SI8911762A - NEW ORGANIC NITRATES AND PROCEDURES FOR THEIR PREPARATION - Google Patents

Abstract

New organic nitrates with a specific composition, consisting of nitrato fatty acids (nitratato alkanoic acids) and sulfur-containing amino acids or peptides, with the general formula 02N—O—CH2—c—(CH2)j 0 ΚΙ I -NR4 O i n -(CH2 )n—C—-(CH2 )Q—C—R (A) i5 (A), in which the symbols have the meanings given in the claims. Based on the general principle of construction - the presence of sulfhydryl groups prevents nitrate tolerance or weakens the already existing tolerance. The compounds are used in medicines for the treatment of circulatory diseases, high blood pressure, heart failure and for the expansion of peripheral blood vessels.

Description

1. TECHNICAL FIELD

The invention relates to the field of organic and pharmaceutical chemistry The invention relates to new organic nitrates and processes for their preparation

2. TECHNICAL PROBLEM

It is an object of the invention to provide novel organic nitrates suitable for the treatment of heart disease and to provide methods for their preparation.

3. BACKGROUND OF THE INVENTION

Organic nitrates (nitric acid esters) have been proven in the treatment of heart disease.

They develop their effects by relieving the heart by reducing pre- and post-loading, as well as by improving the supply of oxygen to the heart by expanding the coronary vessels.

In fact, in recent years it has been established that organic nitrates such as glycerin nitrate have been used in therapy so far *

(GTN), isosorbide-5-mononitrate or isosorbiddinitrate nri by volume and continuous introduction into the body over a relatively short period of time, show a significant decrease in the effect of nitrate tolerance. Numerous experiments indicate that the presence of sulfhydryl groups prevents the formation of nitrate tolerance and diminishes one already formed tolerance.

2.

The mechanism of tolerance generation is today understood as follows:

To the present state of knowledge, the pharmacological action of organic nitro compounds is dependent on the presence of cysteine. With it, organic nitrate forms a common pre-phase, from which the decomposition, among other things, releases the KO radicals that activate the target enzyme, the soluble guanylate cyclase of the smooth muscle cells. Furthermore, by the formation of the subsequent reactions induced by cGMP, relaxation or relaxation occurs. dilatation of blood vessels.

The reactive and short-lived, so far hypothetical intermediate, may be a nitric acid thioester or a single thionitrate. The intramolecular dislocation and subsequent reactions, which have not yet been elucidated, have recently been postulated to form a nitrosothiol, from which nitrogen monoxide is released. nitrite ions. Enzyme-dependent digestion with GSH-reductase may, however, since it leads solely to the formation of nitrite ions, may be irrelevant to its pharmacological action. Non-enzymatic degradation requires, as described, cysteine and thus depletion depending on the dose (depletion of SH-Gruppen pools) so that no longer enough KO is formed as a true guanyl cyclase activator in the long run , and there is a clinically impaired effect.

For the substances according to the invention, it is a specially formulated compound consisting of nitrato fatty acids (nitrato-alkanoic acids) and sulfur-containing or amino acids, respectively. peptides.

It is therefore an object of the present invention to make available new organic nitrates, which by virtue of their general structure - by the presence of sulfhydryl groups - are distinguished by preventing or reducing their existing nitrate tolerance. or tolerance to the resulting nitrate.

3.

4. · DESCRIPTION OF THE SOLUTION

This task is solved by the fact that the compounds have the general formula

(CH ₂ ) _o —CR (A) in which

R is hydroxy, lower-alkoxy, lower-alkoxy, di-lower-alkylamino-lower-alkoxy, acylamino-lower-alkoxy, acyloxy-lower-alkoxy, aryloxy, aryl-lower-alkyloxy, substituted aryloxy or substituted aryloxy-lower-alkoxy is a methyl, halogen or methoxy substituent; amino, loweralkylamino, di-lower-alkylamino, aryl-lower-alkylamino, hydroxyamino or amino acid residue via a peptide bond, hydrogen, alkyl of 1 to 6 carbon atoms, substituted lower-alkyl, wherein the substituent is halogen, hydroxy, lower- alkoxy, aryloxy, amino, lower-alkylamino, acylamino, acyloxy, arylamino, mercapto, lower-alkylthio, arylthio,

R and R denote hydrogen or lower alkyl, * 3

R ^J hydrogen or arrays alkyl,

R ^ hydrogen, lower-alkyl, phenyl, methoxyphenyl, phenyl-lower-alkyl, methoxy-phenyl-lower-alkyl, hydroxyphenyl-lower-alkyl, hydroxy-lower-alkyl, alkoxy-lower-alkyl, aminonyl alkyl, acylamino-lower-alkyl, mercapto - lower-alkyl or lower-alkylthio-lower-alkyl,

4.

C

R is lower-alkylthiol, its S-acyl compounds, in particular

S-acetate, S-propionate, S-butyrate, S-capronate, S-karylate, 't

S-pivalate, S-benzoate; lower-alkyl-S-C-O-R u

lower-alkyl-S-C-N-R, lower-alkylthio-lower-alkyl-carboxylic acids and / or its esters and / or amides, lower-alkylthio-lower-alkyl,

R and R may be attached to each other to form one thiolactone,

R and R ^ may be attached to each other to form an ester or amide,

R ^ and R ^ can be bonded to each other to form one alkylene bridge with 2 to 4 carbon atoms, one alkylene bridge with 2 to 3 carbon atoms and a sulfur atom, one alkylene bridge with 3 to 4 carbon atoms, containing a double bond or an alkylene bridge as above, substituted by hydroxy, lower-alkoxy, lower-alkyl or di-lower-alkyl, m, nio have a number of values 0 to 10, and pharmaceutically acceptable salts thereof.

According to a further embodiment of the process, the nitrate fatty acid components have a series length of - Οθ; they may be of the normal sequence, branched, racemic or optical isomers.

The amino acids cysteine, methionine or homocysteine are primarily used.

Suitable amino acids have a stereochemical L-form.

5.

Cysteine and / or methionine may be in the form of their methyl, ethyl or propyl esters.

