HUT74732A - Use of xanthine derivatives for reducing the pathological hyper-reactivity of eosinophilic granulocytes, novel xanthine compounds and process for producing them - Google Patents

Description

to reduce its increased reactivity, a new xanthine and a process for their preparation

The present invention relates to the use of tertiary 1- (hydroxyalkyl) -3alkylxanthines for the preparation of a pharmaceutical composition for the treatment of diseases associated with an abnormal reactivity of eosinophilic granulocytes, to novel xanthine compounds substituted as described above, and to a process for the preparation thereof.

Hyperactive eosinophilic granulocytes are at the center of the pathogenesis of certain pulmonary, cardiac, and cutaneous disorders that are specifically associated with the atopic form.

Atopic form includes diseases associated with an allergic predisposition that are triggered by exogenous, non-infectious substances (environmental allergens) based on a specifically altered immune condition. In principle, allergic diseases can affect all important organs of the human body and manifest themselves in many different forms, such as arthralgia, asthma, erythema exsudativum multiforme, cataract, inflammation of the kidneys, inflammation of the nose or the vein. Wochenschr. (1993) 105/23: 661-668].

In clinical practice, immune responses mediated by Immunoglobulin E (IgE) (Type I allergies) are predominant in the form of anaphylaxis, allergic pulmonary asthma, allergic rhinitis and conjunctivitis, allergic urticaria, allergic gastritis and cataracts. In addition, materials from the natural environment (such as pollen from grasses, ta kh, etc.)

Λ

There is a genetic predisposition to react to spores, house dust and house mites, animal hair or food) in the form of an immediate atopic antibody (reagin) type hypersensitivity. The prevalence rate among the population is currently 10% (Pschyrembel, Klinisches Wörterbuch, Walter de Gruyter Verlag, 255, 1986, p. 148), but this proportion is constantly increasing, especially in industrialized countries.

There is concern that, despite intense efforts to improve diagnostic and therapeutic options, increasing morbidity worldwide is leading to increased mortality, such as bronchial asthma (Deutsche Apotheker Zeitung (1993) 133/18: 1635- 1636]. Thus, asthma, which is characterized by a progressive inflammatory process that causes reversible airway damage, is the only chronic disease in industrialized countries that causes death due to inadequate therapy (Therapiewoche (1993) 43/7: 340-341).

In the current state of science, the pathogenetic process is centered on chronic inflammation, in which a large number of immune-competent cells are involved in the release of pro-flanimation mediators. However, it should be assumed that in the acute phase of inflammation, so-called early or immediate reactions, especially basophilic granulocytes and mast cells (mastocytes) are involved, while progressive tissue damage and loss of function

-3 ···· · · · · · · · · · · · · · · · · · · · · · · · · · · · eosinophilic granulocytes and probably neutrophilic granulocytes play a key role [Münch. med. Wschr. (1993) 135/5: 52).

Basophilic granulocytes and mast cells, also called histamine cells, not only histone upon activation of Ig-E produced by B lymphocytes at specific high affinity receptors on the cell surface, but not only by histamine. many other inflammatory mediators are released. These include a broad spectrum of proteases, ipid mediators, including platelet activating factor (PAF), pro-taglandins and leukotrienes, and cytokines (Immunopharmacology (1994) 27: 1-11).

These mediators are largely vascular and bronchoconstrictive, increase secretion, and interfere with hemostatic regulation. They may also be attributed to the chemotactic properties that enable them to mobilize additional cells involved in the inflammatory process, including granulocytes responsible for late reactions, which, once activated by degranulation, also select inflammatory mediators, thereby continuing the inflammatory process. and they become chronic.

The eosinophilic granulocyte is a potent effector cell with pronounced leukocyte-specific properties such as chemotaxis, adherence, phagocytosis, release of granuloproteins, and lipid mediators and

Production and selection of -4-reactive oxygen species, which explains the dominant role in the pathogenetic process of allergic inflammatory reaction [Dt. Arzteblatt (1992) 84/43, Al: 3574-3585].

The post-allergen exposure process is initiated by recruitment of eosinophils from bone marrow and invasion of antigen-affected tissue, leading to local eosinophilia and subsequent cellular activation. In this pathophysiological process, various immunocompetent cells, such as Th2-type T-helper cells, macrophages, neutrophils, mast cells and the eosinophils themselves, are involved, which produce a range of appropriate factors for differentiation, prof1iferation, migration and activation.

These include eosinophil-selective interleukin-5 (IL-5) produced by immunomodulatory cytokines, such as Thelpers cells, which control the differentiation and proliferation and functional activation of eosinophilic granulocytes, with granulocyte / macrophage-specific granulocyte-activating (GN-CSF) as well as chemotactic factors responsible for migration and activation such as PAF and leukotriene B4 (LTB4).

Nevertheless, the complement cleavage product, C5a, also has a strong chemotactic and cellular stimulatory effect on eosinophils.

The activated eosinophilic granulocytes are mediators for granular proteins, lipid mediators and cytotoxic oxygen.

·· · · · reacts by synthesizing and degrading metabolites.

The pro-inflammatory lipid mediators include, in particular, leukotriene C4 (LTC4), thromboxane A2 (TXA ₂ ) and, repeatedly, PAF., Which enhance vascular permeability, induce vasoconstriction and bronchial obstruction, and exudate. (Pharmazie in unserer Zeit (1992) 21/2: 61-70).

Among the protein mediators are the enzymatic eosinophilic peroxidase (EPO) and the non-enzymatic base protenins which play a particularly important role in the degradation processes, such as the Major Basic Protein (MBP), the eosinophilic cationic protein (ECP) and the eosinophil. derived neurotoxin / eosinophil protein X (Eosinophil-Derived Neurotoxin / Eosinophil Protein X, EDN / EPX). Their versatile biological properties are distinguished by their cytotoxic activity, which extends over a wide range of cells ranging from parasites to bronchial epithelial cells, nerve cells, myocardial cells and tumor cells. Therefore, together with selected reactive oxygen metabolites, they play a pivotal role in the tissue damage associated with progressive loss of function during allergic inflammatory reactions.

In addition, they stimulate the release of histamine from mast cells, thereby releasing a new early phase attack as circulatory vitiosus.

In addition, high-toxicity protein mediators are also · ·

-6diagnostic significance because of increased ECP levels in serum and other body fluids of atopic patients such as bronchoalveolar lavatories, sputum and nasal secretions, as well as the binding of these proteins to the affected tissues because of the severity of the disease, this parameter appears to be suitable for objectively measuring disease activity and for assessing the success of treatment after an intervention (Therapiewoche (1991) 41/45: 2946-2947).

The pathogenetic relationships described in atopic diseases make it clear that the focal allergic inflammatory process with early and late phase reactions is a complex interplay between immune cells and their inflammatory mediators, and therefore a therapeutic response is expected to be a multifunctional agent immediately. blocks and is able to permanently inhibit the recruitment of eosinophils and, above all, granulocyte recruitment in a chronic late reaction [Pharmazeutische

Ze itung (1992)

137/5:

249-258; Agents and Actions (1991) 32/1 + 2: 24-33].

Surprisingly, we have found that 1,3-dialkylxanthines of formula I having a tertiary hydroxy group on the alkyl group at the 1-position fulfill the above-mentioned requirement for a therapeutic agent for the treatment of atopic diseases. Λ

implications.

