WO2000067731A2

WO2000067731A2 - Medicinal substance utilization for preventing nitrate tolerance

Info

Publication number: WO2000067731A2
Application number: PCT/DE2000/001413
Authority: WO
Inventors: Michael Stoeter; Gerd König; Eberhard Bassenge
Original assignee: Alpharma-Isis Gmbh & Co.Kg
Priority date: 1999-05-05
Filing date: 2000-05-05
Publication date: 2000-11-16
Also published as: AU5802000A; BG105955A; PL352024A1; EP1173214A2; DE19920775A1; WO2000067731A3

Abstract

The invention describes the utilization of medicinal substances for preventing nitrate and/or nitrate cross-tolerance and pathological phenomena accompanying the latter.

Description

USE OF MEDICINAL PRODUCTS TO AVOID NITRATOLERANCE

FIELD OF APPLICATION OF THE INVENTION The present invention relates to the use of medicaments for avoiding nitrate and / or nitrate cross tolerance as well as pathological phenomena associated therewith.

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. 2370437), isosorbide-5-mononitrate (ISMN) (DE-OS-2221080, DE-OS-2751934, DE-OS-3028873, DE-PS-2903927, DE-OS-3102947, DE-OS-3124410, EP- A1- 045076, EP-A1-057847, EP-A1-059664, EP-A1-064194, EP-A1-067964, EP-A1-143507, US-PS-3886186, US-PS-4065488, US-PS- 4417065, US-PS-4431829), Isosorbide Dinitrate (ISDN) (L. Goldberg, Acta Physiolog. Scand. 15, 173 (1948)), Propatyl Nitrate (Medard, Mem. Poudres 35: 113 (1953)), Trolnitrate (FR- PS-984523) or Nicorandil (US-PS-4200640) and similar compounds are vasodilators, some of which have been focussing on the indication of angina pectoris or ischemic heart disease for decades

(IHK) find the widest therapeutic use (Nitrangin®, Pentalong®, Monolong®, etc.). The pharmaceutical processing of organic nitrates into pharmaceuticals

Preparations for the treatment of angina pectoris or ischemic heart disease are 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, in particular the explosive properties known to be attached to the organic nitrates, which require attention to special safety precautions and special processing technologies, the chosen form of application, the desired duration of action and the avoidance of drug-auxiliary incompatibilities are of particular importance. For medicinal products with the indication angina pectoris or ischemic heart disease, peroral, parenteral, sublingual or transdermal application in the form of tablets, dragees, capsules, solutions, sprays or plasters is described above (DD-A5-293492, DE-AS-2623800 , DE-OS-3325652, DE-OS-3328094, DE-PS-4007705, DE-OS-4038203, JP application 59/10513 (1982)). In addition to the long-known applications, it has a nitrosating effect