SH-group of cysteines can be esterified with lower alkane carboxylic acid with 2 to R carbon atoms

According to one particularly advantageous further embodiment of the process the protected compounds have the following chemical formulas:

N- (2-Kitratoacetyl) -cysteine ethyl ester

N- (2-Nitratoacetyl) -S-acetyl-cysteine ethyl ester

N- (2-nitroacetyl) -S-propionyl-cysteine ethyl ester

N- (2-Nitratoacetyl) -S-pivaloyl-cysteine ethyl ester

N- (2-Nitratoacetyl) -methionine methyl ester

N- (2-Nitratopropionyl) -cysteine

N- (2-Nitratopropionyl) -cysteine ethyl ester

N- (2-Kitratopropionyl) -methionine ethyl ester

N- (2-Nitratobutyryl) -cysteine

N- (2-Nitratobutyryl) cysteine ethyl ester

N- (2-Nitratobutyryl) -S-acetyl-cysteine ethyl ester

N- (2-Nitratobutyryl) -S-butyryl-cysteine ethyl ester

N- (2-Kitratobutyryl) -methionine-ethyl ester

N- (2-Nitratoisobutyryl) -cysteine

N- (2-Nitratoisobutyryl) -cysteine ethyl ester

N- (2-Nitratoisobutyryl) -S-benzoyl-cysteine ethyl ester N- (2-Nitratoisobutyryl) -S-acetyl-cysteine ethyl ester N- (2-Nitratoisobutyryl) -S-pivaloyl-cysteine ethyl ester <N- (2 -Nitratoisobutyryl) -raethionine ethyl ester N- (3-Nitratobutyryl) -cysteine

N- (3-Nitratobutyryl) -cysteine ethyl ester

N- (3-Nitratobutyryl) -S-acetyl-cysteine ethyl ester

N- (3-Nitratobutyryl) -S-propionyl-cysteine ethyl ester N- (3-Nitratobutyryl) -methionine ethyl ester N- (3-Nitratopivaloyl) -cysteine

N- (3-Nitratopivaloyl) -cysteine ethyl ester

6.

N- (3-Nitratopivaloyl) -cysteine ethyl ester-S-carbonate

N- (3-Nitratopivaloyl) -S-acetyl-cysteine ethyl ester

N- (3-Nitratopivaloyl) -S-propionyl-cysteine ethyl ester

N- (3-Nitratopivaloyl) -S-butyryl-cysteine ethyl ester

N- (3-Mtratopivaloyl) -S-isobutyryl-cysteine ethyl ester

N- (3-Nitratopivaloyl) -S-pivaloyl-cysteine ethyl ester

N- (3-Nitratopivaloyl) -S-benzoyl-cysteine ethyl ester

N- (3-Nitratopivaloyl) -methionine ethyl ester

N- (3-Nitratopivaloyl) -methionine

N- (3-Nitratopivaloyl) -homocysteine thio lactone

N- (2-Nitratohexanoyl) -cysteine ethyl ester

N- (2-Nitratohexanoyl) -S-propionyl-cysteine ethyl ester

N- (3-Nitratohexanoyl) -cysteine ethyl ester

N- (3-Nitratohexanoyl) -methionine methyl ester

N- (12-Nitratolauroyl) -cysteine h- (12-Nitratolauroyl) -eisteine ethyl ester

N- (12-Nitratolauroyl) -S-acetyl-cysteine

N- (12-Kitratolsuroyl) -S-pivaloyl-cysteine

According to another embodiment of the invention, the medicaments comprise one and / or one mixture of compounds of the invention.

Θνί drugs can be used to treat diseases of the bloodstream, for example as coronary dilators, as agents for the treatment of high pressure, heart failure, for the peripheral blood vessels to spread, including the blood vessels of the brain and blood vessels of the kidney.

Finally, the compounds may be prepared in a manner known per se, wherein the nitrate fatty acids or their reactive derivatives are condensed with the amino group of one amino acid or. of one peptide. The compounds obtained may optionally undergo, at a further stage of the reaction, alkylation or acylation of the side chain.

7.

inactive derivatives according to the invention of nitratomatic acids used are, for example, acid halides, acid anhydrides, activated amides or activated esters. Preferred are chlorides.acids, acid azides, symmetric acid anhydrides, activated esters, and mixed anhydrides with organic or inorganic acids.

The condensation reactions of nitrate fatty acids with amino groups of amino acids can be carried out in one inert solvent and in the presence of a condensing agent that interferes with the formation of acid amide bonds, one carbodiimide, such as N, N'-dicyclohexylcarbodiimide, one imine compound, one imine compound such as diphenylketene-N-cyclohexyl imine or pentamethylene-ketene-K-cyclohexyl imine or a single phosphate or phosphite, such as triethylphosphite, ethylpolyphosphate or isopropylpolyphosphate over a period of 1 - 48 hours, at temperatures of -10 ° C to the reflux temperature of the solvent used.

The examples clarify the invention, however, which is not limited to them.

EXAMPLE 1

Preparation of N- (3-nitratobutyryl) -cysteine ethyl ester

1. Careful work

Saponification of 3-bidroxybutyric acid ethyl ester

13.2 g (0.1 mol) of 3-hydroxybutyric acid ethyl ester (Aldrich) was treated with 4.0 g (0.1 mol) of NaOH dissolved in 100 ml of water. The reaction is complete when the solution becomes homogeneous.

The treatment was carried out in such a way that the resulting solution was acidified with 10 ml of concentrated HCl and extracted twice with 100 ml of ethyl acetate each. After that, the solution is evaporated on a rotary evaporator, leaving a low viscosity oil.

The yield was 8.81 g (theoretical: 10.4 g) of 3-hydroxybutyric acid.

2. Careful work

Nitration of 3-hydroxybutyric acid

8.81 g (0, 8 mol) of 3-hydroxybutyric acid and 50 mg of urea are dissolved at 5 ° 0 in 50 ml of acetic acid, 6.27 ml (0.15 mol) of HNO4 are added dropwise, and thereafter 14.17 ml (0.15 mol) under cooling. The reaction mixture was stirred overnight.

The treatment was carried out in such a way that the resulting solution was treated with 200 ml of ice water and extracted with ethyl acetate. The organic phase is extracted with NaHCO3. The NaHCO3 phase was acidified with concentrated HCl and extracted with ethyl acetate. After that, the solution is evaporated on a rotary evaporator, leaving a low viscosity oil.

