Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
2119669
~he invention relates to phenylglycinamide~ of hetero-
cyclically substituted phenylacetic acid derivatives,
processes for their preparation and their use in medica-
ments, in particular as hypotenRive and antiatherosclero-
tic agents.
It is known that renin, a proteolytic enzyme, eliminate~
the decapeptide angiotensin I from angioten~inogen in
vivo, and the angiotensin I is in turn degraded in the
lung, the kidneys or other tiff~ues to give the hyperten-
sive octapeptide angiotensin II. The variou~ effects ofangiotensin II, such as, for example, vasoconstriction,
Na' retention in the kidney, aldosterone releaæe in the
adrenal gland and increase in tone of the sympathetic
nervous system act synergistically in the sense of a
blood pressure increase.
Moreover, angiotensin II has the property of promoting
the growth and the replication of cells such as, for
example, cardiac muscle cells and smooth muscle cells,
these growing and proliferating in an increased manner in
various disease states (for example hypertension, athero-
sclerosis and cardiac insufficiency).
Apart from inhibition of renin activity, a possible
starting point for intervention in the renin-angiotensin
system (RAS) is the inhibition of the activity of the
angiotensin-converting enzyme (ACE) and the blockade of
.
Le A 29 546
21196fi9
angiotensin II receptors.
The present invention relates to new phenylglycinamide~
of heterocyclically substituted phenylacetic acid deriva-
tives of the general formula (I)
A'~ ~ D ~ R3
~,CO- N~CO NR4R5
R1 ,,
in which
A represents straight-chain or branched alkyl or
alkenyl each having up to 8 carbon atoms, or
represents cycloalkyl having 3 to 8 carbon atoms,
B represents hydrogen, halogen or perfluoroalkyl
having up to 5 carbon atoms,
D represents a group of the formula -CH2-OR6 or -CO-R',
in which
R6 denotes hydrogen or straight-chain or branched
alkyl having up to 8 carbon atoms,
R' denotes hydrogen, hydroxyl or straight-chain or ~ :
branched alkoxy having up to 8 carbon atoms,
LQ A 29 546 - 2 --
: ~ .,: . , , : , : .: :: :-: , .: : . : - : : .
.
2119669
E represents hydrogen, halogen, nitro, hydroxyl,
trifluoromethyl, trifluoromethoxy, straight-chain or
branched alkyl, alkoxy or alkoxycarbonyl each having
up to 6 carbon atoms, cyano or carboxyl,
5 L represents hydrogen or straight-chain or branched
alkyl having up to 4 carbon atoms,
R1 represents cycloalkyl having 3 to 8 carbon atoms, or
represents ~traight-chain or branched alkyl having
up to 8 carbon atoms, which is optionally
substituted by cycloalkyl having 3 to 8 carbon
atoms,
R2 represents hydrogen or straight-chain or branched
alkyl having up to 6 carbon atoms,
R3 represents hydrogen, hydroxyl, halogen or straight-
chain or branched alkoxy having up to 6 carbon
atoms,
R4 and Rs are identical or different and represent hydro-
gen, phenyl, pyridyl, cycloalkyl having 3 to 8
carbon atoms, or straight-chain or branched alkyl
having up to 8 carbon atoms, which is optionally
substituted by hydroxyl, pyridyl, carboxyl,
straight-chain or branched alkoxycarbonyl having up
to 6 carbon atoms or by a group of the formula
-NR~R9,
Le A 29 546 - 3 -
... .
2119669
in which
R8 and R9 are identical or different and denote
hydrogen or straight-chain or branched alkyl
having up to 4 carbon atoms
or
R4 and Rs, together with the nitrogen atom, form a 6-
membered saturated heterocycle having up to 3
heteroatoms from the group consisting of S, N and 0,
and their salts.
The compounds of the general formula (I) accordin~ to the
invention can also be present in the form of their saits.
In general, salts with organic or inorganic bases or
acids may be mentioned here.
In the context of the present invention, physiologically
acceptable salts are preferred. Physiologically accep-
table salts of the new phenylglycinamides of hetero-
cyclically substituted phenylacetic acid derivatives can
be salts of the substance~ according to the invention
with mineral acids, carboxylic acids or sulphonic acids.
Particularly preferred salts are, for example, those with
hydrochloric acid, hydrobromic acid, sulphuric acid,
phosphoric acid, methanesulphonic acid, ethanesulphonic
acid, toluenesulphonic acid, benzenesulphonic acid,
naphthalenedisulphonic acid, acetic acid, propionic acid,
Le A 29 546 - 4 -
21196fi9
lactic acid, tartaric acid, citric acid, fumaric acid,
maleic acid or benzoic acid.
Physiologically acceptable salts can also be metal or
ammonium salts of the compounds according to the inven-
tion which have a free carboxyl group. Those particularly
preferred are, for example, sodium, potas~ium, magnesium
or calcium salts, and also ~mmonium salts which are
derived from ammonia, or organic amines such as, for
example, ethylamine, di- or triethylamine, di- or trieth-
anolamine, dicyclohexylamine, dimethylaminoethanol,
arginine, lysine or ethylenediamine.
The compounds according to the invention can exist in
stereoisomeric forms which either behave as image and
mirror image (enantiomers), or which do not behave as
image and mirror image (diastereomers). The invention
relates both to the enantiomers or dia~tereomera or to
their respective mixtures. Like the diastereomers, the
racemic forms can be separated into the stereoisomerical-
ly uniform constituents in a known manner. tcf. E.L.
Eliel, Stereochemistry of Carbon Compounds, McGraw Hill,
1962].
