HRP20041078A2 - Pentafluorosulfanyl-benzoylguanidine method for the production thereof and its utilization as medicament or diagnostic agent and medicament containingsame - Google Patents

Description

The invention relates to pentafluorosulfanyl-benzoylguanidines of formulas I and II

[image]

where

R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, alkoxy with 1, 2, 3 or 4 C atoms, F, Cl, Br, I, CN, NR10R11, -Op-(CH2)n-(CF2 )o-CF3 or -(SOm)q-(CH2)r(CF2)3-CF3;

R10 and R11 are independently hydrogen, alkyl with 1, 2, 3 or 4 C atoms or -CH2-CF3;

m is zero, 1 or 2;

n, o, p, q, r and s are independently zero or 1;

R2 hydrogen, F, Cl, Br, I, -CN, -SO2CH3, -(SOh)z-(CH2)k-(CF2)-CF3, alkyl with 1, 2, 3, 4, 5 or 6 C atoms, cycloalkyl with 3, 4, 5, 6, 7 or 8 C atoms, in which 1, 2, 3 or 4 hydrogen atoms can be replaced by fluorine atoms;

h is zero, 1 or 2;

z zero or 1;

k zero, 1, 2, 3 or 4;

1 is zero or 1; or

R2 is -(CH2)t-phenyl or -O-phenyl, which is unsubstituted or substituted with 1, 2 or 3 residues selected from the group consisting of F, Cl, Br, I, -Ou-(CH2)V-CF3 , 1, 2, 3 or 4 C-atom alkoxy, 1, 2, 3 or 4 C-atom alkyl and -SO2CH3;

t is zero, 1, 2, 3 or 4;

u is zero or 1;

v is zero, 1, 2 or 3; or

R2 is -(CH2)w-heteroaryl, which is unsubstituted or substituted with 1, 2 or 3 residues selected from the group consisting of F, Cl, Br, I, -Ox-(CH2)y-CF3, alkoxy with 1, 2, 3 or 4 C atoms and alkyl with 1, 2, 3 or 4 C atoms, -SO2CH3;

w is zero, 1, 2, 3 or 4;

x is zero or 1;

y zero, 1, 2 or 3;

R 3 and R 4 are independently hydrogen or F;

as well as their pharmaceutically acceptable salts.

Preference is given to compounds of formulas I and II, in which

R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, alkoxy with 1, 2, 3 or 4 C atoms, F, Cl, NR10R11, -O-CH2-CF3 or -SOm-(CH2)r-CF3 ;

R10 and R11 are independently hydrogen, alkyl with 1, 2, 3 or 4 C atoms or -CH2-CF3;

m is zero, 1 or 2;

r is zero or 1;

R2 hydrogen, F, Cl, -SO2CH3, -(SOh)z-(CH2)k-CF3, alkyl with 1, 2, 3 or 4 C atoms, cycloalkyl with 3, 4, 5, 6 or 7 C atoms, in in which 1, 2, 3 or 4 hydrogen atoms can be replaced by fluorine atoms;

h is zero, 1 or 2;

z is zero or 1;

k is zero, 1, 2, 3 or 4; or

R2 is phenyl or -O-phenyl, which is unsubstituted or substituted by is or 2 residues selected from the group consisting of F, Cl, -Ou-(CH2)V-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3;

u is zero or 1;

v is zero, 1, 2 or 3; or

R2 is heteroaryl, which is unsubstituted or substituted with 1 or 2 residues selected from the group consisting of F, Cl, -Ox-(CH2)y-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3;

x is zero or 1;

y is zero, 1, 2 or 3;

R 3 and R 4 are independently hydrogen or F;

as well as their pharmaceutically acceptable salts.

Particular preference is given to compounds of formulas I and II, in which

R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms,

methoxy, ethoxy, F, Cl, NR10R11, -O-CH2-CF3 or -SOm-(CH2)r-CF3;

R10 and R11 are independently hydrogen, alkyl with 1, 2, 3 or 4 C atoms or -CH2-CF3;

m is zero, 1 or 2;

r is zero or 1;

R2 is hydrogen, F, Cl, -SO2CH3, -(SOh)z-(CH2)k-CF3, alkyl with 1, 2, 3 or 4 C atoms, cycloalkyl with 3, 4, 5, 6 or 7 C atoms, in which 1, 2, 3 or 4 hydrogen atoms can be replaced by fluorine atoms;

h is zero or 2;

z is zero or 1;

k zero or 1; or

R2 is phenyl or -O-phenyl, which is unsubstituted or substituted by 1 or 2 residues selected from the group

consist of F, Cl, -O-(CH2)v-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3;

v is zero, 1, 2 or 3; or

R2 is heteroaryl, which is unsubstituted or substituted with 1 or 2 residues selected from the group consisting of F, Cl, -O-(CH2)y-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3;

y is zero, 1, 2 or 3;

R 3 and R 4 are hydrogen; as well as their pharmaceutically acceptable salts.

It is particularly advantageous if in compounds of formula I and/or II R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, F or Cl. It is also particularly advantageous if in the compounds of formula I and/or II R2 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, F, Cl or -O-phenyl, which is unsubstituted or substituted as stated above.

If the substituents R1 to R4 contain one or more centers of asymmetry, then they can independently have both S and R configurations. The compounds may exist as optical isomers, as diastereomers, as racemates or as mixtures thereof.

The proposed invention includes all tautomeric forms of compounds of formulas I and II.

Alkyl residues can be straight or branched. This also applies when they carry substituents or when they occur as substituents on other residues, for example in fluoroalkyl residues or alkoxy residues. Examples of alkyl residues are methyl, ethyl, n-propyl, isopropyl (= 1-methyl-ethyl), n-butyl, isobutyl (= 2-methylpropyl), sec-butyl (= 1-methylpropyl), tert-butyl f= 1 ,1-dimethylethyl), n-pentyl, isopentyl, tert-pentyl, neopentyl and hexyl. Preferred alkyl radicals are methyl, ethyl, n-propyl and isopropyl.

In alkyl radicals, one or more, for example 1, 2, 3, 4 or 5 hydrogen atoms can be substituted with fluorine atoms. Examples of such fluoroalkyl radicals are trifluoromethyl, 2,2,2-trifluoroethyl and pentafluoroethyl. Substituted alkyl residues can be substituted in any position.

Examples of cycloalkyl radicals are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl cycloheptyl or cyclo-octyl. In cycloalkyl radicals, one or more, for example 1, 2, 3, or 4 hydrogen atoms may be substituted with fluorine atoms. Substituted cycloalkyl residues can be substituted in any position.

Phenyl residues may be unsubstituted or mono- or poly-substituted, for example mono, doubly or triply, with the same or different residues. If the phenyl radical is substituted, it preferably carries one or two identical or different substituents. This also applies to substituted phenyl radicals in groups such as for example phenylalkyl or phenyloxy. In monosubstituted phenyl residues, the substituent can be in position 2, position 3 or in position 4. Disubstituted phenyl can be substituted in position 2,3, in position 2,4, in position 2,5, in position 2,6, in position 3.4 or in position 3.5. In trisubstituted phenyl residues, the substituents can be in the 2, 3,4 position, in the 2, 3,5 position, in the 2,4,5 position, in the 2,4,6 position, in the 2,3,6 position, or in position 3,4,5.

Heteroaryl residues are aromatic ring compounds, in which one or more ring atoms are an oxygen atom, a sulfur atom or a nitrogen atom, for example 1, 2 or 3 nitrogen atoms, 1 or 2 oxygen atoms, 1 or 2 sulfur atoms, or combinations of different heteroatoms . Heteroaryl moieties may be linked via any position, for example via position 1, via position 2, position 3, position 4, position 5, position 6, position 7 or position 8. Heteroaryl residues may be unsubstituted or mono- or polysubstituted, for example singly, doubly or triply substituted, with the same or different residues. This also applies to heteroaryl radicals as for example in the heteroarylalkyl radical. Heteroaryl means, for example, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, indazolyl, quinolyl, isoquinolyl, phthalazinyl, quinoxalinyl, quinazolinyl, and quinolinyl.