Substances are their use for the treatment and prevention of diseases 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-A1 -92/18002). The use of certain organic nitric acid esters as endothelial protective agents (DE-A1 -4410997) and as agents for the treatment of erectile dysfunction (WO-A1 -96/32118) have recently been described. The known organic nitrates (nitric acid esters) have a number of therapeutic disadvantages. For example, the so-called nitrate tolerance can be observed, ie the decrease in the nitrate effect at high doses or when long-acting nitrates are applied. Side effects such as headache, dizziness, nausea, weakness, reddening of the skin and the risk of a greater drop in blood pressure with reflex tachycardia are also documented (Mutschier, drug effects, Wissenschaftliche Verlagsgesellschaft mbH, Stuttgart, 1991). The metabolism of GTN and other organic nitrates has been extensively investigated (Taylor et al., Prague. Drug Metab., 10 (1987), 207). It was found, among other things, that organic nitrates have a pronounced oxidative effect on compounds carrying thiol groups (Boschan et al., Chem. Rev. 55, 485 (1955); Taylor et al., Progress in drug metabolism, Vol. 10, 207 ( 1987); Feelisch et al., Methods in Nitric Oxide Research, John Wiley & Sons, Chichester, 1996)). Furthermore, it is generally accepted and scientifically well documented that organic nitrates inevitably trigger counter-regulatory processes via the NO mechanism, for example the formation of angiotensin II in the vascular wall, which form large amounts of superoxide radicals when the endothelial enzyme NADH synthase is activated. which themselves have a strong oxidative effect and immediately react with the NO released from organic nitrates. According to popular belief, the state of nitrate tolerance is determined by a massive accumulation of superoxide radicals, which lead to the formation of peroxynitrite and thus inactivation of NO according to the reaction scheme ON ^' + 0 ₂ ^' → ONOO. This view is reinforced by the fact that nitrate tolerance can be counteracted to a limited extent by the administration of antioxidants such as ascorbic acid (vitamin C) or vitamin E (Münzel et al., The physiology and pathophysiology of the nitric oxide / superoxide System, Vascular Endothelium (GVR Born, CJ Schwartz edt., P. 205-220, Schattauer Verlag, New York (1997); Harrison, J. Clin. Invest. 100 (9) (1997), 2153; Miyamoto et al., Proc. Soc. Exp. Biol. Med. 211 (1996), 366. Despite these findings, it has not hitherto been possible to use organic therapies Successful combination of nitrates with antioxidants, even though their effect is based on the actual physical and therefore pharmacologically harmless substance NO. Münzel et al. (J. Clin. Invest. 98 (1996), 1465) prevented GTN-induced nitrate tolerance by using hydralazine, an inhibitor of membrane-bound NADH oxidase. However, the actual mechanism underlying nitrate tolerance is still unclear (Nitroglycerin VIII, Mehmel (ed.), Verlag W. de Gruyter (1996)). Miyamoto et al. (Proc. Soc. Exp. Biol. Med. 211 (1996), 366) found that the xanthine oxidase inhibitors AHPP, allopurinol and alloxanthine with normal NO reactivity of vessels to a species-dependent minimal amplification (AHPP) or almost no amplification ( Allopurinol) of the NO-mediated vasorelaxation, whereby there was even increased superoxide radical formation with allopurinol. In line with current opinion, they concluded that xanthine oxidase was a regulatory protein for the NO effect by means of superoxide formation and the associated NO consumption. From a stoichiometric point of view (oversupply of NO with nitrate administration and in the state of nitrate tolerance), however, this explanation is implausible, because complete inactivation of the NO by the superoxide formed by the xanthine oxidase, as postulated to explain the nitrate tolerance, cannot take place (Ellis et al., Europ. J. Pharmacol. 356 (1998), 41). In addition, peroxynitrite itself breaks down again into superoxide and NO radicals and thus serves independently as a source of NO (Moro et al., Proc. Natl. Acad. Sei. 91 (1994), 6702; Moro et al., Br. J Pharmacol 116 (1995), 1999). In addition, the xanthine oxidase in turn can release NO from organic nitrates under hypoxic conditions (Millar et al., FEBS Lett. 427 (2) (1998), 225; Millar et al., Biochem. Biophys. Res. Commun. 249 (1998), 767). In addition, NO itself inhibits xanthine oxidase (Fukahori et al., Free Rad. Res. 21 (4) (1994), 203; Cote et al., Am. J. Physiol. 271 (1996), L869).

The systems of xanthine oxidase, also called xanthine oxidoreductase in more recent literature, have been extensively investigated (Fridovich, J. Biol. Chem. 245 (1970), 4053; Battelli et al., FEBS Lett. 113 (1980), 47; Parks et al ., Acta Physiol. Scand. Suppl. 1986; 548: 87-99; Parks et al., Am. J. Physiol. 254 (Gastrointes. Liver Physiol. 17) (1988), G768; Kooij, Histochem. J. 26 (1994), 889; Hille, Chem. Rev. (1996), 2757) and various enzyme inhibitors (Kooij, Histochem. J. 26 (1994), 889). The galenical processing of xanthine oxidase inhibitors such as allopurinol or oxipurinol (DE-A1- 3839826) is also known. DISCLOSURE OF THE INVENTION The object of the invention is to counteract nitrate and / or nitrate cross tolerance as well as pathological phenomena associated with this by using suitable medicinal substances.