Preferred compounds of the general formula (I) are those
in which
A represents straight-chain or branched alkyl or
alkenyl each having up to 6 carbon atoms, or repre-
Le A 29 546 - 5 -
2il9669
sents cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl or cycloheptyl,
B represents hydrogen, fluorine, chlorine, bromine or
perfluoroalkyl having up to 4 carbon atoms,
D represents a group of the formula -CH2-OR6 or -Co-R7,
in which
R6 denotes hydrogen or straight-chain or branched
alkyl having up to 6 carhon atoms,
R7 denotes hydrogen, hydroxyl or straight-chain or
branched alkoxy having up to 6 carbon atoms,
E represents hydrogen, fluorine, chlorine, bromine,
trifluoromethyl, carboxyl or straight-chain or
branched alkyl, alkoxy or alkoxycarbonyl each having
up to 4 carbon atoms,
15 L represents hydrogen or methyl
Rl represents cyclopropyl, cyclopentyl, cyclohexyl or
cycloheptyl, or
represents straight-chain or branched alkyl having
up to 4 carbon atoms, which is optionally substitu-
ted by cyclopropyl, cyclopentyl, cyclohexyl or
cycloheptyl,
R2 represents hydrogen or straight-chain or branched
Le A 29 546 - 6 -
2~ 19669
alkyl having up to 4 carbon atoms,
R3 represents hydrogen, hydroxyl, fluorine, chlorine,
bromine or ~traight-chain or branched alkoxy having
up to 4 carbon atoms,
R4 and Rs are identical or different and repre~ent hydro-
gen, pyridyl, cyclopentyl, cyclohexyl, phenyl or
straight-chain or branched alkyl having up to 6
carbon atoms, which is optionally substituted by
hydroxyl, pyridyl, carboxyl, straight-chain or
10branched alkoxycarbonyl having up to 4 carbon atoms
or by a group of the formula -~R~R9,
in which
R8 and R9 are identical or different and denote
hydrogen or methyl,
or
R4 and R5, together with the nitrogen atom, form a mor-
pholine, piperazine or piperidine ring,
and their salts.
; Particularly preferred compounds of the general formula
(I) are those
in which
Le A 29 54~ - 7 -
:
. ,
211~669
A represents str~ight-chain or branched alkyl or
alkenyl each having up to 4 carbon atoms, or
repre~ents cyclopropyl, cyclopentyl or cyclohexyl,
B represents hydrogen, fluorine, chlorine or perfluo-
roalkyl having up to 2 car~on atoms,
D represents a group of the formula -CH2 oR6 or -Co-R7,
in which
R6 denotes hydrogen or straight-chain or branched
alkyl having up to 4 carbon atoms,
R7 denotes hydrogen, hydroxyl or straight-chain or
branched alkoxy having up to 4 carbon atoms,
E represents hydrogen, fluorine, chlorine, bromine,
trifluoromethyl, trifluoromethoxy or methyl
L represents hydrogen or methyl,
R1 represents cyclopentyl, cyclohexyl or cycloheptyl,
or
repre~ents straight-chain or branched alkyl having
up to 3 carbon atoms, which is optionally substitu-
ted by cyclopentyl, cyclohexyl or cycloheptyl,
R2 represents hydrogen, methyl or ethyl,
R3 represents hydrogen, hydroxyl, fluorine, chlorine,
Le A 29 546 - 8 -
2119669
.
bromine or methoxy,
R4 and Rs are identical or different and represent hydro-
gen, pyridyl, cyclohexyl, phenyl or straight-chain
or branched alkyl having up to 4 carbon atoms, which
is optionally substituted by hydroxyl, pyridyl,
carboxyl, straight-chain or branched alkoxycarbonyl
having up to 3 carbon atoms, amino or dimethylamino,
or
R~ and Rs, together with the nitrogen atom, form a mor-
pholine, piperazine or piperidine ring,
and their salts.
Processes for the preparation of the compounds of the
general formula (I) accordin~ to the invention have
additionally been found, characterized in that
[A] either compounds of the general formula (II~
N ~
A ~ N'l`D
E ~.
~ CO2H
R~
in which
A, B, D, E and Rl have the abovementioned meaning, : :
Le A 29 546 - 9 - :~.
2ll96fig
are reacted with compounds of the general formula (III)
~ R3
~ L (I~
HNR~ CO-NR4R5
in which
L, R2, R3, R4 and Rs have the abovementioned meaning,
or
[B] compounds of the general formula (IV)
A'~ ~ D' ~ R3
,C~ NP~2 C2-X
R,
in which
L, A, P., Rl, R2 and R3 have the anovementioned meaning,
D' represents the -CH2-O~- group
and -~
X represents Cl-C4-alkyl, :
Le A 29 546 - 10 -
21196fi~
are reacted with amines or ammonia of the general formula
tv)
HNR4Rs ( ~ ),
in which
R4 and R5 have the a~ovementioned meaning,
in inert ~olvents, in the presence of a base and of an
auxiliary,
a~d in the case where R2, R4 and/or Rs ~ ~, an alkylation
follows,
and the substituents A, B, D~ and E are converted by
customary methods, such as, for example reduction,
and if appropriate the i~omerY are separated, and in the
case of the preparation of the salts reacted with an
appropriate ba e or acid.
The process according to the invention can be illustrated -~
by way of example by the following reaction ~cheme~
tA~ M~thaIlesulphonyl Lhlositle.
~c DMF, N~2H5)3
H3C~CH2)3 N CHO C~Hs
f~ 2.) lizNlCO-NHz
~~/ 4 (N,N-L)imcthy1~ o)py~idine ~ :
Le A 29 546
. . .-,
.... , .. , -
. ~: - .. -, . .~, ~ ,
2119669
N ~C~
H3C-~CH2)3--~N CHO l~d
~CO--N~CO-NHz
~B]
H~C-~CH2~3 J~
~ CO--~ ~ GO~H2CH~ + NH3
~3C-~CH2~3 ~
~ 1
1~5CO--NH CONH2
Suitable solvents for the process are the customary
Le A 29 546 - 12 -
K~'
2119669
organic solvents which do not change under the reaction
conditions. These preferably include ethers such as
diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl
ether, or hydrocarbons such a~ benzene, toluene, xylene,
hexane, cyclohexane, or mineral oil fractions, or haloge-
nohydrocarbons such as dichloromethane, trichloromethane,
tetrachloromethane, dichloroethylene, trichloroethylene
or chlorobenzene, or ethyl acetate, triethylamine, pyri-
dine, dimethyl sulphoxide, dimethylformamide, hexamethyl-
phosphoramide, acetonitrile, acetone or nitromethane. Itis also possible to use mixtures of ~he solvents men-
tioned. Dimethylformamide and tetrahydrofuran are prefer-
red.
The bases employed for the process according to the
invention can in general be inorganic or organic bases.