1- (5-Hydroxy-5-methylhexyl) -3-methylxanthine is described in WO 87/00523. There, it has been suggested for the treatment of peripheral and cerebral blood flow disorders and mitochondrial myopathies, without suggesting that the compound can be used to reduce the pathological hyperresponsiveness of eosinophilic granulocytes and thus to treat atopic diseases.

Although many xanthine derivatives are known to possess bronchospasmolytic activity by phosphodiesterase inhibitory activity and thus are useful in the prevention and symptomatic treatment of acute bronchospasm induced by early mediated asthma, these compounds do not provide a cure for atopic diseases. they do not affect the underlying cause of the disease, the chronic reaction to eosinophils-mediated chronic inflammation. Theophylline is the most prominent representative of this class of compounds.

Recently, some 8-substituted 1,3-dialkylxanthines (EP 389,286; WO 92/11260), 1,3,7-trialkylxanthines (EP 421,587), and 7-sulfonated 1,3- dialkylxanthine and 1,3,3,7,8-tetrahydroxy xanthine (WO 92/11260), which reduce the number of eosinophils in the blood in artificially induced eosinophilia in animal experiments. However, they have not been shown to inhibit the development of pathologically

-8 series of eosinophils, which determine the course of the disease, primarily in the tissues affected by the allergic inflammatory process, so that the therapeutic utility of these compounds has not been demonstrated.

In contrast, the compounds of formula (I) have the property of inhibiting the early phase reaction and inhibiting not only the recruitment of eosinophils but also the pathological hyperresponsiveness of the target tissue to cellular mediators relevant to the course of the disease. , and selectively disable the effector function of high-grade inflammatory cells at the center of chronic disease.

EP 544 391

1,3,7-trialkylated xanthines, pentoxifylline (3,7-dimethyl-1- (5-oxo-hexyl) -xanthine), propentophylline [3-methyl-1- (5-oxo-hexyl) -7- propyl xanthine] and torbafillin [7-methyl] -1- (5-hydroxy-5-methylhexyl) -3-methylxanthine] have been suggested for topical treatment of psoriasis and atopic dermatitis, but without reference to that 1) these xanthine derivatives are also effective for non-topical use, or 2) could be applied topically or even non-topically to other diseases of the atopic form.

The invention thus provides at least one compound of formula (I) and / or a physiologically tolerable salt of the compound of formula (I) and / or a compound of formula (I).

a stereoisomeric form of a compound of formula

r! is methyl or ethyl,

R is C 1-4 alkyl and

X is hydrogen or hydroxy and n is 1-5 for use in the preparation of a pharmaceutical composition for reducing pathological hyperactivity in eosinophilic granulocytes. The compound of formula (I) is particularly useful for the prevention and treatment of atopic diseases such as anaphylaxis, allergic pulmonary asthma, allergic rhinitis and conjunctivitis, allergic urticaria, allergic gastrointestinal or atopic dermatitis.

Preferably, compounds of formula I are used wherein R is methyl or ethyl.

It is also preferred to use compounds of formula I wherein R ¹ and R ¹ are independently methyl or ethyl, X is hydrogen or hydroxy, and n is 3-5.

The use of 1- (5-hydroxy-5-methylhexyl) -3-methylxanthine is particularly preferred.

The invention further relates to novel compounds of formula (I) and / or a physiologically tolerable salt of a compound of formula (I) and / or a stereoisomeric form of a compound of formula (I) wherein

R 1 is methyl or ethyl,

R ^{6 is C} 1 -C 4 alkyl,

X is hydrogen or hydroxy, and n is 1-5, except 1- (5-hydroxy-5-methylhexyl) -3-methylxanthine.

Also preferred are compounds of formula I wherein R is methyl or ethyl, wherein R * is simultaneously non-methyl when X is hydrogen and n is 4.

Also preferred are compounds of formula I wherein X is hydrogen, and R 1 and R 2 are both non-methyl when n is 4.

Finally, the compounds of formula I in which:

R 1 is methyl,

R 4 is methyl or ethyl,

X is hydrogen and n is 1-5; furthermore, R 1 is not methyl when n is 4.

The present invention is hereinafter

-11 ·· · · · · ·

Relates to an analogous process for the preparation of compounds of formula (I), the method of which is described in WO

International Patent Application Publication No. 87/00523 is in principle described. Preferably, a 3,7-disubstituted xanthine derivative of formula (II)

OR ^a

wherein R is C 1 -C 4 alkyl and R ^a is an easily removable leaving group such as a hydrolytically cleavable methoxy, ethoxy, propoxy or butoxymethyl group, or a benzyl or diphenylmethyl group which can be removed by reduction, wherein the phenyl ring is unsubstituted or substituted, preferably in the presence of a basic condensing agent or a salt thereof

a) an alkylating agent of formula III, (in)

HO R ¹

In the formula -12a, R 1, X and n are as defined above and Z is a halogen atom, preferably a chlorine, bromine or iodine atom, or a sulfonic acid ester or a phosphoric acid ester group, an (IV)

converting to a 1,3,7-trisubstituted xanthine of the general formula wherein R 1, R 2, R ^a , X and n are as defined above or when X is hydrogen,

b) a ketone compound of formula V,

H ₃ C-CO- (CH ₂ ) _n -Z (V) wherein n and Z are as defined above for a compound of formula (VI)

1,3,7-trisubstituted xanthine, then one

With a methyl or ethyl metal compound of the formula R 1 -M, preferably with methyl or ethyl lithium (R 1 -Li) or the corresponding Grignard compound (R 1 -MgHal), with the reactive alkylation of a carbonyl group

• ί »·· •« ··· * «

and n is as defined above or when X is hydrogen and is methyl,

c) a carboxylic acid ester of the formula, (C _{r C4)} Alki lO-CO- (CH _{2) n} -Z (VIII) wherein n and Z (VIII)

one (IX) general (IX)

It is converted to 1,3,7-trisubstituted xanthine, followed by two equivalents of methyl metal compound, preferably methyl lithium or CHg-MgHal, by double reductive alkylation of the ester group of formula (X)

(X) ···· · «··

Converting to a 1,3,7-trisubstituted xanthine wherein R ² , R ^a and n are as defined above and then R ^{a is} leaving the intermediate of formula (IV), (VII) or (X). xanthine of formula (I).

The 3,7-disubstituted xanthines of formula (II) and the alkylating agents of formulas (III), (V) and (VIII) used herein as starting materials are largely known or are known in the art (see for example WO 87/00523 ) I can easily produce. For example, the obtained tertiary general formula (III) alcohol organometallic synthesis, when not sterically inhibited by Hal _{(CH2) n-CO-,} -CH ₂ -X haloketone with a so-called build-up during the reaction of the carbonyl group by reductive alkylation with an r An alkyl metal compound of the formula -M, wherein M is a metal, in particular magnesium, zinc or lithium, or optionally an alkylmagnesium halide of the formula R 1 -MgHal (Grignard compounds) or an alkyl of the formula R ⁴ -Li lithium compound can be reacted under normal conditions. A similar reaction of a halo ketone of the formula (Hal) (CH8) n-CO-R6 and a methyl magnesium halide or methyl lithium also leads to the compounds of formula (III) wherein X is hydrogen. For the preparation of compounds of structure (III) in which a hydrogen atom R ⁴ is methyl and X is a convenient way to serve as an omega-halo-alkanoic acid alkyl ester [Hal (CH2) n-COO-alkyl] with two equivalents • · · ·

Also with a -15-methyl-metal compound where the ester is converted to a tertiary alcohol by introducing two methyl groups on the keto group. Similarly, the omega-hydroxycarboxylic acid ester can be converted to the diol without the protecting of the hydroxy group, for example in the form of tetrahydropyranyl- (2) or methoxymethyl ether, or optionally in the form of a lactone as a cyclic ester, from which the selective esterification of the primary hydroxyl group with a sulfonic acid or a phosphoric acid halide or anhydride yields the active alkylating agent (III).

