WO1995026725A1

WO1995026725A1 - Pharmaceutical preparations and medicaments for the prevention and treatment of endothelial dysfunction

Info

Publication number: WO1995026725A1
Application number: PCT/DE1995/000421
Authority: WO
Inventors: Eike Albrecht Noack; Georg Kojda
Original assignee: Isis Pharma Gmbh
Priority date: 1994-03-30
Filing date: 1995-03-28
Publication date: 1995-10-12
Also published as: HU9602671D0; IS4365A; HU220165B; NO964102L; DE4410997A1; BG63073B1; EP0752858A1; BG100930A; AU698359B2; DE19580261D2; LT96148A; JPH09510979A; LV11666B; SI9520047A; LV11666A; LT4310B; FI963883A0; CZ283696A3; CA2186783A1; NO964102D0

Abstract

The present invention describes the use of compounds which release or transfer nitrogen monoxide, of endogenous nitrogen monoxide formation stimulators, and of guanylate cyclase stimulators for preventing, treating and eliminating endothelial dysfunctions and diseases associated with or caused by said dysfunctions. The invention further describes the use of said compounds for preparing pharmaceutical products for said areas of application.

Description

2

Pharmaceutical preparations and drugs for the prevention and treatment of endothelial dysfunction

description

Field of application of the invention

The invention presented here relates to the use of nitrogen monoxide releasing and / or transmitting compounds, stimulators of endogenous nitrogen monoxide formation and stimulators of guanylate cyclase for the prevention, treatment and elimination of endothelial dysfunctions as well as diseases which are caused by or associated with endothelial dysfunctions. According to the invention, the provision of pharmaceutical preparations for the named indications is made possible at the same time.

Known technical background

Organic nitric acid esters such as glycerol trinitrate (GTN) (Murrel, Lancet: 80, 113, 151 (1879)), pentaerythrityl tetranitrate (PETN) (Risemann et al., Circulation, Vol. XVII, 22 (1958), US Pat. No. 2,370,437) , Isosorbide-5-mononitrate (ISMN) (DE-OS 22 21 080, DE-OS 27 51 934, DE-OS 30 28 873, DE-PS 29 03 927, DE-OS 31 02 947, DE-OS 31 24 410, EP-PS 45 076, EP-PS 57 847, EP-PS 59 664, EP-PS 64 194,

EP-PS 67 964, EP-PS 143 507, US-PS 3 886 186, US-PS 4 065 488, US-PS 4 417 065, US-PS 4 431 829), isosorbide dinitrate (ISDN) (L. Goldberg, Acta Physiolog. Scand. 15, 173 (1948)), propatyl nitrate (Medard, Mem. Poudres 35: 113 (1953)), trolnitrate (FR-PS 984 523) or Nicorandil (US-PS 4,200,640) and similar compounds Vasodilators, some of which have had the broadest therapeutic use for decades in the indication of angina pectoris or ischemic heart disease (IHK) (Nitrangin®, Pentalong®, Monolong®, Isoket®, Elantan® and others). Comparable and improved pharmacological activity when used in the above-mentioned indication areas have organic nitrates of a newer type such as SPM 3672 (N- [3-nitratoρivaloyl] -L-cysteine-ethyl ester) (US Pat. No. 5,284,872) and its derivatives The use of organic nitrites such as isoamyl nitrite as coronary dilators has long been known (Brunton, Lancet: 97 (1867)). Other nitrogen monoxide releasing or transferring compounds such as thionitrites, thionitrates, S-nitrosothiols or nitrosoproteins (Harrison et al., Circulation 87: 1461-1467 (1993)) as well as substituted furoxanes (1,2,5-oxadiazole-2-oxides, furazan-N-oxides) (Feelisch et al., Biochem. Pharmacol. 44: 1149-1157 (1992) or substituted sydnonimines, in particular molsidomine (DE-AS 16 95 897, DE-AS 25 32 124, DD-PS 244 980) are also described as potent coronary dilators All of these substances are capable of acting themselves or in the form of their pharmacologically active metabolites, for example the molsidomine metabolites "SIN 1" and "SIN-" 1 A "(Noack, Nitroglycerin VII, Walter de Gruyter & Co., Berlin 1991, 23-28) and their derivatives and structural analogues (Noack and Feelisch, Molecular mechanism of nitrovascular bioactivation; in" Endothelial Mechanisms of Vasomotor Control "(ed. Drexler et al.), Pp. 37-50, Steinkopff Verlag, Darmstadt, FRG (1991)), to emit or transfer nitrogen monoxide in vivo.

