CS48892A3 - Combination of active compounds comprising sydnonimine and hirudin - Google Patents

Description

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A combination of active ingredients consisting of sydnonimine and hirundin

FIELD OF THE INVENTION The present invention relates to the combination of active ingredients existing from sydnonimine and hirudin and its use.

Background Art

Sydnonimines are compounds whose pharmacology has long been known (Chemie in unserer Zeit, 13th year, page 51 (1984), in particular antithrombotic effects are described in addition to vasodilatory effects). Cardiovasc. Pharma -col. 1989, 14 / Suppl. 11 /, page 129 /. Sydnonimines can be used as a prophylaxis and for the treatment of cardiac and blood circulation diseases and are marketed as such.

Hirudin is a polypeptide that can be isolated from the medicament (Hirudo medicinalis) and is known as inhibitorrorabine (Die Pharmazie 36.653 / 1931). More recently, sufficient amounts of recombinant hirudingen technology (MUnch) can be made. copper. Síschr. 127, 16/1985 // However, a large number of synthetic hirudins are known.

It has now surprisingly been found that the antithrombotic effects of sydnonimine and hirudin may be enhanced.

SUMMARY OF THE INVENTION Accordingly, the present invention relates to a combination of active ingredients consisting of sydnonimine and hirudin. Sydnonimine refers to all pharmacologically active sydnonimines. In particular, they are sydnonimines included in the following publications: DE-OS-162O5O1; DE-OS-1670127; -2- DE-OS-1695897; EP-A-23343; EE-A-59356; EP-A-76952, EP-A-210474; ΕΣ-Α-276710; EP-A-312773; EP-A-324408; EP-A-346684; EP-a-346694; EP-A-367036; EP-A-406659; and EP-A-406661.

Preferred sydnonimines are those corresponding to general formula (I)

(1) salts, wherein the compound or their pharmacologically tolerated amino group of the formula

R 2 represents hydrogen, alkyl, cycloalkyl, arylalkyl, ayyl-X-alkyl, alkoxyalkyl, alkylthioalkyl or alkenylthioalkyl; -3- 3 7

Ir is hydrogen or -COR; R4 is alkyl, cycloalkyl, alkyl-X-alkyl, arylalkyl, hydroxyalkyl;

c A R? is hydrogen or one of R; R is hydrogen or methyl; T 1 is aryl, 1 or 3 halogen atoms and / or 1 to 3 alkyl radicals and / or 1 to 3 alkoxy radicals and / or nitro radicals is a monosubstituted, di-substituted or trisubstituted aryl radical or OR or has one from the meanings of R; X is NR4, RSO2R8, NCO2alkyl, S / O / n, O, CH2 or a single bond; n is 0, 1 or 2; R 8 is alkyl, aryl, alkylaryl or dialkylamino.

Alkyl radicals and alkenyl radicals may have a straight or branched chain and preferably have 1 to 6 C atoms, most preferably 1 to 4 C atoms. This is also true when there are linkages with other radicals, such as alkoxy, arylalkyl, and the like. hem. Examples of alkyl radicals are methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl. butyl.

The cycloalkyl preferably has 5 to 7 C atoms and in particular is cyclopentyl and cyclohexyl.

The aryl preferably has 6 to 10 G atoms, and most preferably is -O-naphthyl or 4-naphthyl or phenyl.

Arylalkyl is preferably benzyl and phenylethyl.

Aryl-X-alkyl is preferably phenoxymethyl or phenoxyethyl

The aryl radicals R may be monosubstituted, disubstituted or trisubstituted, but a maximum of 2 nitro groups -4- such as 2-methyl-4,6-dinitrophenyl and 2-chloro-6-methyl 4-nitrophenyl may be present in the triple substitution Halogen substituents for aryl radicals include, for example, chlorine or bromine atoms. In particular, R @ 1 aryl radicals may be mentioned: methylphenyl (= tolyl), nitrophenyl and chlorophenyl, especially 4-nitrophenyl and 4-chlorophenyl. Preferred R1 radicals are morpholine, 3,3-dimethylthiomorpholine, cis-2,6-dimethylpiperidine, and 1,1-dioxo-3,3-dimethylthiomorpholine. 2 of R is preferably hydrogen. R? is preferably hydrogen, ethoxycarbonyl, propionyl or prop-anisoyl.

