HU196364B - Process for producing aliphatic thioethers and pharmaceutical compositions containing them as active components - Google Patents

Abstract

Thioethers of formula (I) and their salts are new: R1 = 1-3C alkyl, opt. substd. by OH which can be esterified; R2 = 5-15C opt. unsatd. aliphatic gp.; R3 = OH, alkoxy or opt. substd. amino; X = bond, CH2 or opt. N-acylated primary aminomethylene; the S atom and the O atom of OH are trans to each other. - Specifically the gp. -S.CH2.X.CO.R3 is the residue of mercaptoacetic acid; 3-mercaptopropionic acid; N-(3-mercapto propionyl)glycine; L-cysteine (opt. N-acylated by trifluoroacetyl) or a gp. R4-Cys-Gly-R3'; R3' = OH or 1-4C alkoxy; R4 = H, CF3CO or gamma-glutamyl.

Description

The present invention relates to a process for the preparation of a compound of formula I:

R 'is (C 1 -C 3) -alkyl or (C 1 -C 3) -hydroxyalkyl whose hydroxyl group is esterified with (C 1 -C 4) -alkanecarboxylic acid or protected by a tetrahydropyranyl group,

R ² is a C 3 -C 15 saturated aliphatic group or a C 8 -C 12 unsaturated aliphatic group,

R ³ is hydroxy, C 1 -C 4 alkoxy or -NH-CH ₂ -CO-R ³ , substituted amino group of formula (-Rb) wherein R ³ , hydroxy or C 1 -C 4 alkoxy and -X- is a single bond, methylene or> CH-BH-R 4 (-XO) optionally N-acylated primary amino group in which

R ^{4 is} hydrogen or an acyl group of a polyhalo (C 2 -C 5) alkanoic acid with the oxygen atom of the hydroxy group in a relative trans configuration relative to the sulfur moiety, and the alkali metal or aliphatic ammonium salts of such compounds having salt-forming properties.

The spatial representation in the above formula (I) is understood to mean that the first row of symbols is above the representation plane and the third row is below (or vice versa), which corresponds to the opposite configuration of both central carbon atoms (RS) - (SR). of the formula according to the Kahn-Ingold-Prelog Convention.

The present invention also relates to a process for the preparation of pharmaceutical compositions containing the compounds of formula (I) as an active ingredient.

They are useful, for example, in the alleviation and elimination of disease states in which the compounds of the present invention exhibit a pronounced leukotriene antagonizing effect, as in various types of allergies, particularly asthma.

Several years ago it was shown that isolates from biological materials of various origins, known as SRS (slow-reacting substance of anaphylaxis), were already known in immunological studies: see HR Morris et al., Natúré 285, 1045-1060 (May 1980). ) and L. Ocrning, S. Hanunarström and B. Satnuclsson, Proc. Natl. Acad. Sci. USA 77, (4), 2014-2017 (1980), are identical to the study of arachidonic acid melabolism. leukotrienes. so e.g. it is clear from both of the aforementioned work that the active ingredient labeled as SRS-A, which is most likely to be the primary mediator of the hypersensitivity reactions that occur immediately after bronchoconstriction in asthma, leukotriene D (see below for the LTD formula). Similarly effective is leukotriene C, whose spatial structure has recently been established by total synthesis [E. J. Corey et al., (1980) J. Am. Chem. Soc. 102 (4): 1436-1439.

The structural backbone of Icukotrienck is generally a polyunsaturated, non-branched, eicosanoic acid having the characteristic 1-, 5- and 6-position stability as shown in Formula (a) above.

(Here, spatial representation is understood to mean that the entire olefin chain is in the plane of representation, and the dashed lines drawn by the arrow are above the plane and the dotted lines are below it.)

In terms of their physiological properties, leukotrienes are generally distinguished by the fact that they cause a characteristic contraction in a variety of smooth muscles. Such an effect from a health point of view is generally undesirable and, accordingly, the search for suitable maize antagonists in this field is at the forefront of research.

Surprisingly, it has now been shown that the compounds of formula (I) according to the invention, although they share a number of common structural features with known leukotrienes, exhibit pronounced antagonistic activity against them. Thus, they have a clear demonstration of leukotriene antagonism in various in vitro assays.

For example, they inhibit smooth muscle contraction induced by leukotriene D ₄ (L _{1 '} D ₄ ; see formula a) in the concentration range tested (0.1-25 μΜ / Ι). This is the so-called. Experimentally, LTD ₄ antagonism can be established as follows. Segments of a ileum from 303 to 400 g of teugcrimal piglets; and incubated in an organ bath in Tyrodeok at 38 ° C, saturated with a gas mixture containing 95% oxygen and 5% carbon dioxide at a load of 1 g. In the segments, synthetic leukotriene D ₄ gytl (as potassium salt) was induced (contraction) and recorded isotonic. The degree of inhibition induced by the test substance is determined after a two minute preincubation and is expressed as IC ₅₀ , the concentration which reduces the test contraction to 50%. LTD ₄ antagonism can also be detected in vivo by standard bronchoconstriction in guinea pigs. (Test methods are described in the Appendix after the examples.)

In another assay, compounds of general formula I inhibit leukotriene B ₄ (LTR ₄ ) -induced aggregation of rat peritoneal leukocytes at concentrations of 1-100 μΜόΙ / l. In the experiment, Wistar rats (400-600 g) were sacrificed 24 hours after injection of 16 ml of 12% sodium caseinate solution, the cells were washed from the peritoneum with buffered Eagles minimal essential medium (E-MEM), and 0.5-0.5. ml of cell suspension (10 ⁷ cells in 1 ml E-MEM) was added to the plate of a plate aggregomel and heated to 37 ° C with constant agitation (800-900 rpm). Four minutes after the addition of the test substance (2 μΐ), aggregation was induced by the addition of 2 μΐ LTB ₄ (1 ng / ml final concentration) and recorded. The concentration of test substance that reduces control aggregation (LTB ₄ alone) to 50% is called IC ₅₀ .

Unexpectedly, the compounds of formula (I) also inhibit other physiologically important enzyme systems. Thus, inhibition of human leukocyte phospholipase A ₂ can be observed at ca. In the concentration range of 0.5 to 50 μΜόΙ / Ι. (The experiment is described in more detail in the appendix after the examples.) Inhibition of human platelet-derived phospholipase C can also be observed for approx. In the concentration range of 1-100 μΜόΙ / l (the experiment is described in the Appendix after the examples).

196,364

The above methods are used to detect allergy and in vitro allergy. anti-inflammatory properties can also be demonstrated in animal experiments in vitro. Thus, the local antiinflammatory activity can be demonstrated, for example, by the method of G. Tonelli and L. Thibault (Endocrionology 77, 625 (1965)), ca. In a dose range of 1-100 mg / ml by testing for inhibition of crotil ear induced croton oil in normal rats.

Due to these valuable pharmacological properties, the compounds of the formula I can be used for therapeutic purposes whenever the allergenic effect of leukotrienes leads to disease and this effect must be reduced or eliminated. As a result, they are useful, for example, in allergic conditions and diseases, particularly asthma, but also in charcoal, obstructive pulmonary disease, and cystic fibrosis. At the same time, due to their anti-inflammatory activity, they are also useful as anti-inflammatory agents, especially as external (topical) skin phlogistics for the treatment of inflammatory dermatoses of all origins, such as mild skin irritations, contact dermatitis, rashes and burns, and mucosal phlogistics. for the treatment of mucositis of the eyes, nose, lips, mouth and genital and anal areas. They can also be used as sunscreens. The strong inhibitory effect on various blood factors indicates that the compounds of formula I can be used therapeutically in the indication range of thrombosis and coagulation.

As mentioned above, there is a general analogy between the structure of the compounds of formula I of the invention and the leukotrienes, in particular in the obligatory trans configuration of vicinal sulfur and oxygen atoms and in the introduction of -S-CH ₂ -XCO-R ^3. in the entire structure of the mercaptoalkanoic acid residue (A) (in particular, it is typically in the form of a cysteine peptide). However, they are significantly different from leukotrienes in that

OH

I

R ₂ -CH = CH-CH-C-CH ₂ -R ₁ , (B)

I

Their olefinic group of formula H lacks the characteristic terminal carboxyl group. In these vcgyiiletck, unlike leukotrienes, the influence of the number, nature and spatial structure of the multiple bonds and the total length of the olefinic group (B) on the compounds is insignificant, and the absolute configuration of the two symmetric carbon atoms discussed above is not critical. , such as high-activity N- [S-5 (E), 6 (S) -5-hydroxy-7,9-trans-11-cis-eicosrien-6-yl-N-trifluoroacetylcysteinyl See, for example, the sodium salt of glycine which has the absolute configuration of the 5 and 6 carbon atoms of the hydrocarbon chain as opposed to the natural leukotrienes.

In the general formula (1), R ¹ is preferably a C 1-3 alkyl group such as methyl, propyl and above all ethyl, or a C 1-3 hydroxyalkyl group, preferably a? -Hydroxyalkyl group, in particular? -hydroxyethyl, in which the hydroxyl group may be not only free but also esterified with C 1-4 alkanecarboxylic acid or protected with tetrahydropyranyl. The detected hydroxy group is preferably esterified with acetic acid.

Preferably, the aliphatic group represented by R ² is a non-branched group, e.g. an alkyl group having from 5 to 15 carbon atoms, preferably from 7 to 12 carbon atoms, in particular the heptyl, nonyl, undcylc dodecyl group, or a suitable mono- or poly-unsaturated C8-C12 group having one, two or three unsaturated bonds, Thus it contains triple bonds, but above all double bonds in any cis or trans configuration of choice, in any combination. These unsaturated bonds are preferably preferably located near the sulfur atom, that is, they are conjugated to the first double bond at the [beta] position relative to the carbon atom bearing the sulfur atom. This type of preferred R ² groups include straight chain C8-12 alkenyl Ι, 1,3-1,3,6-alkadienyl and alkatrienyl groups, in particular the 1-heptenyl, 1-octenyl -, 1-nonenyl, 1-decenyl, 1-undecenyl and 1-dodecynyl, and 1,3-ocladicynyl, 1,3-decadicnyl, 1,3-dodcadicnyl, and 1, 3,6-dodecatrienyl group in which the double bonds can be individually in the cis or trans configuration and in any combination.

The R ³ substituent of formula (I), together with the adjacent -CO- crononyl group, forms a free or functionally substituted carboxyl group; in the case where R ^{3 is} hydroxy, it forms the carboxyl group of a free carboxylic acid with the carbonyl group, when R ³ forms an carboxylic acid ester, especially an alkoxy group having up to five carbon atoms, in particular methoxy, and when R ^{3 is} an amino group, NH-CH ₂ - -CO-R b) where R 1 is hydroxy or C 1-4 alkoxy.

On the one hand, -X- as defined in the introductory part can be a single CC bond and thus forms, with adjacent groups, a m-captoacetic acid residue of the formula -S-CH ₂ -CO-R ³ ; in this case, it is particularly preferred that R ³ is hydroxy. Second, X- is an optionally acylated on the nitrogen atom of aminomethylene groups may be of the R ⁴ -NH-CH (-X _p -) corresponds to the formula wherein R ⁴ is hydrogen or a polyhaloC 2-5 an acyl group of a carbon-containing alkane carboxylic acid, for example chlorinated or fluorinated C 2 -C 5 alkane carboxylic acids such as trifluoroacetic acid.

The above-described aminomethylene moiety, together with adjacent moieties, forms an optionally acylated cysteine moiety of the general formula (-A _o ), wherein R ³ cs R ^{4 has} the above general and preferred meanings wherein L-cysteine is removed at its naturally occurring asymmetric carbon configuration. In this case, R ³ is preferably hydroxy, C 1-4 alkoxy, or a glycine group optionally esterified with a C 1-4 alkanol attached to the nitrogen atom, and especially R ⁴ is hydrogen, trifluoroacetyl or γ-glutamyl. (also in salt form).

Most of the compounds of formula (1), by their individual nature, may also exist in salt form. These

Of the 196,364 having sufficient acidity, especially those having a free carboxyl group, may form salts with bases, in particular inorganic bases, preferably physiologically tolerable alkali metal salts, in particular sodium and potassium salts.

Compounds of formula (I) which have sufficient basicity, such as esters and amides of amino acids, may form acid addition salts, in particular physiologically tolerable salts, with conventional pharmaceutically acceptable acids; of the inorganic acids, in particular the halogenated hydrocarbons such as hydrochloric acid and sulfuric and phosphoric acids, and the pyrophoric acid, in particular the sulfonic acids such as aromatic sulfonic acids such as benzene or -toluenesulfonic acid, embonic acid and sulfonyl acid, or lower alkanesulfonic acids such as methanesulfonic acid, cyananesulfonic acid, hydroxyoctanesulfonic acid and ethylene disulfonic acid, but also aliphatic, alicyclic, aromatic or heterocyclic carboxylic acids such as, succinic acid, glycolic acid, lactic acid, malic acid, tartaric acid, citric acid, fumaric acid, maleic acid, hydroxy-maleic acid, oxalic acid, pyruvic acid, phenylacetic acid, benzoic acid, p-aminobenzoic acid, anthranilic acid, p-hydroxybenzoic acid, salicylic acid salicylic acid and ascorbic acid. Compounds of general formula (I) which possess both basic and acidic functional groups, such as the free carboxyl and amino groups, may also exist as internal salts.