9.

The yield was 9.4 g (theoretically 11.9 g) of 3-nitratobutyric acid.

3. Careful work

Preparation of N- (3-nitratobutyryl) -cysteine ethyl ester

16.6 g (0.11 mol) of 3-Nitratobutyric acid were dissolved in 100 ml of dichloromethane. At 15 ° G, 17.9 g (0.12 mol) of cysteine ethyl ester were added slightly to the introduction. Subsequently, 24.7 g (0.12 mol) of dicyclohexylcarbodiimide (DCC) dissolved in 80 ml of dichloromethane are added dropwise at 15 ° C with the introduction of Np. At the end of the reaction, the dicyclohexyl carbamide formed was treated and the solution was washed with 150 ml of 0.1 N HCl. Then the solution is evaporated on a rotary evaporator.

Purification of the substance is carried out by preparative column chromatography and recrystallization from ethanol / n-hexane.

Yield: 6.88 g (theoretical: 30.83 g).

M.p .: 77.8 ° C.

EXAMPLE 2

Preparation of H- (3-nitratobutyryl) -methionine ethyl ester

6.35 g (0.043 mol) of 3-Kitratobutyric acid, 7.47 g (0.043 mol) of methionine ethyl ester and the top of the dimethylaminopyridine (DMAP) spatula were dissolved with stirring and cooled to 10 ° 0 in 100 ml of dichloromethane. 10.31 g (0.05 mol) of DCC was dissolved in 80 ml of CHpClp and, with the simultaneous introduction of nitrogen, was slowly added dropwise. After the reaction is complete, the solution is washed, washed with NaHCO3 and then with HCl. The solution was evaporated on a rotary evaporator, leaving behind oil.

Purification is performed by preparative column chromatography or cold crystallization.

The yield is 1.95 g (theoretical 12.05 g) of K- (3-nitratobutyryl) methionine ethyl ester as a colorless oil.

EXAMPLE 3

Preparation of N- (3-nitratopivaloyl) -cysteine ethyl ester

1. Careful work

Preparation of nitratopivalic acid methyl ester

25.0 g (0.19 mol) of hydroxypivalic acid methyl ester and 0.12 g of urea are dissolved at room temperature in 250 ml of CH 2 Cl 2 and cooled to 5 ° C with stirring. This was added dropwise with stirring of 23.8 g (0.38 mol) of HNO4 (100%), so that the temperature did not exceed 10 C. After that, it was cooled to 5 C and then dropped.

38.6 g (0.38 mol) of acetic anhydride, so that the temperature does not exceed 10 ° C. It is stirred for 15 minutes with cooling in an ice bath, after which it is slightly warmed to room temperature and stirred overnight at room temperature. This mixture was added gently with stirring to 500 ml of ice water. The CH2 Cl2 phase was separated by washing once with 100 ml of distilled H 2 O, 100 ml of saturated aqueous NaHCO 3 and once again with 100 ml of dist. H ^ O. The CH2 Cl2 -Extract was then evaporated on a rotary evaporator at a bath temperature of a maximum of 40 [deg.] C. under vacuum with a water jet until dry. The light yellow oily residue was distilled in a vacuum obtained with an oil vacuum pump at 60 ° C of bath temperature as a low-viscosity colorless oil.

Yield: 31.5 g 5 * 94.0% of theory.

2. Careful work

Preparation of nitratopivalic acid

14.0 g (0.350 mol) of NaOH was dissolved in the dist. H ^ O and cool

11.

at about 10 ° C. To this was added with stirring a solution of 31 »0 g (0.175 mol) of nitratopivalic acid methyl ester in 250 ml of methanol, whereby the reaction mixture turned yellowish and the temperature increased to about 25 ° C. After stirring for 90 minutes, the mixture was neutralized with 29.5 ml (0.35 mol) of 37% HCl and methanol was completely distilled off on a rotary evaporator. The aqueous phase was extracted twice with 200 ml of methylene chloride each. The combined methylene chloride extracts were washed once with 50 ml of distilled H 2 O and the methylene chloride phase was evaporated to dryness on a rotary evaporator. The colorless oily residue was dissolved in 100 ml of ethyl acetate and re-evaporated on a rotary evaporator to dryness, leaving a solid white residue from which the solvent residues were removed on vacuum obtained with an oil pump (0.4 Torra) at a bath temperature of about 40 ° C for 15 minutes on a rotary evaporator. A solid white residue of 25.44 g (89.1% of theory) was dissolved in 100 ml of boiling n-hexane and treated with 2 m diisopropyl ether. After cooling to room temperature and adding grafting crystals, the product crystallizes. The product was left at 0 ° C for 72 hours, the crystals were processed and after washing twice with 10 ml n-hexane each, they were dried in a vacuum oven to constant weight at room temperature at about 2 Torr.

M.p .: 54.2 ° C.

Yield: 23.66 g = 82.9% of theory.

*}. Careful work

Preparation of N- (3-nitratopivaloyl) -cysteine ethyl ester

10.7 g (71.7 mol) of the L-cysteinethyl ester of the base was dissolved at room temperature under the N ^ -atmosphere with stirring in 200 ml. .

methylene chloride. To this was added 11.4 g (70.0 mmol) of crystalline nitroxypivalic acid and dissolved at room temperature with stirring. Into this mixture? with stirring and under the atmosphere of '12.

of nitrogen, a solution of dicyclohexyl urea (DCC) in 50 ml of methylene chloride was added dropwise at room temperature for 15 minutes, with the temperature rising to about 35 ° C. After further stirring, white dicyclohexyl urea is separated. The mixture was cooled to room temperature and stirred overnight under a nitrogen atmosphere. Dicyclohexyl urea is then poured into a glass beaker and washed once with 50 ml of C ^ C ^. The combined methylene chloride solutions were washed once with 100 ml of 1 n HCl once and twice with 100 ml of dest. ^ 0 (under ^ -atmosphere) and then evaporated on a rotary evaporator at a bath temperature of about 40 ° C and at a water-jet vacuum at 550 mbar to about 20 mbar initially. Light brown oil is obtained.