The object of the invention is achieved by the use of antagonists of purine derivative metabolizing enzymes to prevent or reduce nitrate and / or nitrate cross tolerance and of pathological phenomena associated with them. An antagonist for the purposes of the present invention is a substrate which competes with purine derivatives or an inhibitor of the enzyme. The terms competitive or non-competitive inhibitor are also suitable for describing the term antagonist. A particular and preferred embodiment of the invention is the use of antagonists of xanthine oxidase to prevent or reduce nitrate and / or nitrate cross-tolerance and of pathological phenomena associated therewith, wherein the antagonist can have the meaning described above. Allopurinol or oxipurinol are e.g. B. two typical already therapeutically used inhibitors of xanthine oxidase. Other suitable compounds are in particular in Janero et al. (Life Sci. 44 (1989), 1579) and Kooij (Histochem. J. 26 (1994), 889), reference being made to these references.

Surprisingly, it turned out that the nitrate tolerance by the procedure according to the invention, for. B. by inhibition of xanthine oxidase by means of allopurinol or its active metabolite oxipurinol (alloxanthin) can be antagonized and the reactivity of the considered vessels, which no longer adequately react to NO in the state of nitrate tolerance, could be restored to nitrogen monoxide. It was unexpected for the person skilled in the art that the state of the extinct (not present) NO effect can be eliminated and the effect of exogenously supplied NO can be restored with continuous administration of nitrate. For the first time, it was possible to recognize from the use according to the invention that it is not the superoxide radical formation which is etiologically responsible for the development of the nitrate tolerance, but probably an increased purine catabolism, caused by activation of the xanthine oxidase. The resulting lack of adenosine triphosphate (ATP) and guanosine triphosphate (GTP) prevents NO from developing its physiological effect inside the cell: Both for the guanylate cyclase and for the Ca ²⁺ -ATPase there is a lack of substrate, which leads directly to a reduced availability of the Second Messengers cyclical GMP (cGMP), as well as reduced transport of cytosolic calcium into the cellular storage organelles. Another unexpected result was that activation of the xanthine oxidase occurred in the nitrate tolerance state. The results according to the invention here, namely that the pharmacological inhibition of xanthine oxidase with allopurinol and derivatives leads to an improvement in the nitrate effect, thus run counter to the previous view. For the first time, it was possible to solve an application problem that has existed for over a hundred years since therapeutic use of organic nitrates, which opens up new perspectives for this class of compounds, since they can now be used in a variety of different ways in terms of dosage, frequency of use, side effects and interactions, as well as the associated improved effectiveness. For example, they can now be given to the permanent treatment of hypertension in normal or higher doses. In all cases, there is the advantage of not having to resort to more expensive accompanying or individual medications, such as sydnonimine derivatives, calcium channel blockers, ACE inhibitors or the like, to a greater extent.

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.

embodiments

example 1

2083 g of an 8% glycerol trinitrate adsorbate on polyvinyl alcohol (PVA) (12% vinyl acetate content) are exposed to a temperature of 60 ° C. for 30 minutes, then they are mixed with 1583 g PVA (12% vinyl acetate content), 1250 g allopurinol and 84 g magnesium stearate and pressed into tablets with an average mass of 400 mg.

Example 2

Tablets containing 20 mg, 40 mg or prolonged-release tablets containing 40 mg or 60 mg isosorbide mononitrate and at the same time 100 mg allopurinol are blistered, provided with instructions for use and spatially combined in a packaging unit. Example 3

Slow-release tablets containing 20 mg, 40 mg, 60 mg, 80 mg or 120 mg isosorbide dinitrate and 100 mg allopurinol at the same time are blistered, provided with instructions for use and spatially combined in one packaging unit.

Example 4

Tablets containing 20 mg pentaerythrityl tetranitrate and 100 mg allopurinol are blistered, provided with instructions for use and spatially combined in a packaging unit.

Example 5

Tablets containing 50 mg of pentaerythrityl tetranitrate and 300 mg of allopurinol are blistered, provided with instructions for use and spatially combined in a packaging unit.