Of particular note are those compounds of formula I wherein the whole of the above-mentioned mercapto-alkanoic acid group is represented by one of the formulas (A1) to (A-6), with "natural" L-series amino acid groups being preferred. . Also included are corresponding compounds wherein the carboxyl group is in the form of a C 1 -C 4 alkyl ester or, in particular, an alkali metal salt.

In particular, the compounds of formula I described in the examples should be highlighted.

The thioethers of the invention may be prepared in a manner known per se, for example, by introducing an unsaturated aliphatic trans-cpoxide ring of formula (II) as defined in the introduction, wherein R ¹ and R ² are as defined above and on the oxirane ring. the two hydrogen atoms of each other having a trans orientation relative to one another and in which a hydroxy group, optionally present, may be in protected form with a mercaptoalkanoic acid corresponding to the group (A) as defined above, in particular R ³ and -X- is as defined above, which may be protected in the form of an amino group, optionally present, or a salt thereof or a derivative containing a modified carboxy group, and if necessary or desired, the hydroxyl and / or amino group (s). protecting group (s) and / or one the ester compound is saponified into the free acid or a salt thereof and, if desired, the resulting free salt-forming compound is converted into its salt or the resulting salt into the free compound.

The reaction is carried out under conditions well known in the art. -20 ° C to + 50 ° C, preferably room temperature, and in particular in the presence of a basic medium such as an amine, especially a tertiary, aliphatic, arylaliphatic or saturated heterocyclic amine such as a trialkylamine (e.g. triethylamine) or ethyldiisopropylnmin), dialkyl-benzylamine (e.g. N, N-dimethyl-benzylamine), Ν,,-dialkylaniline (e.g. Ν, Ν-dimethylaniline), or N-methyl or N-ethylpiperidine or N, N'-dimethylpiperidine. The reaction is usually carried out in an indifferent organic solvent such as a lower alkanol, e.g. methanol or ethanol.

If the starting material, especially the compound of formula 11, has a free hydroxyl group in the R * substituent, it may be protected during the reaction, so that it can be etherified. Preferably, the hydroxyl protecting group * 3 is easy to cleave, in particular acidolysis. ^Anther than those in general, especially in the peptide and szícroidkcmiában are well-known; in addition, tert-butyl, in particular tetrahydropyranyl ether (THP-ether) protecting groups are particularly preferred. These protecting groups can be removed after the main reaction (i.e., condensation of the epoxide with mercaptocarboxylic acid) by the release of the hydroxyl group in a generally known manner: pi. by treatment with an organic acid such as formic acid, acetic acid, oxalic acid or trihydrochloric acid, or a mixture thereof, in the presence of water and / or an inert organic solvent such as lower alkanol (e.g. methanol or ethanol) and cyclic ether (e.g. in the presence of tetrahydrofuran or dioxane).

If the mercapto-carboxylic acids used as starting materials contain a free amino group, they may preferably be in the protected, especially acylated, form during the main reaction. Preferably, amino protecting groups which are readily removable by acidolysis are used, as are generally well known, particularly in peptide chemistry, and the conditions for their removal are well known. Of particular interest among the amino protecting groups is the trifluoroacetyl group which, after the main reaction, may remain in the final product of the invention or, if desired, be subsequently cleaved. Cleavage of the N-trifluoroacetyl group is carried out in a known manner, preferably by hydrolysis, especially under basic conditions such as alkali metal carbonates (e.g. sodium or potassium carbonate) or dilute alkali bases (e.g. sodium or potassium hydroxide), in the presence of water, a water miscible organic solvent such as lower alkanol (e.g. methanol or ethanol) or a cyclic ether (e.g. tetrahydrofuran or dioxane), e.g. 0-80 hőmerseklc-cn ° C, preferably at a slightly elevated temperature, from 50 to 6 (i ° C. 1 Ia of the ester groups are hydrolyzed products, such as R ¹ OH of formula alkilcsaportban an acylated hydroxy group, or (A ) an esterified carboxyl group in a mercaptoic acid group, these are simultaneously saponified under these conditions.

The main action binary (addition to epoxide) is preferably m-nitro-acetic acid in an ester form, preferably in the form of a C 1 -C 4 alkyl ester (such as methyl or ethyl ester); if the final product of the invention is to be prepared as the free acid or its salt, the resulting ester must be hydrolyzed. The hydrolysis is carried out under conventional conditions, such as those described above for the base-catalyzed hydrolytic reaction of the N trifluoroacetyl group.

196 364 cleavage. But under milder conditions, especially at lower temperatures (preferably room temperature), equivalents of stoichiomelic amounts of alkali are available with a shorter reaction time, optionally under analytical control, e.g. by thin-layer chromatography, the ester group can be selectively cleaved while retaining the N-trifluoroacetyl group, but in most cases it is cleaved simultaneously.

The starting materials for the addition process according to the invention are either known per se or known analogous methods can be prepared in a manner known per se, e.g. important mercaptocarboxylic acids of formula (III) are described (e.g., E. J. Corex et al., Tetrahedron Letters 1980, 3143), and other analogous acids are available in the same manner from appropriate known starting materials. For the preparation of cysteine derivatives, it is preferable to use analogous known cysteine glycols and the disulfide bond is subjected to conventional reductive cleavage or is suitably protected as a cysteine derivative, e.g. trityl or acetylaminomethyl-protected mercapto group.

The unsaturated trans-epoxide compound of formula (III) used as starting material can be prepared, in particular, by the same methods as used for the synthesis of leukotrienes. In a typical general synthesis route e.g. starting from an unsaturated aliphatic aldehyde (alkanal) of formula IV wherein R 'is as defined above, wherein the free hydroxyl group present in R ¹ is protected as an ether, e.g. in a form as described above. This compound is condensed with formyl methylene triphenyl phosphorane (or an equivalent reagent value), and the corresponding α, β-unsaturated aldehyde (V) is formed 2-trans-alkene of formula, wherein R ¹ is as defined above and the free hydroxyl group which may be present in R ¹ is protected as an ester. This compound is finally epoxidized with aqueous hydrogen peroxide in a manner known per se, preferably under mildly alkaline conditions (e.g. in the presence of alkali carbonates), to give a trans epoxide, 2- (RS) -, 3- (SR) of formula (VI). ) -epoxy-alkane is formed in which R ¹ is as defined above and is protected in the form of an ester of the free hydroxyl group which may be present in R ¹ . This epoxy aldehyde is converted to the desired unsaturated aldehyde, e.g. the compound of formula (II) as defined above, wherein a free hydroxyl group which may be present in R ^{1 is in a} protected, esterified form, may be condensed with a corresponding known alkylidene triphenylphosphorane! with condensation. For polyunsaturated epoxides, e.g. (II) compounds of formula in which R contains ^two, one or more double bonds, is an indirect alternative serves the epoxy adehidet formula (VI) with an unsaturated chain ylidene phosphorane first formyl-methyl-trifcnil place of the Wittig reaction -phosphorane or γ-triphenylphosphorus anilidene croton aldehyde (4-trienylphosphoroanilidene-2-trans-butenal) is extended by two and four carbon atoms (1 or 2 double bonds) respectively 4 (RS) -, 5 (RS) -epoxy-2-alkenylated, respectively. 6 (RS) -, 7 (RS) -epoxy-2,4-alkadiene with a simple saturated alkylidene triphenylphosphorane or a less complicated alkylidene triphenylphosphorane to the desired polyunsaturated epoxide [e.g. condensed into a compound of formula II). j

If individual diastereomers are desired, preference is given to reacting the individual diastereomers of a starting material on the preferred horses with either a racemic or optically inactive starting material with stereoselective reaction conditions or an optically active reagent, preferably a diastereomer or a diastereomer. with the aid of optically active auxiliaries, it can be separated into an optically individual diastereomer.

However, from a stereochemical point of view, both the addition of the formation components II and III according to the invention and the preparation of the starting materials should preferably be carried out in such a way that the starting materials are always stereochemically homogeneous, preferably stereoselectively. using optically active reagents and / or auxiliaries, and stereochemically homogeneous products obtained from the reaction mixtures are isolated immediately after the reaction. so e.g. the cis and trans double bond isomers which may be formed in the preparation of unsaturated starting materials must be separated immediately from each other, for which conventional physical separation methods, in particular chromatography, are suitable. In the main section, the epoxide of formula II is used as the individual trans-stereoisomer, but is used in racemic form (as cz is usually formed by epoxidation of an olefin); the mercapto-alkanoic acid of formula (III), when optically active, is preferably used as an individual antipode (which is common in cysteine and its derivatives) - this solution allows both optically active diastereomers to be formed simply by known physical methods, e.g. . separating by chromatography; when an optically inactive mercapto-alkanoic acid is used, it is inevitable that the methods of cleaving the antipodes with optically active excipients, e.g. formation of salts with optically active bases. All suitable separation methods are known per se and may be repeatedly or sequentially combined.

Since there is a close relationship between the free compounds and the new compounds in the form of their salts, the corresponding salts, respectively, under the former and the stones in the free chemical or their salts, respectively. free compounds.

The invention also relates to embodiments of the process wherein the starting material is a compound which can be obtained in any step of the process and the missing step is carried out, or a starting material is used in the form of a salt or formed under the reaction conditions.

Preferably, starting materials and reaction conditions are selected so as to obtain the compounds described above as particularly advantageous.

The present invention also relates to the preparation of pharmaceutical compositions and medicaments which are a compound of formula (I) according to the invention or a pharmaceutically acceptable

They contain 196 364 salts. The pharmaceutical compositions according to the invention are particularly intended for topical administration and, above all, for administration by inhalation, e.g. in the form of an aerosol, a micropowdered powder or a fine spray solution - are available to the mammal and contain the active ingredient either alone or in combination with a pharmaceutically acceptable carrier.

For topical use of pharmaceutical compositions, e.g. for skin care - lotions and creams are emulsions and ointments in liquid or semisolid oil-in-water or water-in-oil (preferably containing a preservative). Particularly suitable for the treatment of the eye are granules containing the active ingredient in aqueous or oily solution and ointments, which are preferably prepared in sterile form. Aerosols and sprays (similar to those described below for use in the treatment of airways), coarse powders for rapid inhalation through the nostrils, and above all nasal formulations containing the active ingredient in aqueous or oily solution, are suitable for nasal treatment; lozenges suitable for the topical treatment of the oral cavity, containing the active ingredient in the form of a mass of sugar and gum arabic or tragagant, to which may be added flavoring agents, and lozenges which contain the active ingredient in an inert mass, e.g. containing by weight of gum arabic and gelatine and glycerol or sugar.

As pharmaceutical compositions, they are suitable for administration in the form of aerosols or sprays, e.g. solutions, suspensions or emulsions of the active ingredient of formula (I) according to the invention in a suitable pharmaceutically acceptable solvent, in particular ethanol and water, or a mixture thereof. They may also contain other pharmaceutical excipients, such as nonionic or anionic surfactants, emulsifiers or stabilizers, as appropriate, and other types of active ingredient, and are preferably admixed with a propellant gas at a pressure higher than the inert gas or particularly at atmospheric pressure, below normal room temperature (e.g., about -30 to +10 ° C), as at least a partially fluorinated polyhalogenated lower alkane, or a mixture of such liquids. Pharmaceutical compositions which are used primarily as intermediates or kit products in the form of solids for the preparation of the appropriate medicament generally have an active ingredient of ca. 0.1 to approx. 10% by weight, especially approx. 0.3 and approx. 3% by weight. For the preparation of a pharmaceutical formulation in ready-to-use form, such pharmaceutical composition is filled into a suitable holding container such as a bottle and a pressurized bottle, which may be sprayed with a suitable device for this purpose. are fitted with a valve. Preferably, the valve is constructed as a metering valve which, in use, permits a predetermined amount of liquid, corresponding to a predetermined dose of the active ingredient. Alternatively, the final dosage form can be prepared by separately filling and mixing the appropriate amount of the pharmaceutical composition and propellant as a stock solution. The administration of the compound of formula (1) to be administered and the frequency of administration always depend on the individual potency of the individual compound. duration of action, the disease or condition to be treated depends on the severity of the symptom, '' as well as the sex, age, weight <s of the mammal being treated, and the individual response. In general, the recommended daily dose of a compound of general formula (I) of the invention is about 75 kg for a 75 kg mammal (primarily humans). 10 cs approx. 500 mg, preferably between 25 and approx. It is preferably in the range of 250 mg and may be administered in several daily doses as needed.