Yield: 21.2 g = 102.9% of theory.

The substance was purified by crystallization from ethanol / hexane in the cold.

Yield: 13.42 g = 65.1% of theoretical K- (3-nitratopivaloyl) cysteine ethyl ester as a light pink oil.

4. Careful work

Preparation of N- (3-nitratopivaloyl) -S-acetyl-cysteine ethyl ester

In 10.3 g (35.0 mmol) of N- (3-nitratopivaloyl) -cysteine. The ethyl ester in 70 ml of dichloromethane was added dropwise to a cold stirring solution of 4.3 g (42.0 mmol) of acetic anhydride in 10 ml of dichloromethane. After that, a cold solution of 5.0 g (49.0 mmol) of triethylamine in 20 ml of dichloromethane was added to the hay.

After completion of the reaction, the mixture was washed with 1 n HCl, 10% aqueous sodium bicarbonate solution and water. The dichloromethane extract was evaporated to dryness on a rotary evaporator. 11.6 g of a light yellow oily product are obtained from which crystallization from ethanol / water in the cold with the addition of grafting crystals yields 7.8 g of the crystalline product (66.3% of theory)

T.t .: z. 7 ° C

13.

4. Operation phase / variant 1

Preparation of N- (3-nitratopivaloyl) -S-butyryl-cysteine ethyl ester

If 6.7 g (42.0 mmol) of butyric anhydride are used, instead of the step of step 4 of the acetic anhydride described above, 13.0 g of a light yellow oily product is obtained in the same reaction and treatment, from which, as as described in step 4, cold crystallization obtained

9.7 g of crystalline product (= 76.2% of theory).

M.p .: <5 ° C.

4. Work phase / variant 2

Preparation of N- (3-nitratopivaloyl) -S-pivaloyl-cysteine ethyl ester

If 7.8 g (42.0 mmol) of pivalic acid anhydride are used, instead of the step 4 of the acetic anhydride described above, 14.1 g of a light yellow oily product is obtained in the same reaction and from which, as as described in step 4, crystallization yielded 10.5 g of the crystalline product (= 79.5% of theory).

M.p .: 45 ° C.

4. Operation phase / variant 3

Preparation of N- (3-nitratopivaloyl) -cysteine ethyl ester-S-carbonate

If 4.3 g (42.0 mmol) of hydrochloric acid ethyl ester are used, rather than in step 4 of the acetic anhydride described above, it is obtained by the same reaction and treatment,

11.5 g of a light yellow oily product from which, as described in step 4, crystallization gives 9.5 g of the crystalline product (= 74.1% of theory).

M.p .: 36 ° C.

14.

EXAMPLE 4

Preparation of N- (3-nitratopivaloyl) -methionine ethyl ester

12.4 g (70.0 mmol) of L-methionine ethyl ester base were dissolved at room temperature under stirring in 250 ml of methylene chloride. To this was added 11.4 g (70.0 mmol) of crystalline nitratopivalic acid and dissolved with stirring at room temperature. A solution of 14.8 g (71.7 mmol) of N-Ndicyclohexyl carbamide (DCC) in 50 ml of methylene chloride was added dropwise to this mixture under stirring and nitrogen for 15 minutes at which temperature the temperature was increased to 35 °. C. After further stirring, the white dicyclohexyl urea precipitates. The mixture was cooled to room temperature and stirred overnight under a nitrogen atmosphere. The DCC-urea was then filtered through a glass beaker and washed once with JO ml CHpC ^. The combined methylene chloride solutions were washed once with 100 ml of 1 n HCl and twice with 100 ml of dest, H ^ O (under Nr, atmosphere) and then evaporated on a rotary evaporator at a bath temperature of about 40 ° C and under a water-jet vacuum. initially from 550 mbar to about 20 mbar. A light yellow oil is obtained.

A

Yield: 24.9 g = 110.3% of theoretical crude N- (3-nitratopivaloyl-L-methionine ethyl ester).

The crude product is purified by preparative column chromatography.

Yield: 17.6 = 78.0% of theoretical L- (3-nitratopivaloyl) methionine ethyl ester as a colorless oil.

EXAMPLE 5

N- (12-Nitratolauro yl) -S-acetyl-cysteine

15.

1. Work phase

Preparation of 12-nitratolauric acid

54.1 g (0.250 mol) of 12-hydroxylauric acid and 0.3 g of urea were dissolved with gentle heating in 1.3 L of CHCl 3 and cooled with stirring at 20 ° C. With stirring it is lightly removed

23.6 g (0.375 mol) of HNO4 (100%), with the temperature increasing to 27 ° C. Thereafter, it was cooled to 20 ° G and quenched with stirring and cooling, 38.3 g (0.375 mol) of acetic anhydride, maintaining a temperature limit of 25 ° C. It is stirred at room temperature overnight. Finally it is washed five times with 0.5 1 dest. HpO. The NaCl2-dried and charcoal-dyed CHCl2 phase was evaporated to dryness on a rotary evaporator at a bath temperature of 50 [deg.] C. under a water-jet vacuum. The oily residue of 60.8 g was dissolved in 500 ml of boiling n-hexane and kept at 0 ° 0 overnight after cooling to room temperature. The crystallized product was treated and washed twice with 50 ml n-hexane each. Finally, the product was dried in a vacuum oven at room temperature and under about 2 Torr to constant weight.

M.p .: 29 ° C.

Z *

Yield: 39.4 g = 60.3% of theory.

2. Careful work

Preparation of 12-nitratolauric acid chloride

2.6 l of (10 mmol) nitratolauric acid were dissolved in 50 ml of methylene chloride, and 4.44 g (35 'mmol) of oxalyl chloride were added dropwise to 50 ml of methylene chloride with stirring at room temperature. Scratches overnight. Finally, the product is evaporated to dryness on a rotary evaporator.

Yield: 3 g = 93.2% of theory.

