NO317993B1 - Benzoylguanidine Derivatives, Pharmaceutical Preparations, Methods for Preparing and Using Them in the Preparation of Medications - Google Patents

Description

The present invention relates to benzoylguanidine derivatives, pharmaceutical preparations, methods for their production and their use in the production of medicines.

The present invention relates to new benzoylguanidine derivatives with the general formula I,

in which

Ri can mean pyrrolyl, furyl, pyridyl;

phenyl unsubstituted or once substituted with a branched or straight chain C 1 -C 4 alkyl group, a branched or straight chain C 1 -C 4 alkoxy group, a trifluoromethyl group or halogen, or with a pyrrolyl group;

optionally in the form of the individual tautomers or optionally enantiomers and their mixtures as well as in the form of the free bases or the corresponding acid addition salts with pharmacologically acceptable acids.

Preferred in the sense of the present invention are compounds of the general formula I, in which

R 1 may mean an unsubstituted phenyl ring or a phenyl ring, which may be substituted with fluorine or a methyl, trifluoromethyl, a methoxy group or with a pyrrolyl residue, or.

Particularly preferred are the following compounds:

4-(4-(2-pyrrolylcarbonyl)-1-piperazinyl)-3-trifluoromethanesulfonate and 4-(4-(4-fluorophenylcarbonyl)-1-piperaziny])-3-trifluoromethyl-benzoylguanidine-methane

Halogen stands - unless otherwise stated - for fluorine, chlorine, bromine, iodine and preferably for fluorine, chlorine or bromine.

Compounds of this type are already known from German Offenlegungsschrift 196 01 303.8.

According to the present invention, a method for the preparation of compounds of the general formula I is described

characterized by reacting 4-(1-piperazinyl)-3-trifluoromethylbenzoic acid ester of the general formula II with a compound of the general formula Ul, in which Q means a leaving group substitutable with the piperazine nitrogen, possibly in the presence of auxiliaries, preferably carbonyldiimidazole, and slurries the resulting benzoic acid derivative of the general formula IV

in a suitable - preferably anhydrous - solvent - preferably dimethylformamide - and mixed with the mixture of a solution or suspension of a base - preferably sodium hydride in a suitable, anhydrous solvent - preferably dimethylformamide - with a guanidine salt - preferably guanidine hydrochloride - and isolating the reaction product and optionally forms the desired acid addition salt with a pharmacologically acceptable acid.

The present invention further relates to a pharmaceutical preparation, characterized by a content of a compound according to one of claims 1 to 4 and their acid addition salts in addition to usual excipients and carriers.

It is further described the use of compounds of the general formula I, their acid addition salts for the preparation of a medicament for the therapeutic treatment of ischemia.

As a result of their action, such compounds are useful as inhibitors of the cellular Na<+>/H<+> exchange as active substances in pharmaceuticals or can find use as intermediates for the production of such active substances. The compounds according to the invention work against arrhythmias, which for example occur with hypoxia. They are also applicable for diseases that are associated with ischemia (examples: cardiac, cerebral, gastrointestinal - such as mensenteric thrombosis/embolism -, pulmonary, renal ischemia, ischemia in the liver, ischemia in the skeletal muscles). Corresponding diseases are, for example, coronary heart disease, heart attack, angina pectoris, stable angina pectoris, ventricular arrhythmias, subventricular arrhythmias, heart failure - further for support in bypass operations, for support in open heart operations, for support in operations that necessitate an interruption of the blood supply to the heart and for support during heart transplants - embolism in the pulmonary circulation, acute or chronic renal failure, chronic renal insufficiency, myocardial infarction, reperfusion damage due to renewed blood flow to brain areas after dissolution of cartilage blockages and acute and chronic blood flow disorders of the brain. Here, the aforementioned compounds are also useful in combination with thrombolytic agents such as t-PA, streptokinase and urokinase.

