CH644842A5 - FATTY ACID AMIDES THEIR PRODUCTION AND USE AS PHARMACEUTICALS. - Google Patents

Description

The invention relates to compounds of the formula I O

II

A-C-NH-R2 (I)

in the

A (I) denotes the C7 to C23 tail of an unsaturated fatty acid which contains 1 to 4 ethylenically unsaturated sites, or (II) denotes a corresponding radical in which the ethylenically unsaturated sites -CH = CH— by cyclopropanyl groups

CH2 -CH-CH-

are replaced, and

R2 represents a radical of formula II, III or IV R_. , R,

| 3 /

-CH- (CH0)

R,

2 g

(II)

R,

-CH-CH.

(IV)

"N!

in the 8th

R4 has the meaning mentioned above,

R7 represents Q to C8-alkyl or benzyl, and Rs is hydrogen, Cj to Cs-alkyl or benzyl,

provided that when R2 is a radical of formula (II) or (III), A has the meaning described in (II).

2. Compounds according to claim 1, in which R2 represents a radical of formula (IV).

3. Compounds according to claim 2, in which A has the meaning (I).

where g is 0 or 1,

50 R4 hydrogen, fluorine, chlorine, bromine or C, to C3 alkyl or alkoxy,

Rs is hydrogen, fluorine, chlorine or Ci to C3 alkyl or alkoxy, and

R3 is hydrogen, Cx to Cs-alkyl or a radical of the formula VI

- (chi / Y

means in the 60 j 0 or 1,

Y is hydrogen, fluorine, chlorine, bromine or Cx to C3-alkyl or alkoxy, and

Y represents hydrogen, fluorine, chlorine or C1 to C3 alkyl or alkoxy; . . R

f ^

(in)

R6

in the

3rd

644 842

R5 has the meaning mentioned above, and R6 is hydrogen, fluorine, chlorine, bromine, Q to C3-alkyl or alkoxy or a radical of the formula VII

(VII) 5

means in the

B - CH2 - or —O -,

R,

f is 0 or 1, and

R4 has the meaning mentioned above;

COOR?

-CH-CH,

I / O - * -

(IV)

!

Rr i3 - (CH2) n - (CH = CH) 0 - (CH2) p -

CH3— (CH2) r— (CH = CH — CH 2) s— (CH 2) t—

20th

in the "ß

R4 has the meaning mentioned above,

R7 represents Cj to Cs-alkyl or benzyl, and Rs is hydrogen, Ct to Cs-alkyl or benzyl,

provided that when R2 represents a radical of formula II or III, A has the meaning described under (II).

In certain compounds according to the invention, A has the meaning given under (I), is referred to as A 'and then represents a C7 to C23 tail of a fatty acid with 1 to 4 ethylenically unsaturated sites. The fatty acids A'— COOH themselves contain 8 up to 24 carbon atoms and 1 to 4 ethylenically unsaturated positions. These acids are preferably unbranched and are selected from the range of natural fatty acids, which means that they have an even number of carbon atoms. A 'is therefore preferably 30 unbranched and contains an odd number of carbon atoms.

Particularly preferred radicals A 'are those of the formula IX, as radicals A'j and of the formula X, as radicals A'2,

(IX)

CH

where n is 1 to 10,

o is 1 to 4, and p is 3 to 9,

provided that n + 2 x o 4- p does not exceed 22, 40

(X)

in which r 1 to 4,

s 2 to 4, and 45

Is 11 to 7,

provided that r + 3 x s + t does not exceed 22.

Preferred radicals of the formula IX are those in which n + 2 x 0 + p represents an even number, especially those in which n = 5 or 7, o = 1 and p = 7. Preferred radicals of formula X are 50 in which r = 1 or 4, s = 2 to 4 and t = 2 or 6.

Since the compounds in which A has the meaning A 'contain one or more unsaturated sites, they have isomeric forms. The invention is not limited to any particular isomeric form. Compounds in which the hydrogen atoms have the cis configuration at the ethylenically unsaturated sites are preferred.

Particularly preferred residues A 'are those of palmitoleic, oleic, petroselinic, vaccene, elaostearic, parinaric, gadoleinic, cetoleinic, linoleic, linolenic, and arachidonic acids, especially of oleic, linoleic, 60 Linolenic, arachidonic and palmitoleic acid.

