hydroxyethyl) oxazolidine g of BOC-ACHPA-OC 2 H, (preparation according to J. Med.

Chem. 28, 1779 (1985)), 500 mg of p-toluenesulfonic acid and 7.2 ml of dimethoxypropane are heated at 80 0 C in 160 ml of toluene under argon for 2 h. The mixture is then concentrated. The residue is added dropwise at 0 C to a suspension of 2 g of LiAlH 4 in 200 ml of THF. After as 2.5 h at 0 C, 100 ml of 5 strength NaHS04 solution are -015 added and the mixture is extracted three times with EA.

O"o The combined organic phases are washed with satd. NaHCO 3 o solution. After drying using Na 2

SO

4 the mixture is Sc concentrated and chromatographed on silica gel (E16).

o R(f) 0.1 (E16); MS (DCI) 342 Example 26 cis-ACC-Phe-His- (S-cyclohexymeylmethyl-2S-hydroxy-5- (Npropyl-2-imidazolyl) )pentrlamide The title com'pound was prepared by the method described in Example 18 from the compound in Example MS (FAG) 717 (M+1) h il opon a rprdb temto ecie

Claims (5)

1. A compound of the formula I R2OH R in which Ra denotes (C 3 -cycloalkyl, (C 4 -C 10 -bicycloalkyl, (CO-C 1 2 -tricycloalkyl, (C 3 -Ca) -cycloalkyl- (C 1 -C 6 alkyl, (C 4 -bicycloalkyl- -alkyl and (C.-C 1 2 -tricycloalkyl- -alkyl, in which in each case the cycloalkyl, bicycloalkyl and tricycloalkyl 0 '0,substituents can be substituted by one or two identical or different radicals from the series comprising F, Cl, Br, I, hydroxyl, (Cl-C 6 )-alkoxy, 0a (Cl-C6) -alkyl carboxyl, -alkoxycarbonyl., 0 0 of 0 0 carbamoyl, carboxymethoxy, amino, (C 1 -monoalkyl- a amino, (C 1 -C&)-'dialkylamito, amino- (C 1 -C 6 -alkyl, C 6 -alkylamino- (C 1 alkyl1, di- (Cl-C 6 -alkylamino- (Cl-CO)-aJlkyl, amidino, hydroxamino, hydroximino, hydrazono, imino, guanidino, (C-C 6 -alkyloxy- o 00 0 sulfonyl, (Cl-C 0 -alkyloxysulfenyl, trifluoromethyl, 00(Cl-CA) -a lkoxyc arbonyl amino or (C 6 -Cj 2 -aryl- (C 1 -C 4 00 alkoxycarbonylamino; W stands for -0-Ca- or -SO 2 R2 Rdenotes hydr~ogen, (C 1 -C 1 )-alkyl, -cycloalkyl, 8(C 4 -C 7 -cycloalkyl- (C 1 -C 4 -alkyl, (C 6 ,-ClA) -aryl or (Ce-C,4 -aryl- (Cl-C 4 alkyl, R 3 denotes hydrogen, (C- 1 0 -alkyl, (C-Cl) -aryl, (Ce-Cj4) -aryl- (Cl-CA) -alkyl, hydroxyl or amino, R 4 denotes a radical of the formulaII (CH 2 ),-CHR 5 -D (II) -34 where R 5 stands for hyrdrogen, (C 1 -C 7 -alkyl, (C,-C 5 alkoxy, (C 1 -C 5 dkyithic, -alkylamino, hydroxyl, azido, fluorine, chlorine, bromine or iodine, D denotes a radical Het, where Het denotes a 5- to 7-meinbered heterocyclic ring which can be fused to benzene, aromatic, partially hydrogenated or completely hydro- genated, which can contain one or two identical or different radicals from the group N, 0, S, NO, SO or SO 2 as heteroatoms and which can be substituted by one or two identical or dif- ferent radicals from the group comprising (Cl-C 4 ~-ly,(C 1 -C 4 -akxhydroxyl, halogen, 0 amino, mono- or di- (C 1 -C 4 -a lkyl amino or CF., and 0m denotes 0, 1, 2, 3 or 4; 0 0 000 0 A and B independently of one another denote an amino acid linked at the nitrogen terminus to Ra-W or A and at the carbon terminus to B or NH-CHR 2 CHOH-CHR3-R 4 froim $1 0*00 0000 the series comprising phenylalanine, histidine, o tyrosine, tryptophan, methionine, leucine, iso- leucine, asparagine, aspartic acid, p-2-thienylala- 0000 nine, p-3-thienylalanine, f-2-furylalanine, p-3- furylalanine, lysine, ornithine, valine, alanine, 2,4-diaminobutyric acid, arginine, 4-chiorophenyl- 00 0ID 0 alanine, methionine sulfone, methionine suif oxide, 00 2-pyridylalanine, 3-pyridylalanine, cyclohexyl- 0 0 alanine, cyclohexyiglycine, im-methyihistidine, 0- methyltyrosine, 0-benzyltyrosine, 0-tert. -butyl- tyrosine, phenyiglycine, 1-naphthylalanine, 2- naphthylalanine, 4 -nitrophenylalanine, norvaline, pi-