The active compounds of formula (I) of the present invention are useful in the alleviation or reduction of the condition or symptom of a mammal, particularly a human, which is due to an allergenic effect of leukotrienes and particularly occurs in asthma. This application, respectively. A suitable treatment is characterized by treating the patient's body or body part with an anti-allergic amount of a compound of formula I, either alone or in the form of a medicament, in particular a pharmaceutical composition for inhalation. By "an allergy effective amount" is meant an amount of the active ingredient sufficient to significantly inhibit leukotriene-induced contraction.

The following examples further illustrate the invention without limiting its scope. All temperatures are in degrees Celsius. The amino acids, as components of the compounds described, are in the "natural" L-configuration.

/. example

N- {S- [5 (S, R), 65R, S) -5-hydroxy-7-trans, 9-trans,

11-cis, 14-cis-ejcatetraen-6-yl] -N-trifluoroacetylcysteine} glycine methyl ester and individual 5 (S) -, 6 (R) - and 5 (R) - , 6 (S)-diastereomers

103 mg (0.36 mmol) of 5 (S, R) -, 6 (S, R) -5,6-cpoxy-7-trans, 9-trans, 11-cis, 14-cis-ejcatetraene, 154 mg (0 (54 mmol) of a solution of N- (N'-trifluoroacetylcysteinyl) glycine methyl ester and triethylamine (145 mg, 1.44 mmol) in methanol (0.5 mL) was stirred at room temperature for 4 hours and with methanol (2 mL). diluted. Reverse phase chromatography in 2: 1 acetonitrile / water gives 212 mg (100%) of the diastereomeric mixture which is separated by subsequent high pressure chromatography on a Zorbax ODS RP column with 85:15 methanol / water and ca. . in equal proportions. Under these circumstances, the 5 (S) -, 6 (R) diastereoisomer has a shorter retention time than the 5 (R) -, 6 (R) diastereoisomer.

UV (in methanol): 5 (S), 6 (R) -isomer:

λ nax = 271, 281, 290 nm (ε ₂₈₁ = 40,600).

5; R), 6 (S) -isomer: _H x A = 271, 280, 290 nm.

IR (CH ₂ Cl ₂ ) is essentially the same for both diastereoisocarbons: 3570 (wide), 3370 (broad), 3000, 2950, 2920, 2850, 1742, 1720, 1680, 1518, 1460, 1435, 1380-1350, 1208. 1165, 1000 cm ¹ .

[α] D [5 (S), 6 (R) -isomer]: + 59 ± 1 ° C (1.4% in CHCl ₃ ).

-611

196,364

The racemic 5 (S, R), 6 (S, R) -5,6-epoxy-7-trans, 9-trans, 11-cis, 14-cis-eicosatetraene used as starting material can be prepared as follows:

A solution of pentanal (51.4 mL) and triphenylphosphoranilidene acetaldehyde (147 g) in chloroform (480 mL) was heated at reflux for 5 days, the solvent was distilled off under reduced pressure, and the resulting residue was stirred with 400 mL of 3: 1 hexane / ether. Crystalline precipitated triphenylphosphine oxide is suction filtered, the filtrate is concentrated in vacuo and the residue is distilled in vacuo. 24.5 g (45.2%) of 2-trans-heptenal are obtained in the form of a colorless oil.

MP: 60-65 C / 24 mbar.

Dissolve 2-trans-heptanol (2.27 g) in methylene chloride (24 ml) and methanol (44 ml) with stirring at 0 ° C. Add 2.04 ml of 30% aqueous H ₂ O ₂ and add 100 mg of solid potassium carbonate. The resulting reaction mixture was stirred for 3 hours at 0 ° C. Finally, 100 ml of methylene chloride are added and washed twice with 25-25 ml of pH 8.0 phosphate buffer. The aqueous portions were subsequently extracted with 50 ml of methylene chloride. The combined organic extracts were dried over magnesium sulfate and concentrated under reduced pressure. The crude product thus obtained is chromatographed on Merck Kieseigel 60 (85 g), eluting with methylene chloride, to give a total of 1.59 g (62%) of 2 (R, S), 3 (S, R) -2,3-eopoxyheptanol. obtained in the form of a colorless oil.

[Rf (silica gel; toluene / ethyl acetate 4: 1) = 0.59;

(! CH ₂ C ₂₎ IR: 2950, 2925, 2850, 2820, 1721, 1463,

1430, 1378, 1210, 850 cm ^-1 ).

To a solution of 640 mg of 2 (R, S), 3 (S, R) -2,3-epoxy-heptanol in 10 ml of methylene chloride at room temperature for 1 hour, 1.65 g of γ-triphenylphosphoranilidene crotonaldehyde in 15 ml of methylene chloride solution was added dropwise and the resulting reaction mixture was stirred for an additional hour at room temperature. For work-up, the reaction mixture is concentrated under reduced pressure and the residue is chromatographed on a silica gel column (40 g) in toluene / ethyl acetate (19: 1). A mixture of cis and trans isomers is obtained which is dissolved in 5 ml of low iodine crystalline methylene chloride for isomerization and allowed to stand at room temperature for 4 hours. The solution is then concentrated in vacuo and chromatographed as described above. Yield: 511 mg (57%) of 6 (S, R), 7 (S, R) -6,7-epoxy-2-trans, 4-trans-undecadienal as a yellowish oil.

UV (ethanol): A _max = 276 nm;

e = 29,900;

IR (CH ₂ Cl ₂ ): 2950, 2920, 2850, 2800, 2720, 1678,

1640, 1600, 1460, 1163, 1120, 1007, 985 cm ^-1 ;

Rf [Kieseigel; toluene / ethyl acetate (4: 1)] = 0.56.

378 mg of 3-cis-nonen-1-yl triphenylphosphonium toluenesulfonate [I. Ernest, A.J. Main, R. Menassé, Tetrahedron Letters 23, 167 (1982)], 4.2 ml of tetrahydrofuran and 1.26 ml of hexamethylphosphoric triamide at -78 ° C under an atmosphere of 0.31 ml of 20 butyl lithium solution in hexane dropwise and the resulting solution was stirred at -78 ° C for an additional 30 minutes. The resulting triphenylphosphoric anilidene-1-cis-nonenylaliosis at -78 ° C was 110 mg of 6 (S, R), 7 (S, R) -6,7-epoxy-2-trans, 4-trans- undecadienal A solution of knife in 1.0 mL of tetrahydrofuran was added dropwise and the resulting reaction mixture was stirred at -78 ° C for 30 minutes. For work-up, the reaction mixture was partitioned between 100 mL of ether and 30 mL of 8.0 µl phosphate buffer, and both phases were treated with ether or ether. extraction with buffer solution. The combined ethereal phases were dried over magnesium sulfate and concentrated in vacuo to give 421 mg of an oily crude product. This was mixed with 2-3 mL of a 3: 1 mixture of hexane-ether, the crystalline triphenylphosphine oxide was separated and the filtrate was stored. The resulting residue (195 mg) was chromatographed on an alumina column (10 g) prepared in hexane containing 0.5% triethylamine using the same eluent. 90.6 mg (57%) of 5 (S, R), 6 (S, R) -5,6-epoxy-7-trans, 9-trans, 11'-cis, 14-cis-cis -cyanate are obtained. in the form of a yellowish oil.

UV (methanol): λmax - 270, 280, 291 nm; ε _20Ο = _59,600 ;

IR (CH ₂ Cl ₂ ): 3000. 2900, 2850, 1450, 1370, 992, 963 cin '.

Example 1A

N- {S- [5 (S), 6 (S) -5-Hydroxy-7-trans-, 9-trans-, β-cis, 14-cis-ejcatetraen-6-yl] -N-trifluoro- acetyl-cysteinyl] glycine methyl ester [from optically individual epoxy-olefin]

130 mg (0.450 mmol) of 5 (S), 6 (S) -5,6-epoxy-7-trans, 9-trans, 11-cis, 14-cis-ejecatetraene, 260 mg (0.902 mmol) of N- (N). A solution of trifluoroacetylcysteinyl) glycine methyl ester and 250 ml (1.8 mmol) of triethylamine in 1.0 ml of methanol was stirred under argon at room temperature for 3.5 hours and then diluted with a small amount of methanol on a 12 RP sheet. (Opti-UPC ₁₂ , NATECH AG, Bennvil, Switzerland, 20x20 cm) is chromatographed in acetonitrile: water (1: 1). The title compound (216 mg) thus obtained is chromatographed again on a high pressure RP column (Zorbax ODS) in methanol: water = 87:13.

122 mg of pure title compound are obtained which is identical in all respects to the diastereoisomer with a shorter retention time (see Example 1).

Aniax ⁼ 271, 281,290 nm, (ε ₂₈₁ = 40,600);

[a] ² D = + 59 ° ± 1 ° C (1.4% in ₃ CIlCl).

The optically active starting material, i.e., 5 (S), 6 (S) -5,6-epoxy-7-trans, 9-trans, 11-cis, 14-cis-ejection, is prepared as follows:

Lithium aluminum hydride (g) was dissolved in ether (400 mL) and 16.9 g (2 mL) of 2-heptinol dissolved in 200 mL of ether was added dropwise over 30 minutes with stirring and the resulting reaction mixture was refluxed overnight. The excess LiAlH * was quenched by addition of 40 mL of ethyl acetate while cooling in an ice-water bath and the resulting reaction mixture was taken up in ether and cold 1N sulfuric acid. The acidified (pH = 2) aqueous phase

-713

196,364 were re-extracted with ether, the combined organic extracts dried over magnesium sulfate and concentrated in vacuo. The residue (18 g) was distilled under reduced pressure to give 13.2 g of 2-trans-7-heptanol as a colorless oil.

Mp: 71.5 ° C-2 ° C / 13 mbar.

2.28 g (20 mmol) of 2-trans-heptanol are dissolved in 10 ml of methylene chloride and added at -20 ° C to a solution of 5.94 ml of tetrakisopropyl orthotitanal and

It contains 4.12 g of dictyl ester of L - (+) - tartaric acid dissolved in 210 ml of methylene chloride, followed by addition of 9.75 ml of a 4.1 M solution of t-butyl hydroperoxide in 1,2-dichloroethane. The resulting reaction mixture was allowed to stand overnight at -20 ° C. After addition of 8 ml of dimethyl sulfide, the mixture is stirred at -20 to -23 ° C for 45 minutes, then 50 ml of 10% aqueous L - (+) - tartaric acid solution is added and 30 minutes at -20 ° C. stirring without cooling for a minute. The organic layer was separated, washed with water (100 mL) and, after drying over magnesium sulfate, concentrated under reduced pressure. The residue is dissolved in 150 ml of ether, stirred at 0 ° C for 30 minutes with 60 ml of 1N sodium hydroxide, the aqueous phase is separated off, extracted again with ether and the combined organic extracts are brine. Shake. The organic layer was dried over magnesium sulfate and the solvent was distilled off in vacuo to give 2.3 g of 2 (S), 3 (S) -2,3-epoxy-leptanol as a colorless, unstable oil. This will be further processed immediately.

A solution of 1.2 g of 2 (S), 3 (S) -2,3-cpoxy-heptanol in 28 ml of methylene chloride was added at room temperature to a freshly prepared solution of 70 ml of niethylene containing 5.5 g of chromium trioxide and 8.76 g of pyridine. chloride solution and the resulting reaction mixture is stirred for 30 minutes more. The dark reaction mixture was decanted from the precipitated material, washed with methylene chloride (160 mL) and the combined organic extracts washed with 80 mL of pH 8.0 phosphate buffer. After drying over magnesium sulfate and evaporation under reduced pressure, the crude product is chromatographed on 90 g of Merck Kieselgel in toluene / ethyl acetate (4: 1). 464 mg of 2 (R), 3 (S) -2,3-epoxy-heptanol are obtained in the form of a colorless oil.

[α] D = + 101 ± 1 ° C (1.225% in CHCl ₃ );

IR (CH ₂ Cl ₂ ): 2950, 2925, 2860, 2815, 2730, 1722,

1462, 1432, 1380, 1360, 1230, 1156, 850 cm ^-1 .

804 mg of γ-triphenylphosphoranilidene crolonaldehyde are dissolved in 10 ml of methylene chloride and the solution is added dropwise at room temperature over 1 hour with 260 mg of 2 (R), 3 (S) -2,3-epoxyheptanol in 10 ml of methylene chloride and the resulting reaction mixture was then stirred for 1.5 hours. The solution was then concentrated under reduced pressure for ca. Concentrate to 2 mL and immediately chromatograph on a 20 g Kieselgel 60 column with toluene / ethyl acetate (19: 1). 120 mg of a mixture of cis, trans and trans, trans isomers are obtained, which is dissolved in 2 ml of methylene chloride with 2 g of iodine and allowed to stand at room temperature for isomerization. After 2.5 hours, the solution was concentrated in vacuo and chromatographed again as described above to afford 103 mg of pure, optically active 6 (S), 7 (S) -6,7-epoxy-2-trans, 4-trans-undecadienol. It is a yellow oil which crystallizes at -20 ° C.