In other compounds according to the invention, A has the meaning given under (II), is referred to as A "and then represents a (C7 to C23) radical whose ethylenically unsaturated positions - CH = CH - by the cyclopropanyl unit 65

CH2 -CH-CH-

are replaced. Apart from the cyclopropanyl radical, the A "radicals are preferably unbranched. Particular radicals A" are those of the formula XI, indicated as radicals A "j.

CH2

CH-CH2

CH-

.- (CH2) B

(XI)

CH3— (CH2) U—

in the u 1 to 15,

v is 1 to 2, and w is 1 to 13,

provided that if v = 1, u + w = 3 to 19 and if v = 2, u + w is 2 to 16.

Particularly preferred A'V residues are those of the formula XI in which

1. if v = 1, u + w = 7 to 19,

if v = 2, u + w = 4 to 16,

2. if v = 1, u + w = an odd number,

if v = 2, u + w = is an even number,

3. if v = 1, u = 5 or 7 and w = 6, and if v = 2, u = 4 and w = 6.

The preferred radicals A "are derived from mono- or di-unsaturated natural fatty acids, for example from palmitoleic or oleic acid (v = 1) and from linoleic acid (v = 2).

The compounds with radicals A "also have isomerism. However, the invention is not restricted to any particular isomeric form. Compounds in which the hydrogen atoms which are on the tertiary carbon atoms of the cyclopropyl radicals and which have the cis configuration are preferred .

Since compounds with a single cyclopropyl radical in A "have a smaller number of asymmetric carbon atoms than those with several radicals, the purification of the first is generally easier to carry out and such compounds are preferred when purity is an important factor.

In a group of compounds which is indicated as la, A = A 'and R2 is a radical of the formula IV. When R7 = alkyl, it is preferably unbranched and specifically means ethyl. If R4 represents an alkyl group, this is preferably methyl. If it is preferably an alkoxy group, it is especially methoxy. When R4 is halogen, it is especially fluorine or chlorine. If R4 has a meaning other than hydrogen, it is preferably in the 5-position of the indole nucleus. However, R4 is preferably hydrogen. R8 also preferably denotes hydrogen.

In further groups of compounds, A = A ", and R2 represents a radical of the formula II (compound Ib) or a radical of the formula III (compound lc) or a radical of the formula IV (compound ld).

The preferred meanings of these compounds are the same as for the mentioned compounds in which A = A '.

Group la compounds are preferred.

The invention also relates to a process for the preparation of the compounds of the formula I, characterized in that a compound of the formula XII

H2N-R2

(XII)

in which Rj and R2 have the meanings mentioned above, with an acid of the formula XIII

A-COOH (XIII)

in which A has the meaning already mentioned, or is implemented with its functional derivatives.

The acylation can be carried out in a manner known for converting amines to their corresponding amides, e.g. with mixed anhydrides, wherein the amine of formula XII with an anhydride of formula XHIa

O

O

A-C-O-C-OR,

(XHIa)

644 842

4th

where A is as defined above and R12 is Ci to C6-n-alkyl.

This procedure is appropriate at temperatures from -10 to + 35 ° C in an inert organic medium such as a chlorinated hydrocarbon, e.g. Methylene dichloride. Another 5 method is the amine of formula XII with an acid chloride of formula XHIb

O

II

To implement A-C-Hal (XlIIb) io, in which A has the meaning mentioned above and Hai is chlorine or bromine.

This embodiment can be implemented very well at temperatures of 10 to 50 ° C, preferably at 20 to 30 ° C in an inert medium and in the presence of an acid acceptor.

The compound of formula XII can be used as an acid addition salt in acylation. Excess reagents, if they are liquid under the reaction conditions, can serve as the reaction medium.

• • 20

The isomeric form of the starting products is not influenced by the acylation according to the invention. Products are therefore obtained which have the corresponding isomeric form.

The products of formula I obtained can be isolated and purified in ways known per se.

The compounds of formula XII are either known or can be prepared in a manner known per se from known starting materials.