2-benzo(b]thienylalanine, p-3-benzo(bjthienyl- alanine, 2-f luorophenylalanine, 3-f luorophenyl- alanine, 4-fluorophenylalanine, norleucine, cysteine, S-methylcysteine, 1,2,3, 4-tetrahydroiso- -44 quinoline-3-carboxylic acid, homophenylalanile, DOPA, 0-dimethyl-DOPA, N-methylhistidine, 2-amino-

4-(2-thienyl)-butyric acid, 2-amino-4.-(3-thienyl)- butyric acid, 3- (2-thienyl) -serine, (Z )-dehydro- phenylalanine, -dehydrophenylalanine, 1, 3-dioxo- lan-2-ylalanine, N-pyrrolylalanine and 3- or 4- pyrazolylalaie, and~ their physiologically tolerable salts. 2. A compound as claimed in claim 1, wherein R' and W are as defined in claim 1, R denotes hydrogen, (C-Col-alkyl,(C-)ccoly, (C 4 -C 7 -cycloalkyl- (Cl-C 4 -alkyl, -aryl or 0 a R 0 R 3 denotes hydrogen, (Cl-C 1 0 -alkyl, (C6-C 1 4 -aryl (C.-C 14 -aryl- (Cl-C 4 -alkyl hydroxyl or amino, -0 0 R~ denotes a radical of the formula II, in which stands for hydrogen, (C 1 -C 4 )-alkyl, (Cl-C 4 alkoxy, (C 1 -C 4 -alkylthio, (C.-CO )-alkylamino, 0 0 hydroxyl, azido, fluorine, chlorine, bromine or 0 iodine, 00 a D is def ined as the radical Het in claim l and mn denotes 0, 1 or 2; 00 4 A and B independantly of one another denote a bivalent radical from the series comprising phenylalanine, 0 histidine, tyrosine, tryptophan, methionine, leucin- e, isoleucine, asparagine, aspartic acid, o3-2-- thienylalanine, p-3-thienylalanine, p-2-furylalan- mne, p-3-furylalanine, lysine, ornithine, valine, alanine, 2,4-diaminobutyric acid, arginine, 4- chlorophenylalanine, methionine suifone, iethionine sulfoxide, 2-pyridylalanine, 4-pyridylalanine, 3- pyridylalanine, cyclohexylalanine, cyclohexyJlglycin- e, im-methylhistidine, 0-methyltyrosine, O-benzyl- tyrosine, 0-tert. -butyltyrosine, phenylglycine, I- .4 -36- naphthylalanine, 2-,naphthylalanine, 4-nitrophenylal- anine, norvaline, fl-2-benzo[bjthienylalannie, fl-3- benzo thienylalanine, 2-f luorophenylalanine, 3- fluorophenylalanine, 4-f luorophenylalaiine, nor- leucine, cysteine, S-mw-thylcysteine, 1,2,3, 4-tetrah- ydroisoquinoline-3-carboxylic acid, homophenylalani- ne, DOPA, 0-dimethyl-DOPA, N-methylhistidine, 2- amino-4-(2-thienyl) -butyric acid, 2-amino-4-(3- dehydrophenylalanine, -dehydrophenylalanine, 1,3- dioxolan-2-.ylalanine, N-pyrrolylalanine and 3- or 4-pyrazolylalanine, and their physiologically tolerable salts. 