These preferably include alkali metal hydroxides such as,
for example, sodium hydroxide or potassium hydroxide,
alkaline earth metal hydroxides such as, for example,
barium hydroxide, alkali metal carbonates such as sodium
carbonate or potassium carbonate, alkaline earth metal
carbonates such as calcium carbonate, or alkali metal or
alkaline earth metal alkoxides such as sodium methoxide
or potassium methoxide, sodium ethoxide or potassium
ethoxide or potassium tert-butoxide, or organic amineC
(trialkyl(C,-C6)amine~) such as triethylamine, or hetero-
cycles æuch as 1,4-diazabicyclo[2.2.2]octane (DABC0),
1,8-diazabicyclo[5.4.~]undec-7-ene (DBU), pyridine,
4-(N,N-dimethylamino)pyridine, methylpiperidine or mor-
pholine. It is also possible to employ alkali metals such
Le A 29 546 - 13 -
.. ~ . ., . ,, . , . ~ .
21196fi9
as sodium or their hydrides such as sodium hydride as
bases. Sodium hydride, potassium carbonate, triethyl-
amine, pyridine, potas6ium tert-butoxide and 4-(N,N-di-
methylamino)pyridine are preferred.
In general the base is employed in an amount from 0.05 to
10 mol, preferably from 1 mol to 2 mol, relative to 1 mol
of the compound of the formula (III)~
The proce6s according to the invention is in general
carried out in a temperature range from -30C to ~100C,
preferably from -30C to +60C.
The process according to the invention is in general
aarried out at normal pressure. ~owever, it is also
possible to carry out the process at elevated pressure or
at reduced pressure ~for example in a range from 0.5 to
5 bar).
The amidation is in general carried out in one of the
abovementioned solvents, preferably in N,N-dimethylforma-
mide.
The amidation is in general optionally carried out via
the activated stage of the acid halides or mixed
anhydrides, which can be prepared ~rom the corresponding
acids by reaction with thionyl chloride, phosphorus
trichloride, phosphorus pentachloride, phosphorus tribro-
mide or oxalyl chloride or methanesulphonyl chloride.
Le A 29 546 - 14 -
2119669
The amidation or acylsulphonamidation is in general
carried out in a temperature range from -50C to +80C,
preferably from -30C to +20C, and at normal pressure.
In addition to the abovementioned base~, suitable bases
for this reaction are preferably triethylamine and/or
dimethylaminopyridine, DBU or DABC0.
The base i8 employed in an amount from 0.5 mol to 10 mol,
preferably from 1 mol to 5 mol, relative to 1 mol of the
appropriate carboxylic acid. -
Acid-binding agents which can be employed for the
amidation or sulphoamidation are alkali metal or alkaline
earth metal carbonates such as sodium carbonate or potas-
sium carbonate, alkali metal or alkaline earth metal
hydroxides such as, for example, sodium hydroxide or
potassium hydroxide, or organic bases such as pyridine, ~ -
triethylamine, N-methylpiperidine, or bicyclic amidine~
such as 1,5-diazabicyclot4.3.0]non-5-ene (DBN) or 1,5-di-
azabicyclo[5.4.0]undec-5-ene (DBU). Triethylamine is
preferred.
::'
Suitable dehydrating reagents are carbodiimides æuch as,
for example, diisopropylcarbodiimide, dicyclohexylcar- -
bodiimide or N-(3-dimethylaminopropyl)-N'-ethylcarbodi~
imide hydrochloride or carbonyl compounds such as car-
bonyldiimidazole or 1,2-oxazolium compounds such as ~ - -
2-ethyl-5-phenyl-1,2-oxazolium-3-sulphonate or propane-
phosphonic anhydride or isobutyl chloroformate, or
Le A 29 546 - 15 -
2119669
benzotriazolyloxy-tris-(dimethylamino)phosphonium hexa-
fluorophosphate or diphenyl phosphoramidate or methane-
sulphonyl chloride, if appropriate in the presence of
bases such as triethylamine or N-ethylmorpholine or
N-methylpiperidine or dicyclohexylcarbodiimide and N-hy-
droxysuccinimide.
The acid-binding agents and dehydrating reagents are in
general employed in an amount from 0.5 to 3 mol, prefer-
ably from 1 to 1.5 mol, relative to 1 mol of the cor-
responding carboxylic acids.
The reduction of alkoxycarbonyl compounds or aldehydes tothe corresponding alcohols i8 in general carried out
using hydrides, such as lithiumaluminium hydride or
sodium borohydride, preferably using lithiumaluminium
hydride in inert solvents such as ethers, hydrocarbons or
alcohols or mixtures thereof, preferably in ethers such
as, for example, diethyl ether, tetrahydrofuran or diox-
ane, or alcohols such as ethanol, in the case of the
aldehydes preferably using sodium borohydride in ethanol,
in a temperature range from 0C to +150C, preferably
from +20C to +100C, at normal pressure.
The alkylation is in general carried out in one of the
abovementioned solvents using alkylating agents such as,
for example, (Cl-C~)-alkyl halides, ~ulphonic acid esters
or substituted or un~ubstituted ~C~-C6)-dialkyl or
(Cl-Cl0)-diaryl sulphates, preferably methyl iodide,
p-toluenesulphonic esters or dimethyl sulphate.
Le A 29 546 - 16 -
2119669
The compounds of the general formula (II) are new and can
be prepared
by reacting compounds of the general formula (VI~
l-'H2C ~
~CO2 W (Vl),
R1
in which
E and R1 have the abovementioned ~eaning,
T repre~ent~ a typical leaving group such as, for
example, chlorine, bromine, iodine, to~ylate or
mesylate, p.referably bromine,
and ~ - -
W represents ætraight-chain or branched (Cl-C~)-alkyl, . -~
first with imidazoles of the general formula (VII)
A ~ ~ D ~vn~,
H -~
'
in which
Le A_29 546 - 17 -
21~96fi9
A, B and D have the abovementioned meaning,
in one of the abovementioned inert solvents, if appropri-
ate in the presence of a base, and under a protective gae
atmosphere.
and in a last ~tep hydrolysing the esters according to
customary methods.
A suitable solvent for the reaction with the compounds of
the general formula (IV) i8 preferably dimethylformamide.
The bases employed are preferably sodium hydride, potas-
sium carbonate, triethylamine, pyridine and potassium
tert-butoxide.