Compounds of formula XIIIc

A'-COOH (XIIIc) in which A 'has the meaning mentioned above are also known or can be obtained in a manner known per se from known starting materials.

Also compounds of the formula XHId 35

A "—COOH (XHId)

in which A "has the previously mentioned meaning are known and can be prepared, for example, by reacting the ethylenically unsaturated sites in the corresponding compounds of the formula XIIIc in 40 cyclopropanyl groups. This can be carried out in a manner known per se, for example by reaction with Methylene iodide according to the method of Simmons-Smith ("JACS", 81.4256 [1959]). To prepare compounds with a single cyclopropanyl group, the starting acids can be in the ice or 45 trans form. The resulting product then has the appropriate shape. Accordingly, mixtures of ice and trans forms will lead to the corresponding product mixtures.

If an acid containing cyclopropanyl groups or its derivative is reacted with a compound of the formula XII having asymmetrical carbon atoms, the product of the formula I obtained is formed as a mixture of diastereoisomers which

if desired, can be separated in a manner known per se, e.g. by chromatography or crystallization. However, the acid containing cyclopropanyl groups can also first be separated into its 55 antipodes and then reacted, as a result of which the corresponding isomeric product can be obtained in a relatively pure form. For the production of acids with two or more cyclopropanyl groups, starting from olefinic acids with a corresponding number of double bonds, the “o Simmons-Smith reaction leads to a mixture of diastereoisomers which, if desired, is separated before the further reaction.

The compounds of the formulas XHIa and XIHb can be prepared in a manner known per se, expediently in situ. 65

The compounds of the formula I according to the invention are pharmacologically active and can be used in particular for regulating the cholesterol level in the blood vessel walls of mammals. Therefore, they are particularly indicated as a remedy for atherosclerosis, which means that they are useful in the prophylactic treatment of atherosclerosis and in the regulation of atherosclerotic symptoms by the accumulation of cholesterol-role esters in the blood vessel walls. Regulatory capacity can be demonstrated using known test methods by reducing the total cholesterol level in cultured cells by the test compound and comparing it with the level in untreated cells, e.g. is described below:

A. Cell culture

Smooth muscle cells from the rhesus monkey, which the blood vessel, e.g. the aortic wall, by the method of K. Fischer-Dzoga et al. ("Experimental and Molecular Pathology", 18, 162-176 [1973]) are routinely grown in 75 cm2 tissue culture bottles in a minimum Eagle essential medium enriched with 10% fetal bovine serum. For the test procedure a bottle is selected which has an almost merged cell structure. The culture cells are removed from the bottle surface by mild enzymatic treatment with Pronase. After centrifuging and decanting the enzyme solution, the cell beads are resuspended in a suitable amount of the medium so that the desired number of 60 mm 5 ml of the diluted cell suspension is pipetted onto each dish, after which the dishes are labeled with the cell type, the date and the number of the culture bottle and in an atmosphere containing about 5% CO 2 at 37 ° C. incubated in a high humidity incubator. As soon as the cultures flow together, use started testing the connections.

For this purpose, these are routinely dissolved in 100% ethanol. An equivalent amount of ethanol is also added to the control groups. The tissue culture dishes are randomly distributed in groups. An amount of 5% by volume of hyperlipidemic rabbit serum (HRS) is supplied to a group as a control. 5% HRS 1 mg test compound per 100 ml medium is added to the other groups, after which all dishes are placed in the incubator again for 24 h. All actions up to the final incubation are carried out sterile. After incubation, the dishes are viewed with a phase contrast microscope (Zeiss Axiomat®) and the state of the cultures is noted, such as the size, number and structure of the cytoplasmic inclusions and the morphology of the cells. The media are removed from the cultures, after which a 0.9% NaCl solution is added. The cells are transferred from the bottles into conical, calibrated centrifuge tubes using rubber spatulas. The cells are washed by suspending them three times in an isotonic saline solution and centrifuging for 10 minutes with a force of 800 g and separating them from the above liquid.

B. Cell Extraction

A suitable amount of isopropyl alcohol of about 1 ml / mg protein is added to the cell beads. The sample is then sonicated with a microsensor (140 x 3 mm), which is connected to a Bronwell Biosonik IV apparatus, for 10 s Ultrason and centrifuged for 15 min. The clear liquid above is decanted and used for cholesterol analysis.