Examples of heteroaryl radicals include, in particular, 2- or 3-thienyl, 2- or 3-furyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, 4- or 5-imidazolyl, 1-, 3-, 4- or 5-pyrazolyl, 1,2,3-triazol-1-, -4 or -5-yl, 1,2,4-triazol-1-, -3- or -5-yl, 1- or 5-tetrazolyl , 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 1/2,3-oxadiazol-4-yl-5-yl, 1,2,4-oxa-diazol-3-yl -5-yl, 1,3,4-oxadiazol-2-yl or -5-yl, 2-, 4-or 5-thiazolyl, 3-, 4- or 5-isothiazolyl, 1,3,4-thiadiazol- 2- or -5-yl, 1,2,4-thiadiazol-3-or-5-yl, 1,2,3-thiadiazol-4- or -5-yl, 2-, 3- or 4-pyridyl, 2-, 4-, 5- or 6-pyrimidinyl, 3- or 4-pyridazinyl, pyrazinyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 1-, 2-, 4- or 5-benz-imidazolyl, 1-, 3-, 4-, 5-, 6- or 7-indazolyl, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl, 2-, 4-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-quinolinyl, 2-, 3-, 5-, 6-, 7- or 8-quinoxalinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazines1. Also included are the corresponding N-oxides of these compounds, so for example 1-oxy-2-, 3- or 4-pyridyl.

Particularly favorable heteroaromatics are 2- or 3-thienyl, 2- or 3-furyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, 4- or 5-imidazolyl, 2-, 3-, 4 -, 5-, 6-, 7- or 8-quinolyl, 1-, 3-, 4- or 5-pyrazolyl, 2-, 3- or 4-pyridyl, 2- or 3-pyrazinyl, 2-, 4- , 5- or 6-pyrimidinyl and 3- or 4-pyridazinyl.

The invention further relates to a process for the preparation of a compound of formula I and II and/or its pharmaceutically acceptable salt, which is characterized in that the compound of formula III or IV

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in which R1 to R4 have the given meanings, and L is a leaving group which can be easily nucleophilically substituted, reacts with guanidine.

Activated acid derivatives of formulas III and IV, in which L represents an alkoxy-, preferably a methoxy group, a phenoxy group, a phenylthio-, a methylthio-, a 2-pyridylthio group, a heterocycle with nitrogen, preferably a 1-imidazolyl group, are conveniently obtained in a known manner from chloride basic carboxylic acids (formulas III, IV; L = Cl), which in turn can be produced in a known manner from basic carboxylic acids (formulas III, IV; L = OH), for example with thionyl chloride.

In addition to carboxylic acid chlorides of formulas III and IV (L = Cl), further activated acid derivatives of formulas III and IV can also be obtained in a known manner directly from basic benzoic acid (formulas III, IV; L = OH), as methyl esters of formula III and IV with L = OCH3 treatment with gaseous HCl in methanol, imidazolide of formula III and IV treatment with carbonyl-diimidazole [L = 1-imidazolyl, Staab, Angew. Chem. Int. Ed. English 1, 351-367 (1962)], mixed anhydrides of formulas III and IV with Cl-COOC2H5 or tosyl chloride in the presence of triethylamine in an inert solvent, and it is also possible to activate benzoic acid with dicyclohexyl-carbo-di-imide (DCC) or with O-[(cyano(ethoxycarbonyl)-methylene)-amino]-1,1,3,3-tetramethyluronium-tetrafluoro-borate ("TOTU") [Proceedings of the 21st European Peptide Symposium, Peptides 1990, Editors E. Giralt and D. Andreu, Escom, Leiden, 1991]. A number of suitable methods for the preparation of activated carboxylic acid derivatives of formulas III and IV are given with reference to J. March, Advanced Organic Chemistry, Third Edition (John Wiley & Sons, 1985, p. 350).

The conversion of the activated carboxylic acid derivative of formulas III and IV with guanidine is primarily carried out in a known manner in a protic or aprotic polar, but inert, organic solvent. Methanol, isopropanol or THF at 20°C up to the boiling point of that solvent can prove to be good for the reaction of benzoic acid methyl ester (III, IV; L = OCH3) with guanidine. For most reactions of III and IV with salt-free guanidine, it is useful to work in an aprotic inert solvent, such as THF, dimethoxyethane, dioxane.

However, for the reaction of compounds III and IV with guanidine, water can also be used as a solvent with the use of a base, such as NaOH.

If L represents Cl, it is convenient to work with the addition of an acid-binding agent, for example in the form of excess guanidine, to bind the hydrohalic acid.

The invention also includes intermediate products of formula V

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wherein R 5 is hydrogen or (C 1 -C 4 )-alkyl, and wherein the methyl group may be in the 2-position or in the 3-position of the aromatic ring.

Pentafluorosulfanyl-benzoylguanidines I and II are generally weak bases and they can bind acid to form salts. Suitable acid addition salts are all pharmacologically tolerable acids, for example halides, especially hydrochlorides, lactates, sulfates, citrates, tartrates, acetates, phosphates, methylsulfonates, p-toluenesulfonates.

Compounds I and II are substituted acylguanidines and they inhibit cellular sodium-proton antiporters (Na+/H+- Exchanger, NHE).

Compared to known compounds, the compounds according to the invention are characterized by an exceptionally high efficiency in inhibiting Na+/H exchange, as well as improved ADMET properties. Due to the xenobiotic structure (especially due to the introduction of "unnatural/naturally foreign" SF5 substituents), the possibility of metabolic attack is hindered. Among other things, this leads to longer S9 stability (stability in the liver, stability to enzymatic attack) and to a longer half-life in vivo. At the same time, resorption is not significantly affected and the high biological availability of acylguanidine is maintained.

Contrary to the acylguanidines described in the literature, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts described here do not exhibit any undesirable and unfavorable saliduretic properties.

Due to the NHE-inhibitory properties, the compounds of the formula I and/or II and/or their pharmaceutically acceptable salts are suitable for the prevention and treatment of diseases caused by the activation or activated NHE, as well as secondary diseases caused by injuries caused by NHE.

Since NHE inhibitors act predominantly through their influence on the regulation of the pH value in the cell, they can generally be advantageously combined with other compounds that regulate the pH value in the cell, whereby as participants in the combination come into consideration inhibitors of the enzyme group carbonic anhydrase, inhibitors of bicarbonate system, as sodium-bicarbonate cotransporters (NBC) or sodium-dependent chloride-bicarbonate exchangers (NCBE), as well as NHE inhibitors with an inhibitory effect on other NHE subtypes, because the pharmacologically important effects of NHE inhibitors on pH regulation described here values can amplify or modulate.

The use of compounds according to the invention refers to the prevention and treatment of acute and chronic diseases in veterinary medicine and in human medicine.

Thus, the NHE inhibitors according to the invention are suitable for the treatment of diseases caused by ischemia and reperfusion.

Due to their pharmacological properties, the compounds described here are suitable as antiarrhythmic drugs.

Due to their cardioprotective component NHE inhibitors of formula I and/or II and/or their pharmaceutically acceptable salts are particularly favorable for the prophylaxis of heart attacks and for the treatment of heart attacks as well as for the treatment of angina pectoris, whereby they also prevent or greatly reduce the pathophysiological processes in the occurrence of injuries caused by ischemia, especially when causing cardiac arrhythmias caused by ischemia. Due to their protective effect against pathological hypoxic and ischemic conditions, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts according to the invention, due to the inhibition of the cellular mechanism of Na+/H+ exchange, can be used as a medicine for the treatment of all acute or chronic injuries caused by ischemia or diseases caused primarily or secondarily by it.

This also applies to their use as medicine in surgical procedures. Thus, these compounds can be used in organ transplants, where these compounds can be used to protect the organ in the donor before and during organ retrieval, to protect the extracted organ, for example during processing or its disposal in physiological fluid baths, as well as during transfer into the body of the recipient.

The compounds according to the invention are also a valuable drug with a protective effect during angioplasty operations, for example on the heart, as well as on peripheral organs and blood vessels.

The compounds according to the invention have been shown to be remarkably effective drugs against life-threatening arrhythmias. Ventricular flutter stops and the physiological sinus rhythm of the heart is restored.

Since NHE 1 inhibitors not only effectively protect human tissue and organs, especially the heart, against injury caused by ischemia and reperfusion, but also against the cytotoxic effects of drugs, such as especially those used in cancer therapy and autoimmune disease therapy, combined administering said compounds with compounds of formula I and/or II and/or their pharmaceutically acceptable salts is suitable for inhibiting cytotoxic, especially cardiotoxic side effects. In addition, with the reduction of cytotoxic effects, especially cardiotoxicity, due to simultaneous treatment with NHE 1 inhibitors, the dose of the cytotoxic therapeutic can be increased and/or the treatment with such a drug can be prolonged. The therapeutic benefit of such cytotoxic therapy can be significantly increased by combination with NHE inhibitors.