In general, the base i8 employed in an amount from
0.05 mol to 10 mol, preferably from 1 mol to 2 mol,
relative to 1 mol of the compound of the formula (IV).
The process according to the invention is in general
carried out in a temperature range from -30C to +100C,
preferably from -10C to +60C.
The process according to the invention i8 in ge~eral
carried out at normal presæure. However, it is also
possible to carry out the process at elevated pressure or
at reduced pressure (for example in a range from 0.5 to
5 bar).
Le A 29 546 - 18 -
2119669
Suitable bases for the hydrolysis are the customary
inorganic bases. These preferably include alkali metal
hydroxides or alkaline earth metal hydroxides such as,
for example, lithium hydroxide, sodium hydroxide, pota~-
sium hydroxide or barium hydroxide, or alkali metalcarbonates such as sodium carbonate or potassium car-
bonate or sodium hydrogen carbonate, or alkali metal
alkoxides such as sodium methoxide, sodium ethoxide,
potassium methoxide, pota~sium ethoxide or pota~ium
tert-butoxide. Lithium hydroxide, sodium hydroxide or
potassium hydroxide is particularly preferably employed.
Suitable solvents for the hydrolysis are water or the
organic solvents customary for hydrolysis. Theqe prefer-
ably include alcohols such as methanol, ethanol,
propanol, isopropanol or butanol, or ethers such as
tetrahydrofuran or dioxane, or dimethylformamide or
di~ethyl sulphoxide. Alcohols such as methanol, ethanol,
propanol or isopropanol are particularly preferably used.
It i~ also possible to employ mixtures of the solvents
mentioned.
The hydrolysis is preferably carried out using acids such
as, for example, trifluoroacetic acid, acetic acid,
hydrochloric acid, hydrochloric acid/dioxane, hydrobromic
acid, methanesulphonic acid, sulphuric acid or perchloric
acid, particularly preferably using trifluoroacetic acid
or hydrochloric acid/dioxane.
The hydrolysis i8 in general carried out in a temperature
Le A 29 546 - 19 -
2119669
range from 0C to +100C, preferably from +20C to +B0C.
In general, the hydrolysis is carried out at normal
pressure. However, it is also possible to work at reduced
pressure or at elevated pressure (for example from 0.5 to
5 bar).
When carrying out the hydrolysis, the ba~e is in general
employed in an amount from 1 to 3 mol, preferably from 1
to 1.5 mol, relative to 1 mol of the ester. Molar amounts
of the reactants are particularly preferably used.
When carrying out the reaction, the carboxylates of the
compounds according to the invention are formed in the
first ~tep as intermediates which can be isolated. The
acids according to the invention are obtained by treating
the carboxylates with customary inorganic acids. These
preferably include acids such a~, for example, hydrochlo~
ric acid, hydrobromic acid, sulphuric acid, phosphoric
acid or trifluoroacetic acid. It has al80 proven advanta-
geous in this case in the preparation of the carboxylic
acids to acidify the basic reaction mixture from the
hydrolysis in a second step without isolation of the
carboxylates~ The acids can then be isolated in a cus-
tomary manner.
The compounds of the general formulae (III), ~V) and
(VII) are known.
The compounds of the general formula (VI) are for the
'
:
Le A 29 546 - 20 - -
21~366~
most part new and can be prepared by reacting the cor-
responding 4-methyl compounds in the ~ense of a sub~titu-
tion, for example by halogenation in the pre~ence of one
of the abovementioned bases and/or auxiliarie~ and of one
of the solvents.
~he compound~ of the general formula (IV) are new per ~e
and can be prepared, for example, by reaction of the
compounds of the general formula (II) with compounds of
the general formula (VIII)
,
R3 -~
L,~ (V~l),
HN-R2 ~C02-X -
in which
L, R2, R3 and X have the abovementioned meaning,
as described in tA]. -
The compounds of the general formula (YIII) are known in
~ome cases or can be prepared by customary method6.
The compoundæ of the general formula (I) according to the
invention exhibit an unforeseeable, useful ~pectrum of
pharmacological action.
The compounds according to the invention have a specific
A II-antagonistic action, since they competitively
~ - -
Le A 29 546 - 21 - -~;
211g669
inhibi~ the binding of angiotensin II to the receptor~.
They suppress the vasoconstrictory and aldosterone ~ecre-
tion-stimulating effects of angiotensin II. They moreover
inhibit the proliferation of ~mooth muscle cells.
They can therefore be employed in medicaments for the
treatment of arterial hyperten~ion and atherosclerosis.
They can moreover be employed for the treatment of coron-
ary heart di6eases, cardiac insufficiency, di~orders of
the brain function, ischaemic cerebral di~ea~es, peri-
pheral circulatory disorders, functional di~orders of the
kidney and adrenal gland, bronchospastic diseases and
respiratory tract diseases having a vascular component,
sodium retention and oedemas.
Inve~ti~ation of the inhibition of the contraction in-
duced by aaonists
Rabbits of either sex are stunned by a blow to the back
of the head and exsanguinated, or in some cases anaes-
thetized with Nembutal (about 60-80 mg/kg i.v.) and
sacrificed by opening the thorax. The thorax aorta is
removed, freed from adhering connective tissue, divided
into ring segments 1.5 mm wide and individually trans-
ferred under an initial loading of about 3.5 g to 10 ml
organ baths containing Krebs-Hen~eleit nutrient ~olution,
which is temperature-controlled at 37C and aerated with
95% 2/5% CO2, of the following composition: 119 mmol/l of
NaCl; 2.5 mmol/l of CaCl2 x 2H2O; 1.2 mmol/l of K~2PO~;
10 mmol/l of glucose; 4.8 mmol/l of KCl; 1.4 mmol~l of
Le A 29 546 - 22 -
2119669
MgS04 x 7H20 and 25 mmol/l of NaHC03.
The contraction~ are detected isometrically by Statham
UC2 cells by means of bridge amplifiers (ifd Mulheim or
DSM Aalen) and digitalized and a~ses~ed by means of A/D
converters (System S70, Xeithley Munich). Agonist do e
response curves (DRC) are carried out hourly. With each
DRC, 3 or 4 individual concentrations are applied to the
baths at a 4 min interval. After the end of the DRC and
subsequent washing-out cycles (16 time~ in each case
about 5 sec/min with the abovementioned nutrient
solution), a 28-minute rest or incubation phase follows,
during which the contraction~ a~ a rule reach the star-
ting value again.