3. Careful work

Preparation of NI (12-Nitratolauroyl) -cysteine

Under a nitrogen atmosphere, 6.06 g (50 mmol) of L-cysteine in 300 ml of DMF was added. 5.60 g (20 mmol) of 12-nitratolauric acid chloride in 50 ml of dichloromethane are added dropwise. Since no clear solution is obtained, it is heated to 60 ° C. Finally add 100 ml of dest. HpO and stirred overnight at room temperature. After that, it is diluted with 300 ml of dist. HpO and extracted four times with 200 ml of ethyl acetate each. The organic phase was dried over NapSO4 and then evaporated. The residue was taken up in 100 ml of ether and left to crystallize overnight at 0 ° C. White crystals are obtained.

M.p .: 74-75 ° C.

Yield: 4.1 g of K- (12-nitratolauroyl) -cysteine.

4. Careful work

Preparation of N- (12-nitro tolauroyl) -S-acetic 1-cysteine

1.82 g (5 mmol) of N- (12-nitratolauroyl) -cysteine were placed under a nitrogen atmosphere in 20 ml of ethyl acetate. It was then cooled to 0 ° C and 2.5 ml of acetic anhydride was added dropwise. Then, 1.52 g (15 mmol) of triethylamine, which was dissolved in 5 ml of ethyl acetate, was added dropwise at -5 ° C. The reaction solution was washed with ^ and evaporated to dryness,

Melting point: at room temperature, oil.

Yield: 2 g = 98.4% of theory.

17.

EXAMPLE 6

Preparation of R- (12-nitratolauroyl) -cysteine ethyl ester g (26.8 mmol) The cysteine ethyl ester base is dissolved in 50 ml of methylene chloride and 2.8 g (10 mmol) of 12-nitratolauric acid chloride, which is dissolved in 50 ml of methylene chloride and stirred overnight. Precipitated cysteine ethyl ester. The HCI was removed and the solvent was removed on a rotary vacuum evaporator. The oily residue (6 g) was dissolved in 100 ml of ether and left in the refrigerator at 0 ° C overnight. The precipitated product is treated.

M.p .: 59-60 ° C.

Λ

Yield: 1.6 g = 40.0 theoretical.

EXAMPLE 7

Preparation of N- (2-nitratopropionyl) -cysteine ethyl ester

Careful work

Preparation of nitrate lactic acid ethyl ester g (0.28 mol) of lactic acid ethyl ester was dissolved in 300 ml of dichloromethane. After addition of 100 mg of urea, 22.5 g (0.56 mol) of 100% nitric acid were added dropwise at 5-10 ° C. The solution was cooled to 0 ° C. Then 52.8 ml (0.56 mol) of acetic anhydride was added dropwise so that the temperature did not exceed 5 ° C. The solution was allowed to stand at room temperature overnight and then washed with 250 ml of water. The organic phase was separated and dried over sodium sulfate. After filtration, dichloromethane is pre-distilled. The resulting oily residue is treated with distillation.

Yield: 30.34 g = 66.4% of theory

T.klj. 34 ° C (0,25 Torra)

18.

2. Careful work

Preparation of nitratomaric acid g (L, 18 ml) of nitratomaric acid ethyl ester was dissolved in 80 ml of dioxane. The solution was treated with 30 ml of water and 2 g (0.02 mol) of sulfuric acid and refluxed for 19 hours. The solution was evaporated to a volume of about 50 ml and then diluted with 300 ml of water. The pH is adjusted by adding sodium bicarbonate to 7-8. The untransformed ester is removed by extraction with dichloromethane.

The aqueous phase was adjusted to pH 1 with hydrochloric acid, extracted three times with 150 ml of ethyl acetate each. The extracts were combined and dried over sodium sulfate. After filtration, ethyl acetate is completely removed by rotary evaporator.

Yield: 14.6 g of a colorless oil = 59.2% of theory

3. Careful work

Preparation of N- (2-nitratopropropionyl) -cysteine ethyl ester g (0.13 mol) of nitratomaric acid and 18.9 g (0.13 mol) of cysteinethyl ester was dissolved in nitrogen atmosphere at 10-15 ° E in 200 ml of dichloromethane. At 15 - 20 ° C a solution of

28.6 g (0.14 mol) of N-N-dicyclohexylcarbodiimide and 75 ml of dichloromethane. After 1 hour, the precipitated N-Ndicyclohexyl urea was filtered and washed with 75 ml of dichloromethane.

The filtrate was extracted twice with 50 ml of 0.1 n hydrochloric acid each. The organic phase is completely evaporated on a rotary evaporator. The crystalline crude product (22.4 g) was recrystallized from 100 ml ethanol / n-hexane (1: 1).

Yield: 7.6 = 22.6% of theory

M.p .: 92.8 ° C

19.

Pharmacological test system 1

The effect of new organic nitrates on the bloodstream parameters of a waking dog for evidence of nitrate activity.

The purpose of the test is to determine how new organic nitrates after intravenous or oral administration affect different blood flow parameters in a waking dog. All experiments were performed on trained dogs of the Beagle breed; blood flow parameters were described using one arterial catheter-tinmanometer and one in the V. Jugularis introduced submerged catheter (Einschwemmkatheter). To describe the effect on the arterial system, systolic arterial pressure (SAP systolic arterial pressure), mean arterial pressure (MAP mean arterial pressure), and diastolic arterial nritis (DAP diastolic arterial pressure) blood pressure (BP) and heart rate (HR heart rate) were measured ) <Peripheral resistance (TRP total periphere resistance) and arterial elongation (COMPL) were therefore calculated. The low pressure system is described by 'CVP central venous pressure' and pulmonary arterial pressure (PAP pulmonal arterial pressure). Isosorbide-5-mononitrate (ISM-5) was used as the reference solvent.

The following figures 1 and 2 explain graphically the spectrum of action of organic nitrates according to the invention.

In Figure 1, the effects of the orally and intravenously administered ISM-5 are presented. After both applications of the ISM-5 downhill, a little systolic blood pressure, at medium pressure, almost does not work, the elasticity of the bellows increases significantly and the pressures in the low pressure system decrease.

Figure 2 shows, the corresponding effects of K- (3-nitratopivaloyl) methionine ethyl ester (Nitrato-Piv-Meth-Et) in the corresponding

20.

parts of the bloodstream. Here again, there is a comparison between intravenous and oral administration for good bioavailability (BioverfUgbarkeit).