= -28 + 1 C (0.735% in CllCl ₃ ).

90 mg of the latter compound in 2 ml of abs. dissolved in tetrahydrofuran at -78 ° C, condensed with a solution of triphenylphosphoranilidene-3-cis-nonene, 338 mg, in an appropriate solution of the corresponding phosphonium toluene sulfonate, which is solubilized in butyl lithium and processed as described for the racemic product (see Example 1). 132 mg (91%) of 5 (S), 6 (S) -5,6-epoxy-7-trans, 9-trans, 11-cis, 14-cis, 2-cis-ethacetraene are obtained in the form of a yellowish oil.

_[Tt t ² = -23 ° + 1 (0.81% in CHCl _3).

Example IB

N- {S- (5 (S), 6 (R) -5-Hydroxy-7-trans, 9-trans, 11-cis, 1'-cis-cis-calsactrac-6-yl-cis (cinyl)) glycine potassium salt mg N- {S- [5 (S), 6 (R) -5-hydroxy-7-trans, 9-trans, 11-cis, 14-cis-ejcatetraen-6-yl] -N-trifluoro The acetyl-cystinyl-glycine methyl ester was dissolved in tetrahydrofuran (3.8 mL) and methanol (3.8 mL) and added to a solution of potassium carbonate (276 mg) in water (11.5 mL) and stirred at room temperature under argon for 44 hours. Finally, it is concentrated under reduced pressure to about 3 ml and transferred to a reversed phase plate by chromatography on acetonitrile / water (1.1) to give 40 mg of the title compound which is kept in 5.0 ml of ethanol at -78 ° C.

UV (ethanol): λ _max = 271, 280, 290 bm;

280 ⁼ 48,600.

Example 1C

Potassium salt of N- {S- [5 (R), 6 (S) -5-hydroxy-7-trans, 9-trans, 11-cis, 14-cis-ejcetetraen-6-yl] -cysteinyl} -glycine

In the same manner as in the previous Example IB, 32 mg of N- {S- [5 (R), 6 (S) -5-hydroxy-7-trans, 9-trans, 11-cis, 14-cis] were prepared. 8.5 mg of the title compound of ejection cortra-6-yl) -N-trifluoroacetylcystienyl] -glycine were stored in ethanol at -78 ° C.

UV (ethanol): _.lambda.max = 270, 280, 290 nm;

ε ₂ 8ο = 49,000.

Example 2

N- (S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-heptanadecin-6-yl-N-trifluoroacetyl-cis-ylcyl) glycine methyl ester

480 mg of 5 (RS), 6 (R, S) -5,6-epoxy-7-cis-heptadecene (E1) and 660 mg of N- [N-trifluoroacetylcysteinyl] glycine-815

196,364

-mcli-ester [E. .1. To a solution of Corey cl al., Telrahedron Lelt 1980, 3143], 6 ml of methanol was added 0.78 ml of triethylamine. The solution was stirred at room temperature under argon for 16 hours, the solvent was evaporated in vacuo and the residue was purified by chromatography on silica gel with dichloromethane / ethyl acetate (85:15). The title compound is also obtained in the form of an oil. ι

IR (C1I ₂ CI _2): 3400, 2940, 2870, 1755, 1740, 1690, 1530 cm⁻¹.

In a similar manner, the compounds of the following Examples were prepared:

Example 3

N- [S-5 (RS), 6 (S, R) -5-hydroxy-7-cis-undecen-6-yl-N-1-fluoroacetylcystinyl] -glycine methyl ester ntg 5 (RS), From 6 (RS) -5,6-cpoxy-7-cis-undcene (E2), 486 mg of N- (N-trinylacetylcystycinyl) glycine incl.

IR (CH ₂ Cl ₂ ): 3400, 2960, 2940, 2880, 1750, 1730,

1640, 1525 cnr '.

*.

Example 4

N- (S-5 (FI)), 6 (SR) -5-hydroxy-7-cis-tridecene-6-yl-N-trifluoroacetyl-cysteinyl-N-glycine methyl ester

0.5 g of 5 (RS), 6 (RS) -5,6-epoxy-7-cis-tridecene (E3) and 0.8 g of N- (N-trichloroacetylcysteinyl) glycine inclyl ester .

IR (CH ₂ Cl ₂ ): 3400, 2980, 2940, 2870, 1760, 1740,

1700, 1535 cm ^-1 . Yield: 78%.

Example 5

S-5 (RS), 6 (SR) -5-hydroxy-7-cis-tridecene-6-yl-cysteine methyl ester

0.5 g of 5 (RS), 6 (RS) -5,6-epoxy-7-cis-tridecene (E3) and 0.87 g of cysteine methyl ester hydrochloride.

IR (CII ₂ Cl ₂ ): 3600, 3400, 2970, 2950, 2880, 1750,

1445 cm ^- '. Yield: 53%.

Example 6

S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-1-piperidin-6-ylmethylcaploacetic acid, ethyl ester

0.72 g of 5 (RS), 6 (RS) -5,6-epoxy-7-cis-tridecene (E3) and 0.43 g of methyl mercaptoacetic acid.

IR (CH ₂ Cl ₂ ): 3600, 2980, 2940, 2880, 1745, 1475,

1445 cm ^-1 . Yield: 80%.

Example 7

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-pentadecen-6-yl-N-trifluoroacetyl-cis-cinyl] -glycine-methyl-ester

0.5 g of 5 (RS), 6 (RS) -5,6-epoxy-7-cis-pentadecene (E4) and 0.71 g of N- (N-trifluoroacetylcysteinyl) glycine methyl- észlcrből.

IR (CII ₂ Cl ₂ ): 3400, 2970, 2950, 2870, 1760, 1740,

1695, 1530 cm ^-1 . Yield: 76%.

Example 7a

S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-pentladccn-6-yl-N-trifluoroacellyl cysteine methyl ester

0.5 g of 5 (RS), 6 (ES) -5,6-epoxy-7-cis-pentadecene (E4) and 0.58 g of N-trifluoroacetylcisleine methyl ester.

R f = 0.6 (silica gel, ethyl acetate / licxane (3: 2)). Yield: 41%.

Example 8

S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-pentadeccen-6-yl-cis-cinyl-incyl ester

0.5 g of 5 (RS), 6 (RS) -5,6-cpoxy-7-cis-pentanedecene (E4) and 0.77 g of cysteine methyl ester hydrochloride.

IR (CH ₂ Cl ₂ ): 3600, 3400, 2970, 2940, 2870, 1750,

1475, 1445 cm ^-1 . Yield: 64%.

Example 9

Methyl S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-pentadecene-6-yl-mercaptoacetic acid

0.9 g of 5 (RS), 6) RS) -epoxy-7-cis-butylcutadecene (E4) and 0.47 g of 2-transcapto-acetic acid methyl ester.

IR (CH ₂ Cl ₂ ): 3600, 2970, 2990, 2870, 1750, 1475,

1445 cm ^-1 . Yield: 83%.

Example 10

S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-heptanadecin-6-yl-cisleinyl methyl ester Ig-5 (RS), 6 (RS) -5,6-epoxy-7- cis-bcptadecene (E1) and 1,4 cysteine methyl ester hydrochloride.

IR (C ₁₂ Cl ₁₂ ): 3600, 3400, 2940, 2S70, 1750, 1475 cm -1. · Yield. 61%.

Example 11

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-eicosen-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

1 g of 5 (RS), 6 (RS) -5,6-cpoxy-7-cis-cytosine (E5) in 1 g of N- (N-trifluoroacetyl-cystinyl) glycine methyl ester.

IR (CII ₂ Cl ₂ ): 3400, 2930, 2860, 1750, 1735, 1690,

1530 cm ^- '. Yield: 89%.

-917

196 364/2. example

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-triclosen-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

0.6 g of 5 (RS), 6 (RS) -5,6-cpoxy-7-cis-tricosose (E6) and 0.58 g of N- (N-trifluoromethyl-cysticyl) glycine mclil -észtcrből.

IR (CH ₂ Cl ₂ ): 3400, 2940, 2870, 1760, 1740, 1700,

1530 cm ^-1 . Yield: 87%.

Example 13

N- {3- [4- (RS), 5 (SR) -4-Hydroxy-6-cis-tetradecen-5-ylthio-1-propionyl} glycine methyl ester

300 mg of 4 (RS), 5 (RS) -4,5-epoxy-6-cis-tetradcene (E7) and 275 mg of N- (3-mercaptopropionyl) glycyl methyl ester [A. Okuyama et al., Chem. Pharm. Bull. 30, 2453 (1982)]. Yield: 63%.

IR (CH ₂ Cl ₂ ): 3450, 2970, 2940, 2870, 1760, 1690,

1525 cm ^- '.

Example 14

N- [S-4 (RS), 5 (SR) -4-Hydroxy-6-cis-tetradecen-5-yl-N-trifluoroacetylcysteine] glycine methyl ester

0.7 g of 4 (RS), 5 (RS) -4,5-epoxy-6-cis-trcradccene (E7) and 1.2 g of N- (N-trillluoroacetylcysteinyl) glycine methyl ester .

IR (CH ₂ Cl ₂ ): 3400, 2980, 2940, 2870, 1760, 1740,

1700, 1530 cm ^-1 . Yield: 45%.

Example 15

N - {- 3- (4 (RS), 5 (SR) -4-Hydroxy-6-cis-nonadecen-5-ylthio] -propionyl] -glycine methyl ester

250 mg of 4 (RS), 5 (RS) -4,5-cpoxy-6-cis-nonadcceil (E8) and 175 mg of N- (3-methylcaptopropionyl) glycine methyl ester.

IR (CH ₂ Cl ₂ ): 3450, 2980, 2940, 2870, 1760. 1690,

1530 cm ^- '.

Example 16

N- [S-4 (RS), 5 (SR) -4-Hydroxy-6-cis-nonadcene-5-yl-N-trifluoroacetylcysteinyl] glycine inethyl ester

500 mg of 4 (RS), 5 (RS) -4,5-epoxy-6-cis-nonadecene (E8) and 20 mg of methyl N- (N-trifluoracetylcis-lcinyl) glycine.

IR (CH ₂ Cl ₂ ): 3400, 2970, 2940, 2870, 1760, 1740,

1700, 1530 cm ^- '. Yield: 99%.

Example 17

N- [S-4 (RS), 5 (SR) -4-Hydroxy-6-cis-eicosene-5-yl-N-trifluoroacetyl-cysteinylglycine ester ester

680 mg of 4 (RS), 5 (RS) -4,5-epoxy-6-cis-eicosene (E9) cs 830 mg of N- (N-trityloylacetylcysteinyl) glycine inclyl ester.

IR (CH ₂ Cl ₂ ): 3400, 2940, 2870, 1760, 1740, 1700,

1530 cm ^- '. Yield: 40%.

/ Example 7a

N- {3- [4- (RS), 5 (SR) -4-Hydroxy-6-cis-eicosene-5-ylthio] -propionyl} -glycine methyl ester

680 mg of 4 (RS), 5 (RS) -4,5-epoxy-6-cis-cis-cosene. (E9) and 720 mg of N - (? - mercaptopropionyl) glycine melyl ester.

IR (CH ₂ Cl ₂ ): 3450, 2450, 2880, 1760, 1690, 1525,

1225 cin ^- '. Yield: 34%.

Example 17b

S- [4 (RS), 5 (SR) - '1-yldioxy-6-cis-cis-cosyl-5-yl] -N-trifluoroacetylcystinomethyl ester

From 680 mg of 4 (RS), 5 (RS) -4,5-epoxy-6-cis-eicosene (E9), 620 mg of N-trifluoroacrylcystinomethylmethyl ester.

IR (CII ₂ Cl ₂ ): 3420, 2940, 2870, 1760, 1740, 1540,

1220, 1180 cm ^-1 . Yield: 45%.

Example 18

N- [S-6 (RS), 7 (SR) -6-Hydroxy-8-cis-eicosene-7-yl-trichloroacellyl-ciscininyl] -glycine methyl ester

0.54 g of 6 (RS), 7 (RS) -6,7-epoxy-8-cis-eicosene (E10) and 0.53 g of N- (N-trifluoroacetyl-cysteinyl) glycine methyl ester .

IR (CM ₂ Cl ₃ ): 3400, 2940, 1870, 1760, 1740, 1700,

1530 cm ^- '.

Example 18a

S- [6 (RS), 7 (SR) -6-Hydroxy-8-cis-eicosene-7-yl) -cysteine methyl ester

0.54 g of 5 (RS), 7 (RS) -6,7-epoxy-S-cis-ejkozcn (E10) and 0.45 g of cysteine methyl ester.