The pharmaceutical processing of the organic nitrates or nitrites and other nitrogen monoxide liberating or transferring compounds into pharmaceutical preparations for the treatment of angina pectoris or ischemic heart disease is generally known. It is carried out in accordance with the working methods and rules which are generally familiar to the pharmaceutical expert, the choice of the technologies to be used and the pharmaceutical auxiliaries used being based primarily on the active ingredient to be processed. Questions of its chemical-physical properties, the chosen form of application, the desired duration of action and the avoidance of drug-excipient incompatibilities are of particular importance. Peroral, parenteral, sublingual or transdermal application in the form of tablets, dragees, capsules, solutions, sprays or plasters has been described for medicinal products with the indication angina pectoris or ischemic heart disease (DD-PS 293 492, DE-AS 26 23 800, DE-OS 33 25 652, DE-OS 33 28 094, DE-PS 40 07 705, DE-OS 40 38 203, JP application 59/10513 (1982)).

In addition to the long-known applications of nitrosating substances, their use for the treatment and prevention of diseases is described which are caused by pathologically increased concentrations of sulfur-containing amino acids in body fluids. These disease states, caused by congenital or acquired defects in the metabolism of these amino acids and which are characterized by increased blood and urine concentrations of said amino acids (homocystinuria), are summarized under the term homocysteinemia (WO 92/18002).

The anti-ischemic effectiveness of the organic nitrates and the other substance classes mentioned above is explained by hemodynamic effects, in particular a heart-relieving effect, which leads to a saving in heart oxygen consumption or corrects the mismatch between the O ₂ supply and demand at the Chamber of Industry and Commerce . The cause is a preferred expansion of the venous capacity vessels (venous pooling) or reduction in preload and a direct coronary dilatation effect, particularly in the area of coronary stenoses. This may have a favorable influence on post-stenotic perfusion (positive steal effect), since the organic nitrates in atherosclerotic vascular areas are obviously more potent than in healthy vascular sections (Kojda et al., Endothelium 1 (Suppl.): Abstr. 299, p p76 (1993)), particularly in the area of coronary stenoses. This purely hemodynamic effect is mediated by radical nitrogen monoxide, NO-, which is released uniformly from all nitrovasodilators despite the very different chemical structure of the compounds. The bioactivation pathways that ultimately lead to the provision of NO on-site, i.e. in the endothelial cell and smooth muscle cell of the vessel, are very different (Noack and Feelisch, Molecular mechanism of nitrovascular bioactivation; in "Endothelial Mechanisms of Vasomotor Control" (ed. Drexler et al.), Pp. 37-50, Steinkopff Verlag, Darmstadt, FRG (1991)). This could be clarified beyond doubt by direct NO measurement using different techniques in recent years (method Noack et al., Neuroprotocols 1: 133-139 (1992)). NO has a vasodilatory effect by activating the soluble guanylate cyclase. This stimulates the formation of cGMP from GTP. cGMP in turn leads to various

Phosphorylation reactions (eg on protein kinases) which promote intracellular Ca storage (Karczewski et al., Z. Kardiol. 79 (Suppl. 1): 212 (1990)). The relaxation of the intracellular free Ca ²⁺ level then occurs. It has been known since 1987 that the endothelium derived relaxing factor (EDRF) is identical to NO or a substance containing NO (Palmer et al., Nature, 327: 524-526 (1987); Ignarro et al., Proc. Natl. Acad. Sci. 84: 9265-9269 (1987)) and has an important meaning for local blood circulation.

The endothelial cells form a complete monolayer in the area of the inner wall of the blood vessels. This results in a total surface area of approximately 800 m ² for the adult human being with a weight that corresponds to that of the human liver with 1.5 to 2 kg. From today's perspective, the functions performed by endothelial cells relate to two things: a mechanical and a functional one. On the one hand, they exercise a kind of barrier function with the aim of preventing blood components such as low-density lipoproteins (LDL) from penetrating into the lumen-like vessel wall (intima). On the other hand, they have an endocrine function. Different stimuli lead to an increased synthesis of bioactive substances such as EDRF / NO and prostaglandin-I ₂ (PGI ₂ ), with which the function of the flowing cells (Pohl and Busse, Eur. Heart J .: 11 (Suppl. B) 35-42 (1990)), regional hemodynamics (Furchgott, Circ. Res. 53: 557-573 (1983)) and the structural structure of the vessel wall (Di Corleto, Exp. Cell Res. 153: 167- 172 (1984) ) are fundamentally influenced. At the same time, this explains casually that if the endothelium is damaged, for whatever reason (endothelial damage caused by Hypercholesterolemia (TJ Verbeuren et al., Circ. Res. 58: 552-564 (1986), endothelial damage in the post-infarct phase (MR Sigreid et al., Circ. 86 (Suppl. 1): 21 (1992)), to a pathological This affects endothelial function, which can have different consequences, including regional vasoconstriction or vasospasm and remodeling or growth processes in the vascular wall, which are regarded as initial processes of atherogenesis.