The disubstituted xanthine derivative of formula (II) and the alkylating agent of formula (III), (V) or (VIII) are reacted in the usual manner in a dispersion or solvent which is inert to the reaction components. Suitable solvents are, in particular, dipolar aprotic solvents such as formamide, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, tetramethylurea, hexamethylphosphoric triamide, dimethylsulfoxide, acetone or butanone; but alcohol such as methanol, ethylene glycol and its mono- and di (C 1-4 alkyl) ether, ethanol, propanol, isopropanol and various butanols; hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons such as methylene chloride or chloroform; pyridine, as well as mixtures of said solvents or mixtures thereof with water.

The alkylation reaction is preferably a basic condenser.

-16 times. For this purpose, for example, alkali or alkaline earth metal hydroxides, carbonates, hydrides, alcoholates and organic bases such as trialkylamines, such as triethyl or tributylamine, quaternary ammonium or phosphonium hydroxides, and crosslinking agents may be used. resins with fixed, optionally substituted ammonium or phosphonium groups. However, the xanthine derivatives may also be reacted directly in the form of their individually prepared salts, such as their alkali metal, alkaline earth metal or optionally substituted ammonium or phosphonium salts. In addition, the disubstituted xanthine compounds are preferably in the form of a biphasic or intermediate phase, in the presence of a so-called phase transfer, in the presence of the aforementioned inorganic condensing agent and in the form of their alkali metal or alkaline earth metal salts. Suitable, generally commercially available phase transfer catalysts include tetra (C 1-4 alkyl) and methyl trioctylammonium and phosphonium, methyl, iniristyl, phenyl and benzyltri (C 1-4 alkyl). and cetyltrimethylammonium and C 1 -C 12 alkyl and benzyl triphenylphosphonium salts, whereby compounds containing a larger and symmetric cation are usually more effective. The above procedure is generally carried out at 0 ° C and at the reflux temperature of the reaction medium, preferably between 20 ° C and 130 ° C, optionally

at elevated or reduced pressure, but usually at ambient pressure, where the reaction time may be less than one hour or more.

The organoleptic conversion of xanthine having the 1-position functional group of formula (VI) or (IX) is in principle similar to that described for the preparation of the tertiary alcohol of formula (III) as alkylating agent. Thus, for example, the ketone (VI) and the ester (IX) are, for example, with alkyl potassium, sodium, lithium, magnesium, zinc, cadmium, aluminum and tin. it can be reductively alkylated. Recently used alkyl titanium and zirconium compounds (D. Seebach et al., Angew. Chem. 95: 12-26 (1983)) may also be used. Because of the high reactivity of the alkyl metal compounds of sodium and potassium to the side reactions is neglected and the alkyl metal compounds of zinc and cadmium are relatively less reactive, in general alkyl lithium and alkyl magnesium (Grignard) compounds are preferred.

Highly nucleophilic organometallic compounds are highly sensitive to hydrolysis and oxidation. Their safe handling therefore requires work in an anhydrous medium, possibly in an inert gas atmosphere. Conventional solvents or dispersants are preferably those suitable for the preparation of alkyl metal compounds. Such solvents include, in particular, ethers having one or more ether oxygen atoms, such as diethyl ether, dipropyl, and the like.

-18dibutyl or diisoamyl ether, 1,2-dimethoxyethane, tetrahydrofuran, dioxane, tetrahydropyran, furan and anisole, and aliphatic or aromatic hydrocarbons such as petroleum ether, cyclohexane, benzene, toluene, xylene, diethylbenzene and the like. tetrahydronaphthalene; however, tertiary amines such as triethylamine or dipolar aprotic solvents such as hexamethylphosphoric triamide and mixtures of these solvents can also be used successfully. The reaction of the carbonyl compounds of formula (VI) and (IX) with the Grignard compounds of the formula R 1 -MgHal can also advantageously be carried out by combining the organometallic compound with an ether and the α-ketone or ester with methylene chloride or 1,2-. dichloroethane in solution. It is also often desirable to add inagnesium bromide, which can enhance the nucleophilic nature of the organometallic compound by virtue of its participation in a complex cyclic transition state.

The reaction of the ketone or ester with the organometallic compound is usually carried out at a temperature of -20 ° C to 100 ° C, preferably 0 ° C to 60 ° C, or at room temperature without external cooling, whereby the alkyl metal compound is generally used in a slight excess. Thereafter, the reaction is generally completed by refluxing for a short period of time, usually from a few minutes to several hours. An aqueous solution of ammonium chloride or dilute acetic acid is preferably used to decompose the alcohol formed.

The elimination of ^a leaving group of ^the compounds of formulas (IV), (VII) and (X) during the formation of xanthine (I) according to the present invention is well-known and well-known protecting group developed under standard conditions, in particular for alkaloid and peptide synthesis. technology.

Thereafter, the optionally substituted benzyl or diphenylmethyl group in the phenyl ring is preferably reduced reductively. In particular, in addition to the chemical reduction of benzyl compounds with sodium in liquid ammonia, it is preferable to catalytically hydrogenate the two aralkyl groups mentioned above in the presence of a noble metal catalyst, where ammonium formate is often suitable as the hydrogen donor to replace the molecular hydrogen. The reaction medium for this reaction is usually the addition of a small amount of carbon alcohol, optionally formic acid or ammonia; an aprotic solvent such as dimethylformamide or glacial acetic acid in particular; but a mixture of these with water may also be used. Suitable hydrogenation catalysts are preferably palladium on carbon and palladium on activated carbon or barium sulphate, while the presence of other noble metals such as platinum, rhodium and ruthenium often leads to side reactions due to competing inaghydrogenation and should therefore be used only conditionally. The hydrogenolysis is conveniently carried out at a temperature of 20 ° C to 100 ° C and at atmospheric pressure, or preferably at a slight excess pressure of up to 10 bar, usually for a few minutes and several hours.

-20 reaction time required.

The 1,3,7-trisubstituted (IV), (VII) and (X) Formula xanthines bearing an alkoxymethyl group ⁱⁿ place of R, Ο, Ν acetal form, as a result, in normal circumstances and they can be easily disassembled by acid hydrolysis

Preferred groups include methoxyethoxy, propoxy and butoxymethyl. The reaction is preferably carried out in a dilute mineral acid such as hydrochloric or sulfuric acid, optionally with glacial acetic acid, dioxane, tetrahydrofuran or a small amount of carbon alcohol as a solubilizer. Occasionally, perchloric acid or organic acids such as trifluoroacetic acid, formic acid and acetic acid may be used together with catalytic amounts of mineral acids. Cleavage of the ether group may, in principle, be effected by means of a Lewis acid such as zinc bromide and titanium tetrachloride in anhydrous media, preferably methylene chloride or chloroform. For the cleavage in mineral acid solution, the reaction temperature is chosen so that no significant dehydration occurs in the tertiary hydroxyalkyl group; therefore, the temperature of 60 ° C should not normally be exceeded.