R f = 0.19 (CH ₂ Cl ₂ / ethyl acelate 85:15). Yield:

%.

Example 19

N- [S-5 (RS), 6 (SR) -5-Hydroxy-1-tetrahydropyranyl-2-oxy-7-cis-octadecen-6-yl-N-trifluoroacetyl-cysteinyl] glycine methyl ester

0.7 g of 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyl10

-1019

196,364

-2-oxy-7-cis -ocadadecene (Eli) and 0.6 g of N- (N-trifluoroacetyl-cysteinyl) glycine methyl ester.

IR (CH ₂ C ( ₂ ): 3400, 2940, 2870, 1740, 1700, 1530 cm -1 ⁾ . Yield: 98%.

Example 19a

S-5 (RS), 6 (SR) -5-Hydroxy-1-tetrahydropyranyl-2-oxy-7-cisoctadecen-6-yl mercaptoacetic acid methyl ester g 5 (RS), 6 (RS) ) -5,6-epoxy-1-tetrahydropyranyl-2-oxy-7-cis-octadecene (E1) and 0.35-mercapto-acetic acid methyl ester.

IR (CH ₂ Cl ₂ ): 3600, 3500, 2940, 2870, 1745 _cm -1. Yield: 81%.

Example 20

N- (S-5 (RS), 6 (SR) -5-Hydroxy-1-tetrahydropyranyloxy-7-cis-eicosene-6-yl-N-trifluoroacetyl-cysteinyl) -glycine methyl ester

0.8 g of 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyloxy-7-cis-eicosene (E12) and 0.64 g of N- (N-trifluoroacetyl-cysteinyl) glycine methyl ester.

IR (CH2Cl2): 3400, 2940, 2860, 1760, 1740, 1700,

1530 cm ^-1 . Yield: 99%.

Example 20a

S-5 (RS), 6 (SR) -5-Hydroxy-1-tetrahydropyranyl-2-oxy-7-cis-ejosan-6-yl mercaptoacetic acid methyl ester

0.8 g of 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyl-2-oxy-7-cis-eicosene (E12) and 0.4 g of mercaptoacetic acid methyl ester .

IR (CH ₂ CI ₂₎ = 3600, 3500, 2940, 2870, 1745 cm ^- '.

Yield: 56%.

Example 21

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7-trans-9-cis-nonadecadien-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

0.55 g of 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cisnonadecadiene (E13) and 0.62 g of N- (N-trifluoroacetylcysteinyl) glycine methyl ester.

R f = 0.3 (3: 2 hexane: ethyl acetate). Yield: 46%.

Example 21a

N- (-3- (5 (RS), 6 (SR) -5-hydroxy-7-trans-9-cis-nonadecadieno-yl) thio-propionyl) glycine methyl ester

0.55 g of 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-nonadecadiene (E3) and 0.53 g of N- (3-mercaptopropionyl) glycine methyl ester.

IR (CH ₂ C1 _2): 3600, 3450, 2940, 2870, 1760, 1525 cm ^- · '. Yield: 39%.

Example 21b

S- (5 (RS), 6 (SR) -5-Hydroxy-7-trans-9-cis-nonadckadicn-6-yl) -cystcin-n-ethyl ester

0.55 g of 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-nonadecadiene (E3) and 0.40 g of cysteine methyl ester.

IR (CH ₂ Cl ₂ ): 3600, 3400, 2940, 2870, 1750 cm -1.

Yield: 82%.

Ic example

S- (5 (RS), 6 (SR) -5-Hydroxy-7-trans-9-cis-nonadecadien-6-yl] -mercaptoacetic acid methyl ester

0.55 g of (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-nonadecadiene (E13) and 0.32 g of mercapto-c-acetic acid m.c.

IR (CH ₂ Cl ₂ ): 3600, 2940, 2870, 1745 cm -1. Yield: 39%.

Example d

S- [5 (RS), 6 (SR) -5-Hydroxy-7-trans-9-cis-nonadecadien-6-yl] -mercaptoacetic acid

0.55 g of 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-nonadacadiene (E3) and 0.30 g of mercaptoacetic acid.

R f = 0.3 (CH ₂ Cl ₂ / methanol 15: 1). Yield: 71%.

Example 2le

N- [S-5-hydroxy-7-trans-9- cis ejkozadien-6-yl-N-trifluoroacetyl-ciszteinilj-glycine methyl ester; diastereomers 5 (R), 6 (S) [A] and 5 (S), 6 (R) [B] (by chromatography)

0.6 g of racemic 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-eicosadiene (E13a) and 52 g of N- (N-trifluoroacetylcysteinyl) - from glycine methyl ester in 88% yield, the mixture of the two diastereoisomers was ca. It is obtained in a 1: 1 ratio, which is chromatographed on silica gel (7: 3) with hexane / ethyl acetate to give the individual forms [A] and [B] of the title compound.

(A) -isomer R (CH ₂ Cl ₂ ): 3630, 3400, 2940, 2870, 1760,

1700, 1535 cm -1.

[B] -isomer R (CH ₂ Cl ₂ ): 3630, 3400, 2940, 2870,

1760, 1695, 1535 cm -1.

Example f

N- [S-4 (RS), 5 (SR) -4-Hydroxy-6-trans-8-cis-nonadcadien-5-yl-N-trifluoromethyl-cysteinyl] -glycine methyl ester

0.6 g of 4 (RS), 5 (RS) -epoxy-6-trans-8-cis-nonadecadiene (E13b) and 0.52 g of N- (N-trifluoroacetyl-cysteinyl) glycine methyl ester .

IR (CH ₂ Cl ₂ ): 3391), 2930, 2860, 1755, 1730, 1690,

1525, 1215, 1170 cm -1. Yield: 27%.

-1 121

196,364

Example 22

N- [S-5-hydroxy-l-tetrahydropyranyl-2-oxy-7-trans-9- cis-octadecadienoic-6-yl-N-trifluoroacetyl-cysteinyl} -glycine methyl ester; diastereomers 5 (R), 6 (S) [A] and 5 (S), 6 (R) [B] (by chromatography)

0.6 g of Racine from 5 (RS), 6 (RS) -5,6-cpoxy-1-tetralidopyranyl-2-oxy-7-trans-9-cisoctadecadiene (E1) and 0.52 g of N- (N) Trifluoroacetyl cysteinyl) glycine methyl ester was obtained in 78% yield, each diastereomer being ca. 1: 1 ratio. This was separated into the title compound [A] and [B] by chromatography on silica gel (1: 1) with hexane / ethylene acetate. Both compounds show essentially the same spectrum:

IR (CH ₂ C _2): 3400, 2940, 2870, 1760, 1740, 1695,

1530 cm -1.

the example

S- [5 (RS), 6 (SR) -5-Hydroxy-1-telrabidropyranyl-2-oxy-7-trans-9-cis-octadecadien-6-yl] -mercaptoacetic acid methyl ester

0.9 g of 5 (RS), 6 (RS) -cpoxy-1-l-tetrahydropyranyloxy-7-trans-9-cis-ocladecadecin [E14] and 0.45 g of methacaptoacetic acid methyl ester.

IR (CH ₂ Cl ₂ ): 3600, 3460, 2940, 2870, 1745 cm -1.

Yield: 68%,

Example 23

N- [S-5-hydroxy-1-tetrahydropyranyl-2-oxy-7-trans-9-cis-eicosadien-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester: 5 (R), 6 (S) - [A] and 5 (S), 6 (R) - [B] diastereomers (by chromatography)]

0.5 g of racemic 5 (RS), 6 (RS) -5,6-epoxy-1-tetrabidropyranyl-2-oxy-7-trans-9-cis-eicosadiene (E15) and 0.4 g of N- (N). Trifluoroacetylcysteinyl) glycine methyl ester (73% yield), the two diastereoisomers were ca. A 1: 1 mixture is obtained which is separated by chromatography (silica gel, 1: 1) on silica gel, eluting with the title compound (AJ and [B]).

IR (CH ₂ Cl ₂ ): 3400, 2940, 2870, 1760, 1740, 1700,

1530 cm -1.

Example 23a

S-5 (RS), 6 (SR) -5-Hydroxy-1-tetralidopyranyl-2-oxy-7-trans-9-cis-eicosadien-6-yl] -mercaptoacetic acid methyl ester

0.91 g of 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyl-7-trans-9-cis-eicosadiene (E15) and 0.45 g of mercaptoacetic acid methyl ester .

IR (CH ₂ Cl ₂ ): 3600, 3470, 2940, 2870, 1745 cm -1.

Yield: 68%.

Example 24

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7,9-trans-11-cis-lindexadecatriene-6-yl-N-trifluoroacetyl-cysteinyl] glycine methyl ester

1.1 g of 5 (RS), 6 (Rs) -5,6-epoxy-7,9-trans-11-cis-lexadecatriene (EI6) and 1.3 g of N- (N-trifluoroacetylciscincin) glycine methyl ester.

IR (CH ₂ Cl ₂ ): 3600, 3400, 2980, 2950, 2880, 1740,

1700, 1530 cm ^-1 . Yield: 64%.

This diastereomeric mixture was chromatographed on silica gel with hexane / ethyl acetate 4: 1 to separate the two optically uniform forms: first, the 5 (S), 6 (R) diastereomer was eluted, followed by the 5 (R), 6 (S) diastereomer. .

Example 25

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7,11-cis-9-trans-licyxadecatrienyl-N-trifluoroacetylcysteinyl] glycine methyl ester

0.5 g of 5 (RS), 6 (RS) -5,6-epoxy-7,11-cis-9-trans-hexadecatriene (E17) and 0.62 g of N- (N-trifluoroacetyl) ci: ; ztcinil) glycine methyl észlerből.

IR (CH ₂ Cl ₂ ): 3400, 2970, 2950, 2880, 1740, 1700, 530 cm -1. Yield: 91%.

This diastereomeric mixture was separated by chromatography on silica gel (2: 1) hexane: ethyl acetate to give each of the two optically uniform forms: first the diastereoisomer 5 (S), 6 (R) was eluted, [? 4 ° C, followed by 5 (R), 6 (S) diastereomer, _[Q j = -41.4 ± 2.9 ° C;

both values were measured in chloroform solution at a concentration of 0.255% (g / v), respectively. 0.35% (g / v).

Example 25a

N- [S-5 (RS), 6 (SR) -5-hydroxy-7,9-trans-1-cis-octadecatrien-6-yl-N-trifluoroacetylcysteine] glycine methyl ester

'), 5g5 (RS), 6 (RS) -5,6-cpoxy-7,9-trans-11-cisoctadecatriene (E17a) and 0.62 g of N- (N-trifluoroacetylcysteinyl) glycine methyl -észterbő !.

IR (CH ₂ Cl ₂ ): 3610, 3400, 2980, 2950, 2870, 1760,

1740, 1695, 1530 cm -1. Yield: 44%.

Example 26

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7,9-trans-11-cis-eccathenyl-6-yl-N-trifluoroacetyl-cysteinyl] glycine methyl ester

0.54 g of 5 (RS), 6 (RS) -5,6-epoxy-7,9-trans-11-cis-eicosacrylene (E18) and 0.58 g of N- (N-trifluoroacetylcysteinyl) -g! için methyl ester.

-1 223

196,364

IR (CH ₂ Cl ₂ ): 3610, 3400, 2970, 2940, 2870, 1760,

1740, 1700, 1530, 1220, 1175 cm ^-1 . Yield:

%.

Chromatography on silica gel with hexane / ethyl acetate (3: 2) separates this disomeric mixture into individual optically uniform forms; in the first fractions the 5 (S), 6 (R) -distributor is followed by N-IS-5 (R), 6 (S) -5-hydroxy-7,9-trans-11-cis-ejcatriene 6-yl-N-trifluoroacetyl-cysteinyl] glycine methyl ester, which, like its diastereomer, exhibits the same spectral properties as the diastereomeric mixture.

Both optically individual compounds may be prepared from the corresponding optically uniform 5,6-epoxides by condensation with N- (N-trifluoroacetylcysteinyl) glycine methyl ester as follows:

Example 26a

N- [S-5 (R), 6 (S) -5-Hydroxy-7,9-trans-11-cis-eicosacrylen-6-yl-N-trityluoroacetyl-cis-cinyl] -glycine methyl ester

A solution of 2.35 g of 5 (R), 6 (R) -5,6-epoxy-7,9-trans-11-cis-eicosacrene in 28 ml of methanol under argon, 2.48 g of triethylamine and 2.53 g of N - (N-trifluoroacetylcysteinyl) glycine methyl ester was added and the mixture was stirred for 16 hours at 20 ° C. The volatiles were removed under water jet vacuum and the residue was chromatographed on silica gel. Elution with 3: 2 hexane / ethyl acetate gave the title compound as a colorless oil.

IR (CH ₂ Cl ₂ ): 3610, 3400, 2940, 2870, 1760, 1740,

1700, 1530, 1220, 1175 cm -1. Yield: 41%.