The substance ft- (3-nitratopivaloyl) -cysteine ethyl ester also shows good bioavailability and one typical nitrate course of action.

The invention demonstrates that both test substances have an effect comparable to that of ISM-5 and have good bioavailability.

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FIG. 1 ~ r "T"

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160 240 300

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120 -

180

360 [MINI

ISM-5 5.12 · οΑβ p.o., and Ethanol pain + gem · M 3

240

300

** ι-, -

Ι- <~ Ir ~

ΙΟ-

Η-4-l · --i — f-J - I - i-i · * ϊ * -, -______

100

Ig ΘΟ .Fig. 3 —ι-r120 1Β0 -, -, -,

240 300 360 (MINI

Abb. 1

SV [ml]

TPA iamHa / l / ainl

BO -

-

0—1 p -—-- r - · -> - _r - ------! - | ...........- -i

O BO 120 1B0 240 300 360 (HINJ

FIG. 4

ISM-5 5.12 ao / kfl p.o., and Ethanol paan -f sem. N - 3

200-,

160 -

120 -

BP [mmHg] Hfl [l / min]

ao40-ADP-- I

-m ', ^ Η-Η-Η-Η + Η-1 1' · ' ¹ ' ¹ lili i - ^ - ^ -ι-ΐ4-Η -} - _? 4Η4-Ρΐ-Ί4-1-4Η4-Ρ-Ι-ί-Ρ | 0-1, - _r

60

FIG. 5

120 -,. ,,

160 240 - _r —— · j

300 360 [MINI

Abb. 1

ΙΟ-,

CVP

CO [mmHg] [1 / alnl a6-

JSM-5, 5.12 Bfl / kfl I.V. · 1Π NeCl 0.9% mean + sem. N - 3. --HDDI

PAP CmmHg]

Coapl Cal / 40mmHg]

20Lu-H-H-h-h-H-H-H-H-I '

10.

0-J

K ⁴ t · —H- - «- 0 60

FIG. 7

-I - * - p ........... j-Γ ——........ Γ-— -..... 1

120 ISO 240 300 360 (MIN)

100 — r SV (al) _

TPfl CmmHg / 1 / αΙπ]

Abb. 1 -

-

Oi4 —r

I

120

t ....... ¹ r '........— i-1

ISO 240 300 360

FIG. 8 ΧΜΧΝ)

ISM-5.

mean + sem. N 3

S.12 ng / kg I.v «. ln NaCl 0.9Χ

200 —j BP [ramHg] Hfl [1 / nin]

120 -

160-

f I-l · -!; - J—

H

1 — i — i — 1 — i — I i — i — I — i — i — i

400-4 “T” T “—ΓΟΟ —r—

120 "T"

150

ISO —and 210

FIG. 1 [MIN]

Abb. 2 —j CVP [ramHg] CO [1 / ratn] a

The scoreboard is now 0-1

H + i-4-1-i-i-Ϊ-i-J-i

O 30 Fig. 2 —T —r “

120 "T"

150 r ¹¹¹

160 —I 210 [MIN]

Nltroxy-PlV “Meth-2t, masn + sem. N · 8

8.63 mg / kg IV, m OMSO 100Χ

Abb. 2

50 — ι -

-

Pap [noHg]

Compl [ml / 40ramHg]

-hhH-4

- t - i - _f —s— t-ί-l · —1— I -

ri ri ao 50

FIG. 3 -, -, - _r - · p —... „,

120 150 150 210

Abb. 2

100-,

SV [ml]

TPA [mmHg / l / aln]

BOSO—

4020I --- I-.ri-d -> - and —I-ri

M-I I I I I I

0-4, · .— - ι, - ', —j,

30 50 90 120 150 150 210

FIG. 4 MM

Nitroxy-Piv ~ Meth “Et mman * sem. N - 5

5.53 mg / kg l.v., and DMSO 100Χ

200-, BP [mmHol HR Cl / minl

OJ, - _r

60

FIG. 5 —j I 'I -! -, _t

120 ISO 240 300 360 [MIN]

Abb. 2

2OJ

CVP co [mmHfl]

Cl / nin]

H-l-f '

Hh-i-i- i -i i i i o

FIG. 6 "T" and ^-

120 —r “

ISO —r—

240

-h - r—

300 — and 360 [MINI

Nitroxy-Piv-Moth-Ht, pean 4- ge ™ * N · 5

8.63 Bfl / kfl p.0 "In DMSO 100"

50-, ΡΑΡ [nmHo]

Coapl [®l / 40ramHfl]

υΗΤ-Η-Η'Η-Η'Η'Ή'ί ·

4030 ™ '*' l · -I- i .J. ₊ 4 - itii - Hl · 4H - HI- 4- -1- + ' ⁱ - P -I- -i Il · -I- if -h -I- -iiP * - * * · ^{? -I}

0-4 _r -. -,

O 60

FIG. 7 -, -, -, - | - ........ |

120 180 240 300 360 (MINI

Abb. 2

100-,

60SV [acre] TPH -

40n 14-1 1 H-H-H

20 --- 60 120 -, - 1—

160 240

Fig · θ -, - 300 360 (MINI

Nltroxy-PlV “Meth ~ et, MM + sem, N S

8.63 «p / kO Ρ · 0 ·, ln DMSO 100Χ

29.

Pharmacological test system 2

The effect of new organic nitrates on the increase in coronary flow. on isolated perfused hearts to determine residual tolerance.

The aim of the following experiments was to investigate the effect and retention of tolerance of novel nitrate compounds on isolated perfused hearts in rats. For this purpose, one rat heart was isolated and prepared as a “Working Heart”.

The heart fulfills in this test system one defined work on circular blood flow, from which one defined oxygen consumption and coronary flow result. The action of nitrate compounds in this model can be measured on a pharmacologically induced increase in coronary flow.

On the isolated heart of rats it works, resistance of coronary vessels is selected as a parameter for determining nitrate activity. The heart of rats weighing about 1 g was perfused over the left ventricle with a plasma-like solution containing nutrients and saturated with oxygen. The left ventricle pumps the solution against one defined pressure in the aorta. In proportion to physiological conditions, one part of this solution flows through the coronary blood vessels for ^c heart satisfaction only. For defined cardiac output, this part, from which coronary resistance can be calculated, is constant. The addition of a single nitrate or other coronary dilator pharmaceutical preparations has the effect of reducing this coronary resistance.