The compounds of formula I are deprotonated at the 7-position to form salts and solvates with basic agents. Pharmaceutically acceptable alkali and alkaline earth metal salts and salts with organic bases such as ethylenediamine or basic amino acids lysine, ornithine and arginine are preferably used for this purpose. The present invention thus provides the 1,3-dialkyl-21- of formula (I).

also includes pharmaceutically acceptable salts and / or solvates of xanthines.

The tertiary 1- (hydroxyalkyl) -3-alkyl xanthines of formula (I) contain an asymmetric carbon atom when X is hydroxy or X is hydrogen and r is hydrogen. is ethyl. These compounds thus exist in stereoisomeric forms. The invention also relates to pure stereoisomeric compounds and mixtures thereof.

The novel xanthine compounds of formula (I) of the present invention are particularly suitable for use as an active ingredient in a medicament, in particular for the prophylactic and curative treatment of atopic forms of diseases caused by pathological eosinophilic hyperresponsiveness due to their valuable pharmacological properties. and thus the treasure trove is greatly enriched. These compounds may be administered alone, for example in the form of a microcapsule, in admixture with one another, or in combination with suitable carriers.

Accordingly, the present invention also provides a medicament comprising as active ingredient at least one compound of formula (I), except for 1- (5-hydroxy-5-methylhexyl) -3-inet 1-xanthine.

It is a further object of the present invention to provide a pharmaceutical composition for oral, rectal, topical, parenteral, or inhalation administration in the treatment of a disease which is a compound of formula (I).

-22 in which the reactivity of eosinophil granulocytes is pathologically increased. Suitable solid or liquid galenical formulations include granules, powders, tablets, dragees, (micro) capsules, syrups, emulsions, suspensions, milks, creams, ointments, gels, aerosols, drops or injectable solutions in the form of ampoules, excipients such as carriers, disintegrants, binders, coatings, swellers, lubricants or lubricants, flavors, sweeteners, or solubilizing agents are generally employed.

Commonly used excipients are magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatin, starch, vitamin, cellulose and derivatives thereof, animal and vegetable oils, polyethylene glycol and solvents such as sterile water, alcohol , glycerol and polyhydric alcohols.

The pharmaceutical compositions are preferably formulated and administered in dosage units, each of which contains the active compound of the formula I in a fixed dose. For solid dosage units such as tablets, capsules and suppositories, this dosage will be up to 1000 mg, preferably 100-600 mg, and 300 mg, preferably 20-200 mg, for ampoule injection solutions.

For the treatment of an adult patient, depending on the efficacy of the compound of formula (I), a daily dose of 100-2000 mg of active ingredient, preferably 300-900 mg, may be administered orally * ·

-23, and 10-500 mg, preferably 20-200 mg, for intravenous administration. However, depending on the circumstances, higher or lower daily doses may be used. The daily dose may be administered as a single dose unit, or in several smaller dosage units, and in divided doses given at regular intervals.

Finally, the xanthine derivatives of formula (I) may be used in the preparation of the above-mentioned galenical formulations, if necessary with other suitable active ingredients such as antihistamine, anticholinergics, 13g- ^m4 - ^me mics, phosphodiesterases, phospholipase A2 and lipoxygenase inhibitors, leukotriene antagonists, corticosteroids, chromo-glycic acid, nedocromil and cyclosporin A.

The structure of the compounds described below and summarized in Table 1 was confirmed by elemental analysis and IR and 1 H-NMR spectra.

Example 1

1- (2-Hydroxy-2-methylpropyl) -3-methylxanthine (a) 1-Chloro-2-hydroxy-2-methylpropane

To a stirred solution of 44.9 g (0.6 mol) of methyl 1-magnesium chloride in 20% tetrahydrofuran and 200 ml of dry diethyl ether at 0 to 5 ° C was 46.3 g (0.5 mol). A solution of l-chloro-2-propanone in 50 ml of anhydrous diethyl ether was added dropwise. The mixture was then stirred for 1 hour

····

After stirring for 24 hours at room temperature and refluxing for an additional hour, the resulting tertiary alkanolate is quenched by the addition of 50% aqueous animonium chloride solution, the ether phase is separated off and the aqueous phase is extracted with ether. The combined ethereal extracts were washed successively with aqueous sodium hydrogen sulfite solution and sodium bicarbonate solution, then with a little water, dried over sodium sulfate, filtered, evaporated under reduced pressure, and the liquid residue was fractionally distilled.

Yield: 31.1 g (57.3%).

Fp. 125-127 ^! C

C ₄ H ₉ ClO (MW = 108.6)

Similarly, the compound can be prepared from chloroacetic acid methyl or ethyl ester in two molar amounts of methyl magnesium chloride in 60% yield.

b) 7-Benzyl-1- (2-hydroxy-2-methylpropyl) -3-methylxanthine

25.6 g (0.1 mol) of 7-benzyl-3-methylxanthine, 15.2 g (0.11 mol) of potassium carbonate and 11.9 g (0.11 mol) of tertiary alcohol prepared in step a) A mixture of 500 ml of dimethylformamide was heated at 110-120 ° C for 8 hours with stirring, then filtered hot and concentrated under reduced pressure. The residue is taken up in chloroform, washed with 1N sodium hydroxide solution, then with water, neutralized, dried, the solvent is distilled off under reduced pressure and the solid residue is recrystallized from ethyl acetate with petroleum ether.

-25 Yield: 26.6 g (81.0%)

Mp 115-117'C ^ 17 ^^ 20 ^ 4θ3 (MW = 328.4)

Elemental Analysis: Calculated: C: 62.18%; H: 6.14%; N: 17.06%; found: C: 62.60%; H: 6.18%; N, 17.00%. The compound can also be prepared by that 7-benzyl-3-methyl-

xanthine was first converted to 7-benzyl-3-methyl-1- (2-oxo-propyl) -xanthine and 7-benzyl-1-chloro-2-propanone or methyl or ethyl chloro-acetic acid under the reaction conditions described above. 1- (meth (or eth) oxycarbonylmethyl) -3-methylxanthine and then the oxopropyl and alkoxycarbonylmethyl side chains with methylmagnesium chloride in anhydrous diethyl ether, respectively. is reductively methylated.

c) 1- (2-Hydroxy-2-methylpropyl) -3-methylxanthine

The 7-benzylxanthine (13.1 g, 0.04 mol) obtained in step b) was hydrogenated in 200 ml glacial acetic acid in the presence of 1.5 g of 10% palladium on carbon at 60 ° C and 3.5 bar with shaking for 100 hours. After cooling, the mixture is covered with nitrogen, the catalyst is filtered off, the filtrate is evaporated under reduced pressure and the solid residue is recrystallized from ethyl acetate.