Endothelial dysfunction is generally characterized by a restriction or loss of endothelium-mediated physiological vasodilation. At the same time, a reduction or abolition of the NO-mediated vessel relaxation, the NO-mediated vascular protection and the growth processes suppressed by NO in the intima and media can be observed. Endothelial dysfunction is also characterized by proliferative processes in the vascular wall due to increased mitogenesis, increased endothelial adhesion and migration of leukocytes and macrophages, and increased oxidation of low-density lipoproteins (LDL), which are endothelial-damaging. It is regularly observed in pathophysiological conditions in the context of atherosclerosis, hypertension, hypercholesterolemia, diabetes mellitus and heart failure (Creager et al., J. Clin. Invest. 86: 228-234 (1990); Linder et al., Circulation 81 : 1762-1769 (1990); Zeiher et al., Circulation 83: 391-401 (1991)). Hypoxia and low shear forces are also triggering moments for endothelial dysfunction. Among other things, that vasoactive substances such as acetylcholine or serotonin, which normally cause vasorelaxation, cause vasoconstriction due to their direct vasoconstrictive effects on the smooth vascular muscles, which negatively influence the clinical picture (Golino et al., N. Engl. J. Med. 324: 641-648 (1991)). The physiological vasomotor regulation is therefore not only disturbed in the case of endothelial dysfunction, but also the opposite. These changes are even more pronounced with atherosclerotic remodeling of the inner wall of the vessel (Ludmer et al., N. Engl. J. Med. 315: 1046-1051 (1986)).

The endothelium not only contributes with its autocrine and paracrine activity

Keeping the wall of the blood vessel healthy, but also influences the action of exogenous NO-Liberators such as PETN or GTN in that it itself forms EDRF / NO. If, for example, the endothelium is removed from the arterial wall mechanically (in the case of invasive catheter diagnosis or extracorporeally on isolated vascular segments) or if the endothelial NO formation is suppressed by specific inhibitors, the vasodilatory effect of nitrovasodilators such as GTN or PETN is enhanced (Busse et al. , Cardiovasc. Pharmacol. 14 (Suppl. 11): S81-S85 (1989); Kojda et al., J. Vase. Res. 29: 151 (1992A)). Pharmacological inhibition of endothelial NO synthesis leads to the same effect on coronary veins (Kojda et al., Naunyn-Schmiedeberg ^, Arch. Pharmacol. 346: R35 (1992B)). It is known that the action of calcium antagonists, especially those of the 1,4-dihydropyridine type (DHP's), is weakened after removal of the endothelium (Kojda et al., Bas. Res. Cardiol. 86: 254-256 (1991)). Further studies showed that these substances are likely to stimulate endothelial NO formation and release (Günther et al., Basic Res. Cardiol. 87: 452-460 (1992)). Kinins such as bradykinin also develop their biological activity via the increased endothelial formation and release of EDRF / NO (VA Briner et al., Am. J. Physiol. 264: F322-F327 (1993); Keim et al., Biochem. Biophys. Res. Commun. 154: 236-244 (1988)).

Statement of the invention

Endothelial dysfunctions are now seen as triggers of common and pathophysiologically significant cardiovascular diseases such as atherosclerosis. Prevention, treatment and elimination of these dysfunctions and the diseases associated with or caused by them are therefore important therapeutic necessities.

It has now been found that the use of nitrogen monoxide-releasing and / or transferring compounds, stimulators of endogenous nitric oxide formation and stimulators of guanylate cyclase, in particular stimulators of soluble guanylate cyclase, for the prevention, treatment and elimination of endothelial dysfunctions and associated with these dysfunctions and / or diseases caused by them is suitable. These endothelial dysfunctions and diseases are primarily endothelial damage caused by hypercholesterolemia, endothelial damage caused by hypoxia, endothelial damage caused by mechanical and chemical noxae, especially during and after drug and mechanical reopening of stenosed vessels e.g. after percutaneous transluminal angiography (PTA) and percutaneous transluminal coronary angiography (PTCA), endothelial damage in the postinfarct phase (endothelial dysfunction at reperfusion) endothelial-mediated reocclusion after bypass surgery, blood supply disturbances in peripheral arteries, as well as cardiovascular diseases such as atherosclerosis, hypertension, including pulmonary and portal hypertension, hypertensive heart disease, diabetic micro- and macroangiopathy, coronary heart disease, heart failure or other diseases that are causally based on endothelial dysfunction.