Example 6

Tablets containing 80 mg of pentaerythrityl tetranitrate and 100 mg of allopurinol are blistered, provided with instructions for use and spatially combined in a packaging unit.

Example 7

Tablets containing a coronary therapeutic and tablets containing a xanthine oxidase inhibitor are blistered separately, provided with instructions for use and combined in a packaging unit in a spatially separate manner.

Example 8

Tablets containing 40 mg of pentaerythrityl tetranitrate and at the same time 40 mg of propranolol hydrochloride are blistered, provided with instructions for use and spatially combined in a packaging unit.

Example 9

Cultures of endothelial cells and smooth vascular muscle cells from pig aorta were incubated natively (control), with 0.1 mM GTN or 0.1 mM GTN and 10 μM, 50 μM, 100 μM or 150 μM oxipurinol for 24 hours at 37 ° C. The subsequent stimulation with 2 μM nitroprusside sodium (SNP) for 3 minutes led to a significantly weakened increase in the intracellular cGMP concentration (48.7% ± 5.9% compared to the control) in the culture pretreated only with GTN, detected by means of a specific radioimmunoassay (Biotrend, Cologne). After simultaneous incubation with 10 μM oxipurinol, the SNP stimulation led to an increase in the cGMP concentration by 56.4% ± 7.5%, under 50 μM oxipurinol by 87.6% ± 9.3% and under 100 μM oxipurinol 93.2% ± 9.1%. In addition, the formation of reactive oxygen species (ROS) was determined by electron spin resonance spectrometry. This was done by measuring the formation of TEMPONE nitroxyl radicals from TEMPONE-H (1-hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine hydrochloride). Compared to the initial value of 25 nM / min / mg protein, stimulation with 0.5 mM GTN led to an 82.6% ± 8.3% increase in ROS formation. Previous incubation with 10 μM oxipurinol for 24 hours resulted in an increase in ROS of 75.3% ± 8.6%, with 50 μM oxipurinol by 35.9% ± 5.3% and with 100 μM oxipurinol by 28.7% ± 4.9%. These experiments demonstrated for the first time that the reduction in intracellular cGMP concentration characterizing a nitrate tolerance is attenuated by inhibition of xanthine oxidase and that the GTN-induced increase in reactive oxygen species can be significantly reduced by xanthine oxidase inhibitors.

Claims

claims

1. Use of antagonists of purine derivative metabolizing enzymes to prevent or reduce nitrate and / or nitrate cross tolerance and pathological phenomena associated with them.

2. Use according to claim 1, characterized in that the antagonist a) is a substrate competing with purine derivatives or b) is an inhibitor of the enzyme.

3. Use of antagonists of xanthine oxidase to prevent or reduce nitrate and / or nitrate cross tolerance and pathological phenomena associated with them.

4. Use according to claim 3, characterized in that the antagonist a) is a substrate competing with purine derivatives or b) is an inhibitor of xanthine oxidase.

5. Use according to claim 4, characterized in that the inhibitor of xanthine oxidase is allopurinol or oxipurinol.

6. Use of antagonists of purine derivative metabolizing enzymes or of antagonists of xanthine oxidase for the production of pharmaceutical agents for preventing or reducing nitrate and / or nitrate cross tolerance and of pathological phenomena associated with them.

7. Pharmaceutical compositions for preventing or reducing nitrate and / or nitrate cross tolerance and pathological phenomena associated therewith, containing a xanthine oxidase inhibitor.

8. Pharmaceutical compositions containing a cardiac / circulatory therapeutic, in particular a coronary therapeutic, on the basis of organic nitrates, preferably pentaerythrityltetra-, pentaerythrityltri-, pentaerythrityldi- or pentaerythritylmononitrate and a xanthine oxidase inhibitor or as a separate preparation for combination use, as a simultaneous preparation. Method for the prevention or reduction of nitrate and / or nitrate cross-toxicity and of pathological phenomena associated therewith, characterized in that pharmaceutical agents according to claim 7 or 8 are used.