Sydnonimines, as claimed, together with anorganic or organic acids form acid addition salts. Suitable acids are, for example, hydrogen chloride, bromo-hydrogen, naphthalenedisulfonic acids, in particular naphthalenedisulfonic acid (1.5) phosphoric acid, nitric acid, sulfuric acid, oxalic acid, lactic acid, tartaric acid, acetic acid, acid salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid isonicotinic acid, methanesulfonic acid, β-toluenesulfonic acid, citric acid or adipic acid. The pharmacologically tolerable acid addition salts are considered to be preferred. Particularly preferred are the hydrochlorides. Examples of the claimed sydnonimines are: N-ethoxycarbonyl-3-morpholinosydnoimine (molsidomine); 3-morpholinosydnonimine hydrochloride (SIN-1); N- [4-methoxybenzoyl] -3- (cis-2,6-dimethylpiperidino) -sydnonimine; <RTI ID = 0.0> 5-K-propionyl-3- (3T3-dimethylthiOTnorpholino) sydnonimine; 3- (3,3-dimethylthioraorfolino) -synnonimine hydrochloride; 3- [ois-2,6-dimethylpiperidino] -synnonimine hydrochloride; 3- / tert. butyl-2-hydroxyethyl-amino-sydnonimine hydrochloride; 3- [tert-butyl- (2-diisopropylaminoethyl) amino] -sydnonimine dihydrochloid; 3- [tert-butyl- (2-morpholinoethyl) amino] -synnonimine hydrochloride (3- [3,3-dimethyl-morpholino] -synnonimine hydrochloride; 3- (2,2-dimethylpiperidino) -sydnonimine hydrochloride; 3- (4-Isopropyl-2,2-dimethylpiperazino) sydnon-imidazole hydrochloride; K-p-anisoyl-3- (4-isopropyl-+, 2-dimethyl-piperazino) sydnonimine dihydrochloride; 3- (2,2,6,6-tetramethyl-4-isopropylpiperazino) sydnonimine dihydrochloride; 3- (2- 6-dimethylpiperidino) -4-hexylsydnonimine hydrochloride; 4-benzyl-3- (2,6-dimethylpiperidino) -synnonimine hydrochloride; 3- (2,6-dimethylpiperidino) -4- (2-phenylethyl) -sydnonimine hydrochloride; 3- (2,6-dimethylpiperidino) -4-phenylthiomethylsydnonimine hydrochloride; K-benzoyl-4-benzylthiomethyl-3- [2-6-diethylpiperidino] -sydnonimine; 3- (2,6-dimethoxypiperidin-4-phenoxymethylsydnonimine hydrochloride); 3- (2-hydroxyethylamino) -sydnonimine hydrochloride; 3-ethylaminosydnonimine hydrochloride; 3-cyclohexylaminosydnonimine hydrochloride; 3- (2-Hydroxycyclohexylamino) -synnonimine hydrochloride; 3- (3,3-dimethyl-1,4-tetrahydrothiazine-1,1-dioxo-4-yl) -sydnonimine hydrochloride; 3- (3,3-dimethylperhydro-1-oxo-1,4-thiazin-4-yl) -sydnonimine hydrochloride; N-p-anisoyl-s-dicyclohexylamino-sydnonimine; N-pivaloyl-3-dicyclohexylaminosydnonimine 3- (2,2-dimethyl-4-methanesulfonylpiperazino) -synnonimine hydrochloride 3- (2,2-dimethyl-4-toluenesulfonylpiperazine) n -disodium imine hydrochloride; 3- (2,2-Dimethyl-4-dimethylaminosulfonylpiperazino) -synnonimine hydrochloride; N-isobutyroyl-3- (2,2-dimethyl-4-methanesulfonylpiperazino) -sydnonimine; 3- / K-tert. 6-butyl-K-methylamino) -synnonimine hydrochloride; 3- [N-tert-butyl-1H-butylamino] sydnonimine hydrochloride.