The optically active epoxide component was prepared using the procedure of Example 1a as follows:

Under anhydrous conditions while stirring

66.3 ml of tetraizopropyl orthotitanate and 38.51 ml of D - (-) - tartaric acid diethyl ester are dissolved in 1.11 methylene chloride at -23 DEG C., 25.7 g of 2-trans-heptanol (successively added). (see example la)) and 140 ml of 3.2 M tert-butyl hydroperoxide in toluene was heated at -20 ° C for 16 hours and 56 ml of 10% aqueous L-tartaric acid solution was added at -23 ° C. drop by drop. After an additional 30 minutes, the mixture was allowed to warm to +20 ° C and stirred until the organic phase was clearly separated. It is stirred with 1 L of 1% aqueous sodium sulfite solution for 1 hour, separated, washed with water, dried over sodium sulfate and evaporated in a water jet vacuum. The residue was dissolved in diethyl ether (1.6 L), cooled to 0 ° C, treated with π-sodium hydroxide solution (675 ml) dropwise and stirred at 0 ° C for 30 minutes. The separated organic layer was washed with brine, dried and evaporated to give 2 (R), 3 (R) -2,3-epoxy-heptanol as a colorless, unstable liquid, which was worked up immediately in the next step.

A solution of 13.3 g of the above compound in 100 ml of methylene chloride is added dropwise over 30 minutes to a suspension of 500 ml of methylene chloride containing 110.1 g of pyridinium chlorochromate and 41.9 g of sodium acetate, while stirring at room temperature. with gentle cooling, keep at 25 ° C. After 3 hours, the reaction mixture was diluted with diethyl ether (500 mL) and filtered over Kieselgel. The filtrate was washed with pH 8 phosphate buffer, dried over sodium sulfate and evaporated. The residue was chromatographed on silica gel with petroleum ether (Fp 3045 ° C) and diethyl ether (3: 2) to give 2 (S), 3 (R) -2,3-cpoxy-heptane as a colorless liquid; the product exhibits similar spectral properties to its 2 (R), 3 (S) -antipode (see Example 1a).

A solution of 6.7 g of 2 (S), 3 (R) -2,3-epoxyethane in 250 ml of methylene chloride is added dropwise at 20 [deg.] C. to a solution of 20.85 g of γ-triphenyl- phosphorus anilidene croton aldehyde in 200 ml of methylene chloride and stirred for an additional 1 hour at 20 ° C. The reaction mixture was diluted with hexane (240 mL) and ethyl acetate (120 mL), filtered over silica gel and concentrated. The residue was taken up in an equal volume of hexane and ethyl acetate, stirred for 15 minutes, filtered again over Kieselgel and concentrated. The resulting mixture of cis, trans, and trans, trans isomers was dissolved in 200 ml of methanol for isomerization, 200 mg of iodine was added and allowed to stand at 20 ° C for 3 hours. The solution was then washed with aqueous sodium thiosulfate solution and water, dried over sodium sulfate and evaporated, and the residue was chromatographed on silica gel. Elute with 4: 1 hexane / ethyl acetate to give the desired 6 (R), 7 (R) -6,7-epoxy-2,4-trans-undecadicinal in the form of a yellowish oil.

[α] 20 = -21.1 + 1.3 ° C (0.75 g / v% in chloroform), the spectral properties of which correspond to those of the antipodes 6 (S), 7 (S) (see Example 1a).

5.15 g of nonyl triphenylphosphonium bromide are dissolved in 50 ml of tetrahydrofuran, cooled to -78 ° C, stirred and 6.85 ml of a 1.6 M solution of butyl lithium in toluene are added. After 30 minutes at -78 ° C, 15.1 g of hexamethylphosphoric triamide and 1.52 g of 6 (R), 7 (R) -6,7-epoxy-2,4-trans-undecadienal are added dropwise in succession. A solution of 10 ml of tetrahydrofuran was heated at -78 ° C for an additional 15 minutes and allowed to warm to 0 ° C. The reaction mixture was mixed with pH 8 phosphate buffer and extracted with ether. The combined ether extracts were stabilized with a few drops of triethylamine, dried over sodium sulfate and freed of volatile constituents under vacuum at 20 ° C. The residue was stirred with a small amount of ether and separated from the desired solid triphenylphosphine oxide by filtration. The final traces of triphenylphosphine oxide from his hair were removed by filtration through a silica gel column, prepared by washing with a 4: 1 mixture of ether-hexane to which 2% triethylamine was added.

From the filtrate, the solvent was distilled off to give the desired 5 (R), 6 (R) -5,6-epoxy-7,9-trans-11-cis-eicosecrene in the form of fine yellow crystals.

M.p. 31-32 ° C.

Example 26b

N- [S-5 (S), 6 (R) -5-Hydroxy-7,9-trans-11-cis-cis-cis-cis-6-yl-N-trifluoroacetyl-cisleicl] glycine methyl ester was prepared in the same manner. as in Example 26a, but starting from 5) S), 6 (S) -5,6-epoxy-7,9-trans-11-cis-eicosecrene. The compound given in the title13

-1 325

196 364 corresponds to the product obtained by chromatography of diastereomeric mixtures (see above).

The starting material 5 (S), 6 (S) -5,6-epoxy-7,9-trons-11-cis-cis-cationicene is obtained in a similar manner to its 5 (R), 6 (R) antipode by to convert 6 (S), 7 (S) -6,7-epoxy-2,4-trans-undecadienal (see Example 1a) with a Wittig reagent made in situ from nonyl triphenylphosphonium bromide and butyllithium. 26a. The spectral properties of this compound correspond to those of the 5 (R), 6 (R) -antipodes described in Example 26a.

Example 7

N- [S-5 (RS), 6 (SR) -5-Hydroxy-7,11-cis-9-trans-cis-cinnam-6-yl-N-trinunr-n-methylcystinyl] -glycylmethyl ester

0.94 g of 5 (RS), 6 (RS) -5,6-epoxy-7,11-cis-9-transeikacryriene (E9) and 1.0 g of N- (N-trifluoroacetyl-cysteinyl) glycine methyl észtcrből.

IR (CH ₂ Cl ₂ ): 3400, 2970, 2940, 2870, 1740, 1700,

1535 cm ^-1 . Yield: 35%.

The diastereomeric mixture can be separated by chromatography, as in Example 25, into individual optically uniform diastereomers (eluent: hexane / ethyl acetate 7: 3).

Example 28

N [S-5 (RS), 6 (SR) -5-hydroxy-1-tetrahydropyran! - oxy - 7,9 - trans - 11 - cis - ejcacryrienyl - 6 - yl - N - trifluoroacetylcysteinyl] glycine methyl ester, individual diastereomers 5 (R), 6 (S) - [A] and 5 (S), 6 (R) - [BJ.

0.78 g of racemic 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyloxy-7,9-trans-11-cis-eicosacriene (E20) and 0.63 g of N- (N A 1: 1 mixture of diastereomeric [A] and [B] was prepared from trifluoroacetylcysteinyl) glycine methyl ester in 86% yield and isolated by chromatography on silica gel with hexane / ethyl acetate. : 1 ratio. Both show analog spectrum.

IR (CH ₂ Cl ₂ ): 3400, 2940, 2870, 1740, 1695, 1530

Example 28a

N- [S-5 (RS), 6 (RS) -1-Acetoxy-5-hydroxy-7,9-trans-11-cis-cacosrien-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester, 55% yield, individual diastereomers 5 (R), 6 (S) - [AJ and 5 (S), 6 (R) - [B].

R (= 0.3 [A] isomer, 0.25 [B] isomer (hexane / ethyl acetate 1: 1).

0.78 g of racemic 5 (RS), 6 (RS) -1-acetoxy-5,6-epoxy-7,9-trans-11-cis-eicosecrene (E20a) and 0.63 g of N- (N-trifluoro) -acetyl-cysteinyl) -glycine methyl ester in an approx. A 1: 1 mixture of [A] and [B] isomeric mixtures is obtained, from which the individual forms of the title compound are isolated by chromatography on silica gel (1: 1) with hexane / ethyl acetate.

Example 29 ~ '

N- [S-5 (RS), 6 (SR) -5-hydroxy-7,9-trans -11,14-cis-cis-cortacten-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester ; individual diastereomers 5 (R),

6 (S) - [A] and 5 (S), 6 (R) - [B].

103 mg of racemic 5 (RS), 6 (RS) -5,6-epoxy-7,9-trans11,14-cis-ejcatetraene (E21) and 154 mg of N- (N-trifluoroacetylcysteinyl) glycine of the methyl ester in a yield of 4% yields diastereomers [A] and [B] of ca. A 1: 1 mixture is prepared from which the individual forms of the title compound are isolated by chromatography on silica gel with hexane / ethyl acetate. Both show analog spectrum.

IR (CH ₂ Cl ₂ ): 3360, 2920, 2850, 1740, 1720, 1680,

1520 cm -1.

Example 29a

S-5 (RS), 6 (SR) -5-Hydroxy-7,9-trans-11,14-cis-ejcetetraeu-6-ylmethylcaploacetic acid methyl ester

103 mg of 5 (RS), 6 (RS) -5,6-cpoxy-7,9-trans-1, 1,14cis-ejcatacetraene (E21) and 54 mg of mercaptoacetic acid-methyl-ester.

Rf = 0.5 (5: 1 hexane: ethyl acetate).

Yield: 73%.

Example 30

N- [S-5 (RS), 6 (SR) -5-Hydroxy-1-tetrahydropyranyl-2-oxy-7,9-trans-11,14-cis-ejcatetraen-6-yl-N-trifluoro -acetyl-cysteinyl] -glycine methyl ester; individual diastereomers 5 (R), 6 (S) - [A] and 5 (S), 6 (R) - [B]

0.5 g of racemic 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyloxy-7,9-trans -11,14-cis-ejecatetraene (E22) and 0.42 g of N- From (N-trifluoroacetylcysteinyl) glycine methyl ester in a yield of 40% with an approx. A 1: 1 mixture of diastereomers [A] and [B] is obtained, from which the individual forms of the title compound are isolated by chromatography on silica gel (1: 1) with hexane / ethyl acetate. Both show the same spectrum.

IR (CH ₂ Cl ₂ ): 3400, 2940, 2880, 1760, 1740, 1700,

1530 cm ^-1 .

Example 31

N- [S-5 (RS), 6 (SR) -1-Acetoxy-5-hydroxy-7,11,14-cis-9-trans-eccatosetraen-6-yl-N-trifluoroacetyl-cysteinyl] - Glycine methyl ester: individual diastereomers 5 (R), 6 (S) - [A] and 5 (S), 6 (R) - [B]

490 mg of racemic 5 (RS) -acetoxy-5,6-epoxy-7,11,14-cis-9-trans-ejcatetraene (E23) and 400 mg of N- (N-trifluoroacetyl-cysteinyl) glycine methyl ester (49% yield) in an approx. A 1: 1 mixture of diastereomers [A] and [B] is obtained, from which the individual forms of the title compound are isolated by chromatography on silica gel (1: 1) with hexane / ethyl acetate. Both show the same spectrum.

-1 427

196,364

IR (CH ₂ Cl ₂ ): 3400, 2940, 2870, 1740, 1700, 1530 cm ^-1 .

Diastereomer [A]: [α] D = -36.6 ± 2 ° C (0.5 g / v% in chloroform).

Diastereomer [BJ: ⁰ - + 63.0 ± 2 ° C (0.5 g / v% in chloroform).

Subsequent cleavage of the hydroxyl protecting group

Example 32

N- (S-5 (RS), 6 (SR) -1,5-Dihydroxy-7-cis-ejosan-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

1.2 g of the title compound 1-tetrahydropyranyl ether (see Example 20) are dissolved in 70 ml of a mixture of 4 volumes of acetic acid, 2 parts of tetrahydrofuran and 1 part of water and the solution is stirred for 6 hours at 45 ° C. C. The solvent was evaporated in vacuo, the residue was extracted several times with toluene and the extracts were concentrated in vacuo. The residue was chromatographed on silica gel (15: 1) with dichloromethane / hexane to give the title compound.

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2870, 1760, 1740,

1700, 1530, 1220, 1180 cm ^-1 . Yield: 84%.

Example 33

N- [S-5 (RS), 6 (SR) -1,5-Dihydroxy-7-cisoctadecen-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

The title compound was prepared according to the procedure of Example 32 from 0.7 g of the corresponding 1-tetrahydropyranyl ether (see Example 19).

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2870, 1760, 1740,

1700, 1535, 1220, 1180 cm ^-1 . Yield: 80%.

Example 33a

Methyl S-5 (RS), 6 (SR) -1,5-Dihydroxy-7-cis-octadecen-6-yl-mercaptoacetic acid

The title compound was prepared according to the procedure of Example 32 from 0.7 g of the corresponding 1-tetrahydropyranyl ether (see Example 19a).