In addition, the NHE1 inhibitors of formula I and/or II and/or their pharmaceutically acceptable salts according to the invention can be used in overproduction of thyroid hormone, thyrotoxicosis which is harmful to the heart, or in external supply of thyroid hormone. The compounds of formula I and/or II, and/or their pharmaceutically acceptable salts, are therefore suitable for improving therapy with cardiotoxic drugs.

In accordance with their protective effect against ischemia-induced injuries, the compounds of the invention are also suitable as a drug for the treatment of ischemia of the nervous system, in particular of the central nervous system, where they are suitable, for example, for the treatment of stroke or cerebral edema.

The compounds of formula I and/or II and/or their pharmaceutically acceptable salts are also suitable for the therapy and prophylaxis of diseases and disorders caused by hypersensitivity of the central nervous system, in particular for the treatment of diseases of the epileptic circle form, centrally caused conical and tonic spasms, mental states of depression, sick fear and psychosis. The NHE inhibitors described here can be used alone or in combination with other antiepileptic or antipsychotic active substances, or with carbonic anhydrase inhibitors, for example with acetazolamide, as well as with other NHE inhibitors or sodium-dependent chloride-bicarbonate exchangers (NCBE ).

Furthermore, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts can also be suitably used to treat forms of shock, such as allergic, cardiogenic, hypovolemic and bacterial shock.

The compounds of formula I and/or II and/or their pharmaceutically acceptable salts can also be used for the prevention and treatment of thrombotic diseases, because they, as NHE inhibitors, can also inhibit platelet aggregation by themselves. Furthermore, after ischemia and reperfusion, they inhibit or reduce the occurrence of excessive release of inflammation and clotting mediators, especially Willebrand factor and thrombogenic selectin proteins. This can reduce and exclude the pathogenic effect of important thrombogenic factors. Therefore, the NHE inhibitors of the proposed invention can be combined with further anti-coagulation and/or thrombolytic active substances, such as for example recombinant or natural tissue plasminogen activator, streptokinase, urokinase, acetyl-salicylic acid, thrombin antagonists, factor Xa antagonists, drugs which act fibrinolytically, thromboxane receptor antagonists, phosphodiesterase inhibitors, factor Vila antagonists, clopidogrel, ticlopidine, etc. The combined use of the proposed NHE inhibitors with NCBE inhibitors and/or with carbonic anhydrase inhibitors, such as for example with acetazolamide, is especially advantageous.

The compounds of formula I and/or II and/or their pharmaceutically acceptable salts, used according to the invention, are further characterized by a strong inhibitory effect on cell proliferation, for example on the proliferation of fibroblast cells and on the proliferation of smooth muscle cells of blood vessels. Therefore, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts can be considered as valuable therapeutics for diseases in which cell proliferation is the primary or secondary cause, and therefore they can be used as antiatherosclerotics, agents against chronic kidney failure, against cancer.

It could be shown that cell migration is inhibited with the compounds according to the invention. For this reason, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts come into consideration as valuable therapeutics for diseases in which cell migration is a primary or secondary cause, such as cancer diseases with a strong tendency to metastasize.

The compounds of the formula I and/or II and/or their pharmaceutically acceptable salts are characterized by retardation or prevention of fibrotic diseases. They are therefore suitable as an outstanding agent for the treatment of cardiac fibrosis, as well as pulmonary fibrosis, liver fibrosis, renal fibrosis and other fibrotic diseases.

They can therefore be used to treat hypertrophy and hyperplasia of organs, for example the heart and prostate. They are therefore suitable for the prevention and treatment of heart failure (congestive heart failure CHF) as well as for the treatment and prevention of hyperplasia, i.e. hypertrophy of the prostate.

Since NHE is significantly elevated in essential hypertensives, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts are suitable for the prevention and treatment of high blood pressure and cardiovascular disease.

In doing so, they can be administered alone or with a suitable participant in the combination and formulation for the treatment of high blood pressure and cardiovascular disease. Thus, for example, one or more thiazide-acting diuretics, loop diuretics, aldosterone and pseudoaldosterone antagonists, such as hydrochlorothiazide, indapamide, polythiazide, furosemide, piretanide, torasemide, bumetanide, amiloride, triamterene, spironolactone or epleron, can be combined. The NHE inhibitors of the proposed invention can further be used in combination with calcium antagonists, such as verapamil, diltiazem, amlodipine or nifedipine, as well as with ACE inhibitors, such as for example ramipril, enalapril, lisinopril, fosinopril or captopril. Further favorable participants in the combination are also β-blockers such as metoprolol, albuterol, etc., antagonists of the angiotensin receptor and its receptor subtypes such as losartan, irbesartan, valsartan, omapatrilat, gemopatrilat, endothelin antagonists, renin inhibitors, adenosine receptor agonists, inhibitors and activators potassium channels such as glibenclamide, glimepiride, diazoxide, cromokalim, minoxidil and their derivatives, activators of mitochondrial ATP-sensitive potassium channels (mitoK (ATP) channel), Kv 1.5 inhibitors, etc.

NHE1 inhibitors of formula I and/or II and/or their pharmaceutically acceptable salts have been shown to have significant antiphlogistic activity and can therefore be used as anti-inflammatory agents. In doing so, the release of inflammatory mediators is inhibited. Therefore, these compounds can be used alone or in combination with antiphlogistics in the prevention or treatment of chronic and acute inflammatory diseases. Steroidal and non-steroidal anti-inflammatory drugs can usefully be used as participants in the combination.

In addition, compounds of formula I and/or II and/or their pharmaceutically acceptable salts have been found to exhibit a beneficial effect on serum lipoproteins. It is generally recognized that excessive fat in the blood, so-called hyperlipoproteinemia, is an important risk factor for the development of arteriosclerotic changes in blood vessels, especially coronary heart disease. Therefore, for the prophylaxis and regression of atherosclerotic changes, the reduction of elevated lipoproteins in the serum is extremely significant. In addition to the reduction of total cholesterol in the serum, the reduction of specific atherogenic lipid fractions of that total cholesterol, especially low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL), is particularly significant, because these lipid fractions represent an atherogenic risk factor. Conversely, high-density lipoproteins have been attributed a protective function against coronary heart disease. Accordingly, hypolipidemics must be able to reduce not only total cholesterol, but especially VLDL and LDL cholesterol fractions in the serum. NHE1 inhibitors have now been found to exhibit valuable therapeutically useful properties in terms of their effect on serum lipids. Thus, they significantly reduce elevated concentrations of LDL and VLDL in the serum, which are observed, for example, due to increased dietary intake of food rich in cholesterol and lipids, or in pathological changes in metabolism, for example in genetically determined hyper-lipidemia. They can therefore be used for prophylaxis and for the regression of atherosclerotic changes, whereby they exclude the causal risk factor. This includes not only primary hyperlipidemias, but also certain secondary hyperlipidemias, such as those that occur, for example, in diabetes. The compounds of formula I and/or II and/or their pharmaceutically acceptable salts further lead to a clear reduction of the infarction caused by the anomalies of metabolism and in particular to a significant reduction of the size of the infarction thus caused and the degree of its severity. Therefore, the mentioned compounds can be usefully used for the preparation of a drug for the treatment of hypercholesterolemia; for the preparation of a medicine for the prevention of atherogenesis; for the preparation of a medicine for the prevention and treatment of atherosclerosis, for the preparation of a medicine for the prevention and treatment of diseases caused by an increased amount of cholesterol, for the preparation of a medicine for the prevention and treatment of diseases caused by endothelial dysfunction, for the preparation of a medicine for the prevention and treatment of hypertension caused by atherosclerosis, for the preparation of a drug for the prevention and treatment of thrombosis caused by atherosclerosis, for the preparation of a drug for the prevention and treatment of ischemic injuries and post-ischemic reperfusion injuries caused by hypercholesterolemia and endothelial dysfunction, for the preparation of a drug for the prevention and treatment of cardiac hypertrophy and cardiomyopathy and congestive heart failure (CHF) caused by hypercholesterolemia and with endothelial dysfunction, for the preparation of a drug for the prevention and treatment of coronary spasms of blood vessels and myocardial infarction caused by hypercholesterolemia and endothelial dysfunction, for the preparation of a drug for the treatment of the mentioned ailments in combination and with substances that lower high blood pressure, primarily with angiotensin-converting enzyme inhibitors (ACE inhibitors) and angiotensin-receptor antagonists. A combination of NHE inhibitors of formula I and/or II and/or their pharmaceutically acceptable salts with an active substance that lowers the amount of fat in the blood, preferably with an HMG-CoA reductase inhibitor (for example lovastatin or pravastatin, the latter being attributed a hypo-lipidemic effect and thus, it increases the hypolipidemic properties of NHE inhibitors of formula I and/or II and/or their pharmaceutically acceptable salts, proved to be the most favorable combination with an enhanced effect and with the use of a smaller amount of active substance.