Nitric oxide releasing and / or transmitting compounds, stimulators of endogenous nitric oxide formation as well as stimulators of guanylate cyclase in the sense This invention includes, inter alia, compounds which act directly or indirectly on the guanylate cyclase, compounds which can be released as indirect stimulators of the guanylate cyclase being those which can release guanylate cyclase stimulants or otherwise increase their enzyme-effective concentration and / or have an antagonistic effect against inhibitors of guanylate cyclase or their enzyme-effective concentration humiliate. As indirect stimulators of the guanylate class, compounds are used which are suitable for increasing endogenous NO formation or release, such as calcium channel blockers, in particular those of the 1,4-dihydropyridine type, for example nifedipine, felodipine, nimodipine, amlodipine and others. The use of compounds which are capable of increasing the endothelial kinine content is also suitable. Above all, these are stimulators of the kinin receptors such as kinins or substances having an analogous effect, stimulators of endothelial kinin formation and inhibitors of kinin degradation, in particular inhibitors of the angiotensin converting enzyme (ACE inhibitors) such as captopril, enalapril, moexipril, ramipril and related active substances. The use of compounds that are generally effective

Developing release and / or transmission of endogenous or exogenous nitrogen monoxide is the particularly preferred embodiment of the present invention. Substance classes and compounds that are particularly suitable here are organic nitrates, in particular glycerol trinitrate, pentaerythrityl tetranitrate, isosorbide-5-mononitrate, isosorbide dinitrate, mannitol hexanitrate, inositol hexanitrate, propatyl nitrate, trol nitrate,

Nicorandil, newer nitrates such as SPM 3672 and their physiologically compatible derivatives, organic nitrites such as isoamyl nitrite, thionitrites, thionitrates, S-nitrosothiols such as S-nitroso-N-acetyl-D, L-penicillamine, nitrosoproteins, nitrogen monoxide-liberating furoxane derivatives, especially nitrogen monoxide derivatives, nitrogen monoxide derivatives, nitrogen monoxide derivatives , Mesocarb and their analogues, nitrosyl complex compounds, especially iron

Nitrosyl compounds such as nitroprusside sodium and nitrogen monoxide itself. Since the nitrogen monoxide release and / or transfer often takes place in vivo via pharmacologically active metabolites, these are in principle also suitable for use in the sense of the present invention. At the same time, the use of physiologically compatible derivatives of all the compounds mentioned above is possible. Above all, common addition compounds, salts or enzymatically or hydrolytically cleavable compounds such as esters, amides and the like are possible variations. The selection of the respective active ingredient is based on general pharmacological principles and the therapeutic requirements which are known to the person skilled in the art. In addition to the desired pharmacological effect, the state of health, the stage of the disease, the physical condition, the known effects and side effects, contraindications, the frequency of treatment, the duration of use, drug interactions and parallel drug use must also be taken into account. The dosage is in each case in therapeutic doses, which are based on those in which the respective active ingredients are already used for known indications. The daily total dose can be up to 500 mg depending on the active ingredient. In general, daily doses up to 350 mg will be sufficient. Dosage and dosing interval should be chosen so that therapeutic plasma levels that are as constant as possible are built up. The compounds used according to the invention can themselves or as part of a galenical preparation, as a single active ingredient or in combination with one another or with known cardiovascular therapeutics, for example ACE inhibitors, antiatherosclerotics, antihypertensives, beta-blockers, cholesterol-lowering agents, diuretics, calcium-antagonists, coronary dilators, lipid-lowering agents, peripheral Vasodilators, platelet aggregation inhibitors or other substances, also used as cardiovascular therapeutics, combined, can be used. The preparation of galenical preparations is carried out according to the working methods and rules generally familiar to the pharmaceutical expert, the choice of the technologies to be used and the galenic auxiliaries used being based primarily on the active substance to be processed. Questions of its chemical-physical properties, the chosen form of application, the desired duration of action, the place of action and the avoidance of drug-auxiliary incompatibilities are of particular importance. It is therefore up to the person skilled in the art to select pharmaceutical form, auxiliary substances and production technology in a trivial manner on the basis of known substance and process parameters. The pharmaceutical form in question should be designed so that it contains the respective active ingredient in order to achieve constant therapeutic plasma levels, which makes it possible to distribute the daily dose to 1 to 2 in the case of release-controlled systems and to up to 10 individual doses in the case of other dosage forms. Continuous application using long-term infusion is also suitable.