If, therefore, a constant concentration of organic nitrates is added to the hearts, after one initial decrease in resistance over a period of 20 minutes, a partial decrease in activity occurs. Substances also show a coronary dilator effect in this model, which is not accompanied by a decrease in activity.

Even after 60 minutes, the maximum decline in coronary resistance is still fully present. The test substances are compared

30.

at equimolar doses with 10 ^{4 4} M nitroglycerintrinitrate. Continuous infusion of nitroglycerin causes a rapid increase in coronary flow by 7.6 - 1.88 ml / min (- SD). During 20 minutes the flow decreases by 55.9%. On further perfusion, the action of nitroglycerin proceeds unchanged. Row nitrates show in this experimental model also an increase in coronary flow, which is actually accompanied by only a very small decrease in activity. This result indicates that the described new compounds do not show any suppression of tolerance as common nitrates.

Table 1: Effect of new organic nitrates compared with nitroglycerin on coronary flow in isolated rat perfused hearts. x - SEM, n = 7

Maximum Coronary Flow Increase (ml / min ³ ^)

100 pM Nitroglycerin 7.6 - 0.71

56.0

100 µM N- (3-Ritratopivaloyl) -cysteine ethyl ester 6.6 ί 0.88 5.2

100 R- (3-Ritratopivaloyl) -methioninethyl ester 8.3 - 0.92 7.o ί SEM = standard deviation of the mean n = number of standard deviations

Finally, the action of N- (3-nitratopivaloyl) cysteine ethyl ester (Nitrato-Piv-Cy-Et) on the hearts of guinea pigs (guinea pigs) is characterized. Nitrato-Piv-Cy-Et brings on Working

Λ '

Heart model to the increase of coronary flow depending on concentrations, already in one very low dosage range. A 25% increase in flow was achieved with 380 g of nitrate-Piv-Cy-Et / l perfusion medium corresponding to 1.3 Mol / 1. The appropriate concentration for glyceroltrinitrate (GTN) at 5 mg / l is higher than the minimum factor of 12. According to torn, nitrato-Piv-Cy-Et is particularly a blood vessel active compound. The attenuation of the coronary dilator effect as an expression of a single tolerance development could not be detected at any one hour dosing. perfusion. Therefore, it can be concluded that nitrate-Piv-Cy-Et, as opposed to GTN, does not cause any vessel tolerance.

Nitrato-Piv-Cy-Et results in isolated guanylate cyclase from concentration-dependent enzyme activation corresponding to a single increased cGMP formation per unit time (compare Fig.). The peculiarity of this compound, therefore, is in comparison with conventional organic nitro compounds in thorns, that in vitro activation also occurs in the absence of cysteine. This immediately explains the observation, that nitrato-Piv-Cy-Et and chemical compounds e.g. the so-called Working Heart Models ** do not cause any tolerance, one finding, which is of particular great practical importance for the long-term clinical use. For the half-maximal activation (ΕΕ ^ θ) of guanylate cyclase, the required concentration is about 200 mol / l. The comparable values for GTN (in the presence of 5 mmol / l cysteine) range from about 80 mol / l.

Claims (8)