Thus, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts lead to effective protection against endothelial injuries of various genesis. With this protection of blood vessels against endothelial dysfunction syndrome, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts are a valuable drug for the prevention and treatment of coronary spasms of blood vessels, diseases of peripheral blood vessels, especially intermittent claudication, atherogenesis and atherosclerosis, left atrial hypertrophy and dilated cardiomyopathy, and thrombotic diseases.

In addition, the benzoylguanidines of formula I and/or II and/or their pharmaceutically acceptable salts have been found to be useful in the treatment of non-insulin dependent diabetes (NIDDM) syndrome, thereby suppressing insulin resistance. At the same time, to enhance the antidiabetic efficiency and quality of action, it may be advantageous to combine these compounds according to the invention with a biguanide, such as metformin, with an antidiabetic sulfonylurea, such as glyburide, glimepiride, tolbutamide, etc., with a glucosidase inhibitor, with PPAR agonists , such as rosiglitazone, pioglitazone, etc., with insulin preparations of different forms of administration, with a DB4 inhibitor, with an insulin sensitizing agent or with meglitinide.

In addition to acute antidiabetic effects, the compounds of formula I and/or II and/or their pharmaceutically acceptable salts act against the occurrence of late diabetic complications and can therefore be used as a drug for the prevention and treatment of diabetic late injuries, such as diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, diabetic cardiomyopathy and other diseases that appear as consequences of diabetes. In doing so, they can also be usefully combined with anti-diabetic drugs described in the treatment of NIDDM. A combination with a favorable form of insulin administration could be particularly significant.

In addition to the protective effects against acute ischemic events and the following also acute burdening reperfusion events, the NHE inhibitors of formula I and/or II and/or their pharmaceutically acceptable salts according to the invention also show direct, therapeutically useful effects against diseases and disorders of the entire mammalian organism which are associated with the phenomena of the aging process, which takes place chronically and independently of acute conditions of insufficient blood supply, and which also appear under normal, non-ischemic conditions. Those pathological phenomena caused by aging, which appear during a long period of aging as morbidity, severe incurable disease and death, the treatment of which has been made possible by new NHE inhibitors, are diseases and disorders that are largely caused by age-related changes in organs and their functions necessary for life and in an aging organism they gain increasing importance.

Diseases that are associated with dysfunction due to aging, the occurrence of organ wear and tear, are, for example, insufficient excitability and the ability of blood vessels to respond to contraction and relaxation reactions. This decline in the ability of blood vessels to react to constrictor and relaxant impulses due to age, which is an essential process of the cardiac circulation system and thus life and health, can be significantly improved or limited with NHE inhibitors. An important function and measure for properly maintaining the responsiveness of blood vessels is the blocking, or retardation, of advanced endothelial dysfunction caused by age, which can be significantly improved with NHE inhibitors. The compounds of formula I and/or II, and/or their pharmaceutically acceptable salts, are therefore very suitable for the treatment and prevention of advanced age-related endothelial dysfunction, especially intermittent claudication.

An example of a further measure that characterizes the aging process is the decline in heart contractility and the decline in the heart's adaptability to the required heart pump power. This decrease in the ability to act, as a result of the aging process, is in most cases related to heart dysfunction, which is caused by, among other things, the accumulation of connective tissue in the heart tissue. This accumulation of connective tissue is characterized by an increase in the weight of the heart, enlargement of the heart and limitation of heart function. It was surprising that such aging of the heart organ could be almost completely inhibited. The compounds of formula I and/or II, and/or their pharmaceutically acceptable salts, are therefore very suitable for the treatment and prevention of congestive heart failure (CHF).

While the subject of patent protection in previous patents and patent applications is the treatment of various forms of cancer that have already occurred, now it was extremely surprising that by inhibiting proliferation, not only already formed cancer can be treated, but that with NHE inhibitors, it can be prevented and highly significantly delayed as well incidence of cancer due to age. The discovery that the appearance of age-related diseases of all organs, and not only certain forms of cancer, was interrupted, or highly significantly delayed, is particularly worth mentioning. The compounds of formula I and/or II, and/or their pharmaceutically acceptable salts, are therefore very suitable for the treatment and prevention of cancer forms that are caused by aging.

Now it has been established, not only above the statistical normal measure, a highly significant delay in the onset of age-related diseases of all investigated organs, including the heart, blood vessels, liver, etc., but also a highly significant delay in senile cancer {colon cancer and lung cancer in older). In addition, in a surprising way, there is also a prolongation of life to an extent that could not be achieved with any other group of drugs, that is, with any natural substances. This unique effect of HNE inhibitors, in addition to the application of the active substance on humans and animals, also allows these HNE inhibitors to be combined with other gerontologically useful active substances, measures, substances and natural substances, which are based on a different mechanism of action. Classes of such active substances that are used in gerontological therapy are: especially vitamins and substances that act as antioxidants. Since there is a correlation between caloric load, i.e. food intake and the aging process, a combination with dietary measures, for example with appetite suppressants, is also possible. It is also possible to imagine a combination with drugs to lower blood pressure, such as ACE inhibitors, angiotensin receptor antagonists, diuretics, Ca2+ antagonists, etc., or with drugs to normalize metabolism, such as cholesterol-lowering agents. The compounds of formula I and/or II, and/or their pharmaceutically acceptable salts, are therefore very suitable for preventing age-related tissue changes and for prolonging life while maintaining a high quality of life.

The compounds according to the invention are effective inhibitors of cellular sodium-proton antiporters (Na/H-Exchanger), which are elevated in numerous diseases (essential hypertension, atherosclerosis, diabetes, etc.) also in such cells that are easily accessible for measurements, such as for example erythrocytes, platelets or leukocytes. The compounds used according to the invention are therefore suitable as a prominent and simple scientific tool, for example in their use as a diagnostic for determining and differentiating certain forms of hypertension, but also atherosclerosis, diabetes and diabetic late complications, proliferative diseases, etc.

What is further claimed is a drug for human medicine, veterinary medicine or for phytoprotective use containing an effective amount of a compound of formula I and/or II and/or a pharmaceutically acceptable salt thereof, together with pharmaceutically acceptable carriers and additives, alone or in combination with other pharmacologically active substances or medicines.

Thereby, drugs containing at least one compound of formula I and/or II, and/or a pharmaceutically acceptable salt thereof, can be administered orally, parenterally, intravenously, rectally, percutaneously or by inhalation, the preferred method of administration depending on the respective the appearance of the disease. In this sense, compounds I and/or II can be used alone, or together with pharmaceutical aids in veterinary and human medicine. The drug contains the active substance of formula I I and/or II, and/or its pharmaceutically acceptable salt, generally in an amount of 0.01 mg to 1 g per dosage unit.

Thanks to his professional knowledge, the expert will know which excipient is suitable for the desired pharmaceutical formulation. In addition to solvents, gelling agents, suppository bases, tablet excipients and other carriers of the active compound, for example, antioxidants, dispersants, emulsifiers, antifoaming agents, flavor correctors, preservatives, to promote dissolution or dyes.

To prepare a formulation for oral use, the active compounds are mixed with additives suitable for this purpose, such as carriers, stabilizers or inert diluents, and processed by conventional methods into forms suitable for application, such as tablets, coated tablets, hard gelatin capsules or aqueous, alcoholic or oily solutions. Gum arabic, magnesium oxide, magnesium carbonate, potassium phosphate, lactose, glucose or starch, for example corn starch, can be used as inert carriers. At the same time, preparations can be produced in the form of dry or wet granules. Vegetable oils or animal oils, such as for example sunflower oil or cod liver oil, are suitable for use as oil carriers or as solvents.