According to the invention, the named compounds can be administered primarily orally, intravenously, parenterally, sublingually or transdermally. The respective pharmaceutical preparation is preferably provided in liquid or solid form. Solutions are suitable for this, in particular for the preparation of drops, injections or aerosol sprays, furthermore suspensions, emulsions, syrups, tablets, film-coated tablets, dragées, capsules, pellets, powders, pastilles, implants, suppositories, creams, gels, ointments, plasters or other transdermal Systems. The pharmaceutical preparations contain customary galenically usable, organic or inorganic carriers and auxiliaries which are themselves chemically indifferent to the respective active ingredients. Suitable for this are, but are not limited to, water, salt solutions, alcohols, vegetable oils, polyethylene glycols, gelatin, lactose, 95/26725

Amylose, magnesium stearate, talc, highly disperse silicon dioxide, paraffin, fatty acid mono- and diglycerides, cellulose derivatives, polyvinylpyrrolidone and the like. The preparation can be sterilized and, if necessary, with auxiliary substances such as fillers, binders, lubricants, mold release agents, lubricants, disintegrants, humectants, adsorbents or counter-disintegrants, preservatives, stabilizers, emulsifiers, solubilizers, salts for influencing the osmotic pressure , Buffer solutions, coloring, fragrance, aroma or sweeteners may be added. The pharmaceutical expert will make a suitable selection based on known substance parameters to avoid drug-excipient incompatibilities.

With the presented invention, a new therapeutic possibility is opened, pathological situations such as hypoxia, high serum cholesterol, increased blood pressure, diabetes, post-stenotic reperfusion e.g. for myocardial infarction and mechan. and former noxae, among others, that promote endothelial dysfunction, counteract or completely prevent the development of endothelial dysfunction. The therapeutic use of suitable compounds, irrespective of the galenical preparation, therefore allows, for the first time, the prevention and topical therapy of cardiac and vascular diseases of this etiology, such as atherosclerosis and the resulting complications. These include coronary artery disease, vascular stenosis and circulatory disorders of the peripheral arteries, micro- and macroangiopathies in the context of diabetes mellitus etc. Surprisingly, the compounds identified above show an independent endothelial protective effect, which is independent of the previously known, especially the purely hemodynamic and anti-ischemic properties, e.g. the organic nitrates or their effectiveness in hypercysteinemia. Their use can therefore bring these pathological processes to a standstill or even reverse them as long as they are not yet irreversible. It is therefore an unexpected, novel effect component that has not been described so far and was not expected in this form.

The following examples are intended to explain the invention in more detail with regard to its nature and its implementation, but without restricting its scope. Practical examples

example 1

Experiments on a pharmacological in vivo model (New Zealand rabbit)

In animal experiments, cholesterol feeding is suitable for producing endothelial dysfunction within weeks to months, which allows the effects of pharmaceuticals to be investigated and quantified (Jayakody et al., Can. J. Physiol. Pharmacol. 63: 1206-1209 (1985 ); Verbeuren et al., Circ. Res. 58: 552-564 (1986); Freiman et al, Circ. Res. 58: 783-789 (1986)).

Groups of 9 female New Zealand rabbits each were fed a standard diet or a cholesterol-enriched (0.75%) diet (40 g / kg / day) over a period of 15 weeks. Cholesterol feeding led to an increase in the plasma level from 69.8 + 10.4 to 907.1 + 85.5 mg / dl and caused atherosclerotic lesions in the area of the aorta, which were stained with Sudan IV using computer-aided laser scanning technology were quantified. The aortic changes included an area of 73.3 + 1.9% on the aortic arch, an area of 46.3 ± 2.5% on the thoracic aorta and 49.6 + 3.5% in the area of the abdominal aorta (Fig. 2, Control).

Example 2

The atherosclerotic vessels showed no change in the willingness to contract to phenylephrine, but the function of the endothelium-mediated vasorelaxation after administration of 1 μM acetylcholine was changed in comparison to the controls (standard diet) in a manner that can best be described as endothelial dysfunction. The segments of the thoracic aorta of the cholesterol-fed animals (+) show a significantly weaker sensitivity to acetylcholine than the aortic segments of the control animals (Fig. 1). The degree of measured endothelial dysfunction correlated directly with the respective severity of the atherosclerotic lesions (r = 0.67, p <0.0001) (Fig. 3). These data show that endothelial dysfunction developed after feeding with cholesterol.