During the reperfusion of an ischemic heart (e.g. after an Angina pectoris attack or a myocardial infarction) irreversible damage can occur to cardiomyocytes in the affected region. The compounds according to the invention work i.a. a. in such a case cardioprotective.

In the field of ischemia, the prevention of damage to grafts must also be taken into account (e.g. as protection for the graft - such as liver, kidney, heart or lung - before, during and after implantation, as well as during the storage of the grafts), which can act in connection with the transplant. The compounds also act as a protective drug during angioplasty operations on hearts and peripheral vessels.

In essential hypertension and diabetic nephropathy, the cellular sodium-proton exchange is elevated. The compounds according to the invention are therefore suitable as inhibitors of this. replacement for preventive treatment of these diseases.

The compounds according to the invention are further distinguished by a strong inhibitory effect on the proliferation of cells. Therefore, the compounds are interesting as drugs in diseases in which cell proliferation plays a primary or secondary role and can be used as a remedy against cancer, benign tumors, or for example prostatic hypertrophy, atherosclerosis, organ hypertrophies and hyperplasias, fibrotic diseases and diabetic tendon complications.

Furthermore, it is known that compounds of this type can have a beneficial influence on the blood level of the serum lipoproteins.

Surprisingly, it was now found that the compounds of the general formula 1, in relation to the benzoylguanidine derivatives already known from the state of the art, have the advantage that, in addition to an unexpectedly higher effect, they entail the advantage of oral availability.

The active substances of the general formula I can be used as an aqueous injection solution (e.g. for intravenous, intramuscular or subcutaneous application), as tablets, as suppositories, as an ointment, as a patch for transdermal application, as an aerosol for inhalation use through the lungs or as a nasal spray.

The active ingredient content in a tablet or suppository is between 5 and 200 mg, preferably between 10 and 50 mg. For inhalation, the single dose is between 0.05 and 20 mg, preferably between 0.2 and 5 mg. In the case of a parenteral injection, single doses are between 0.1 and 50 mg, preferably between 0.5 and 20 mg. If necessary, the mentioned doses can be given several times a day.

In the following, some examples of pharmaceutical preparations with the active ingredient are given:

The solution can be sterilized using standard procedures.

The solution listed above is suitable for nasal application in a spray or in combination with a device that produces an aerosol with a particle size preferably between 2 and 6 (im for application through the lungs.

Capsules for inhalation

The compounds with the general formula 1 are in micronized form (particle size essentially between 2 and 6 µM), filled in hard gelatin capsules, optionally with the addition of micronized carrier substances, such as lactose. For inhalation, ordinary devices for powder inhalation serve. In each capsule, e.g. between 0.2 and 20 mg of the active substance with the general formula I and 0 to 40 mg of lactose filled in.

The preparation is preferably filled in an aerosol container with a dosing valve, the individual dose is measured so that a dose of 0.5 mg is delivered. For the other dosages of the specified range, preparations with a higher or lower proportion of active ingredients are used appropriately.

The ingredients are processed in the usual way into an ointment.

The method of preparation for the compounds according to the invention is generally known from the state of the art and the preparation takes place as indicated above.

The present invention is illustrated through the following examples:

Examples:

4- Fluoro- 3- trifluoromethyl- benzosvremetv] ester

35.4 g (170 mmol) of 4-fluoro-3-(trifluoromethyl)-benzoic acid in 250 ml of methanol are mixed under ice-cooling at 5° C. during 25 minutes with 68 ml of SOCb. After the addition is finished,

the reaction mixture was further heated over a period of 3 h at reflux. The reaction solution is cooled to room temperature and evaporated in vacuo. The oily residue is taken up in 200 ml of diethyl ether, extracted with water, saturated NaHCOa solution and again with water. The combined organic phases are dried over magnesium sulfate and evaporated in vacuo.