PATENT APPLICATIONS A process for the preparation of new organic nitrates as well as for the preparation of their pharmaceutically acceptable salts, characterized by a complexion, to form a compound of the corresponding fatty acid nitrate of general formula (A) O! L (A) R ¹ R in the form of its free acid, its reactive acid halide, azide, ester and / or anhydride, condenses with an amino group of a compound of the general formula (B) R ' R ⁴ ( ^CH 2) n • (ch ₂ ) _o (B) to give a compound of general formula (I) R0 R ' R ⁴ O ₂ N — O— € H ₂ -C— (CH ₂ ) _m R ² -N— (CH ₂ ) _n —C— (CH ₂ ) _q R ⁵ -R (I) which is optionally alkylated on the side set with some Cp C ^ -group, wherein in the general formulas (A), (B) and (I) the meaning of i -35R = hydroxy, C 1 -C 4 -alkoxy, amino, C 1 -C 4 -alkylamino, di-C 1 -C 4 -alkylamino, C 1 -C 6 -alkyl 1 am and no-C 1 -C C 1 -alkoxy, di-C 1 -C 6 -alkyl 1 amino-C 1 -C 3 -alkoxy and, acyloxy-C 1 -C 4 -alkoxy, arylC 1 -C 6 -alkoxy, substituted aryl-1-alkoxy, wherein the substituent is methyl, halogen or methoxy or amino acid residues selected from the group of cysteine, methionine or homocysteine coupled by peptide bonds = hydrogen, C 1 -C 4 -alkyl 1, amino, acylamino, hydroxy or acyloxy, r 2 = hydrogen or C-C - a T ki 1 »r3 = hydrogen, = hydrogen or C ^ -C ^ j-al ki 1, r5 = mercapto-Ci-C ^ j-al ki 1, Cj-Cg-al canoes 1-C 1 -C 4 -alkyl 1, especially alkylene-S-pivalate, alkylene 1-S-butyrate, alkylene-propionate, alkylene-S-acetate, alkylene-S-hexanoate, alkylene-co-tanoate, alkylene 1 -S-benzoate, alkylene-S-succinoyl ester, alkylene thiocarbonate, alkylene thio-carbamate, alkylene thio-lower alkyl, lower alky 1-thio-lower alky 1-carboxylic acid and / or its esters and / or its amides, m, n, o = numerical values 0 and / or 1-10 R and r5 may be linked together to form a thiolactone R and R ^ may be attached to each other to form an ester or amide R ^ and R ² * may be bonded to each other to form an alkylene bridge with 2 to 4 carbon atoms, an alkylene bridge with 2 to 3 carbon atoms and one sulfur atom, some alkylene -34 a bridge having 3 to 4 carbon atoms containing one double bond or an alkyl bridge, as above, substituted by hydroxy, lower-alkoxy, lower-alky 1 or di-lower-alky, 2. A process according to claim 1, characterized in that fatty acid nitrate of a species length of C2 is used ϋθ, which may be linear or branched, racemic or optical isomers. Process according to claim 1 or 2, characterized in that cysteine, methionine or homoc i ste n is used as the amino acid. Process according to one of Claims 1-3, characterized in that stereochemical L-form acids are used as amirio acids. Process according to one of Claims 1-4, characterized in that cysteine and / or methionine in the form of methyl, ethyl or propyl ester is used as the amino acid. Process according to one of Claims 1-5, characterized in that a cysteine is esterified on the SH group esterified with alkanecarboxylic acids with 2-8 C atoms. A process for the preparation of compounds according to claims 1-6, characterized in that suitable starting materials are used to give the following compounds corresponding to general formula (I): N- (2-nitratoacetyl) -cysteine ethyl ester N- (2-nitratoacetyl) -S-acetyl-cysteine ethyl ester N- (2-nitratoacetyl) -S-propionyl 1-cysteine ethyl ester N- (2-nitratoacetyl) -S-pival o1-cysteine ethyl ester N- (2-nitratoacetyl) -methionine methyl ester N- (2-nitratopropionyl) -cysteine -35 · Ν- (2-Nitratop ropioni1) -cysteine ethyl ester N- (2-nitratopropionyl) -methionine ethyl ester N- (2-nitratobutyryl) -cysteine N- (2-nitratobutyryl) -cysteine ethyl ester N- (2-nitratobutyryl) -S-acetyl-cysteine ethyl ester N- (2-nitratobutyryl) -S-butyryl-cysteine ethyl ester N- (2-nitratobutyryl) -methionine ethyl ester N- (2-nitratoisobutyryl) -cy stei n N- (2-nitratoisobutyryl) -cysteir ethyl ester N- (2-nitratoisobutyryl) -S-benzoyl-cysteine ethyl ester N- (2-nitratoisobutyryl) -S-acetyl-cysteine ethyl ester N- (2-nitratoisobutyryl) -S-pivaloyl-1-cysteine ethyl ester N- (2- nitratoisobutyryl) -methionine ethyl ester N- (3-nitratobutyryl) -cysteine N- (3-nitratobutyryl) -cysteine ethyl ester N- (3-nitratobutyryl) -S-acetyl-cysteine ethyl ester N- (3-nitratobutyryl) -S-propionyl-cysteine ethyl ester N- (3-nitratobutyryl) -methyl n-ethyl ester N- (3-nitratobutyryl) -homocysteine-thio-1actone N- (3-nitratopivaloyl) -cysteine N- (3-nitratopival oi1) -cysteine ethyl ester N- (3-nitratopivaloyl) -cysteine ethyl ester-S-carbonate N- (3-nitratopivaloyl) -S-acetyl-cysteine ethyl ester N- (3-nitratopivaloyl) -S-propionyl 1-cysteine ethyl ester N- (3 N-(3-nitratopivyl) -S-butyryl-cysteine ethyl ester N- (3-nitratopivyl) -S-isobutyryl-cysteine ethyl ester N- (3-nitratopival) N-(3-nitratopival) N-(3-Nitratopivolyl) -methionine ethyl ester N- (3-nitratopivoyl) -methionine nitratopivaloyl) -S-benzoyl-cysteine ethyl ester N- (3-thratopole and valoi1) -homoc and stei nth o-lactone N- (2-nitratohexanoyl) -cysteine ethyl ester N- (2-nitratohexanoyl) -S-propionyl-cysteine ethyl ester N- (3-nitratohexanoyl) -cysteine ethyl ester N- (3-nitratohexanoyl) -methionine methyl ester N- (12-nitratolauroyl) -cysteine -56 ' N- (12-nitratol-auroyl) -cysteine ethyl ester N- (12-nitratolauroyl) -S ~ acetyl-cysteine N- (12-nitratolauroyl) -S-pivaloyl-cysteines Method for the preparation of medicaments, characterized in that according to claim 1, the compound of general formula (I) is obtained OR ³ o 2N — O — CH ₂ —C— (CH2) _m —C — N— (CH ₂ ) _n I o (I) What they mean about R = hydroxy, C 1 -C 6 -alkyloxy, amino, C 1 -C 6 -alkyl 1 amine, di-C 1 -alkylamino, lower C 2 -alkylamino-lower-alkoxy, di-lower alkyl 1 amino-lower-alkoxy, acyl oxy-2-alkoxy, aryl2-alkoxy, substituted aryl-lower-alkoxy, wherein the substituent is methyl, halogen or methoxy or amino acid residues selected from the group of cysteine, methionine or homocysteine, which are connected to each other via peptide bonds, R1 = hydrogen, C1-C-alkyl, amino, acylamino, hydroxy or aci 1 oxy, R? - hydrogen or C - ^ - C ^ -al kil, R ³ = hydrogen, R ^ = hydrogen or C - ^ - C ^ -alkyl 1, r5 - mercapto-C1-C6 -alkyl 1, C ^ -Cg-al cananoyl-thio-C ^ -C ^ -alkyl, especially alkene 1-S-pival, alkene 1-S-butyrate, alkylene-propionate, alkene 1-S-acetate, alkylene-S-hexanoate, alkylene-coctanoate, alkylene-S-benzoate, alky 1-S-succinoyl ethyl ester , -3 Alkyl 1-thio-carbonate, alkylene-thio-carbamate, alkylene-thionyl-alky, lower-alky 1-thio-lower-alky 1 carboxylic acid and / or its esters and / or its amides, m, n, o = numeric values 0 and / or 1-10 R and r5 may be linked together to form a thiolactone R and R ^ may be bonded to one another to form an ester or amid that r3 and R ^ may be bonded to each other to form an alkylene bridge of 2 to 4 carbon atoms, an alkylene bridge of 2 to 3 carbon atoms and one sulfur atom , an alkylene bridge having 3 to 4 carbon atoms containing one double bond or an alkylene bridge as above substituted with hydroxy, lower-alkoxy, lower-alkyl or di-lower-alkyl. and n / or its pharmaceutically acceptable salts or mixtures thereof, are mixed with additional substances and / or with auxiliary materials customary in the manufacture of medicaments and processed into pharmaceutical preparations.