For subcutaneous or intravenous administration, the active compounds, optionally together with substances customary for this purpose, such as dissolution enhancers, emulsifiers or additional auxiliaries, are brought into solution, suspension, or emulsion. Suitable solvents include, for example, water, physiological salt solution, or alcohol, for example ethanol, propanol or glycerin, and also sugar solutions, such as glucose or mannitol solutions, or also a mixture of these different solvents.

As pharmaceutical formulations that are administered in the form of an aerosol or spray, solutions, suspensions or emulsions of the active compound of the formula I and/or II and/or its pharmaceutically acceptable salts in a pharmaceutically acceptable solvent, such as in particular ethanol or water, or a mixture of such solvents.

If necessary, the formulation may contain additional pharmaceutical aids, such as surfactants, emulsifiers and stabilizers, as well as propellant gas. Such a preparation usually contains an active compound in a concentration of approx. 0.1 to 10, especially from approx. 0.3 to 3 wt. %.

The dosage of the active compound of formula I and/or II upon application, and the frequency of administration depend on the strength and duration of action of the compound used. The dosage used also depends on the nature and severity of the disease being treated, as well as on the sex, age, weight and individual reaction of the mammal being treated.

The average daily dose of the compound of formula I and/or II and/or its pharmaceutically acceptable salt for a patient with severe approx. 75 kg is at least 0.001 mg/kg, for example 0.01 mg/kg, to at most 10 mg/kg, for example 1 mg/kg. In acute disease outbreaks, for example immediately after suffering a heart attack, it may also be necessary to give higher and/or more frequent dosages, eg up to 4 single doses per day. Especially in the case of intravenous administration, for example in the case of a patient with a heart attack in intensive care, up to 700 mg per day may be required and the compounds according to the invention may be given by infusion.

List of abbreviations

ADMET Absorption-Distribution-Metabolism-

Ausscheidung-Toxikologie = absorption-distribution-metabolism-excretion-toxicology

CDId di-imidazol-1-yl-methanone

DIP diisopropyl ether

DIPEA diisopropylethylamine

DME 1,2-dimethoxyethane

DMF N,N-dimethylformamide

EE ethyl acetate (EtOAc)

eq. equivalent

HEP n-heptane

HOAc acetic acid

KOtBu potassium 2-methyl-propan-2-olate

MeOH methanol

mp melting point

MTB tert-butyl methyl ether

Pd(dppf)2 [1,1'-bis-(diphenylphosphino)-ferrocene]-palladium(II)-chloride-methylene chloride complex (1:1)

RT room temperature

TFA trifluoroacetic acid

THF tetrahydrofuran

TMEDA N,N,N',N'-tetramethyl-ethane-1,2-diamine

EXPERIMENTAL PART

Example 1

3-pentafluorosulfanyl-benzoylguanidine hydrochloride

[image]

a) 3-pentafluorosulfanyl-benzoic acid

700 mg of (3-iodophenyl)sulfur pentafluoride (Tetrahedron 56, (2000) 3399) and 300 mg of methyl iodide are dissolved in 20 ml of diethyl ether (anhydrous) and the solution is slowly added dropwise to 155 mg of magnesium in 10 ml of diethyl ether. It is stirred for one hour under reflux, then it is cooled to -10°C and the reaction mixture is exposed to CO2 under normal pressure. It is stirred for 16 hours at room temperature, then the reaction mixture is adjusted to pH = 1 with a dilute aqueous HCl solution and extracted 3 times with 50 ml EE each. It was dried over MgSO4 and the solvent was removed in vacuo. Chromatography on silica gel with DIP/2% HOAc gives 200 mg of a colorless, amorphous powder.

Rf (DIP/2% HOAc) = 0.51 MS (ES+) : 249

b) 3-pentafluorosulfanyl-benzoylguanidine, hydrochloride

30 mg of 3-pentafluorosulfanylbenzoic acid was stirred together with 24 mg of CDI in 5 ml of DMF (anhydrous) for 3 hours at room temperature. Additionally, 69 mg of guanidinium chloride was mixed together with 68 mg of KOtBu in 5 ml of DMF (anhydrous) for 30 minutes at room temperature. The two solutions are then combined and allowed to stand for 18 hours at room temperature. The reaction mixture is diluted with 20 ml of EE and washed 3 times with 5 ml of semi-concentrated aqueous NaHCO3 solution each. It is dried over MgSO4, the solvent is removed in vacuo and the residue is taken up in a 5% aqueous HCl solution. The volatiles were removed in vacuo to give 33 mg of an amorphous solid.

Rf (EE) = 0.30 MS(ES+): 290

Example 2

4-pentafluorosulfanyl-benzoylguanidine, hydrochloride

[image]

a) 4-pentafluorosulfanyl-benzoic acid

2.7 g of (4-iodophenyl)sulfur pentafluoride (Tetrahedron 56, (2000) 3399) is reacted analogously to example 1 a), which gives 630 mg of a colorless, amorphous solid.

Rf (DIP/2% HOAc) = 0.51 MS (ES+): 249

b) 4-pentafluorosulfanyl-benzoylguanidine, hydrochloride

50 mg of 4-pentafluorosulfanyl-benzoic acid is reacted analogously to example 1 b), thereby obtaining 33 mg of the title compound of example 2 as an amorphous powder.

Rf (EE) = 0.30 MS (ES+) : 290

Example 3

4-pentafluorosulfanyl-2-methyl-benzoylguanidine

[image]

a) 4-pentafluorosulfanyl-2-methyl-benzoic acid

3.09 g of TMEDA is dissolved in 30 ml of THF (anhydrous) and 20.5 ml of a 1.3 M solution of sec-butyllithium in cyclohexane is added dropwise at -90°C. Then, at -90°C, a solution of 3.0 g of 4-pentafluorosulfanylbenzoic acid in 20 ml of THF (anhydrous) is added dropwise. It is stirred for another hour at -90°C, then a solution of 5.15 g of methyl iodide in 20 ml of THF (anhydrous) is added dropwise. The temperature is maintained at -80°C. Then it is mixed for another 20 minutes at -78°C, 100 ml of water is injected, adjusted to pH -1 with a diluted aqueous HCl solution and extracted 3 times with 100 ml of MTB. It was dried over MgSO4 and the solvent was removed in vacuo. The residue is then chromatographed on silica gel with DIP/2% HOAc, and 1.60 g of a mixture of 4-pentafluorosulfanyl-benzoic acid and 4-pentafluorosulfanyl-2-methyl-benzoic acid is obtained. This mixture is chromatographed again under the following conditions: Column: Waters X-terra 250x50 mm + pre-column 50x50 mm

Packaging: C18 10 μM Flow: 150 ml/min

Gradient (linear flow)):

Solvent A: water + 2% trifluoroacetic acid

Solvent B: acetonitrile

[image]

900 mg of the title compound is obtained as a colorless solid with a retention time of 21.14 minutes, in addition to 360 mg of 4-pentafluorosulfanylbenzoic acid with a retention time of 20.18 minutes (proved at a wavelength of 220 nm).

Rf (DIP/2% HOAc) = 0.5 MS (Cl+) : 2.63; MS (ES-) : 261

b) 4-pentafluorosulfanyl-2-methyl-benzoylguanidine

910 mg of 4-pentafluorosulfanyl-2-methyl-benzoic acid is dissolved in 25 ml of DMF (anhydrous), 844 mg of CDI is added at room temperature and stirred for 6 hours at room temperature (solution 1). In addition, 1.988 g of guanidinium chloride and 1.947 g of KOtBu were stirred for 30 min in 10 ml of DMF (anhydrous) at room temperature (solution 2). Then solution 2 is added to solution 1 and mixed for 17 hours at room temperature. The reaction mixture is then diluted with 200 ml of MTB and washed once with 100 ml of water. This water is then extracted with 100 ml of MTB. The combined organic phases are then washed once more 3 times with 50 ml of water each and dried over MgSO4. The solvent was removed in vacuo and the residue was chromatographed on silica gel with EE. 600 mg of white crystals are obtained, m.p. 185°C.

Rf (EE) = 0.22 MS(ES+): 304

Example 4

N-[2-chloro-4-pentafluorosulfanyl-benzoyl]-guanidine

[image]

a) 2-chloro-4-pentafluorosulfanyl-benzoic acid

20.0 ml of TMEDA was dissolved in 150 ml of THF (anhydrous) and at a temperature between -80°C and -90°C was mixed with 93.0 ml of a 1.25 M solution of sec. BuLi in cyclohexane. Then, at a temperature between -87°C and -93°C, a solution of 4-pentafluorosulfanyl-benzoic acid (example 2a) in 50 ml of THF (anhydrous) is added dropwise within 35 minutes. It is stirred for another 2 hours at -90°C. and then 38.2 g of 1,1,1,2,2,2-hexachloroethane in 60 ml of THF (anhydrous) is added dropwise at -90°C. Let it warm up to -70°C and then add 100 ml of water. The solvent was removed in vacuo and the residue was chromatographed on silica gel with DIP/2% HOAc. 5.0 g of the desired product is obtained as a partially crystallized oil.