Yield: 29.0 g (77%)

4-( 4- Benzyl- 1 - piperazinevB- 3- trifluoromethyl- benzoic acid methyl ester

7 g (31.5 mmol) of 4-fluoro-3-trifluoromethyl-benzoic acid methyl ester are dissolved in 60 ml of dry dimethyl sulfoxide (DMSO) and mixed with 5.55 g (31.5 mmol) of N-benzylpiperazine and 4.35 g (31.5 mmol) of potassium carbonate. The mixture is stirred for 121 at 90°C. After cooling, the reaction mixture is poured into 200 ml of water and extracted three times with ethyl acetate. The combined organic phases are washed with water and saturated sodium chloride solution, dried over magnesium sulfate and distilled off in vacuum. The residue is chromatographed with an acetic ester/n-heptane mixture on silica gel.

Yield: 3.93 g (33%)

4-( 1 - Piperazinyl)- 3-trifluoromethyl-benzosvreme methyl ester

20.2 g (53.3 mmol) of 4-(4-benzyl-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester is dissolved in 200 ml of methanol and mixed with 2 g of palladium on carbon and hydrogenated over a period of 1.41 at 70°C and below a hydrogen pressure of 5 bar. The solution is filtered off over Cellit and distilled off in a vacuum.

Yield: 14.85 g (97%)

General regulation for the coupling of 4-(l-piperazine-3-trifluoromethyl-benzosvremetvIester with benzos<y>rer: 5 mmol of the corresponding carboxylic acid is dissolved in 30 ml of absolute tetrahydrofuran (THF) and under protective gas at 0° C mixed with 810 mg (5 mmol) of carbonyldiimidazole and stirred for 21 at room temperature (approx. 25 °C). Afterwards, 1.44 g (5 mmol) of 4-(1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester are added and further stirred over a period of approx. 121. The solution is evaporated in vacuo to dryness and taken up with ethyl acetate. After washing with saturated NaHCC>3 solution, saturated NaCl solution and water, the organic phases are dried over MgSO4 and evaporated in vacuo. After crystallization in a suitable solvent or chromatography on silica gel with a suitable eluent, the following compounds are obtained.

1. 4-(4-(3-Methoxyphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 81%

2. 4-(4-(2-Pyrrolylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Crystallization from methanol

Yield: 75%

M.p.: 149°C

3. 4-(4-(4-Fluorophenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 77%

4. 4-(4-(2-Methoxyphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 79%

5. 4-(4-(3-Tyrfluoromethylphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 83%

6. 4-(4-Phenylcarbonyl-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 87%

7. 4-(4-(2-Furylcarbonyl)-1-pipera2inyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 75%

8. 4-(4-(3-Methylphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 79%

9. 4-(4-(4-( 1 -Pyrryl)phenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 87%

10. 4-(4-(2-Pyridylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate/n-heptane (2:1)

Yield: 73%

General regulation for the preparation of acylguanidines from the corresponding carboxylate esters: 5.09 g (127.2 mmol) of 60% NaH in mineral oil are washed twice with ether and decanted from. 200 ml of absolute DMF are added and, under stirring and protective gas, 12.15 g (127.2 mmol) of guanidine hydrochloride are added in small portions. After stirring for one hour, 21.2 mmol of the corresponding methyl ester are added and the solution is further stirred over a period of 2 hours at a

temperature of approx. 120 °C. The reaction mixture is then allowed to cool to room temperature, filtered and the filtrate evaporated in vacuo. After chromatography on silica gel with a suitable

eluent and transfer with ethereal hydrochloric acid or other pharmacologically tolerable acids in the corresponding salts, the following compounds are obtained (in the following structural formulas, the hydrogen atoms are omitted for reasons of clarity, provided they are bound to a carbon or nitrogen atom and provided they are not necessary for the understanding of the invention): 1. Example 4-(4-(3-Methoxyphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl- benzoylguanidine hydrochloride

from 4-(4-(3-methoxyphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester Column chromatography: ethyl acetate/methanol (5:1)

Yield: 71%

Melting point: >200°C

MS: (M+H)<+> = 450 (free base)