004.: 3. A compound as claimed in claim 1 and/or 2, wherein R' denotes (C 4 -C 6 -cycloalkyl and (C 4 -C 6 -cycloalkyl- 4 (C- 2 )-alkyl, in which in each case the cyclo- alkyl substituent can be substituted by one or 42 two identical or differe~nt radicals from the series comprising hydroxy., (Cl-C4)-alkoxy, (C 1 C 2 -alkyl carboxyl, (Cl-C 2 -alkoxycarbonyl, carbamoyl, carboxymethoxy, amino, (C 1 -C 2 )-mono- 0, oo alkylamino, (C-C 2 -dialkylamino, amiy -tC 1 -Cg)- alkyl, (C 1 -C 2 -a lkyl amino- (C 1 -C 2 al kylI di- (C 1 -C 2 alkylamino-(C-C 2 -alkyl, amidino, trifluoro- methyl, (CI-CA) -a2koxycarbonylamIno such as tert .butoxycarbonylamino, (C 6 0-CU) -aryl- alkoxyc arbonyl amino such as benzyloxycarbonyl- amino and rathanesulfonylamino or trifluoro- methanesulfonylamlno; W stands for -Co-; R 2 denotes isobutyl, benzyl or cyclohexylmethyl; R 3 denotes hydrogen or hydroxyl; R 4 denotes a radical of the formula 11# in~ which R 5 stands for hydrogen or fluorine, D stands for a 3- or 4-gyridine radical, a 4- or 5-imidazole radical or a' 2-oxazoline radical, where mentioned can in each case be aubiti.tu'ted by one or two radicals from the group comprising methyl, methoxy, ethyl, propyl, allyl, fluorine, chlor- ine, bromine and CF 3 and m Qd')notes 0, 1. or 2; and A and B independently of one another denote~ a bivalent radical from the series comprising phenylalanine, histidine, tyrosinep, tryptophan, methioniner leucine, isoleucine, asparagine, aspar- tic acid, p-2-thienylalanine, p-3-thienylalanine, p- 2-furylalanine, lysine, ornithine, valine, alanine, o 2,4-diaminobutyric acid, a _ginine, 4-chiorophenyl- alanine, methionine sulfono, methionine sulfoxide, 2-pyridylalanine, 3-pyridylalanine, 4-pyridylala,- nine, cyclohexylalanine, cyclohexylglycine, im- methylhistidine, I0-methyltyrosine, O-benzyltyrrosine, 0-tert butyltyrosine, phenylglycine, 1-naphthylala- nine, 2-naphthylalanine, 4-nitrophenylalanine, 7iorvalinte, norleucine, 1,2,3,4-tetrahydroisoquino- line- 3-carboxylic acid, homophenylalanine, 2-amino- 4-(2-thienyl)-butyric acid and 3- and 4-pyrazo.- 4 ylalanine, and their physiologically tolerable salts. 4. A process for the preparation of a compound of the formula X as claimed in one or more of claims 1 to 3,, which comprises coupling a fragment having a terminal carboxyl group or its reactive derivative with a corres- ponding fragment having a free amino group, optionally removing protective group(s) temporarily introduced f or the protection of other functicnal groups and option- ally converting the compound thus obtained into its physiologically tolerable salts. .4 38 A compound of the forn"ia I as claimed in one or more of claims 1 to 3 for use as a medicament.

6. A compound of the formula I as claimed in one or more of claimr, 1 to 3 for use as a medicament in the treatment of high blood pressure.

7. A pharmaceutical, preparation c a compound of the formiula I as claimed in one or more of claims 1 to 3. DATED THIS 19Lh day of April, 1990 0 0 HOECHST AKTI ENGESELLSCHAF7 WATERMARK PATENT k'x TRADEMARK ATTORNEYS, Znd Floor, Tho Atrium, 290 lBurwood Road, HAWTHORN. VICTORIA 3122. 0 0~ 0