Rf (DIP/2% HOAc) = 0.21 MS(EI): 282 (M+1)+

b) N-[2-chloro-4-pentafluorosulfanyl-benzoyl]guanidine

170 mg of 2-chloro-4-pentafluorosulfanylbenzoic acid was dissolved in 3 ml of DMF (anhydrous) and 127 mg of CDI was added at room temperature. It is stirred for another 2 hours at room temperature and the intermediate imidazolide is obtained.

In addition, 337 mg of KOt-Bu was dissolved in 5 ml of DMF (anhydrous) and a solution of 344 mg of guanidine hydrochloride in 5 ml of DMF (anhydrous) was added dropwise. The solution is stirred for another 30 minutes at room temperature and then the imidazolide solution is added dropwise at room temperature. The reaction mixture was allowed to stand for 16 hours at room temperature and then the solvent was removed in vacuo. The residue is taken up in 50 ml of EE/20 ml of water and washed twice with 20 ml of water, then twice with 20 ml of saturated aqueous NaCl solution. It was dried over MgSO4 and the solvent was removed in vacuo. The residue is chromatographed on silica gel with EE, which gives 90 mg of product (amorphous).

Rf (EE) = 0.13 MS (ES+): 324 (M+1)+

Example 5

N-[2-(4-Fluoro-phenoxy)-4-pentafluorosulfanyl-benzoyl]-guanidine

[image]

a) 2-chloro-4-pentafluorosulfanyl-benzoic acid-methyl ester

4.3 g of 2-chloro-4-pentafluorosulfanyl-benzoic acid is dissolved in 50 ml of methanol and 5.6 ml of SOCl2 is slowly added dropwise at room temperature. It is boiled for 6 hours under reflux, the volatile ingredients are removed in a vacuum, the residue is taken up in 100 ml of toluene and the volatile ingredients are again removed in a vacuum. 4.1 g of colorless oil is obtained.

Rf (HEP/DIP 1:1) - 0.44

b) 2-(4-fluoro-phenoxy)-4-pentafluorosulfanyl-benzoic acid-methyl ester

300 mg of 2-chloro-4-pentafluorosulfanylbenzoic acid methyl ester, 113 mg of 4-fluorophenol as well as 659 mg of Cs2CO3 are dissolved in 1.5 ml of DMF (anhydrous) and stirred at 120°C. Then let it cool, dilute with 10 ml of EE and wash 3 times with 5 ml of water each. It was dried over MgSO4 and the solvent was removed in vacuo. 120 mg of an amorphous solid is obtained, which is chromatographed on reverse phase silica gel.

Flow: 30 ml/min

Gradient: ACN=A; lead + 0.2% TFA=B

0-3 min 5% A; -14 min 95% A; 15-18 min 95% A; -20 min 5% A

Column: XTerra C18 5 μm 30x100 mm

20 mg of an amorphous solid is obtained.

Rf silica gel (HEP/DIP 1:1) = 0.44

c) N-[2-(4-fluoro-phenoxy)-4-pentafluorosulfanyl-benzoyl]-guanidine

270 mg of KOt-Bu as well as 324 mg of guanidine hydrochloride were mixed in 1 ml of DMF (anhydrous) at room temperature for 30 minutes. A solution of 54 mg of 2-(4-fluorophenoxy)-4-pentafluorosulfanylbenzoic acid methyl ester in 1 ml of DMF (anhydrous) is then added. It is then stirred for 2 hours at room temperature, then poured into 10 ml of water, adjusted to pH = 8 with an aqueous HCl solution and extracted 3 times with 5 ml of MTB. The organic phase is washed with another 10 ml of water, then dried over MgSO4 and finally the solvent is removed under vacuum. 20 mg of an amorphous solid is obtained.

Rf (EE) = 0.22 MS (ES+) : 400 (M+1)+

Example 6

N-(2-methyl-5-pentafluorosulfanyl-benzoyl)-guanidine

[image]

a) 1-dichloromethyl-2-nitro-4-pentafluorosulfanyl-benzene

5.4 g of KOtBu was dissolved in 25 ml of DMF (anhydrous) as well as 20 ml of THF (anhydrous) and cooled to -73°C. A solution of 3.0 g of l-nitro-3-pentafluorosulfanylbenzene and 1.1 ml of CHCl3 in 15 ml of DMF (anhydrous) is then added dropwise as quickly as possible (in about 20 minutes), keeping the temperature below -67°C. Stir for one minute at -70°C, then inject 15 ml of glacial acetic acid into 15 ml of methanol and warm to room temperature. The reaction mixture is poured over 200 ml of water and extracted 3 times with 100 ml of CH2Cl2 each. The organic phase is then washed 3 more times with 50 ml of saturated aqueous Na2CO3 solution each. It was dried over MgSO4 and then the solvent was removed in vacuo. The residue was chromatographed on silica gel with EE/HEP 1:10 and 3.0 g of a colorless oil was obtained.

Rf (EE/HEP 1:10) = 0.78

b) 2-methyl-5-pentafluorosulfanyl-aniline 2.0 g of 1-dichloromethyl-2-nitro-4-pentafluorosulfanyl-benzene is dissolved in 25 ml of DME, 200 mg of Pd/C (5%) is added as well as 100 ml of saturated aqueous NaHCO3 solution and hydrate for 54 hours under 5 bar of hydrogen. Then the catalyst is filtered off and the reaction solution is extracted 3 times with 50 ml of MTB. It was dried over MgSO4 and the solvent was removed in vacuo. 1.1 g of colorless oil is obtained.

Rf (DIP) = 0.42

c) 2-bromo-1-methyl-4-pentafluorosulfanyl-benzene

1.6 g of 2-methyl-5-pentafluorosulfanyl-aniline is dissolved in 15 ml of glacial acetic acid and at 5°C, 3 ml of a 48% aqueous solution of HBr is added dropwise. During this, a precipitate is formed. Then, at a temperature between 0°C and 5°C, a solution of 0.52 g of NaNO2 in 3 ml of water is added dropwise within 10 minutes. This diazonium salt solution is then poured into a suspension of 1.2 g of CuBr in 10 ml of a 48% aqueous solution of HBr and 10 ml of water. It is stirred for another 30 minutes at room temperature, then diluted with 50 ml of water and extracted 3 times with 50 ml of DIP each. The organic phase is washed 2 more times with 50 ml of saturated aqueous Na2CO3 solution each. It was dried over MgSO4 and then the solvent was removed in vacuo. 1.6 g of poorly mobile oil is obtained.

Rf (MTB/n-pentane 1:5) = 0.67

d) 2-methyl-5-pentafluorosulfanyl-benzoic acid

200 mg of 2-bromo-1-methyl-4-pentafluorosulfanyl-benzene is dissolved in 6 ml of DMF, 3 ml of tri-n-butylamine and 0.5 ml of water and 129 mg of Cs2CO3 is added. Then 15.1 mg of Pd (OAc)2 as well as 35.3 mg of triphenylphosphine are added and under normal CO pressure it is boiled for 6 hours under reflux. The reaction mixture is diluted with 100 ml of EE and 30 ml of water, adjusted to pH = 2-3 with aqueous HCl solution and extracted twice with 30 ml of EE each. It was dried over MgSO4 and then the solvent was removed in vacuo. Chromatography on the reverse phase gives 17 mg of an amorphous solid.

Chromatography procedure: HPLC, 3 flows; 1 flow with gradient 1, 2 flows with gradient 2.

HPLC: Flow time for both gradients, 1 and 2, is 20 minutes each.

Flow agent: water (double distilled) + 0.2% TFA, acetonitrile (Chromasolv);

flow rate: 30 ml/min.

Column: WatersXterra™ MS C18 5 μm, 30 x 100 mm,

MS: standard 20 min, fractionation: with time.