2. Example 4-(4-(2-Pyrrolylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine methanesulfonate

from 4-(4-(2-pyrrolylcarbonyl)- l -<p>i<p>erazin<y>l )-3-trifluoromethyl<y>l -benzos<y>remethyl ester Column chromatography: ethyl acetate/methanol (5:1 )

Yield: 66%

M.p.: 246°C

MS: (M+H)<+> = 409 (free base)

3. Example 4-(4-(4-fluorophenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine methanesulfonate

from 4-(4-(4-fluorophenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester Column chromatography: ethyl acetate:methanol (5:1)

Yield: 40%

Melting point: 140°C

MS: (M+H)<+> = 438 (free base)

4. Example 4-(4-(2-Methoxyphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine hydrochloride

from 4-(4-(2-Methoxyphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester Column chromatography: ethyl acetate:methanol (5:1)

Yield: 71%

Melting point: 219°C (decomposition)

MS:(M+H)<+> = 450 (free base)

5. Example 4-(4-(3-trifluoromethylphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-

benzoylguanidine hydrochloride

from 4-(4-(3-trifluoromethyl]phenylcarbonyl])-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

Column chromatography: ethyl acetate:methanol (5:1)

Yield: 25%

Melting point: 140°C (decomposition)

MS: (M+H)<+> = 488 (free base)

6. Example 4-(4-phenylcarbonyl-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine hydrochloride

from 4-(4-phenylcarbonyl-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester Column chromatography: ethyl acetate:methanol (5:1)

Yield: 64%

M.p.: 214°C

MS: (M+H)<+> = 420 (free base)

7. Example 4-(4-(2-Furylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine-

methanesulfonate

from 4-(4-(2-Furylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester crystallization from ether

Yield: 19%

Melting point: 190°C (decomposition)

MS: (M+H)+ = 410 (free base)

8. Example 4-(4-(3-Methylphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine methanesulfonate

from 4-(4-(3-methylphenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester

crystallization from methanol/acetic acid

Yield: 76%

M.p.: 199°C

MS: (M+H)+ = 434 (free base)

9. Example 4-(4-(4-(1-pyrrolyl)phenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine dimethanesulfonate

from 4-(4-(4-( 1 -pyrryl)phenylcarbonyl)-1 -piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester Crystallization from methanol

Yield: 48%

Melting point: 150°C (decomposition)

MS: (M+H)<+> = 485 (free base)

10. Example 4-(4-(2-pyridylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine dimethanesulfonate

from 4-(4-(2-pyridylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoic acid methyl ester Column chromatography: acetate: methanol (5:1)

Yield: 34%

Melting point: 115°C (decomposition)

MS: (M+H)+ = 421 (free base)

Pharmacological data:

Inhibition of Na<+>/H<+->exchange in human intestinal cancer cells (HT-29): HT-29 cells are incubated at 37°C, 5% CO2 in growth medium. After 3-5 days, the growth medium is removed, the cells are washed and coated with 7.5 nM BCECF-AM (pH-sensitive fluorescent dye) at 37°C without CO2. After 30 min, the cells are washed and acidified with the following medium: 70 raM Choline chloride, 20 mM NH4Cl, 1 mM MgCl2, 1.8 mM CaCl2, 5 mM glucose and 15 mM HEPES, pH 7.5.

After 6 min incubation at 37°C without CO2, the cells are washed and incubated for 5 min with washing medium: 120 mM Choline chloride, 5 mM KC1, 1 mM MgCl2, 1.8 mM CaCl2, 5 mM glucose and 15 mM MOPS, pH 7.0.

The wash medium is removed and control medium with or without test compound is added: 120 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 1.8 mM CaCl 2 , 5 mM glucose, 15 mM MOPS, pH 7.0.