The residue is dissolved in 0.1N NaOH and used for nitrogen determination according to Lowry et al. ("J. Biol. Chem.", 193, 265, 1951).

C. Analysis

Free cholesterol: The isopropyl alcohol solutions of the sample and blank sample and the solvent isopropyl alcohol itself are treated in the same way. 20 nl of the respective isopropyl alcohol liquids are added to an amount of 0.4 ml of free reagent (Reagent A, see Table 1), which is in a 10 x 75 mm

5

644 842

large glass test tube is located and mixed with it. After leaving the mixture for about 5 minutes at room temperature, it is mixed with 0.8 ml of 0.5N NaOH (Reagent C, Table 1). The fluorescence is measured with an Aminco-Bowman spectrophotofluorescence measuring device, wherein light of 325 nm is emitted and light of 415 nm is emitted. The measurement is carried out in a 1 cm cuvette and with a xenon lamp, an IP28 photomultiplier tube and a slit of 2 mm.

Total cholesterol: The same procedure is used as for the determination of free cholesterol, except that instead of the free reagent, the total reagent (Reagent B, Table 1) is now used and the samples, before adding 0.5N NaOH, first are incubated for a further 20 min at 37 ° C. (Reagent C, Table 1).

Cholesterol analysis, described under C, can also be carried out by another method, as described by Ishikawa et al. in "J. Lipid Res. ", 15,286,1974.

The desired amount of cholesterol esters is now found as the difference between the amounts of total and free cholesterol. If a lower amount of cholesterol is found in the test compound-treated cells than in the untreated cells, the test compound is considered to lower cholesterol esters and is therefore considered active.

Table 1

A. Composition of Reagents for Cholesterol Determination Free Cholesterol Reagent

Sodium phosphate buffer pH 7.0

0.05

mol

Cholesterol oxidase

0.08

U / ml

Horseradish peroxidase

30th

U / ml p-hydroxyphenylacetic acid

0.15

mg / ml

Total cholesterol reagent

Sodium phosphate buffer pH 7.0

0.05

mol

Cholesterol ester hydrolase

0.08

U / ml

Cholesterol oxidase

0.08

U / ml

Horseradish peroxidase

30th

U / ml

Sodium taurocholate

5

mmol

Carbowax-6000

0.17

mmol p-hydroxyphenylacetic acid

0.15

mg / ml

C. Sodium hydroxide solution 0.5N

A daily dose of about 100 to about 5000 mg is recommended for the use mentioned above, which can be administered in portions of 25 to 2500 mg two to four times a day or in a delayed release form.

The compounds are mixed with conventional, pharmaceutically acceptable diluents and carriers and, if appropriate, with other additives, and are preferably administered orally as tablets or capsules.

example 1

a- [(l-oxo-9,12-cis, cisoctadecadieny lamino) J-lH-indole-3-

propanoyl acid ethyl ester

2.5 g of triethylamine and 2.7 g of ethyl chloroformate are successively added to a solution of 7.0 g of linoleic acid in 150 ml of methylene dichloride which has been cooled to −20 ° C. The reaction mixture is stirred for 2 hours, slowly taking on room temperature. First 2.5 g of triethylamine and then 6.7 g of DL-tryptophanethyl ester hydrochloride are added, after which the mixture is stirred for 18 h. The reaction mixture is extracted several times with 2N NaOH and washed with saturated aqueous sodium chloride solution, after which the organic phase is dried over anhydrous sodium sulfate, filtered and evaporated in vacuo. The residue obtained is eluted through silica gel with chloroform as the running liquid. The filtrate contains the title compound, which is obtained as a waxy solid after evaporation.