Example 3

Two further groups of nine white New Zealand rabbits each received pentaerythrityl tetranitrate (PETN) (6 mg / kg / day), whereby the pharmon was incorporated into the press feed, and the animals treated with PETN significantly reduced the atherosclerotic lesions. The extent of these lesions was determined in accordance with Example 1. There was a significantly reduced proportion of atherosclerotic damage in all parts of the aorta (aortic arch: 58.6 ± 2.1%, thoracic aorta 34.7 ± 2.0% and abdominal aorta 39.3 ± 3.1%) (Fig. 2 ).

Example 4

Likewise, after PETN feeding, there was no significant difference in the maximum (1 μM acetylcholine) endothelium-mediated relaxation compared to the animals not fed with cholesterol (endothelial dysfunction), so that these results show a clear protective effect of the nitrovasodilator PETN, since this prevented endothelial dysfunction in the sense of the invention presented here (Figs. 3 and 4; Table 1).

The comparison between Figures 3 and 4 shows that PETN can reduce the extent of atherosclerotic lesions and improve endothelial function. The poorer correlation coefficient in the PETN group indicates that PETN also leads to a dissociation of the close relationship between atherosclerotic lesions and endothelial function.

Table 1 shows the influence of PETN (6 mg / kg / day) on the development of endothelial dysfunction in the thoracic aorta of New Zealand white rabbits, which was induced by feeding on a cholesterol diet (0.75%; 15 weeks). The potency of the endothelium-dependent vasodilator acetylcholine is expressed as the concentration (in - logM; pD2 value) that, when administered cumulatively, antagonized half the action of the vasoconstrictor phenylephrine (the higher this value, the stronger the effect of

Acetylcholine). The maximum dictatorial effect is expressed as the percentage of the effect of the vasoconstrictor phenylephrine that was antagonized at the maximum effective concentration of acetylcholine (1 μM). The endothelial dysfunction induced by cholesterol feeding alone (control) can be recognized by the significantly reduced potency and maximum potency of acetylcholine (*, p <0.05). At the same time

When feeding PETN, the differences can no longer be demonstrated. In addition, PETN significantly (#, p <0.05) improves the potency of acetylcholine and thus the endothelial function after cholesterol feeding, while there is a significant deterioration in endothelial function after standard feeding. Overall, this shows the protective effect of PETN on endothelial function in experimentally induced

Atherosclerosis. The absorption and flooding of PETN in the plasma can also be demonstrated 24 h after the last feeding of the animals using the measured concentrations of the metabolite pentaerythrityl mononitrate (PEMN) in the plasma (Fig. 5). Table 1 :

Control PETN

Standard cholesterol Standard cholesterol

Potency of acetylcholine 6.91 ± 0.02 6.12 ± 0.05 * 6.62 ± 0.06 # 6.47 ± 0.13 # [pD ₂ values]

Max. dictatorial effect 84.8 ± 1.2 60.7 ± 8.5 * 74.7 ± 4.9 * 65.0 ± 4.7 of acetylcholine [%]

Example 5

A typical tablet has the following composition:

Pentaerythrityl tetranitrate ISIS PHARMA 20 mg

Lactose DAB 10 137 mg

Potato starch DAB 10 80 mg

Gelatin DAB 10 3 mg

Talc DAB 10 22 mg

Magnesium stearate DAB 10 5 mg

Silicon dioxide, highly dispersed DAB 10 6 mg

273 mg Example 6

A tablet containing 20 mg pentaerythrityl trinitrate (PETriN) has the following composition:

PETriN 20 mg

Lactose. DAB 10 137 mg

Potato starch DAB 10 80 mg

Gelatin DAB 10 3 mg talc DAB 10 22 mg

Magnesium stearate DAB 10 5 mg

Silicon dioxide, highly dispersed DAB 10 6 mg

273 mg

Example 7

A tablet containing 20 mg pentaerythrityl dinitrate (PEDN) has the following composition:

PEDN 20 mg

Lactose DAB 10 137 mg

Potato starch DAB 10 80 mg G Geellaattiinnee D DAABB 1100 3 mg

Talc DAB 10 22 mg

Magnesium stearate DAB 10 5 mg

Silicon dioxide, highly dispersed DAB 10 6 mg

273 mg Example 8

A tablet containing 20 mg erythrityl tetranitrate (ETN) has the following composition:

ETN 20 mg

Lactose DAB 10 137 mg

Potato starch DAB 10 80 mg

Gelatin DAB 10 3 mg

Talc DAB 10 22 mg

Magnesium stearate DAB 10 5 mg

Silicon dioxide, highly dispersed DAB 10 6 mg

273 mg

Example 9

A tablet containing 20 mg isosorbide mononitrate (ISMN) has the following composition:

ISMN 20 mg

Lactose DAB 10 137 mg

Potato starch DAB 10 80 mg

Gelatin DAB 10 3 mg

Talc DAB 10 22 mg

Magnesium stearate DAB 10 5 mg

Silicon dioxide, highly dispersed DAB 10 6 mg

273 mg Example 10

A tablet containing 20 mg isosorbide dinitrate (ISDN) has the following composition:

ISDN 20 mg

Lactose DAB 10 137 mg

Potato starch DAB 10 80 mg

Gelatin DAB 10 3 mg

Talc DAB 10 22 mg

Magnesium stearate DAB 10 5 mg

Silicon dioxide, highly dispersed DAB 10 6 mg

273 mg

Example 11

A tablet containing 40 mg pentaerythrityl tetranitrate (PETN) and 40 mg propranolohydrochloride has the composition:

PETN 40 mg

Propranolol hydrochloride 40 mg

Lactose 224 mg

Potato starch 80 mg

Gelatin 3 mg

Talc 22 mg

Magnesium stearate 5 mg

Silicon dioxide, finely divided 6 mg

420 mg

5 pages of illustrations / oj

Acetylcholine [~ lθgM]

Illustration 1:

Development of endothelial dysfunction after feeding white New Zealand rabbits on a cholesterol diet (0.75%; 15 weeks). The vasorelaxative effect of the endothelium-dependent vasodilator acetylcholine is shown, and thus the functionality of the endothelium, expressed as a percentage of the pre-contraction remaining at any given concentration, which was triggered by the vasoconstrictor phenylephrine.

Λorlenbogβn T orakalaσrla Abdσmlnaloarta

Figure 2:

Extent of atherosclerotic lesions on the luminal surface of various sections of the aorta after feeding a cholesterol diet (0.75%; 15 weeks) (without control) and simultaneous administration of PETN (6 mg / kg / day). The atherosclerotic lesions were stained with Sudan IV and the percentage of the stained area (based on the total area) was determined using a computer-assisted laser scanning method. PETN causes a significant reduction in the formation of atherosclerotic lesions (p <0.05).

ΛTIlKIlOSKLERσπSCIIE LÄCH E ι% l

Figure 3:

Relationship between the extent of atherosclerotic lesions on the luminal surface of segments of the thoracic aorta after feeding a cholesterol diet (0.75%; 15 weeks) and the maximum relaxation previously determined in the same segment induced by 1 μM acetylcholine (endothelial function). The larger the area of the lesions, the worse the relaxation or endothelial function. ^

ATHEROSCLEROTIC FLAT | % l

Figure 4:

The same representation as in Figure 3 after simultaneous feeding of cholesterol and PETN.

Cholcslcrol Slnndar

Figure 5:

Plasma level of pentaerythrityl mononitrate in the plasma of white New Zealand rabbits after 24 h of food withdrawal before the blood was taken, which preceded the acute test. The standard feed contained pentaerythrityl tetranitrate (150 mg / kg) in both cases and an additional 0.75% cholesterol for the cholesterol group. The concentration of pentaerythrityl mononitrate was determined quantitatively after working up the plasma samples by means of gas chromatography / mass spectroscopy.

Claims

claims

1. Use of nitrogen monoxide releasing and / or transmitting compounds, of stimulators of endogenous nitric oxide formation or of stimulators

Guanylate cyclase for the prevention, treatment and elimination of endothelial dysfunctions and disorders associated with and / or caused by these dysfunctions.

2. Use of nitrogen monoxide releasing and / or transferring compounds, stimulators of endogenous nitric oxide formation or stimulators of

Guanylate cyclase according to claim 1, characterized in that endothelial dysfunctions as well as diseases associated therewith and / or caused by them

Endothelial damage caused by hypercholesterolemia, endothelial damage caused by hypoxia, endothelial damage caused by mechanical and chemical noxae, especially during and after drug and mechanical reopening of stenosed vessels e.g. after percutaneous transluminal angiography (PTA) and percutaneous transluminal coronary angiography (PTCA), endothelial damage in the post-infarction phase (endothelial

Dysfunction in reperfusion), endothelial mediated reocclusion after bypass surgery, circulatory disorders in peripheral arteries as well as cardiovascular diseases such as atherosclerosis, hypertension, including pulmonary and portal hypertension, hypertensive heart disease, diabetic micro and macroangiopathy, coronary artery disease, heart failure.