The height of the 3rd DRC, in a normal ca~e, is used a~
a reference variable for the a~e~sment of the test
substance to be inve~tigated in ~urther runs, which is
applied to the baths in the following DRCs in increa~ing
doses in each case at the start of the incubation period.
Each aorta ring is in this case stimulated for the whole
day, always with the same agonist.
Aqonists and their standard concentrations (ap~lication
volume per individual do e = 100 ~
KCl 22.7; 32.7; 42.7; 52.7 mmol/l
L-Noradrenaline 3 x 10-9; 3 x 10-a;
3 x 10-'; 3 x 10-6 g/ml
Serotonin 10-0; 10-7; 10-6; 10-5 g/ml -
B-HT 920 10-7; 10-6; 10-5; g/ml ;~
Le A 29 546 - 23 -
~ 2119669
Methoxamine 10-7; 10-6; 10-5 g/ml
Angiotensin II 3 x 10-9; 10-~; 3 x 10-~;
10-' g/ml
.,~.,
For the calculation of the ICso (concentration at which
the substance to be investigated cau~e~ a 50% inhibi-
tion), the effect is in each ca~e ba~ed on the 3rd -
submaximal agoni~t concentration.
The compounds according to the invention inhibit the
contraction of the isolated rabbit aorta induced by
angiotensin II in a dose-dependent manner. The contrac-
tion induced by potassium depolarization or other agon-
ist~ was not inhibited or only weakly inhibited at high
concentrations.
Blood DressUre measurements on the anaiotensin II-infused
rat
Male Wistar rats (Noellegaard, Copenhagen, Denmark)
having a body weight of 300-350 g are anae~thetized with
thiopental (100 mg/kq i.p.~. After tracheotomy, a cathe-
ter for blood pressure measuremsnt i~ inserted in the
femoral artery and a catheter for angiotensin II infusion
and a catheter for substance administration are inserted
in the femoral veins. After administration of the gang-
lionic blocker pentolinium (5 mg/kg i.v.), the angioten-
sin II infusion (0.3 ~g/kg/min) is started. As ~oon as
the blood pressure values have reached a stable plateau,
the test substances are either administered intravenously
Le A 29 546 - 24 -
21196~9
~ 318g-7615
or orally as a suspenslon or solution ln 0.5% Tylose.
Determination of the antihvPertensive activitY in conscious
hyPertensive rats
The oral antihypertensive activity of the compounds
according to the lnventlon was tested ln conscious rats using
surgically induced unilateral renal artery stenosls. To do this,
the right renal artery was constricted with a silver clip of 0.18 ~-
mm internal width. In this form of hypertension, the plasma renin
activity is increased in the first six weeks after intervention.
The arterial blood pressure of these animals was -
measured ln a blood-free manner at defined time intervals after
substance administration using the "tail cuff". The substances to
be tested were suspended in a Tylose suspension and administered
intragastrally ("orally") ln various doses by stomach tube. The - -
compounds according to the invention reduce the arterial blood
pressure of the hypertensive rats at a clinically relevant dose.
Additlonally, the compounds according to the invention ~ - -
inhibit the specific binding of radloactive angiotensin II in a
concentration-dependent manner.
','~, ~' ;'"" .
"~ ~
2~l9669
Interaction of the comDounds accordinq to the invention
with the anqiotensin II rece~tor in memhrane fractions of
the adrenal ~land cortex (bovine)
Bovine adrenal gland cortices (AGC), which have been
freshly removed and carefully freed from gland medulla,
are comminuted in sucrose solution (0.32 M) with the aid
of an Ultra-Turrax (Janke & Kunkel, Staufen i.B.~ to give
a coarse membrane homogenate and partially purified in
two centrifugation steps to give membrane fractions.
The receptor binding investigations are carried out on
partially purified membrane fractions of bovine AGC using
radioactive angiotensin II in an assay volume of 0.25 ml,
which specifically contains the partially purified mem-
branes (50-80 ~g), 3H-angiotensin II (3-5 nM), test
buffer solution (50 mM Tris, p~ 7.2), 5 mM MgCl2 and the
substances to be investigated. After an incubation time
of 60 min at room temperature, the unbound radioactivity
of the samples is separated by mean~ of moistened glass
fibre filters (Whatman GF/C) and the bound radioactivity
is measured spectrophotometrically in a scintillation
cocktail after washing the protein with ice-cold buffer
solution (50 mM Tris/~Cl, pH 7.4, 5% PEG 6000). The
analysis of the raw data was carried out using computer
programs to give Kl or IC50 values (Xi: ICso values cor-
rected for the radioactivity used; ICso values: concentra-
tion at which the substance to be investigated causes a
50% inhibition of the specific binding of the radioli-
gand).
Le A 29 546 - 26 -
2119669
Investiaation of the inhibition of the Proliferation of
smooth muscle cells bv the compounds accordinq to the
lnventlon
To determlne the antiproliferative action of the com-
pounds, smooth muscle cells are used which are obtained
from aortas of rats by the media explant technique [R.
Ross, J. Cell. Biol. 50, 172, 1971~. The cells are inocu-
lated in suitable culture dishes, as a rule 96-hole
plates, and cultured in 5% CO2 at 37C for 2-3 days in
medium 199 containing 7.5% FCS and 7.5% NCS, 2 mM L-glut-
amine and 15 mM ~EPES, p~ 7.4. The cells are then ~yn-
chronized by withdrawal of serum for 2-3 days and then
stimulated into growth with serum. Test compounds are
simultaneously added. After 16-20 hours, 1 ~Ci of 3~
thymidine is added and after a further 4 hours the
incorporation of this substance into the TCA-precipitat-
able DNA of the cell~ is determined. To determine the IC50
values, the active compound concentration is calculated
which, on sequential dilution of the active compound,
causes semi-maximal inhibition of the thymidine
incorporation produced by 10% FCS.