Yield: 7.8 g (81.8%)

M.p. 215-217'C ^C 10-14 ^N 4 ° 3 (molecular weight = 238.3)

Elemental Analysis: Calculated: C, 50.41; H: 5.92%; N: 23.52%;

found:

C: 50.10%; H, 5.90%;

N: 23.40%.

• · · · · · ·

-262. example

3-Ethyl-l- (2-hydroxy-2-methylpropyl) xanthine

a) To a suspension of 7-benzyl-3-ethylxanthine (0.5 mole) in 3 ml of methanol (500 ml) was added sodium hydroxide (20 g, 0.5 mole) in water (200 ml) and the mixture was stirred for 1 hour. at 70 ° C for hours and then at the same temperature benzi1 chloride 69.6 g (0.55 mol) was added dropwise, and kept for three hours at 7080 ° C ^e. The reaction mixture was cooled, filtered by suction filtration, the product was washed with water on the filter, dissolved in 1000 ml of 1 N sodium hydroxide solution, filtered, and the filtrate was slowly acidified to pH 9.5 with 4N hydrochloric acid with stirring. The crystals were filtered off from the still warm solution, washed with water to remove chlorine and dried in vacuo.

Yield: 131 g (96.9%)

Mp 217-218'C

C14H14N4O2 (MW = 270.3)

b) 3-Ethyl-1- (2-hydroxy-2-methylpropyl) xanthine

The 7-benzyl-3-ethylxanthine produced in step a) is reacted with the 1-chloro-2-hydroxy-2-methylpropane prepared in Example 1a) in a similar manner to that described in Example 1b) to give 7-benzyl-3-ethyl-1-yl. - (2-Hydroxy-2-methyl-propyl) -xanth is obtained with an molecular weight = 342.4; Yield 46.1%], which was debenzylated by hydrogenolysis according to Example le) to give a crude final product (97.9% yield) which was purified by recrystallization from ethanol.

-27Op. 217-219 ° C

C11H16N4O3 (MW = 252.3)

Elemental Analysis: Calculated: C, 52.37; H, 6.39; N: 22.21%;

Found: C, 52.19; H, 6.29%; N, 21.75%.

Example 3

1- (3-Hydroxy-3-methylbutyl) -3-methylxanthine

a) 1-Chloro-3-hydroxy-3-methylbutane

The compound is formed from methyl magnesium iodide and l-chloro-3-butanone (which can be obtained by addition of hydrogen chloride in methyl ether to diethyl ether) or methyl magnesium chloride and ethyl 3-chloropropionate in methylene chloride. in dichloride as the reaction medium in a similar manner to that described in Example 1a).

Yield: 60-70%

Fp. (18 mbar): 66-68 ° C

C5H11ClO (MW = 122.6)

b) 7-Benzyl-1- (3-hydroxy-3-methylbutyl) -3-methylxanthine

In a manner similar to that described in Example 1b), it is prepared from 7-benzyl-3-methylxanthine and the tertiary alcohol obtained in step a).

Yield: 70%

92-94'C ^ 18 ^ 22 ^ 4 ^ 3 (MW = 342.4)

Elemental Analysis: Calculated: C, 63.14; H, 6.48%; N: 16.36%;

Found: C, 63.10; H, 6.43; N, 16.28%.

-28c) 1- (3-Hydroxy-3-methyl-butyl) -3-methyl-xanthine

In a similar manner to that described in Example le), the product obtained in step b) is debenzylated by hydrogenolysis.

Yield: 87.2%

M.p. 203-205 DEG C. ^ 11 ^ 16 ^ 4 ^ 3 (molecular weight = 252.3).

Elemental Analysis: Calculated: C, 52.37; H, 6.39; N: 22.21%;

Found: C, 52.13; H, 6.52%; N: 22.08%.

Example 4

3-Ethyl-1- (3-hydroxy-3-methyl-butyl) -x-anin

a) 7-Benzyl-3-ethyl-1- (3-hydroxy-3-methyl-butyl) -xanthine

In a manner similar to that described in Example 1b), it is prepared from 7-benzyl-3-ethylxanthine obtained in Example 2a and 1-chloro-3-hydroxy-3-methylbutane obtained in Example 3a.

Yield: 71.8%

133-135 ° C

C19H24N4O3 (MW = 356.4)

b) 3-Ethyl-1- (3-hydroxy-3-methyl-butyl) -xanthine

As described in Example le), the product of step a) is prepared by debenzylation by hydrogenolysis.

Yield: 88.2%

M.p. 241-2343'C ^ 12 ^ 18 ^ 4 ^ 3 (MW = 266.3)

-29 Elemental Analysis: Calculated: C, 54.12; H, 6.81; N: 21.04%;

Found: C, 53.89; H, 6.86%; N: 21.03%.

Example 5

1- (4-Hydroxy-4-methylpentyl) -3-methylxanthine

a) 7-Benzyl-3-methyl-1- (4-oxopentyl) xanthine

In a similar manner to Example 1b, 38.4 g (0.15 mol) of 7-benzyl-3-methylxanthine, 22.4 g (0.162 mol) of potassium carbonate and 26.7 g (0.162 mol) of l-chloro The 4-pentanone-ethylene ketal was reacted with 600 ml of dimethylformamide and the resulting 7-benzyl-1- (4,4-ethylenedioxypentyl) -3-methylxanthine was refluxed with 600 ml of 1N hydrochloric acid without further purification. After boiling for 1 hour, it is subjected to ketal splitting. The reaction mixture was neutralized with concentrated sodium hydroxide solution, the resulting ketone was extracted with chloroform, and the chloroform extract was washed with water, dried over sodium sulfate and then concentrated under reduced pressure.

Yield: 50.4 g (98.7%)

M.p. 104-105 DEG C. ^ 18 ^ 20 ^ 4θ3 (MW = 340.4).

b) 7-Benzyl-1- (4-hydroxy-4-methylpentyl) -3-methylxanthine g (0.12 mole) methyl magnesium chloride as a commercially available solution in 20% tetrahydrofuran; The mixture is cooled to -25 DEG C. and a solution of 34 g (0.1 mol) of the product from step a) is added dropwise while the temperature rises to 20 ° C. Then it is

After stirring for an additional hour at room temperature, the organic layer was separated after the addition of a saturated ammonium chloride solution, the aqueous layer was extracted several times with methylene chloride, and the combined methylene chloride extract was washed with water, dried and evaporated. The solid residue was recrystallized from ethyl acetate.

Yield: 28.3 g (79.4%)

132-133'C ·

C19H24N4O3 (MW = 356.4) c) 1- (4-Hydroxy-4-methylpentyl) -3-inethyl 1-xanthine

As described in Example le), the product of step b) is prepared by debenzylation with hydrogenolysis.

Yield: 65.9%

M.p. 188-189'C ^ 12 ^ 18 ^ 4 ^ 3 (MW = 266.3)

Elemental Analysis: Calculated: C: 54.12%; H, 6.81; N 21st , 04%; found: C: 53.86%; H, 6.88%; N 20th , 93%. Example 6 3-Ethyl-1- (4-hydroxy-4-methyl-1-point 1) -xanthine a) 7-Benzyl-3-ethyl-1- (4- oxopentyl) xanthine In a similar manner to that described in Example 5a) is forward,

starting material is 7-benzyl-3-ethylxanthine from Example 2a).