The term hirudins refers to all pharmacologically active hirudins. Their origin can be natural, synthetic or even gene-technical. Included are compounds that are K- or C-terminally extended to an effective amino acid sequence, as well as pharmacologically active sub-sequences or fragments. In comparison to a native sequence, individual amino acids may be modified or multiple amino acids may be modified or replaced with other amino acids, both naturally occurring amino acids and amino acids that do not occur naturally »

In particular, this term includes hirudins which, as compounds, are disclosed in the following disclosures: DE-A 3,445,532; EP-A-142,860; EP-A-158,564; BP-A158,936; EP-A-168,342; EP-A-171,024; EP-A-173,619; EP-A-193 175; EP-A-200 655; EP-A-209 601; EP-A-225,633, EP-A-227,938; EP-A-236 330; EP-A-252,854; EP-A-273 800, EP-A-324 712; EP-A-345,616; EP-A-364,942.

Particularly preferred hirudins are recombinant desul-fatohirudins (r-hirudins), which are obtained from E. coli, kva-sin or B-subtilis and differ in 2 amino acids from native hirudin. Such r-hirudin is, for example, HBW023 / r- Desulfato-Ile-4-Thyrrolidine Dmgs of the Fu &apos;, 267-280 (1980) In the combination of the active compounds according to the invention, the weight ratio between sydnonimine and hirudin can vary within a wide range .

Particularly preferred is the weight ratio of sydnonimine: hirudin 1 / 0.025 to 40 /, and more preferably the ratio is / is 1: 0.1 to 10. -7-

The active compound combinations according to the invention can be prepared by mixing the individual active compounds in the indicated weight ratios.

It is normally preferred that the active ingredient combination be administered in the form of a pharmaceutical preparation. These preparations may be administered orally, for example in the form of pills, tablets, coated tablets, dragees, hard and soft gelatin capsules, spit, syrups, emulsions or suspensions or aerosol mixtures. . However, foding can also be carried out rectally, for example in the form of suppositories, or perenterally, for example in the form of injection solutions or percutaneously, for example in the form of ointments or tinctures. For example, lactose, corn starch or derivatives thereof, talc, stearic acid or its salts, etc. may be used in the manufacture of pharmaceutical preparations for use in the manufacture of pills, tablets, dragees and hard gelatine capsules. and suppositories are, for example, fats, semisolid liquid polyols, natural and hardened oils, etc. Water, sucrose, invert sugar, glucose, polyols, etc. are suitable as carriers for solutions and syrups. water, alcohols, glycerin, polyols or vegetable oils are suitable.

In addition to the active ingredients and carriers, the pharmaceutical preparations also contain additives such as fillers, builders, disintegrants, lubricants, lubricants, wetting agents, stabilizers, emulsifiers, preservatives, sweeteners, colorants, flavor correctors, or flavoring agents, buffering agents, as well as solvents or dissolving agents, or compositions for depot effect, as well as salts for varying the osmotic pressure, coating compositions or antioxidants. other therapeutically active substances. -8-

Such other therapeutic agents are, for example: β-receptor blockers such as propranolol, pindolol, metoprolol; vasodilators such as carbocbromes; sedatives, such as barbituric acid derivatives, 1,4-benzodiazepines and meprobamate; diuretics such as chlorothiazide; heart ionizing agents such as digitalizing agents; blood pressure lowering agents such as hydralazine, dihydralazine, ramipril, prazosin, cloniden, rauwolfie alkaloids; agents that reduce blood fatty acid levels such as, for example, the nicotibrate, fenofibrate, and prophylaxis preparations such as phenprocoumon. In a particularly preferred embodiment of the present invention, the active compound combination according to the invention is not administered in a mixture, i.e. in one pharmaceutical preparation, but separately from one another, i.e. in two-part pharmaceutical preparations, simultaneously or in succession. the pharmaceutical compositions comprise huclsydnonimine or hirudin but are similar to those described above. In another embodiment of the present invention, there is provided a composition comprising sydnonimine and hirudin and, simultaneously or shortly thereafter, a formulation comprising svdnoniirin or hirudin. Even in this case they correspond to the preparations described above.