The cells are incubated for 4 min at 37°C without CO2 and measured fluorimetrically (Cytofluor 2350). The fluorescence of the dye BCECF is measured at the stimulation wavelengths 485 nm (pH-sensitive) and 440 nm (not pH-sensitive) and at the emission wavelength 530 nm. Cytoplasmic pH is calculated from the ratio of the fluorescence at 485 and 440 nm. The fluorescence ratio is calibrated by measuring the fluorescence signal after equilibration of external and internal pH with Nigericin.

The compounds according to the invention surprisingly show a very good biological availability and long half-lives after oral administration, properties which make them excellently suitable for an oral application.

Pharmacokinetic data:

For the surveys, approx. 200 g heavy male rats (not fasted) were used. For intravenous and oral application, the substances are dissolved in an acidified aqueous solution (pH 3). Single-bolus injections (0.5 mg/kg i.v., 2.5 mg/kg p.o.) are injected into the needle vein (0.2 ml/200g) or added through a cannula into the stomach (1 ml/200 g). The application solutions are analyzed to confirm the applied dose. 0.5 ml Aliquots of blood are taken from the retro-orbital venous plexus under short halothane anesthesia with heparinized glass capillaries according to the following scheme: - after i.v. application: 5 min, 15 min, 30 min, 11,21,41, 61.8 h;

- after oral application: 15 min, 1 1,21,41, 61,81, 241,321.

The samples are centrifuged and the plasma stored for analysis at - 20°C.

Sample preparation takes place through liquid-liquid extraction with an internal standard. The plasma extracts are analyzed by reverse-phase HPLC, coupled with an electrospray tandem mass spectrometer.

The pharmacokinetic data are determined from the corresponding plasma concentrations by compartment-free analysis with the TopFit program (Heinzel, G., Woloszczak, R., Thomann, P. TopFit 2.0 - Pharmacokinetic and pharmacodynamic data analysis system for the PC, Gustav Fisk Verlag, Stuttgart, Jena, New York, 1993).

Claims (1)

1. Benzoylguanidine derivatives of the general formula I, in which Ri can mean pyrrolyl, furyl, pyridyl; phenyl unsubstituted or once substituted with a branched or straight chain C 1 -C 4 alkyl group, a branched or straight chain C 1 -C 4 alkoxy group, a trifluoromethyl group or halogen, or with a pyrrolyl group; optionally in the form of the individual tautomers or optionally enantiomers and their mixtures as well as in the form of the free bases or the corresponding acid addition salts with pharmacologically acceptable acids. 2. Compounds with the general formula I according to claim 1, characterized by R 1 can mean an unsubstituted phenyl ring or a phenyl ring, which can be substituted with fluorine or a methyl, trifluoromethyl, a methoxy group or with a pyrrolyl residue, or 3. 4-(4-(2-pyrrolylcarbonyl)-1-piperazinyl)- 3-trifluoromethyl-benzoylguanidine- methanesulfonate 4. 4-(4-(4-fluorophenylcarbonyl)-1-piperazinyl)-3-trifluoromethyl-benzoylguanidine- methanesulfonate 5. Process for the preparation of compounds of the general formula I characterized by reacting 4-(1-piperazinyl)-3-trifluoromethylbenzoic acid ester with the general formula II with a compound of the general formula 111, in which Q represents a leaving group substitutable with the piperazine nitrogen, optionally in the presence of auxiliaries, preferably carbonyldiimidazole, and suspends the resulting benzoic acid derivative of the general formula IV in a suitable - preferably anhydrous - solvent - preferably dimethylformamide - and mixed with the mixture of a solution or suspension of a base - preferably sodium hydride in a suitable, anhydrous solvent - preferably dimethylformamide - with a guanidine salt - preferably guanidine hydrochloride - and isolating the reaction product and optionally forms the desired acid addition salt with a pharmacologically acceptable acid. 6. Pharmaceutical preparation, characterized by a content of a compound according to one of claims 1 to 4 and their acid addition salts in addition to usual auxiliary and carrier substances. 7. Use of compounds of the general formula I, their acid addition salts for the preparation of a medicament for the therapeutic treatment of ischemia.