Example 2

The following compounds are obtained in an analogous manner if corresponding starting materials are used in almost equivalent amounts:

a) a - [(1-oxo-9-cisoctadecenylamino)] - 1 H-indole-3-propanyl acid ethyl ester (waxy solid);

b) a - [(1-oxo-9,12,15-cis, cis, cisoctadecatrienylamino)] - 1 H-indole-3-propanoyl acid ethyl ester;

c) a - [(1-oxo-9-cishexadecenylamino)] - 1 H-indole-3-propanoyl-acidic ethyl ester;

d) a - [(1-oxo-5,8,11,14-cis, cis, cis, ciseicosatetraenylamino)] - 1H-indole-3-propanoyl acid ethyl ester;

e) ethyl 2- (1-oxo-9-cisoctadecenylamino) -3- (1-methylindolyl) propanoic acid;

f) ethyl 2- (1-oxo-9-cisoctadecenylamino) -3- (1 H-5-methoxyindolyl) propanoyl;

g) ethyl 2- (l-oxo-9-cisoctadecenylamino) -3- (1H-5-chloroindolyl) propanoyl;

h) ethyl 2- (l-oxo-9-cisoctadecenylamino) -3- (l-benzylindolyl) propane-oylate;

i) a - [(l -oxo-9,12-cis, cisoctadecadienylamino)] - 1 H-indole-3-propanoyl acidic n-propyl ester;

j) a - [(l -oxo-9,12-cis, cisoctadecadienylamino)] - 1 H-indole-3-propanoyl acidic n-butyl ester;

k) a - [(1-oxo-9,12-cis, cisoctadecadienylamino)] - 1 H-indole-3-propanoyl acidic n-octyl ester (mp. 66-68 ° C);

1) a [(l -oxo-9,12-cis, cisoctadecadienylamino)] - 1 H-indole-3-propanoyl acid benzyl ester;

m) a - [(1-oxo-9-cisoctadecenylamino)] - 1H-indole-3-propanoyl acidic n-propyl ester;

n) a - [(1-oxo-9-cisoctadecenylamino)] - 1 H-indole-3-propanoyl acidic n-butyl ester;

o) a - [(1-oxo-9-cisoctadecenylamino)] - 1H-indole-3-propanoyl acid n-octyl ester;

p) a - [(l -oxo-9-cisoctadecenylamino)] - 1 H-indole-3-propanoyl acidic benzyl ester;

q) N- (a-methylbenzyl) cis-2'-octylcyclopropanoctanamide; (Mp 44-46 ° C); (made from d (+) - a-methylbenzylamine);

r) a - [(1-oxocis-2'-octylcyclopropanoctylIamino)] - 1 H-indole-3-propanoyl acid ethyl ester;

s) N- (a-methylbenzyl) cis-2'-octylcyclopropanoctanamide; (Mixture of 2 racemates; mp 30-32 ° C) (made from (d, l) -a-methylbenzylamine);

t) N- (o-methylphenyl) cis-2'-octylcyclopropanoctanamide; u) N- [a- (p-methylbenzyl) benzyl] cis-2'-octylcyclopropanoctane amide (mp 78-81 ° C);

v) N- (a-methylbenzyl) cis-2'-hexylcyclopropanoctanamide; w) N- (a-methylbenzyl) trans-2'-octylcyclopropanoctanamide; x) N- (a-methylbenzyl) cis-2'-tetradecylcyclopropane butanamide; y) N- [a- (p-methylbenzyl) -ß- (p-methylphenyl) ethyl] cis-2'-octyl-cyclopropanoctanamide (mp 60-65 ° C);

z) N- (a-methylbenzyl) cis, cis-2 '- [(2-pentylcyclopropyl) methyl] cyclopropanoctanamide (oil mixture of diastereoisomers made from d (+) - a-methylbenzylamine);

aa) a-cis, cis- {1-oxo-2 '- [(2-pentylcyclopropyl) methyl] cyclo-propanoctanamino} -lH-indole-3-propanoyl acidic ethyl ester from D L-triptophanoethyl ester hydrochloride;

ab) N- (a-methylbenzyl) cis, cis-2 '- [(2-pentylcyclopropyl) methyl] cyclopropanoctanamide (mixture of diastereoisomers made from (d, l) -a-methylbenzylamine);

ac) N- (o-methylphenyl) cis, cis-2 '- [(2-pentylcyclopropyl) methyl] cyclopropanoctanamide;

ad) N- [a- (benzyl) -ß- (phenyl) ethyl] cis-2'-octylcyclopropanoctane amide (mp 40-45 ° C);

ae) 2- (1-oxo-9-cisoctadecenylamino) -3- (1 H-5-fluoro-3-indolyl) propionic acid ethyl ester (wax).