Yield: 82.4%

-31Op. 139-141'C * ^ 19 ^ 22 ^ 4θ3 molecular weight = 354.4)

b) 7-Benzyl-3-ethyl-1- (4-hydroxy-4-methylpentyl) xanthine

The product of step a) is reacted with methyl magnesium chloride in a similar manner to that described in Example 5b).

Yield: 81.9%

155-157'C ^ 20 ^ 26 ^ 4θ 3 (molecular weight = 370.5)

Elemental Analysis: Calculated: C, 64.84; H, 7.07%; N: 15.12%;

Found: C, 64.95; H, 7.18%; N, 15.10%.

c) 3-Ethyl-1- (4-hydroxy-4-methylpentyl) xanthine

The compound is prepared in a similar manner to that described in Example le) by debenzylation of the product of step b) by hydrogenolysis.

Yield: 71.3%

Mp 214-216 ° C

C23H20N4O3 (MW = 280.3)

Elemental Analysis: Calculated: C, 55.70; H, 7.19%; N, 19.99%;

Found: C, 55.50; H, 7.20%; N: 20.23%.

Example 7

1- (5-Hydroxy-5-methylhexyl) -3-methylxanthine

Methods for preparing this compound are described in PCT Application Publication No. WO 87/00523.

Example 8

1- (5,6-dihydroxy-5-methylhexyl) -3-methylxanthine

a) 1-Chloro-5,6- (isopropylidenedioxy) -5-methylhexane

To a mixture of 264 g (1.2 mol) of trimethylsulfoxonium iodide and 28.8 g (1.2 mol) of sodium hydride, to which nitrogen is layered, is added dropwise 1000 ml of anhydrous dimethylsulfoxide at 40 DEG C. while stirring. in 10 minutes. After the evolution of gas has ceased (about 2 hours), a solution of 1-chloro-5-hexanone (134.6 g, 1 mol) in dimethylsulfoxide (30 ml) is added dropwise to the reaction mixture over a period of about 1 hour. in 20 minutes. After stirring for 2 hours at room temperature, ice-water (500 ml) was slowly added under ice-cooling and the resulting 1-chloro-5,6-epoxy-5-methylhexane was extracted with diethyl ether. [Yield: 130.5 g (87.8%); GyHg / gClO (molecular weight = 148.6)]. For hydrolytic cleavage of the epoxide ring, the product was stirred in a mixture of 60 ml of water, 600 ml of tetrahydrofuran and 1 ml of 70% perchloric acid for 5 days at room temperature. The reaction mixture was then neutralized with sodium carbonate solution, the tetrahydrofuran was removed by distillation, and the remaining 1-chloro-5,6-dihydroxy-5-methylhexane was extracted with chloroform. [Yield: 124.8 g (85.3%); CylIgClCl (molecular weight = 166.6)]

The nuts are then converted into dioxolane by acid catalysis under 2,2-dimethoxypropane in acetone in the usual manner. Yield: 67.2%

Fp. (0.5 mbar): 84-86'C ^C 10 ^H 19 ^C * * - 2 (molecular weight = 206.7)

b) 1- (5,6-Dihydroxy-5-methylhexyl) -3-methylxanthine

The diol obtained in step a) with 7- (ethoxymethyl) -3-methylxanthine in a similar manner as described in Example 1b) gave a quantitative yield of 7- (ethoxymethyl) -1- [5,6- (isopropylidenedioxy) -5]. is converted to -methyl-hexyl] -3-methylxanthine (C19H3QN4O5, molecular weight = 394.5), and the acidolysis is followed by opening of the dioxolane ring and cleavage of the ethoxymethyl group at the 7-position to give the final product. To this was added xanthine (19.7 g, 0.05 mol) in 1N hydrochloric acid (300 ml) and glacial acetic acid (30 ml) at 70 ° C for 15 hours with stirring, cooled, basified with sodium carbonate and washed with chloroform. It is neutralized with 1N hydrochloric acid and extracted with chloroform. The residue was filtered through a silica gel column with chloroform / methanol (10: 1) and recrystallized from ethyl acetate.

Yield: 11.5 g (77.6%)

Mp 181-182'C

C13H20N4O4 (MW = 296.3)

Elemental Analysis: Calculated: C, 52.69; H, 6.80%; N: 18.91%;

Found: C, 52.46; H, 6.90%; N: 18.66%.

Example 9

1- (5-Hydroxy-5-methyl-heptyl) -3-methylxanthine

7-Benzyl-3-methyl-1- (5-oxo-hexyl) -xanthine was prepared from 7-benzyl-3-methyl-xanthine and 1-chloro-5-hexanone in a similar manner to that described in Example 1b). keto group was then reduced to 34 with ethyl magnesium chloride as in Example 5b).

ethyl acetate, and the resulting 7-benzyl-1- (5-hydroxy-5-methylheptyl) -3-methylxanthine is debenzylated by hydrogenolysis under the conditions of Example le).

Yield: 70.2%

169-170 ° C

C14H22N4O3 (MW = 294.4)

Elemental Analysis: Calculated: C, 57.13; 11: 7.53%; N: 19.03%;

Found: C, 56.90; H, 7.55%; N: 18.96%.

Example 10

3-Ethyl-l- (5-hydroxy-5-methylhexyl) xanthine

7-Benzyl-3-ethyl-xanthine (Example 2a) and 1-chloro-5-hydroxy-5-methylhexane (WO 87/00523) were reacted as in Example 1b) to give 7-benzene in 65% yield. 1-3-Ethyl-1- (5-hydroxy-5-methylhexyl) xanthine (m.p. 212-28 ^ 4θ3 * molecular weight = 384.5) m.p. 112-114 ° C. This compound is debenzylated by hydrogenolysis using ammonium formate as the hydrogen source. To this was added 3.84 g (0.01 mol) of benzyl compound and 1.0 g (0.016 mol) of ammonium formate in 30 ml of ethanol in the presence of 2 g of 10% palladium-on-carbon at 35 ° C for several days while the formate was added until a total amount of 4.4 g (0.07 mol) was added. The mixture is filtered, the filtrate is evaporated, the residue is taken up in sodium carbonate solution, washed with chloroform, the aqueous phase is acidified to pH 4 with hydrochloric acid, the product is extracted with chloroform and the extract is evaporated. The residue was recrystallized from ethyl acetate.

Yield: 2.0 g (67.9%)

180-182 ° C

C14H22N4O3 (MW = 294.4)

Elemental Analysis: Calculated: C, 57.12; II: 7.53%; N: 19.04%;

Found: C, 56.77; H, 7.66%; N: 18.93%.

Example 11

3-Ethyl-l- (5-hydroxy-5-methylheptyl) -xanthine

7-Benzyl-3-ethylxanthine (Example 2a) and 1-chloro-5-hexanone were reacted in a similar manner to Example 1b) to give 7-benzyl-3-ethyl-1- (5-oxo-hexyl) - xanthine (C 20 H 24 N 4 O 3, molecular weight = 368.4; yield: 81.7%), m.p. 123-125 ° C. The keto group was reductively ethylated with ethylagnesium chloride as in Example 5b) to give 7-benzyl-3-ethyl-1- (5-hydroxy-5-methylheptyl) -xanthine (2222 → 30 → 403) molecular weight. = 398.5, yield 86.9%, m.p. 93-94 ° C) was debenzylated by hydrogenolysis as described in Example 10. The final product is recrystallized from ethanol.