IR (CH ₂ Cl ₂ ): 3620, 3460, 2940, 2870, 1745 cm -1. Yield: 37%.

Example 33b

Methyl S-5 (RS), 6 (SR) -1,5-Dihydroxy-7-cis-ejosan-6-yl mercaptoacetic Acid

The title compound was prepared according to the procedure of Example 32 from 0.7 g of the corresponding 1-tetrahydropyranyl ether (see Example 19b).

IR (CH 2 Cl 2): 3620, 3460, 2940, 2870, 1745 cm -1. Yield: 72%.

Example 34

N- (S-5 (S), 6 (R) -1,5-Dihydroxy-7-trans-9-cis-octadecadecn-6-yl-N-trifluoroacrylcis-cinyl) glycine methyl ester

The title compound was prepared according to the procedure of Example 32 from 0.39 g of the corresponding 1-tetrahydropyranyl foodstuff (see Example 22, diastereomeric βJJ).

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2870, 1760, 1740,

1700, 1530, 1220, 1180 cm ^-1 . Yield: 81%.

Example 35

N- [S-5 (R), 6 (S) -1,5-Dihydroxy-7-trans-9-cis-octadecadien-oyl-N-trifluoroacetylcysteinyl] glycine methyl ester

In the same manner as in Example 32, the title compound is obtained from 0.33 g of the corresponding 1-tetrahydrofuranyl ether (see Example 32, diastereomer [A]).

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2870, 1760, 1740,

1700, 1530, 1220, 1180 cm -1. Yield: 79%.

Example 35a

Methyl S-5 (RS), 6 (SR) -1,5-Dihydroxy-7-trans-9-cis-octadecadien-6-yl-mercapto-acetic acid

According to the procedure of Example 32, the title compound is obtained from 0.33 g of the corresponding 1-tetrhydrofuranyl ether (see Example 22a).

IR (CH ₂ Cl ₂ ): 3620, 3460, 2940, 2870, 1745 cm -1. Yield: 68%.

Example 36

N- [S-5 (S), 6 (R) -1,5-Dihydroxy-7-trans-9-cis-eicosadien-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

The title compound was obtained according to the procedure of Example 32 from its corresponding 0.25 g of 1-tetrahydropyranyl ether (see Example 23, diastereomer [B]).

IR (CH ₂ Cl ₂ ): 3640, 3420, 2940, 2880, 1760, 1740,

1700, 1535, 1220, 1180 cm ^-1 . Yield: 91%.

Example 37

N- [S-5 (R), 6 (S) -1,5-Dihydroxy-7-trans-9-cis-ejcosadien-oyl-N-trifluoroacetylcysteinyl] glycine methyl ester

The title compound was obtained according to the procedure of Example 32 from its corresponding 0.24 g of 1-tetrahydropyranyl ether (see Example 23, diastereomer (AJ)).

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2870, 1760, 1740,

1700, 1535, 1220, 1180 cm ^-1 . Yield: 84%.

-1 529

196 364 is an example

Methyl S-5 (RS), 6 (SR) -1,5,5,5-Dihydroxy-7-trans-9-cis-eicosadien-6-yl mercaptoacetic acid

The title compound was obtained according to the procedure of Example 32 from the corresponding 0.25 g of 1-tetrahydropyranyl ether (see Example 23a, diastereomer [B]).

IR (CH ₂ CI _z): 2620, 3440, 2940, 2870, 1745 cm ^-1. Yield: 81%.

Example 38

N- [S-5 (S), 6 (R) -1,5-Dihydroxy-7,8-trans-11-cis-eicosacrien-6-yl-N-trifluoroacetylcysteinyl] glycine methyl- ester

The title compound is obtained from its corresponding 1-tetrahydropyranyl ether (0.56 g) following the procedure of Example 32 (see Example 28, Diastereomer [B]).

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2860, 1760, 1740,

1695, 1535, 1220, 1180 cm ^-1 . Yield: 71%.

Example 39

N- (S-5 (R), 6 (S) -1,5-Dihydroxy-7,9-trans-11-cis-eicosacrien-6-yl-N-trifluoroacetylcysteinyl) glycine methyl -ester

The title compound is obtained from its corresponding 49 mg of 1-tetrahydropyranyl ether according to the procedure of Example 32 (see Example 30, diastereomer [B]).

IR (CH ₂ Cl ₂ ): 3620, 3400, 2940, 2860, 1760, 1740,

1695, 1535, 1220, 1180 cm ^-1 . Yield: 81%.

Example 40

N- [S-5 (S), 6 (R) -1,5-Dihydroxy-7,9-trans-1,14-cis-ejcatetraen-6-yl-N-trifluoroacetyl-cysteinyl] glycine. methyl ester

The title compound was prepared according to the procedure of Example 32 from the corresponding 140 mg of l-tetrachydropyranyl ether (see Example 30, diastereomer [B]).

IR (CH ₂ Cl ₂ ): 3610, 3400, 2930, 2880, 1750, 1725,

1685, 1525, 1215, 1170 cm ^-1 . Yield: 33%.

Example 41

N- [S-5 (R), 6 (S) -1,5-Dihydroxy-7,9-trans-1,114-cis-ejcatetraen-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester

The title compound is obtained from its corresponding 140 µl of 1-tetrahydropyranyl ether according to the procedure of Example 32 (see Example 30, diastereomer (A 1)).

IR (CH ₂ Cl ₂ ): 3610, 3380, 2930, 2860, 1745, 1685,

1525, 1215, 1170 cm ^-1 . Yield: 60%.

Subsequent saponification of the ester group formed

Example 42

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-heptadecn-6-yl-N-trifluoroacetylcysteinyl] glycine

800 mg of the methyl ester of the title compound (see Example 2) are dissolved in 70 ml of methanol. To the solution was added 22 ml of 0.1 N aqueous sodium hydroxide solution and stirred at room temperature for 16 hours. The methanol was evaporated at 20 ° C and 80 ml of acetonitrile were added to the aqueous residue. The solution was filtered through a small amount of Kieselgel and the solvent removed in vacuo at 20 ° C. The residue was extracted several times with chloroform, and the extracts were concentrated in vacuo. It is dried under high vacuum and the residue containing the title compound is pulverized.

IR (CH ₂ Cl ₂ ): 3320, 2980, 2940, 2870, 1730, 1675,

1400 cm ^-1 . Yield: 84%.

Example 43

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-undecen-6-yl-N-trifluoroacellylcystinyl] glycine

The title compound was obtained according to the procedure of Example 41 from the corresponding methyl ester (174 g) (see Example 3).

IR (CH ₂ Cl ₂ ): 3320, 2980, 2940, 2880, 1730, 1675,

1610, 1400 cm ^-1 . Yield: 97%.

Example 44

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-tridecen-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure of Example 41, the title compound is obtained from 800 mg of the corresponding methyl ester (see Example 4).

IR (CH ₂ Cl ₂ ): 3300, 2980, 2940, 2870, 1730, 1670,

1600, 1400 cm ^-1 . Yield: 72%.

Example 45

S-5 (RS), 6 (SR) -5-hydroxy-7-cis-lridecen-6-yl cysteine sodium salt

Following the procedure of Example 41, the title compound is obtained from 300 mg of the corresponding methyl ester (see Example 5).

IR (CH ₂ Cl ₂ ): 2970, 2940, 2860, 1740, 1620, 1430 cm ^-1 . Yield: 67%,

Example 46

Sodium salt of S-5 (RS), 6 (SR) -5-hydroxy-7-cis-tridecen-6-yl mercaptoacetic acid

Obtained according to the procedure of Example 41 in the title

-1 631

196,364 were given 700 mg of the corresponding methyl ester (see Example 6).

IR (CH ₂ Cl ₂ ): 3300, 2960, 2930, 2860, 1600, 1400 cm ^-1 . Yield: 98%.

Example 47

Sodium salt of N- (S-5 (RS), 6 (SR) -5-hydroxy-7-cis-pentadccn-6-yl-N-trifluoroacetylcysteinyl) glycine

Following the procedure described in Example 41, the title compound is obtained from the corresponding methyl ester (710 mg) (see Example 7).

IR (CH ₂ Cl ₂ ): 3300, 2970, 2940, 2860, 1730, 1670,

1610, 1400 cm ^-1 . Yield: 96%.

Example 47a

Potassium S-5 (RS), 6 (SR) -5-Hydroxy-7-cis-pentadccn-6-yl-N-trifluoroacetylcystcin

Following the procedure described in Example 41, the title compound is obtained from the corresponding methyl ester (710 mg) (see Example 7a).

R f = 0.5 (isopropanol / CHCl ₃ / H ₂ O 23: 12: 5). Yield: 47%.

Example 48

S-5 (RS), 6 (SR) -5-hydroxy-7-cis-pentadecen-6-yl cysteine sodium salt

Following the procedure described in Example 41, the title compound is obtained from its corresponding methyl ester (250 mg) (see Example 8).

IR (CH ₂ Cl ₂ ): 2970, 2940, 2870, 1740, 1640, 1590,

1410 cm ^- '. Yield: 94%.

Example 49

Sodium salt of S-5 (RS), 6 (SR) -5-hydroxy-7-cis-pentadecen-6-yl mercaptoacetic acid

In the same manner as in Example 41, the title compound is obtained from 950 mg of the corresponding methyl ester (see Example 9).

IR (CH ₂ C1 _2): 3300, 2960, 2930, 2860, 1600, 1400 cm ^- '. Yield: 98%.

Example 50

S-5 (RS), 6 (SR) -5-hydroxy-7-cis-heptadecen-6-yl cysteine sodium salt

Following the procedure of Example 41, the title compound is obtained from 500 mg of the appropriate methyl ester (see Example 10).

IR (CH ₂ Cl ₂ ): 3300, 2940, 2870, 1600, 1410 cm ^-1 . Yield: 78%.

Example 51

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-ejosan-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure of Example 41, the title compound was obtained from its corresponding methyl ester (1.64 g) (see Example all).

IR (CH ₂ Cl ₂ ): 3300, 2930, 2860, 1730, 1670, 1600,

1400 cm ^-1 . Yield: 96%.

Example 52

Sodium salt of N- [S-5 (SR), 6 (SR) -5-hydroxy-7-cis-triclosen-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure of Example 41, the title compound is obtained from 950 mg of the corresponding methyl ester (see Example 12).

Rf = 0.26 (isopropanol / CI1 ₂ Cl ₂ / water, 23: 12: 5).

Yield: 99%.

Example 53

Sodium salt of N- (3- [4 (RS), 5 (RS) -4-hydroxy-6-eis-triazolin-5-yl-thio-propionyl) glycine

Following the procedure described in Example 41, the title compound is obtained from 200 mg of the corresponding methyl ester (see Example 13).

IR (dioxane): 3460, 1730, 1655, 1605, 1410, 1303,

1040 cm ^- '. Yield: 82%.

Example 54

Sodium salt of N- [S-4 (RS), 5 (SR) -4-hydroxy-6-cis-tetradecen-5-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound is obtained from its corresponding methyl ester (600 mg). (See Example 14.)

IR (CH ₂ C1 _2): 3300, 2980, 2940, 2870, 1720, 1660, 1610, 1560 cm ^- '. Yield: 30%.

Example 54a

Sodium salt of N- [S-4 (RS), 5 (SR) -4-hydroxy-6-cis-nonadecen-5-yl-N-trifluoroacetylcystinyl] glycine

According to the procedure of Example 41, the title compound, m.p. 69-73 ° C, is obtained from 600 mg of the corresponding methyl ester (see Example 16). Yield: 88%.

Example 55

Sodium salt of N- [S-4 (RS), 5 (SR) -4-hydroxy-6-cis-ejosan-5-yl-N-trifluoroacetylcysteinyl] glycine

Obtained according to the procedure of Example 41 in the title

-1 733

196 364 of 420 mg of the corresponding methyl ester (see Example 17).

IR (KBr): 3320, 2930, 1715, 1655, 1620, 1560,

1140 cm ^- '. Yield: 67%.

Example 55a

Sodium salt of N- (3- [4 (RS), 5 (SR) -4-hydroxy-6-cis-cyanocin-5-ylthio] propionyl) glycine

Following the procedure of Example 41, the title compound is obtained from 420 mg of the corresponding methyl ester (see Example 17a).

IR (film): 3320, 2940, 2870, 1610, 1420, 1135 cm -1. Yield: 55%.

Example 55b

Sodium salt of S-4 (RS), 5 (SR) -4-hydroxy-6-cis-ejosan-5-yl-N-trifluoroacetylcysteine

Following the procedure of Example 41, the title compound is obtained from 420 mg of the corresponding methyl ester (see Example 17b).

IR (dioxane): 3480, 1730, 1620, 1415, 1220 cm -1. Yield: 91%.