3. Use of nitrogen monoxide releasing and / or transferring compounds, stimulators of endogenous nitric oxide formation or stimulators of

Guanylate cyclase according to Claims 1 and 2, characterized in that the soluble guanylate cyclase is stimulated.

4. Use of nitrogen monoxide releasing and / or transferring compounds, stimulators of endogenous nitrogen monoxide formation or stimulators of guanylate cyclase according to claim 1 to 3, characterized in that the stimulation of soluble guanylate cyclase takes place directly and / or indirectly.

5. Use of nitrogen monoxide releasing and / or transmitting compounds, stimulators of endogenous nitrogen monoxide formation or stimulators of guanylate cyclase according to claim 1 to 4, characterized in that indirect stimulators of guanylate class are compounds which are capable of releasing stimulants of guanylate cyclase or their enzyme-effective concentration otherwise increase and / or have an antagonistic effect on guanylate cyclase inhibitors or otherwise lower their enzyme-effective concentration.

6. Use of nitrogen monoxide releasing and / or transferring compounds, stimulators of endogenous nitric oxide formation or stimulators of guanylate cyclase according to claim 1 to 5, characterized in that as indirect stimulators of guanylate cyclase

Calcium antagonists, especially those of the 1,4-dihydropyridine type, can be used.

7. Use of nitrogen monoxide releasing and / or transferring compounds, stimulators of endogenous nitrogen monoxide formation or stimulators of guanylate cyclase according to claim 1 to 5, characterized in that nitrogen monoxide releasing and / or transferring compounds are used.

8. Use of nitrogen monoxide releasing and / or transferring compounds, stimulators of endogenous nitrogen monoxide formation or stimulators of guanylate cyclase according to one or more of claims 1 to 5 and 7, characterized in that nitrogen monoxide releasing and / or transferring compounds are organic nitrates, in particular

Glycerol trinitrate (GTN), pentaerythrityl tetranitrate (PETN), isosorbide 5-mononitrate (ISMN), isosorbide dinitrate (ISDN), mannitol hexanitrate, inositol hexanitrate, propatyl nitrate, trolnitrate, nicorandil or SPM 3672 as well as their derivatives, organic nitrite, as well as their physiologically tolerable Nitrite Nitrile, as well as their physiologically tolerable Nitrile Nitrate,

Thionitrite, thionitrate,

S-nitrosothiols such as S-nitroso-N-acetyl-D, L-penicilkmin (SNAP), nitrosoproteins, nitrogen monoxide liberating furoxane derivatives, nitrogen monoxide liberating sydnonimine derivatives, in particular

Mesocarb, molsidomine or their pharmacologically active metabolites, nitrosyl complex compounds such as

Iron nitrosyl compounds, in particular nitroprusside sodium, and nitrogen monoxide (NO) are themselves.

9. Use of nitrogen monoxide releasing and / or transmitting compounds, stimulators of endogenous nitrogen monoxide formation or stimulators of guanylate cyclase according to claim 1 to 5, characterized in that compounds are used as indirect stimulators of guanylate class, which increase the endothelial kinine content.

10. Use of nitrogen monoxide releasing and / or transmitting

Compounds, of stimulators of endogenous nitric oxide formation or of stimulators of guanylate cyclase according to claim 9, characterized in that compounds which increase the endothelial kinine content,

Stimulators of kinin receptors such as kinins or substances having an analogous effect, stimulators of endothelial kinin formation or inhibitors of kinin degradation such as

Are inhibitors of the angiotensin converting enzyme.

11. Use of nitrogen monoxide-releasing and / or transferring compounds, stimulators of endogenous nitric oxide formation or stimulators of guanylate cyclase for the production of pharmaceutical preparations for prevention,

Treatment and elimination of endothelial dysfunctions and diseases associated with and / or caused by these dysfunctions.

12. Use of nitrogen monoxide releasing and / or transmitting compounds, stimulators of endogenous nitrogen monoxide formation or stimulators of guanylate cyclase according to claim 11, characterized in that these compounds with other active substances used for the treatment of cardiovascular diseases, in particular with those from the indication groups the ACE inhibitors, antiatherosclerotics, antihypertensives, beta blockers, cholesterol lowering agents, diuretics, calcium channel blockers, coronary dictators, lipid lowering agents, peripheral vasodilators or platelet aggregation inhibitors can be combined.