Table ~:
Ex. No. ICso tnM]
1 . 0
6 0.2
The new active compounds can be converted in a known
Le A 29 546 - 27 -
21196fi~
~ 31~9-7615
manner into the customary formulatlons, such as tablets, coated
tablets, pills, granules, aerosols, syrups, emulslons, suspensions
and solutions, using inert, non-toxlc, pharmaceutlcally suitable
excipients or solvents. In this case, the therapeutically active
compound should in each case be present in a concentration of
about 0.5 to 90% by weight of the total mixture, i.e. ln amounts
which are sufflcient ln order to achleve the dosage range
indicated.
The formulations are prepared, for example, by extending
the active compounds with solvents and/or excipients, if
approprlate using emulsifiers and/or dlspersants, where, for -~
example, in the case of the use of water as a diluent, organic ~.;
solvents can optionally be used as auxiliary solvents.
The inventlon also extends to a commercial package
containing a compound of the invention, together with instructions
for its use for the treatment of arterial hypertension and
atherosclerosis.
Adminlstration is carried out in a customary manner,
preferably orally or parenterally, in particular perlingually or
intravenously. ~-
In the case of parenteral administration, solutions of
the active compound using suitable liquid excipients can be
employed.
In general, it has proven advantageous in the case of
intravenous administration to administer amounts of about 0.001 to
1 mg/kg, preferably about 0.01 to 0.5 mg/kg of body weight to
achieve effective results, and ln the case of oral administration
the dose is about 0.01 to
28
21~9669
20 mg/kg, preferably 0.1 to 10 mg/k~ of body weight.
In spite of this, it may 60metimes be neces~ary to depart
from the amounts mentioned, in particular depending on
the body weight or the type of administration route, on
individual behaviour towards the medicament, the manner
of its formulation and the time or interval at which
admini6tration takes place. Thu6, in some case~ it may be
sufficient to manage with less than the abovementioned
minimum amount, while in other case~ the upper l; m; t
mentioned must be exceeded. In the case of the admini~-
tration of relatively large amount~, it may be advi~able
to divide the~e into ~everal individual doses over the
course of the day.
Eluents
(A) Petroleum ether:ethyl acetate = 3:7
(B) Dichloromethane:methanol = 8:1
(C) Petroleum ether:ethyl acetate = 1:4
(D) Petroleum ether:ethyl acetate = 1:1
(E) Dichloromethane:methanol = 5:1
(F) Dichloromethane:ethyl acetate = 10:1
(G) Dichloromethane:methanol = 10:1
(H) Petxoleum ether:ethyl acetate = 2:1
(I) Dichloromethane:methanol = 20:1
Le A 29 546 - 29 -
2119669
Startinq Compounds
Exampl¢e I
tert-Butyl 4-methylphenylacetate
H3~ ~
~ ,
co2c(CH3)3
450 g 13 mol) of 4-methylphenylacetic acid, 1.13 1
(12 mol) of tert-butanol and 90 g (0.74 mol) of dimethyl-
aminopyridine are di~601ved in 2 1 of dichloromethane.
After addition of 680 g (3.3 mol) of dicyclohexyl-
carbodi;~;de, dissolved in 400 ml of dichloromethane, the
mixture i~ stirred at 25C for 20 h, the precipitated
urea is filtered off with suction and wa~hed with 200 ml
of dichloromethane, and the organic phase is washed twice
each with 500 ml of 2 N hydrochloric acid and water. The
organic phase i8 concentrated and distilled.
Yield: 408 g (66% of theory)
Boiling point: 73 - 78C/0.2 mm
Le A 29 546 - 30 -
, .
21196fi9
Example II
tert-Butyl 2-cyclopentyl-2-(4-methylphenyl)acetate
~13C~
~ )--
co2~(CH3)3
33.5 g (0.3 mol) of potassium tert-butoxide are initially
introduced at 0C into 100 ml of DMF with exclu~ion of
moisture and 51.6 g (0.25 mol) of tert-butyl 4-methyl-
phenylacetate in 250 ml of DMF are added dropwise. The
mixture i8 stirred at 0C for 30 min and 3~.2 ml
(O.3 mol) of cyclopentyl bromide in 150 ml of DMF are
added dropwise at 5-15C and the mixture is ~tirred at
25C for 20 h. After concentration, the rei~idue iB parti-
tioned between water/diethyl ether, and the ether phase
i5 dried over sodium sulphate and concentrated. The
product crystallize~ out.
Yield: 67 g (97.5% of theory)
Melting point: 51 - 53C ~
.:
-
Le A 29 546 - 31
ExamPle III 2119669
tert-Butyl 2-(4-bromomethyl-phenyl)-2-cyclopentyl-acetate
Br ~
CO2C(C~3)3
27.4 g (0.1 mol) of tert-butyl 2-cyclopentyl-2-(4-methyl-
phenyl~-acietate are dissolved in 200 ml of carbon tetra-
chloride and heated to boiling. After addition of 0.82 gof azobisisobutyronitrile, 18.7 g (0.105 mol) of N-bromo-
succinLmide are added in portions and the mixture is then
refluxed for 1 h, cooled to 0C and ~uccinimide is fil-
tered off. After concentration of the filtrate the pro-
duct precipitate~. It is washed with petroleum ether(40/60) and dried.
Yield: 20 g (57% of theory)
Melting point: 73-76C
:~
Le A 29 546 - 32
.,. . ~ .
2~19669
Example IV
tert-Butyl 2-~4-(2-butyl-4-chloro-5-formyl-imldazol-1-yl-
methyl)phenyl]-2-cyclopentyl-acetate
Cl
H3C ~ N ~ CHO
COzC(CH3)3
1.6 g (O.053 mol) of sodium hydride (80% strength) are
suspended in 50 ml of DMF undex protective gas, 10 g
(0.053 mol) of 2-butyl-5-formy1-4-chloro;m;dazole (prepa-
ration according to EP 324,3773 in 100 ml of DMF are
added dropwise at 0C, then the mixture ic stirred at 0C
for 15 min and 18.9 g (0.053 mol) of tert-butyl 2-(4-bro-
momethylphenyl)-2-cyclopentylacetate in 100 ml of DMF are
added dropwise. The mixture i8 additionally stirred at
0C for 2 h, the solvent i8 evaporated off, the residue
is taken up in diethyl ether, the solid is filtered off
and after concentration the residue is chromatographed on
silica gel 60 using dichloromethane.