The active compound combinations according to the invention can be used in humans in the control of diseases or in their prophylaxis in diseases of the cardiovascular system, in particular as an antithrombotic drug. The dosage can vary within wide limits and is adapted to the individual need in each individual case. An oral daily dosage of about 0.5 to 100 mg is appropriate for oral administration to a human subject. in other forms of administration, the daily dosage is, due to good resorption of the active ingredients, in the range of amounts, i.e. in general also about 0.5 to 100 mg / man. Daily doses are normally divided into multiple doses, for example 2 to 4 sub-doses.

The active compound combinations according to the invention potentiate the antithrombotic effects of sydnonimine and hirudin on each other. This can already be achieved by doses in which neither sydnon-irain nor hirudin itself has a very potent effect.

This effect was found by the following test: Male, about 4 months old domestic pigs were coronary thrombosis produced by the method of Polts et al. Circulation lation 1976.54; 365-70 /. TJ of deeply narcotized animals was chest open; the left descending coronary artery was dissected out and provided with an electromagnetic measuring head that measures blood flow. Still, the coroarynary artery was damaged by a short (1) squeezing vascular clamp away from the measuring head. diameter 1.7 to 2.8 mm, length 2 mm / which narrows the vessel. These pfeque conditions resulted in uniformly cyclic changes in blood flow (CPVzs), due to repeated formation of apholization of thrombi in damaged constricted sites. Thrombi were quantified by determining the frequency / count for 60 min / cyclic changes in blood flow. After irrigation cycles for 60 min, animals were given the following: I. group (n = 6 / N-ethoxycarbonyl-3-morpholinosydnonimine / molsidomine), 0.1 mg / kg intravenously as an injectable dose; -10- Group 2 (η = 6): recombinant desulfatoirudin (r-hirudin) at a total dose of 0.2 mg / kg, divided as intranasate, injected dose of 0.1 mg / kg followed by intravenous continuous infusion of 16.67 ug / kg min / £ 1 mg / kg 60 min /; group 3 / n = 6 /: molsidomin · + r-hirudin, ie 0.1 mg / kg molsidomine iv followed by 0.1 mg infusion In the previous experiment, it was found that single doses of molsidomine and r-hirudin did not act just too anthrobically.

The experiment was terminated 1 hour after the end of the infusion eventually 2 hours after injection of the substances. The cyclic changes in blood flow per hour were determined in all experimental groups before and at the first and second hours after the test substances were injected. Average cyclic frequencies were statistically compared to each other before and after dosing. / Student-t-test for combined random tests /. The p value of 40.05 was a significant difference. The results can be seen in the following table: pb group 1 hour and thrombotic Plow cycles / n / 60 min / 1 for 1 hour 1 16 + 2 19 + 3 / molsidomin / 2 12 + 1 11 + £ 12 + 1 / r-hirudin / 3 12 + 1 8 + 2 2 + 2 * -11- $ no effect; the number of cycles could not be determined, but there was no permanent cleavage / closure / x p 4 0.01 compared to pre-treatment. In all animals, systolic and diastolic peripheral arterial blood pressure (mm Hg) was additionally measured. In Group 2, infusion of r-hirudin did not change blood pressure. Molsidomine (group 1) reduced the systolic blood pressure to a maximum of ol8 + 5µMg / p 4 0.05 /, with diastolic blood pressure decreasing ol0 + 5mmHg at the administered dose. In combination with r-hirudin / group 1 / molsidomin systolic- blood pressure was 21 + 4 mm Eg / p <0.05 / and diastolic blood pressure was insignificantly 11 + 3 µg Hg. This blood pressure-lowering effect of mol-sidomine was reduced in both experimental groups. The results show that the combination of the compounds of the invention results in almost total inhibition of experimental thrombi at this dosage. The antithrombotic effects of these substances with different mechanisms of action are therefore potentiated without leading to an increase in the hemodynamic effect.