Gradient 1: Gradient 2:

0-2.5 min 10% ACN 0-2.5 min 10% ACN

3.0 min 25% ACN 3.0 min 20% ACN

14.0 min 75% ACN 14.0 min 60% ACN

15.0 min 95% ACN 15.0 min 95% ACN

17.5 min 10% ACN 17.5 min 10% ACN

LCMS (ES-) : 261 (M-1)-

e) N-(2-methyl-5-pentafluorosulfanyl-benzoyl)-guanidine

17.0 mg of 2-methyl-5-pentafluorosulfanylbenzoic acid was dissolved in 2 ml of DMF (anhydrous), mixed with 15 mg of CDI and stirred for 4 hours at room temperature.

In addition, 36.5 mg of KOt-Bu and 37.3 mg of guanidine hydrochloride were dissolved in 2 ml of DMF (anhydrous) and stirred for 30 minutes at room temperature. This solution of the guanidine base is then added dropwise to the above imidazolide and left to stand for 16 hours at room temperature. The solvent is removed under vacuum, the residue is taken up in 5 ml of water and adjusted to pH = 9 with an aqueous solution of HCl. It is then extracted 3 times with 10 ml of EE each. It was dried over MgSO4 and then the solvent was removed in vacuo. Chromatography on silica gel with EE gives 7.0 mg of product as an amorphous solid.

Rf (EE) = 0.20 MS (ES+): 304 (M+1)+

NHE inhibition procedure

The IC50 value [nM] of NHE-1 inhibition was determined as follows.

FLIPR assay for determination of NHE-1 inhibitors by measurement of pHi increase in transfected cell lines expressing human NHE-1

The experiment was carried out in the device FLIPR (Fluorescent imaging plate reader) with black-walled microtiter plates with 96 wells and a clear bottom. Transfected cell lines, which express different subtypes of HNE (parental cell line LAP-1 as a result of mutagenesis and then selection does not have any endogenous NHE activity), were seeded a day earlier at a density of approx. 25,000 cells/wells. [Growth medium for transfected cells (Iscove + 10% fetal bovine serum) additionally contained G418 as a selective antibiotic to ensure the presence of transfected sequences.]

The actual experiment was started by removing the growth medium and adding 100 μl/well of loading buffer (5 μM BCECF-AM [2',7'-bis-(carboxyethyl)-5-(i -6)-carboxy-fluorescein, acetoxymethyl ester] in 20 mM NH4Cl, 115 mM choline chloride, 1 mM MgCl2, 1 mM CaCl2, 5 mM KCl, 20 mM HEPES, 5 mM glucose; pH 7.4 [adjusted with KOH]). Then the cells were incubated for 20 minutes at 37°C. This incubation leads to the loading of the cells with a fluorescent dye, whose fluorescence intensity depends on the pHi, and with NH4Cl, which leads to a slight alkalinization of the cell.

[The previous step without the fluorescent dye BCECF-AM is membrane permeable as an ester. Intracellularly, esterases release the actual dye, which does not pass through the membrane].

After this 20-minute incubation, the loading buffer, which contains NH4Cl and free BCECF-AM, is removed by washing three times in a cell washing device (Liquid Columbus) with 400 μl each of washing buffer (133.8 mM choline chloride, 4.7 mM KCl, 1.25 mM MgCl2, 1.25 mM CaCl2, 0.97 mM K2HPO4, 0.23 mM KH2PO4, 5 mM HEPES, 5 mM glucose; pH 7.4 [adjusted with KOH]. Volume retained in wells is 90 μl (possibly 5O-125 μl).With this washing, free BCECF-AM is removed and as a consequence of the removal of external NH4 ions, this leads to an increase in intracellular acidity (pHi approx. 6.3-6.4).

Since the balance of intracellular NH4+ is disturbed with NH3 and H+ with the removal of extracellular NH4+ and with the subsequent immediate passage of NH3 through the cell membrane, the growth process leads to the retention of intracellular H+, which is the cause of the increase in intracellular acidity. Eventually, if it persists long enough, it can lead to cell death.

At this point, it is important that the wash buffer be sodium-free (< 1 mM), as extracellular sodium ions would lead to an immediate rise in pHi due to the activity of the cloned NHE iso-forms.

It is also important that all buffers used (loading buffer, washing buffer, collection buffer) do not contain HCO3- ions, because the presence of bicarbonate would lead to the activation of the bicarbonate-dependent pHi-regulatory systems contained in parental LAP-1. cell line.

Microtiter plates with acidified cells are then transferred (within 20 minutes after acidification) to the FLIPR. In FLIPR, the intracellular fluorescent dye is excited with 488 nm light from an argon laser, and the measurement parameters (laser power, illumination time, and screen of the CCD camera built into FLIPR are chosen so that the average fluorescence signal per well is between 30,000 and 35000 relative fluorescence units.

The actual measurements in FLIPR start with the software managing every two seconds to take one shot with the CCD camera. After ten seconds, an increase in intracellular pH values is caused by the addition of 90 μl of recovery buffer (133.8 mM NaCl, 4.7 mM KCl, 1.25 mM MgCl2, 1/25 mM CaCl2/0.97 mM K2HPO4, 0.23 mM KH2PO4, 10 mM HEPES, 5 mM glucose, pH 7.4 [adjusted with NaOH] using a 96-well pipettor inserted into the FLIPR.

As positive controls (100% NHE activity) wells to which pure recovery buffer was added, negative controls contain (0% NHE activity) washing buffer. In all other wells, recovery buffer with twice the concentration of the test substance is added. Measurement in FLIPR ends after 60 measurement points (two minutes).

The raw data was transferred to the ActivityBase program. With this program, NHE activities were first calculated for each concentration of the test substance and from this the IC50 values for the substances were calculated. As during the entire experiment the course of the increase in pHi is not linear, but in the end decreases due to the decrease of NHE activity at higher pHi values, it is important to select the part in which the increase in the fluorescence of the positive control is linear for the numerical display of the measurement.

[image]

Claims (15)