Yield: 16.2 g (66.7% of theory)
Melting point: 101-102C
Le A 29 546 ~ 33 -
211966~
Example V
2-[4-(2-Butyl-4-chloro-5-formyl-imidazol-1-yl-methyl)-
phenyl]-2-cyclopentylacetic acld
~3C ~N~ HO
~ ,
OD2~ :
2.3 g (5 mmol) of the compound from Example IV are
stirred at 25C for S h in 5 ml of dichloromethane and
5 ml of trifluoroacetic acid. After concentration, the
crude product is chromatographed on silica gel 60 using -
dichloromethane/methanol (100:5). ~. :
Yield: 1.8 g (87.6% of theory) : : :
Melting point: 95-98C ~ ~
.. ~ -: .-
'-: ,':, . ':
: ' ~,
,,
-: -
.
Le A 29 546 - 34 - -
2119669
Exam~le VI
Methyl 2-{2-[4-(2-butyl-4-chloro-5-formyl-Lmidazol-1-yl-
methyl)phenyl]-2-cyclopentyl}-acetamido-2-(2-hydroxy-
phenyl)acetate
~ N ~ CH0
~,C~H CO2CH3 ,,
6.0 g (15 mmol) of the compound from Bxample V are diR-
solved in 180 ml of anhydrou~ tetrahydrofuran and reacted
at 0C with 4.2 ml (30 mmol) of triethylA~lne and 1.26 ml
(16.5 mmol) of methanesulphonyl chloride. After 1 hour,
a solution of 3.92 g (18 mmol~ of 2-hydroxyphenylglycine
methyl ester hydrochloride, 1.82 g (15 mmol) of
4-(N,~-dimethylamino~pyridine and 2.52 ml (18 mmol) of
triethylamine in 60 ml of tetrahydrofuran i8 added and
the mixture i6 additionally stirred for 18 hour~, the
reaction temperature rising to room temperature. The
crude mixture is added to water, adjusted to pH = 2 to
3 using 2 M hydrochloric acid and extracted with ether.
The organic phase i~ dried with magne ium sulphate and
evaporated. After chromatographic work-up on ~ilica gel
60 (Merck, petroleum ether : ethyl acetate - 2:1), 4.03 g
Le A 29 546 - 35 -
, 2119669
(7.1 mmol) of the title compound are obtained.
Rf - 0.18 (dichloromethane:methanol ~ 50:1)
Preparation ExamPles
ExamDle 1
2-{2-[4-(2-Butyl-4-chloro-5-formyl-imidazol-1-yl-methyl)-
phenyl]-2-cyclopentyl}acetamido-2-phenyl-acetamide
a :
~,C ~ o
CONH2
:...................................................................... '.::
1.54 g (3.8 mmol) of the compound from Example V are
reacted in anhydrous N,N-dimethylformamide at -30C with
1.06 ml (7.6 mmol) of triethylamine and 0.48 g (4.2 mmol)
of methanesulphonyl chloride. After 1 hour, 0.69 g
(4.6 mmol) of phenylglycinamide and 0.47 g (3.8 mmol) of
4-(N,N-dimethylamino)pyridine are added and the mixture
is additionally stirred for 24 hours while slowly warming
to room temperature. It is then poured into ether,
treated with water and adjusted to pH = 2 using 1 M
hydrochloric acid. The aqueous phase is reextracted with
ether and the combined organic phases are washed several
times with 0.01 M hydrochloric acid. They are then
extracted several times with aqueous sodium hydroxide
. .:
~.
Le A 29 546 - 36 -
21196fig
solution of pH = 10 and washed with water, the organic
phase is dried with sodium sulphate and the solvent i~
evaporated. The crude product is purified by chromato-
graphy (silica gel 60, Merck 40-63 ~m, petroleum
ether:ethyl acetate = 1:1).
Yield: 1~56 g (2.9 mmol) of product.
Rf = 0.16 and 0.12 (petroleum ether:ethyl acetate = 1:1)
The compounds shown in Table 1 are prepared in analogy to
the procedure of Example 1:
Le A 29 546 - 37 -
2~19B69
~able 1:
Nf
H3C ~N~CHO ~q
~ ~ ,,
l~ co- N CONH2
~1 , '
Ex.No. Diastereomer Rl Rf ( solvent)
4 dia b 0 47 (A)
3 4 dia ~ 0.57 (B)
Example 4 : :
2-[4-(2-Butyl-4-chloro-5-formyl-imidazol-1-yl-methyl)-
phenyl]-2-cyclohexyl-N-phenylglycinamidoacet~ide
H3C ~o 13
~C~Nlf~CONH2
O ~ '
Le A 29 546 - 38 -
.
21196fi9
0.42 g (1 mmol) of 2-[4-(2-butyl-4-chloro-5-formyl-im-
idazol-1-yl-methyl)phenyl]-2-cyclohexylacetic acid is
treated at room temperature in 25 ml of dichloromethane
with 0.2 g (2 mmol) of triethylamine and 0.23 g
(1.5 mmol) of 1-hydroxy-lH-benzotriazole in 5 ml of
tetrahydrofuran and the mixture is cooled to 0C. After
addition of 0.31 g (1.5 mmol) of dicyclohexylcarbodiimide
in 10 ml of dichloromethane and stirring for 30 minutes,
a solution of 0.32 g (1.2 mmol) of phenylglycinamide and
0.1 g (1.2 mmol) of triethylamine in 10 ml of dichlorome-
thane is added and the mixture i8 stirred overnight at
room temperature. For work-up, the mixture is extracted
after addition of dichloromethane, and the combined
organic phases are dried over sodium sulphate, filtered,
concentrated and chromatographed on silica gel 6n (ethyl
acetate/petroleum ether - 1:1).