The following table lists NMR constants of compounds from the previous examples. The NMR determinations

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

644 842

6

mung was carried out in CDC13. The numbers are in parts per million (ppm) and those in parentheses indicate the number of protons; s means singlet, d doublet, t triplet and b broad.

The compounds of the formula I are also generally usable as agents for lowering the cholesterol and cholesterol content in serum. Rabbits who were given 200 mg / kg of the test compound orally in combination with a high choesterol diet over a period of 9 weeks showed a lower cholesterol and cholesterol content io in serum and reduced or even reduced education compared to control animals Absence of a coating in the blood vessels.

For this application, the dosages are the same as those mentioned at the beginning.

E.g.

NMR

1

15.3 (2), d 6.2 (1), s 9.2 (1)

2a

15.3 (4), d 6.1 (1), s 8.8 (1)

2e

15.35 (2), d 6.0 (1), s 3.7 (3) (oil)

2f t 5.4 (2), d 6.1 (1), s 8.4 (1), s 4.85 (3) (wax)

21st

15.35 (4), d 6.1 (1), s 8.5 (1), s 5.1 (2) (oil)

2r b -0.36 (1), d 6.1 (1), s 8.8 (1), s 0.6 (3)

2z b +0.7 (5), b -0.3 (2), d 6.0 (1), s 7.3 (5)

2aa b +0.7 (5), b -0.3 (2), d 6.2 (1), s 8.5 (1) (wax)

2ab b +0.65 (5), b -0.3 (2), d 5.8 (1), s 7.3 (5) (01)

2ae

15.3 (2), d 6.1 (1), s 8.7 (1)

R

Claims (7)

644 842 2nd PATENT CLAIMS 1. Compounds of formula (I) O II A-C-NH-R, (I) in the A (I) denotes the C7 to C23 tail of an unsaturated fatty acid which contains 1 to 4 ethylenically unsaturated sites, or (II) denotes a corresponding radical in which the ethylenically unsaturated sites —CH = CH— by cyclopropanyl groups CH2 -CH-CH- are replaced, and R2 represents a radical of the formula (II), (III) or (IV) -CH- (CH2, g - (= ^ in the gOoderl, R4 is hydrogen, fluorine, chlorine, bromine or C to C3-alkyl or alkoxy, R5 is hydrogen, fluorine, chlorine or Q to C3 alkyl or alkoxy, and R3 is hydrogen, Q to Cs-alkyl or a radical of the formula (VI) (II) Rr 4. A compound according to claim 1, which is a - [(l-oxo-9-cisoc-tadecenylamino)] - 1H-indole-3-propanoyl acid ethyl ester. 5. Compounds according to claim 1, in which A has the meaning (II). 5 6. Compounds according to claim 5, in which R2 represents a radical of formula (II). 7. A compound according to claim 1 which is N- [a- (benzyl) -ß- (phenyl) ethyl] cis-2-octylcyclopropanoloctanamide. 8. Compounds according to claim 1 as an active ingredient against io arteriosclerosis. 9. A process for the preparation of compounds of formula (I), reproduced in claim 1, characterized in that compounds of formula (XII) ] s H2N-R2 (XII) in which R2 has the meanings mentioned in claim 1, with acids of the formula (XIII) A-COOH (XIII) 20 in which A has the meanings mentioned in claim 1, or with its functional derivatives. 10. Medicament containing an active ingredient according to claim 1 and pharmaceutically acceptable diluent and carrier. - <œ2> 3 -GC (VI) 30th means where j is 0 or 1, Y is hydrogen, fluorine, chlorine, bromine or Q to C3-alkyl or alkoxy, and Yt represents hydrogen, fluorine, chlorine or Ci to C3 alkyl or alkoxy; 35 /, 5 ^ (III) in the 6th R5 has the abovementioned meaning and R6 is hydrogen, fluorine, chlorine, bromine, Ct to C3-alkyl or alkoxy or a radical of the formula (VII) _ (B) f_ / W means in the B - CH2— or —O—, f is 0 or 1, and R4 has the meaning mentioned above; COOR- (VII)