1. Pentafluorosulfanyl-benzoylguanidines of formulas I and/or II [image] indicated by that R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, alkoxy with 1, 2, 3 or 4 C atoms, F, Cl, Br, I, CN, NR10R11, -Op-(CH2)n-(CF2)o-CF3 or -(SOm)q-(CH2)r(CF2)s-CF3; R10 and R11 are independently hydrogen, alkyl with 1, 2, 3 or 4 C atoms or -CH2-CF3; m is zero, 1 or 2; n, o, p, q, r and s are independently zero or 1; R2 hydrogen, F, Cl, Br, I, -CN, -SO2CH3, -(SOh)z-(CH2)k-(CF2)-CF3, alkyl with 1, 2, 3, 4, 5 or 6 C atoms, cycloalkyl with 3, 4, 5, 6, 7 or 8 C atoms, in which 1, 2, 3 or 4 hydrogen atoms can be replaced by fluorine atoms; h is zero, 1 or 2; z zero or 1; k zero, 1, 2, 3 or 4; 1 is zero or 1; or R2 is -(CH2)t-phenyl or -O-phenyl, which is unsubstituted or substituted with 1, 2 or 3 residues selected from the group consisting of F, Cl, Br, I, -Ou-(CH2)V-CF3 , 1, 2, 3 or 4 C-atom alkoxy, 1, 2, 3 or 4 C-atom alkyl and -SO2CH3; t is zero, 1, 2, 3 or 4; u is zero or 1; v is zero, 1, 2 or 3; or R2 is -(CH2)w-heteroaryl, which is unsubstituted or substituted by si, 2 or 3 residues selected from the group consisting of F, Cl, Br, I, -Ox-(CH2)y-CF3, alkoxy with 1, 2, 3 or 4 C atoms and alkyl with 1, 2, 3 or 4 C atoms, -SO2CH3; w is zero, 1, 2, 3 or 4; x is zero or 1; y zero, 1, 2 or 3; R 3 and R 4 are independently hydrogen or F; as well as their pharmaceutically acceptable salts. 2. Compound of formula I and/or II according to claim 1, characterized in that R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, alkoxy with 1, 2, 3 or 4 C atoms, F, Cl, NR10R11, -O-CH2-CF3 or –SOm-(CH2)r-CF3 ; R10 and R11 are independently hydrogen, alkyl with 1, 2, 3 or 4 C atoms or -CH2-CF3; m is zero, 1 or 2; r is zero or 1; R2 hydrogen, F, Cl, -SO2CH3, -(SOh)z-(CH2)k-CF3, alkyl with 1, 2, 3 or 4 C atoms, cycloalkyl with 3, 4, 5, 6 or 7 C atoms, in in which 1, 2, 3 or 4 hydrogen atoms can be replaced by fluorine atoms; h is zero, 1 or 2; z is zero or 1; k is zero, 1, 2, 3 or 4; or R2 is phenyl or -O-phenyl, which is unsubstituted or substituted with 1 or 2 residues selected from the group consisting of F, Cl, -Ou-(CH2)V-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3; u is zero or 1; v is zero, 1, 2 or 3; or R2 is heteroaryl, which is unsubstituted or substituted with 1 or 2 residues selected from the group consisting of F, Cl, -Ox-(CH2)y-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3; x is zero or 1; y is zero, 1, 2 or 3; R 3 and R 4 are independently hydrogen or F; as well as their pharmaceutically acceptable salts. 3. Compound of formula I and/or II according to claim 1 or 2, characterized in that R1 represents hydrogen, alkyl with 1, 2, 3 or 4 C atoms, methoxy, ethoxy, F, Cl, NR10R11, -O-CH2-CF3 or -SOm-(CH2)r-CF3; R10 and R11 are independently hydrogen, alkyl with 1, 2, 3 or 4 C atoms or -CH2-CF3; m is zero, 1 or 2; r is zero or 1; R2 is hydrogen, F, Cl, -SO2CH3, -(SOh)z-(CH2)k-CF3, alkyl with 1, 2, 3 or 4 C atoms, cycloalkyl with 3, 4, 5, 6 or 7 C atoms, in which 1, 2, 3 or 4 hydrogen atoms can be replaced by fluorine atoms; h is zero or 2; z is zero or 1; k zero or 1; or R2 is phenyl or -O-phenyl, which is unsubstituted or substituted by 1 or 2 residues selected from the group consist of F, Cl, -O-(CH2)V-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3; v is zero, 1, 2 or 3; or R2 is heteroaryl, which is unsubstituted or substituted with 1 or 2 residues selected from the group consisting of F, Cl, -O-(CH2)y-CF3, methoxy, ethoxy, alkyl with 1, 2, 3 or 4 C atoms and -SO2CH3; y is zero, 1, 2 or 3; R 3 and R 4 are hydrogen; as well as their pharmaceutically acceptable salts. 4. Process for the preparation of the compound of formula I and/or II and/or its pharmaceutically acceptable salts, characterized in that the compound of formula III and/or IV, [image] in which R1 to R4 have the meanings given in claims 1, 2 and/or 3, and L is a leaving group which can be easily nucleophilically substituted, reacts with guanidine. 5. Compound of formula I and/or II and/or its pharmaceutically acceptable salts according to one or more claims 1 to 3, characterized in that it is used as a medicine. 6. The use of a compound of formula I and/or II and/or its pharmaceutically acceptable salts according to one or more claims 1 to 3, characterized in that it is used for the preparation of a medicine for the treatment or prophylaxis of acute or chronic injuries, diseases or indirect consequences of diseases organs and tissues caused by ischemia or reperfusion events, for the treatment or prophylaxis of arrhythmias, life-threatening ventricular flutter, heart attack, angina pectoris, for the treatment or prophylaxis of ischemic conditions of the heart, ischemic conditions of the peripheral and central nervous system or stroke or ischemic conditions of peripheral organs and tissues, for the treatment or prophylaxis of shock conditions, diseases in which cell proliferation is the primary or secondary cause, cancer, metastases, hypertrophy or hyperplasia of the prostate, atherosclerosis or disorders of fat metabolism, high blood pressure, especially essential hypertension, diseases of the central nervous system system, especially diseases that are p a consequence of CNS hypersensitivity, such as epilepsy or centrally induced convulsions, diseases of the central nervous system, especially states of fear, depression or psychosis, for the treatment or prophylaxis of non-insulin dependent diabetes mellitus (NIDDM) or diabetic late injuries, thrombosis, disease resulting from endothelial dysfunction, intermittent claudication, for the treatment or prophylaxis of fibrotic diseases of internal organs, fibrotic liver diseases, fibrotic kidney diseases, fibrotic blood vessel diseases and fibrotic heart diseases, for the treatment or prophylaxis of heart failure or congestive heart failure (e. Congestive Heart Failure), acute or chronic inflammatory diseases, diseases caused by protozoa, malaria and chicken coccidiosis and for use in surgical operations and organ transplants, for the preservation and disposal of transplants for surgical procedures, for the prevention of tissue changes caused by old age, for the preparation of medicine directed against aging or to prolong life, to treat and reduce cardiotoxic effects in thyrotoxicosis or to prepare diagnostics. 7. Use of the compound of formula I and/or II and/or its pharmaceutically acceptable salts according to one or more claims 1 to 3, characterized in that it is used in combination with other drugs or active substances for the preparation of a drug for the treatment or prophylaxis of acute or chronic injuries, diseases or indirect consequences of organ and tissue diseases caused by ischemia or reperfusion events, for the treatment or prophylaxis of arrhythmias, for life-threatening fluttering of the heart ventricle, heart attack, angina pectoris, for the treatment or prophylaxis of ischemic conditions of the heart, ischemic conditions of the peripheral and central nervous system or stroke or ischemic conditions of peripheral organs and tissues, for the treatment or prophylaxis of shock conditions, diseases in which cell proliferation is the primary or secondary cause, cancer, metastasis, hypertrophy or hyperplasia of the prostate, atherosclerosis or disorders of fat metabolism, high blood pressure , especially essential hypertension, disease and of the central nervous system, in particular diseases resulting from CNS hypersensitivity, such as epilepsy or centrally induced convulsions, diseases of the central nervous system, in particular states of fear, depression or psychosis, for the treatment or prophylaxis of non-insulin dependent diabetes mellitus (NIDDM ) or diabetic late injuries, thrombosis, diseases resulting from endothelial dysfunction, intermittent claudication, for the treatment or prophylaxis of fibrotic diseases of internal organs, fibrotic liver diseases, fibrotic kidney diseases, fibrotic blood vessel diseases and fibrotic heart diseases, for the treatment or prophylaxis of heart failure or congestive heart failure (e. Congestive Heart Failure), acute or chronic inflammatory diseases, diseases caused by protozoa, malaria and chicken coccidiosis and for use in surgical operations and organ transplants, for the preservation and disposal of transplants for surgical procedures, for the prevention of tissue changes caused by old age, for the preparation of medicine directed against aging or to prolong life, to treat and reduce cardiotoxic effects in thyrotoxicosis or to prepare diagnostics. 8. Use of the compound of formula I and/or II and/or its pharmaceutically acceptable salts according to claim 7, characterized in that it is used in combination with cardiotoxic and cytotoxic drugs or active substances for the preparation of a drug with reduced cardiotoxic and cytotoxic properties. 9. Use of the compound of formula I and/or II and/or its pharmaceutically acceptable salts alone or in combination with other drugs or active substances according to claim 6 and/or 7, characterized in that it is used for the preparation of a drug for the treatment or prophylaxis of acute or chronic injuries, diseases or secondary consequent diseases of organs and tissues that are caused by ischemia or reperfusion events. 10. The use of the compound of formula I and/or II and/or its pharmaceutically acceptable salts alone or in combination with other drugs or active substances according to claim 6 and/or 7, characterized in that it is used for the preparation of a drug for the treatment of life-threatening ventricular flutter. 11. Use of the compound of formula I and/or II and/or its pharmaceutically acceptable salts alone or in combination with other drugs or active substances according to claim 6 and/or 7, characterized in that it is used for the preparation of a drug for treatment or for prophylaxis metastasis. 12. Use of the compound of formula I and/or II and/or its pharmaceutically acceptable salts alone or in combination with other drugs or active substances according to claim 6 and/or 7, characterized in that it is used for the preparation of a drug for treatment or for prophylaxis fibrotic heart disease, heart failure or congestive heart failure. 13. Medicine for human, veterinary and/or phytoprotective use, characterized in that it contains an effective amount of the compound of formula I and/or II and/or its pharmaceutically acceptable salt according to one or more claims 1 to 3, together with pharmaceutically acceptable carriers and accessories. 14. Medicine for human, veterinary and/or phytoprotective use, characterized in that it contains an effective amount of the compound of formula I and/or II and/or its pharmaceutically acceptable salt according to one or more claims 1 to 3, together with pharmaceutically acceptable carriers and supplements, and in combination with other pharmacologically active substances or drugs. 15. Compounds of formula V [image] characterized in that R 5 represents hydrogen or (C 1 -C 4 )-alkyl.