The preferred combination of active ingredients of the present invention is that the two agents can be used in small doses. This reduces the potential for side effects and combines the use of those patients whose blood circulation does not allow a reduction in blood pressure. The combination according to the invention is characterized by the fact that, in parallel, the treatment and prophylaxis of thrombo-embolic diseases is particularly effective in the treatment of two independent thrombosis mechanisms. First, the effect of thrombin is eliminated with hirudin. This prevents fibrin formation and platelet activation. Thrombin is thought to be the most important platelet activator in arteries / why. Nati. Aoad. Sci USA -12- 1988; 85; 3184-38 /. On the other hand, sydnonimine activates one of the endogenous counter-regulation of platelet stimulation by increasing NO-induced cyclic guanosine monophosphate. This leads to platelet adhesion inhibition and platelet aggregation / Circulation 1989; 79 '· 657 -65 / · Both mechanisms together potentiate each other's antithrombotic effect. EXAMPLES The following examples explain the combination of the active ingredients according to the invention: Example 1 N-ethoxycarbonyl-3-morpholinosydnonimine / molsidomine / and r-hirudin in a 1: 2 weight ratio, Example 2 3-morpholinosydnonimine hydrochloride (SIN-1) and r-hiru-din in a 1: 2 weight ratio. Example 3 N- (4-methoxybenzoyl) -3- (cis-2,6-dimethylpiperidino) -sydnonimine and r-hirudin in a 5: 2 weight ratio. Example 4 N-Propionyl-3- (3,3-dimethylthiomorpholino) - sydnonimine r-hirudin in a 1: 2 weight ratio. Example 5 3- (3,3-Dimethyl-1,1-dioxothiomorpholino) -sydnonimine hydrochloride and r-hirudin in a 1: 1 weight ratio. Example 6 3- (3,3-Dimethylthiomorpholino) -synnonimine hydrochloride and r-hirudin in a 1: 3 weight ratio. Example 7 3- (cis-2,6-dimethylpiperidino) -synnonimine hydrochloride and r-hirudin in a 1: 1 weight ratio.

Claims (9)

-13- CLAIMS »* 2.5 PATENTS A combination of active ingredients consisting of sydnonimine and hirudin. 2. Active substance combination according to claim 1, characterized in that it contains sydnonimine of general formula I, N-RÉ (1), or a pharmacologically tolerable salt thereof, wherein r @ 1 is an amino group of the formula Σ v / R 2 is hydrogen, alkyl, cycloalkyl, arylalkyl, aryl-x-alkyl, alkoxyalkyl, alkylthioalkyl or alkenylthioalkyl, -14- R 4 is hydrogen or -C 0-17, f 4 is alkyl, cycloalkyl, alkyl-X-alkyl, arylalkyl or hydroxyalkyl, c / zl R-7 is hydrogen or one of R, β from R is hydrogen or methyl, R is aryl, 1 to 3 halogen atoms and / or 1 to 3 alkyl radicals and / or or 1 to 3 alkoxy radicals and / or 1 or 2 nitro radicals monosubstituted, disubstituted or trisubstituted aryl p radical or OR or has one of the meanings of R, X denotes NR (n) n or Rs, RCOalkyl, S / O / n, O, CH 2 or is n is a direct bond, n is 0, 1, or 2, and OR is alkyl, aryl, alkylaryl, or dialkylamino. 3. Active compound combination according to claim 2, characterized in that R1 represents a morpholino group, a 3,3-dimethylthiomorpholine group, a cis-2,6-diethylpiperidine group or 1,1-diol-3,3-dimethyl- a thiomorpholino group. Active substance combination according to claim 2 and / or 2 f 3, characterized in that R is hydrogen. Active substance combination according to one or more of the preceding claims, characterized in that R 1 represents hydrogen, ethoxycarbonyl group, propionyl group or p-anisyl group. 6. The active compound combination according to one or more of claims 1 to 5, wherein the hirudin is recombinant desulfatohirudin -15- (r-hirudin). Use of the active compound combination according to one or more of claims 1 to 6 as an antithrombotic agent. * Use according to claim 7, characterized in that sydnonimine and hirudin are administered simultaneously or briefly. 9. A pharmaceutical composition comprising a combination of the active compounds according to one or more of claims 1 to 6 together with pharmaceutically acceptable carriers and additives or optionally one or more other pharmacological active ingredients.