Yield: 66.5%

165-166 ° C

C15H24N4O3 (MW = 308.4)

Elemental Analysis: Calculated: C, 58.42; H, 7.84%; N: 18.17%;

Found: C, 58.30; H: 8.05%; N: 18.33%.

Example 12

1- (6-Hydroxy-6-methylheptyl) -3-methylxanthine • · · «

From 7-Benzyl-3-methylxanthine and 1-bromo-6-hydroxy-6-methylheptane (WO 87/00523) in a similar manner to Example 1b) in 7-benzyl-1- ( 6-Hydroxy-6-methylheptyl) -3-methylxanthine (6-21 ^ 28 ^ 4 ^ 3 »m.p. = 384.5, m.p. 83-85 ° C) was prepared as described in Example le) Debenzylation by hydrogenolysis.

Yield: 82.2%

Mp 166-167 ° C

C14H22N4O3 (MW = 294.4)

Elemental Analysis: Calculated: C, 57.12; H, 7.53%; N: 19.04%;

Found: C, 56.82; H, 7.74%; N: 19.01%.

Example 13

3-Ethyl-l- (6-hydroxy-6-methylheptyl) -xanthine

The reaction of Example 12 was carried out with 7-benzyl-3-ethylxanthine prepared in Example 2a, and the product was debenzylated by hydrogenolysis using ammonium formate in a manner similar to that described in Example 10.

Yield: 72.4%

163-165'C

C15H24N4O3 (MW = 308.4)

Elemental Analysis: Calculated: C, 58.42; H, 7.84%; N: 18.17%;

found:

C: 57.83%; H, 7.64%; N: 18.04%.

····

Table 1 Compounds of formula (I)

Example n X R ¹ R ² melting point ° C 1 1 H ch ₃ ch ₃ 215-217 2 1 H CH3 c ₂ h ₅ 217-219 3 2 H ch ₃ ch ₃ 203-205 4 2 H ch ₃ c ₂ h ₅ 241-243 5 3 H ch ₃ ch ₃ 188-189 6 3 H ch ₃ c ₂ h ₅ 214-216 7 4 H ch ₃ ch ₃ 192-193 8 4 OH ch ₃ ch ₃ 181-182 9 4 H c ₂ h ₅ ch ₃ 169-170 10 4 H ch ₃ c ₂ h ₅ 180-182 11 4 H c ₂ h ₅ c ₂ h ₅ 165-166 12 5 H ch ₃ ch ₃ 166-167 13 5 H ch ₃ c ₂ h ₅ 163-165

Pharmacological tests and results

1. Inhibitory effect on proinflammatory mediators of early phase reaction

Compounds of formula (I) are proinflammatory early-phase mediator with histamine, PAF and leukemia.

The inhibitory effect of -38triene on D4 (LTD4) was investigated in tracheal segments isolated from albino guinea pigs, where the contractions triggered by these mediators served as a measurement parameter.

Freshly prepared organ preparation from a male animal was used for each experiment.

The trachea was disassembled into rings, of which 5-5 tracheal rings were attached to a strand, suspended in an organ bath at 37 ° C and in a carbogen-aerated Tyrode solution at 0.5 g and added with histamine hydrochloride (bath concentration 10 x g). contraction was induced in the absence or control of the test substance (control experiment).

The lungs were cut lengthwise into 2-3 strips, with these strips performed as described above. However, in this case, the voltage applied was 1 g, and the contraction or PAF by adding LTD4 (10 ~ 10 or θ ^- ® g / ml bath concentration) was induced.

Each experiment consisted of examining 6 organ preparations (n = 6).

The effect of the formulations was judged by the ICgQ values, which is the pg / ml concentration which halves the contraction in the control experiment. The data are summarized in Table 2.

• · · · ·

Table 2

Inhibitory effect on proinflammatory early phase mediators

Example number of compound anti-contraction effect (ICcn, μβ / ιηΐ) H i s z t am i n Trachea PAT Lungs ltd ₄ Lungs 4 10 1-3 5 30 3 10 6 30 3 1-3 7 10 - 30 10 - 30 10 - 30 8 10 - 30 6 10 9 10 - 30 10 10 - 30 12 3-10 3-6 6-10 13 10-30 1 Pentoxifene Ilin 30-60 3 30-60 Torbafi Ilin 60 3-6 10 - 30

2. Inhibition of antigen-induced early phase response in pre-sensitized guinea pigs

Albino guinea-pigs of both sexes weighing 180-220 g were sensitized subcutaneously with 1 mg of ovalbunune (0.1% saline) for two consecutive days.

-4020 days later we performed the procedure described by Konzett and Rössler [Arch. exp. Path. und Pharmak. (1940) 195: 75]. To this end, the animals were anesthetized with pentobarbital, artificially ventilated, treated with talconium chloride to eliminate spontaneous respiration, and divided into groups of 6 animals. Intravenous administration of 1 mg / kg of ovalbumin as an antigen in a mediator-induced early asthmatic reaction resulted in a prolonged asthma attack, the intensity of which was determined by the contraction heights in the thoracogram.

Test substances were also administered intravenously 15 minutes after antigen challenge. Instead, the control animals received 0.9% saline. The efficacy of the formulations was determined by the number of animals in the groups in which the asthmatic response of the control animals was reduced by at least 40%. The results are shown in Table 3.

Table 3

Inhibition of antigen-induced early phase response in guinea pigs

Example number of compound Number of protected animals 10 mg / kg 25 mg / kg after intravenous administration of antigen Number% to Number% 2 2/6 33 4/6 67 4 1/6 17 3/6 50

Example number of compound Number of protected animals 10 mg / kg 25 mg / kg after intravenous administration of antigen Number% to Number% 7 2/6 33 4/5 80 8 3/6 50 3/6 50 9 4/6 67 5/6 83 12 3/6 50 4/6 67 Pentoxifene Ilin 1/6 17 2/6 33 Torbaf I Ilin 0/6 0 0/6 0

3. Inhibition of eosinophil activation by late phase mediators

The inhibitory effect of xanthines of formula I on the activation of human eosinophilic granulocytes by the late-phase mediators of IL-5, GM-CSF, C5a and PAF was tested by a lucigenin-dependent chemiluminescent (CL) reaction. For this, the known procedure [Arch. Dermatol. Gap. (1987) 279: 470-477 and J. Invest. Dermatol. (1986) 86: 523-528], obtained from human venous blood of purified eosinophils were pretreated with concentrations of 100 μΜ or clean water [positive control (A)] to be tested at 37 DEG C. for 10 minutes, and then IL-5 (10 ² U / ml ), GM-CSF (10 ³ U / ml), C5a (10 ^{-7 M} ) or PAF (10 ^-6 M), or mixed with water to determine basal activity (B). Measurement based on the CL reaction was performed 30 minutes later at 37 ° C. The residual activity of cells pretreated with the test substances was calculated as a percentage of each positive control using the following formula:

-42CL _X - CL _{b in} the formula

CL cl _a - cl _b is the activity of cells pre-stimulated with xanthine χ 100 pre-treated cells

CL 1 is the activity of cells pretreated with water after stylation, and

CLg is the basic activity of unstimulated water-pretreated cells.

The results of the assay are shown in Table 4.