Yield: 0.31 g (0.56 mmol) 56% of theory
Rf 3 0.52 (dichloromethane/methanol z 9:1)
Le A 29 546 - 39 ~
2119669
Example 5
2- [ 4- ( 2-Butyl-4-chloro-5-hydroxymethyl-imidazol- 1-yl-
methyl ) phenyl J -2 -cyclohexyl-N-phenylglycinamidoacetamide
H~C ~OH ~q
~ ~ -,
l~co. Nll CONH2
0.15 g (0.3 mmol) of 2-r4-(2-butyl-4-chlor~-5-formyl~
imidazol-1-yl-methyl ) phenyl ] -2-cyclohexyl-N-
phenylglycinam~doacetamide are treated at room
temperature in S ml of ethanol with 10 mg of sodium
borohydride and the m~xture is stirred at room tempera-
ture f or 2 hours . For work-up, water is added and the
mixture is adjusted to pH 4-5 using 1 N acetic acid and -
extracted three times with ethyl acetate. The combined
organic phases are dried over sodium sulphate, f iltered
and concentrated.
Yield: 0.16 g (0.29 nunol) 98% of theory
Rf = 0.48 (dichloromethane/methanol = 9:1) ~ -
Le A 29 _ 4Ç - 40 - ~
2119669
Example 6
2-{2-[4-(2-~utyl-4-chloro-5-hydroxymethyl imidazol-1-yl~yl)-
phenyl]-2-cyclopentyl}acetamido2-phenyl-acetamide
~N [~
N~ CO~H2
~1
1.37 g (2.6 mmol) of the compound from Example 1 are
reacted at 20C in 15 ml of ethanol with 0.10 g
(2.6 mmol) of ~odium borohydride. If reaction is i~com-
plete (TLC checking), further ~odium borohydride i~ added
after 2 hours. After a total of 3 hour~, water and ether
are added, excess borohydride is destroyed using hydroch-
loric acid at pH = 2, and the mixture is then adjusted to
pH = 7.5 using 1 M aqueous sodium hydroxide solution and
extracted several times with ether. The organic phase i8
dried using sodium sulphate and evaporated.
Yield: 1.06 g (2.0 mmol)
Rf = 0.78 and 0.75 (dichloromethane:methanol = 5
-
The compounds shown in Table 2 are prepared in analogy to
the procedure of Example 6: i
,
Le A 29 546 - 41 - ~ -~
~` ~
~1196~9
Table 2:
H3 C ~ ~OH
~- \R
Ex.No. Diastereoisomer R~ R2 R3 R4 Rs Rf(solvent)
7 rac dia A ~ H H H H 0.37(A)
8 rac dia B ~ H H ~ H 0.26(A)
9 4 dia ~ H 2-OH H H 0.42 (C)
4 dia ~ H H H H 0.23/0.18 ~A)
r
Le A 29 546 . - 42 ~
,:
Example 11 2113669
2-{2-[4-(2-Butyl-4-chloro-5-formyl-imidazol-1-yl-methyl)-
phenyl]-2-cyclopentyl}-acetamido-2-(2-hydroxyph~nyl)-
acetamide
N--¢
H3C~~N~_ CHO
~C~ NI~CONH2
60 mg (0.11 mmol) of the compound from Examiple VI are
dissolved in 2 ml of anhydrous tetrahydrofuran and reac-
ted with 1 ml of 25~ strength aqueous ammonia solution (6
hours at 50C). The mixture is then adjusted to pH = 7
using 0.1 M sulphuric acid and extracted several times
with ether. The organic phase is dried using magnesium
sulphate and evaporated. After removal of the residual
solvent in a high vacuum, 40 mg (0.07 mmol) of product ~ ~
are obtained. -
Rs ~ 0.21 (petroleum ether:acetic acid = 1:1) ~-~
The compounds showni in Tables 3 and 4 are the separated
diastereomers of Examples 1 and 2 and can be prepared by
separation of these according to customary methods:
Le A 29 546 - 43 -
21196fi9
~able 3:
~CHO
~1;0 NH CONHi,
b
Ex . No . Di aste reo i some r Rf ( solvent ) ~ -
12 dia A û.16 (D)
13 dia B O.12 ~D)
14 dia C O.16 (D)
dia D 0.12 (D)
Le A 29 546 - 44 -
2~19669
Table 4:
H ,C ~ OH [~
~ X
~CC~ tJH CONH2 :.,
b
Ex.No. Diasteroisomer Rf ( solvent )
16 dia A 0.78 (E)
17 dia B 0.75 (~)
18 dia C 0.78 (E)
19 dia D 0.75 (E)
The compounds shown in Table 5 are prepared in analoyy to
the procedure of Example 11:
,- ',
Le A 29 546 - 45 -
: : - ~ , : ~ . -
,: . ~., : : . .. .
211966~
Table 5:
N a
- H3C ~OH [3
~CO N~CON
b
Ex.No. Rs R(solvent)
CH2CO2cH2cH3 0.35 (F)
21 CH2CH2N( C~3 ) 2 0.47 (E)
22 ~N 0.53 (G)
23 ~ 0.75 (E)
24 ~ 0.61 (G)
CH2CH2O~ 0.33 (G)
26 ~ 0.37 (~
.
27 CH3 0.14 (I) - :
The compounds shown in Table 6 are prepared in analogy to
the procedure of Example 6:
::
Le A 29 546 - 46 -
21~9669
Table 6:
cl
H ,C ~~ OH
~CO NHJ<co
Ex . No . L Z Rf ( sol~rent )
28 CH3 NH2 0.32 (I)
The compounds shown in Table 7 are prepared in analogy to ~:
the procedure of Example 1~
Le A 29 546 - 47 -
211g669
Table 7:
H3C l~ ;;3~ ''
~CO N~ :~
Ex . No . L Z R~ ( solvent )
29 C~3 NH2 O 06 (D) -::
Le A 29 546 - 48 - :
2119669
Example 30
<2-{2-[4-(2-Butyl-4-chloro-5-hydroxymethyl-imidazol-1-yl-
methyl)phenyl-2-cyclopentyl~acetamido-2-phenyl-aceta-
mido>-acetic acid
cl ..
N--( ~
N ~)
~CO- NH~CO~\COOH
- b
60 mg (0.1 mmol) of the compound from Example 20 are
dissolved in 2 ml of ethanol and reacted for 1 hour at
22C with 4 ml of aqueous lM sodium hydroxide solution.
The reaction mlxture is diluted with water, the ethanol
component is evaporated and the product is precipitated
by acidifying with 2M hydrochloric acid. The precipitate
is washed with water and dried in a high vacuum over
Sicapent (Merck): 43 mg.
Rf = 0.60 (dichloromethane:methanol = 10:1)
Le A 29 546 - 49 -