Table 4

Inhibition of the activation of these phases by late-phase mediators

Example number of compound Residual activity is A positive IL-5 materials % of control PAF after licking GM-CSF kai is done C5a st imt 7 42 63 19 17 Pentoxifene Ilin 57 70 56 45 Torbaf í Ilin 58 125 51 58

-43 • ···· · · · · · · · · · · · · · · · · ···

4. Inhibition of antigen-induced late-phase reaction

In a chronic, long-term study in guinea pigs challenged with human serum antigen, we investigated the inhibitory effect of the compounds of formula I on the pathologically increased chemotactic infiltration of eosinophilic granulocytes in the abdominal cavity (in vivo) and in eosinophilic granulocytes. Animals treated with each compound (n = 6) were treated with the test substances at an oral dose of 80 mg / kg / day for 15 weeks, while control animals (n = 6) received vehicle (carboxymethylcellulose). After the third week of treatment, all 12 animals were challenged weekly with human serum antigen, and after 48 hours, abdominal rinsing with 50 ml of 5% glucose solution and the number of eosinophils infiltrated and separated by an interrupted Percoll density gradient (> 95% (V ability (98%)) their reactivity to PAF and C5a mediators was determined by actin polymerization by flow cytometry and the Boyden-kanira method by comparing the two groups of animals.

For example, in this experiment, Compound Preparation 7 significantly reduced (p <0.01) the number of eosinophils migrating into the abdominal cavity (34.9 4.8%; x SD) compared to control animals (42.2 5.8%). . In addition, eosinophils from treated animals showed a significant reduction (P <0.05) in chemotactic migration induced by PAF or C5a compared to control animals; actinpoly • ·

The PAF-induced initial phase (<10s) of -44merization was also significantly reduced.

These results clearly demonstrate that the compounds of formula I reduce the antigen-induced, pathological hyperresponsiveness of eosinophilic granulocytes in inflammatory tissues and are therefore particularly useful for the prevention and treatment of atopic form diseases.

Claims (13)

1. At least one general formula (I) is methyl or ethyl, R ^{1 R2} C 1 -C 4 alkyl and is hi-drug. or a hydroxyl group and a value of 1-5 and / or the use of a wearable salt of general formula (I) and / or a stereoisomer of a compound of general formula (I) for the preparation of a composition for reducing a pathological therapeutic hyperreactivity of eosinophilic granulocytes. Use according to claim 1, characterized in that at least one compound of formula (I) O 'is used wherein R is methyl or ethyl. 3. alkalmazáss according to claim 1 or 2, characterized in that with at least one (I) ···· compound or a salt thereof, wherein R and ^h are each independently methyl or ethyl; X is hydrogen or hydroxy and n is 3, 4 or 5. 4. Use according to one or more of claims 1 to 5, characterized in that the compound is 1- (5-hydroxy-5-methylhexyl) -3-methylxanthine or a salt thereof. 5. Use according to one or more of claims 1 to 4 for the treatment and / or prevention of atopic form diseases. 6. Use according to one or more of claims 1 to 4 for the treatment and / or prevention of anaphylaxis, allergic pulmonary asthma, allergic rhinitis and conjunctivitis, allergic urticaria, allergic gastritis and atopic dermatitis. 7. Use according to one or more of claims 1 to 8 for oral, rectal, topical, parenteral or inhalation administration. 8. Use according to claim 7, characterized in that further at least one anti believe amine anticholinergic agent, 02 ~ ^m i ^methyloxy .mu.m, phosphodiesterase, phosphodiesterase-47- ···· · ····· Lipase A £ and 1ipoxygenase inhibitors, PAF and leukotriene antagonists, corticosteroids, chromium · ·································································································· · an effective amount of glycine fatty acid, nedocromil or cyclosporin A is also used. 9. A compound of formula (I) and / or a physiologically acceptable salt of a compound of formula (I) and / or a stereoisomeric form of a compound of formula (I) wherein R ¹ is methyl or ethyl, R ² is C 1-4 alkyl and X is hydrogen or hydroxy, and n its value is 1-5, except for a compound of formula (I) wherein R 1 - n * and R are both methyl, X is hydrogen and n is 4. The compound of claim 9, wherein that (I) ethyl. generally η o s k é p 1 e t be n She R is methyl obsession 11th THE 9 or 10 claim of compound, with characterized in that (I) general wherein X represents bridge- rogénatom. 12th THE 10 and 11 claim of compound , with characterized in that (I) general wherein R ⁴ is a methyl group η R is methyl or ethyl, X is hydrogen and n is 1-5. 13. Method 9-12. A process for the preparation of a compound of formula (I) according to any one of claims 1 to 3, characterized in that it is a compound of formula (II) A 3,7-disubstituted xanthine derivative in which R ² is C 1 -C 4 alkyl and R ^a is a methoxyl, ethoxy, propoxy or butoxymethyl group which is hydrolytically cleavable as an easily removable leaving group or a benzyl or diphenylmethyl group having a reductively removable, substituted or substituted phenyl ring. basic condensation in the presence of a condom or in the form of a salt a) with an alkylating agent of formula III, (1) 1) HO R ^{1 in} the formula R ¹ , X and n are as defined above, and Z is a halogen atom, preferably a chlorine, bromine or iodine atom, or a · · · · -49sulfonic acid ester or phosphoric acid ester group, one (IV) (IV) Was converted to 1,3,7-trisubstituted xanthine, Ra, ^{R2, Ra,} X and n are as defined above, or, in the case where X represents a hydrogen atom, b) a ketone compound of formula V, H ₃ C-CO- (CH ₂ ) _n -Z (V) wherein n and Z are as defined above for a compound of formula (VI) N /> N (VI) The 1,3,7-trisubstituted xanthine is then reacted with an inethyl or ethyl 1-metal compound of the formula R1-M, preferably methyl or ethyl-lithium (R1-Li) or the corresponding Grignard compound (R1-M). -MgHal), in addition to the reductive alkylation of the carbonyl group, is converted to the 1,3,7-trisubstituted xanthine of formula (VII), wherein R ² , R ^a and n are as defined above, or in the case where X is hydrogen and R ^{4 is} methyl, c) with a carboxylic acid ester of formula VIII, (C ₁ -C ₄ ) alkyl 10 -CO- (CH ₂ ) _n -Z (VIII) wherein n and Z are as defined above, a compound of formula IX OR ^a (IX) Converted to 1,3,7-trisubstituted xanthine followed by two equivalents of methyl metal compound, preferably methyl lithium or CHg-MgHal, by double reductive alkylation of the ester group (X) ········································· and then eliminating ^the leaving group R from ^the intermediate of formula (IV), (VII) or (X) to provide the xanthine of formula (I) of the present invention, which may be converted into a pharmaceutically acceptable salt if desired. 14. A pharmaceutical composition comprising: a 9-12. One or more of claims 1 to 13 or 13. An effective amount of at least one compound of formula (I) prepared according to claim 1. 15. A process for the preparation of a pharmaceutical composition according to claim 14, characterized in that at least one of the pharmaceutical compositions according to claims 9 to 12 is produced. A compound of formula (I) according to one or more of claims 1 to 8, which is formulated in a suitable dosage form with pharmaceutically acceptable and physiologically acceptable carriers and excipients, diluents and / or other active substances or excipients.