Example 56

Sodium salt of N- (S-6) RS), 7 (SR) -6-hydroxy-8-cis-ejosan-7-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound is obtained from its corresponding methyl ester (570 mg) (see Example 18).

IR (CH ₂ Cl ₂ ): 3300, 2940, 2870, 1730, 1675, 1620,

1400 cm ^-1 . Yield: 64%.

Example 56a

Sodium salt of S-5 (RS), 6 (SR) -1,5-dihydro'xi-7-cis-octadecen-6-yl mercaptoacetic acid

Following the procedure described in Example 41, the title compound is obtained from 570 mg of the appropriate title compound (see Example 33a).

R f = 0.36 (isopropanol / CHCl ₃ / water, 23:12:15). Yield: 71%.

Example 56b

N- [S-5 (RS), 6 (SR) -1,5-Dihydroxy-7-cis-cis-cinn-6-yl-N-trityluoroacetyl-cysteinyl] -glycine potassium salt

Following the procedure of Example 41, the title compound is obtained from 570 mg of the corresponding methyl ester (see Example 32).

R f = 0.43 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 67%.

Example 56c

Sodium salt of S-5 (SR), 6 (SR) -1,5-dihydroxy-7-cis-ejosan-6-yl-mccapto-c-acetic acid

Following the procedure of Example 41, the title compound is obtained from 570 mg of the corresponding methyl ester (see Example 33b).

R f = 0.56 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 73%.

Example 57

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-trans-9-cis-nonadecadien-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound is obtained from 410 mg of the corresponding methyl ester (see Example 21).

IR (CH ₂ Cl ₂ ): 3300, 2940, 2860, 1730, 1670, 1610,

1400 cm ^- '. Yield: 98%.

Example 57a

Sodium salt of N- {3- [5 (RS), 6 (SR) -5-hydroxy-7-trans-9-cis-nonadecadien-6-ylthio] -propionyl} -glycine

Following the procedure described in Example 41, the title compound is obtained from 410 mg of the corresponding methyl ester (see Example 21a).

R f = 0.56 (isopropanol / CHCl ₃ / water, 33: 12: 5). Yield: 88%.

Example 57b

N-; S-5 (R), 6 (S) -5-Hydroxy-7-trans-9-cis-eicosadien-6-yl-N-trifluoroacetyl-cysteinyl] glycine, potassium salt

According to the procedure of Example 41, 410 mg of the corresponding methyl ester of the title compound is obtained (see Example 21e, diastereomer [A]).

Example 57c

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7-trauma-9-cis-ejcosadien-o-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound is obtained from 410 mg of the corresponding methyl ester (see Example 21e, diastereomer (BJ)).

R f = 0.55 (isopropanol / CHCl ₃ / water, 33: 12: 5). Yield: 91%.

Example 57d

Sodium salt of N- [S-4 (RS), 5 (SR) -4-hydroxy-6-trans-8-cis-nonadecadien-5-yl-N-trifluoroacetylcysteine] glycine

Obtained according to the procedure of Example 41 in the title

-1 835

196 364 of 410 mg of the corresponding methyl ester (see Example 21f).

M.p. '52 -54 ° C. Yield: 89%.

Example 57e

Sodium salt of S-5 (RS), 6 (SR) -1,5-dihydroxy-7-trans-9-cisoctadecadien-6-yl mercaptoacetic acid

Following the procedure described in Example 41, the title compound is obtained from 410 mg of the corresponding methyl ester (see Example 35a).

R f = 0.55 (isopropanol / CHCl ₃ / water, 23.12: 5). Yield: 72%.

Example 7 /

Potassium salt of N- [S-5 (S), 6 (R) -1,5-dihydroxy-7-trans-9-cis-eicosadien-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound was obtained from 410 mg of the corresponding methyl ester (see Example 36).

R f = 0.50 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 44%.

Example 57g

Potassium salt of N- [S-5 (R), 6 (S) -1,5-dihydroxy-7-trans-9-cis-eicosadien-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound was obtained from 410 mg of the corresponding methyl ester (see Example 37).

R f = 0.50 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 62%.

Example 58

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7,9-trans-11-cis-hexadecatrien-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound was obtained from its corresponding methyl ester (1.17 g, see Example 24).

IR (CH ₂ Cl ₂ ): 3300, 2970, 2940, 2880, 1730, 1670,

1620, 1410 cm ''. Yield: 26%.

Example 59

N-] S-5 (RS), 6 (SR) -5-hydroxy-7,11-cis-9-trans-hexadecatrien-6-yl-N-trifluoroacetylcysteinyl] glycine sodium salt

Following the procedure described in Example 41, the title compound was obtained from its corresponding methyl ester (730 mg, see Example 25).

IR (CH ₂ Cl ₂ ): 3300, 2970, 2940, 2880, 1730, 1675,

1625, 1410 cm ^-1 . Yield: 41%.

Example 60

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7,9-trans-11-cis-eicosacrien-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure of Example 41, the title compound is obtained from its corresponding methyl ester (230 mg) (see Example 26).

IR (CH ₂ Cl ₂ ): 3350, 2960, 2930, 2860, 1720, 1675,

1540 cm ^-1 .

Similarly, starting from the appropriate optically individual diastereomers (see Examples 26n and 26b), optically individual products can be prepared. Yield: 82%.

Example 60a

Sodium salt of N- [S-5 (R), 6 (S) -5-hydroxy-7,9-trans-11-cis-cis-cytosrene-6-yl-N-trifluoroacetylcysteinyl] glycine

3.1 g of N- [S-5 (R), 6 (S) -5-hydroxy-7,9-trans-11-cis-cisacrylene-6-yl-N-trifluoroacetyl-cysteinyl] glycine - methyl ester (see Example 26a) in methanol (60 ml) under argon at 0-5 ° C

It is mixed dropwise with 26.8 ml of 0.2 N sodium hydroxide solution, stirred for 20 hours at 20 ° C and concentrated at this temperature in vacuo. After reversed-phase chromatography on Merck RP8 adsorbent, methanol / water (3: 1), the solvent is distilled off in vacuo to give the title compound as a white amorphous powder.

138-140 ° C.

[α] ²⁰ D = +6.4 ° C (methanol: 0.39%).

Yield: 71%.

Example 60b

N- [S-5 (S), 6 (R) -5-Hydroxy-7,9-trans-11-cis-eicosacrien-6-yl-N-trifluoroacetylcysteinyl] glycine, sodium salt

Under the reaction conditions described in Example 60a and with the same amount of reaction partners and auxiliary chemicals, but N- [S-5 (S), 6 (R) -5-hydroxy-7,9-trans-11-cis-eicosrien-6-yl Starting from the methyl ester of -N-trifluoroacetyl-cysteinyl] glycine (see Example 26b), the title compound is obtained as a white amorphous powder. Yield: 55%.

Example 61

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7,11-cis-9-trans-cis-cationic-6-yl-N-trillluoroacetylcystinyl] glycine

Obtained according to the procedure of Example 41 in the title

-1 937

196 364 (370 g, m.p.) of methyl ester (see Example 27).

VR (CH ₂ Cl ₂ ): 3300, 2960, 2430, 2860, 1720, 1660,

1620, 1400 cm ^-1 . Yield: 59%.

Similarly, starting from the appropriate optically individual diastereomers (see chromatographic separation in Example 27), optically individual products, i.e., the 5 (S), 6 (R) -diastereomer [AJ and the 5 (R), 6 (S) -diastereomer [ B] we can win.

Example 61a

N- [S-5 (S), 6 (R) -1-Acetoxy-5-hydroxy-7,11-cis-9-trans-eicosacrien-6-yl-N-trifluoroacetylcysteinyl] glycine potassium salt and N- [S-5 (S), 6 (R) -1,5-dihydroxy-7,11-cis-9-trans-eicosrien-6-yl-N-trifluoroacetylcysteinyl] glycine. potassium

Following the procedure described in Example 41, it is replaced by the corresponding optically individual methyl ester 1-steel (see Example 28) and the crude reaction mixture is separated by reverse phase chromatography (elution with 3: 1 methanol-water). First, the diol compound elutes; yield: 52%, Rf = 0.56; then I-steel; yield 8%, Rf = 0.61 (isopropanol / CIICI ₃ / water, 23 to 12: 5).

Example 61b

N- [S-5 (R), 6 (S) -1-Acetoxy-5-hydroxy-7,11-cis-9-trans-eicosrien-6-yl-N-trifluoroacetylcysteinyl] glycine potassium salt and N- [S-5 (R), 6 (S) -1,5-dihydroxy-7,11-cis-9-trans-eicosrien-6-yl-N-trifluoroacetyl-cysteinyl] glycine. potassium

According to the procedure described in Example 41, the corresponding optically individual methyl ester 1-acetate (see Example 28) was replaced and the crude reaction mixture was separated by reverse phase chromatography (elution with 3: 1 methanol-water). First, the diol compound elutes; yield: 41%; R f = 0.54; then 1-acetate; Yield: 18%, Rf = 0.59 (isopropanol / CHCl3 / water, 23: 12: 5).

Example 61c

Potassium salt of N- [S-5 (R), 6 (S) -5-hydroxy-7,9-trans-11,14-cis-ejcatetraen-6-yl-N-trifluoroacetylcysteinyl] glycine

Following the procedure described in Example 41, the title compound was obtained from 370 mg of the corresponding methyl ester (see Example 29, Diastereomer [A]).

R f = 0.58 (isopropanol / CHCl ₃ / water, 23: 12: 5). mining; 54%.

Example d

Potassium salt of N- (S-5 (S), 6 (R) -5-hydroxy-7,9-trans-11,14-cis-ejcatetraen-6-yl-N-trifluoroacetyl-cysteinyl) glycine

Following the procedure of Example 41, the title compound was obtained from 370 mg of the corresponding methyl ester (see Example 29, Diastereomer (BJ)).

Rf - 0.56 (isopropanol / _CHCl3 / water, 23: 12: 5). Yield: 67%.

this example

Potassium salt of S-5 (RS), 6 (SR) -5-hydroxy-7,9-trans-11,14-cis-ejcatetraen-6-yl mercaptoacetic acid

Following the procedure described in Example 41, the title compound is obtained from 370 mg of the corresponding methyl ester (see Example 29a).

R f = 0.33 (9: 1 CHCl 3 / methanol). Yield: 39%.

Example f

N- [S-5 (S), 6 (R) -1,5-Dihydroxy-7,11,14-cis-9-transcyclotraacen-6-yl-N-trifluoroacetylcysteinyl] glycine potassium salt

Following the procedure of Example 41, the title compound was obtained from 370 mg of the corresponding methyl ester (see Example 31, Diastereomer [B]).

R f = 0.62 (isopropanol / CHCl ₃ / water, 23: 15: 5). Yield: 54%.

Example g

N- (S-5 (R), 6 (S) -1,5-Dihydroxy-7,11,14-cis-9-trans-ejcatetraen-6-yl-N-trifluoroacetyl-cysteinyl) glycine potassium salt

Following the procedure described in Example 41, the title compound is obtained from 370 mg of the corresponding methyl ester. See Example 31, Diastereomer [A]).

R f = 0.56 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 48%.

Post-cleavage of the N-trifluoroacetyl group

Example 62

Sodium salt of N- (S-5 (RS), 6 (SR) -5-hydroxy-7-cis-heptadecen-6-yl-cysteinyl) glycine

N ^c * ^c -trifluoroacetyl derivative of the title compound (see Example 42) 590 mg is dissolved in 15 ml of methanol, the solution was mixed with 1.7 g of sodium carbonate with a solution of 15 ml of water. The resulting suspension was stirred at 60 ° C for 20 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was dissolved in 1: 1 methanol / dichloromethane, filtered again and the filtrate was evaporated in vacuo.

-2 039

196,364 were removed. The residue was subjected to reverse phase chromatography on silica gel with methanol / water (3: 1) to give the desired title compound.

IR (CH ₂ Cl ₂ ): 3300, 2940, 2870, 1680, 1600, 1400 cm ^-1 . Yield: 55%.

Example 63

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-undecen-6-ylcysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 1.74 g of the corresponding N-trifluoroacetyl derivative (see Example 43).

IR (CH ₂ Cl ₂ ): 3300, 2980, 2940, 1730, 1670, 1610,

1400 cm ^-1 . Yield: 47%.

Example 64

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-tridcen-6-yl-cystemyl] -glycine

Following the general procedure described in Example 62, the title compound (320 mg) was obtained from the corresponding N-trifluoroacelyl derivative (see Example 44).

IR (CH ₂ Cl ₂ ): 3400, 2960, 2930, 2860, 1680, 1600,

1400 cm ^-1 . Yield: 54%.

Example 65

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-pentadecen-6-ylcysteinyl] glycine

Following the general procedure described in Example 62, the title compound (400 mg) was obtained from the corresponding N-trifluoroacetyl derivative (see Example 47).

IR (CH ₂ Cl ₂ ): 3380, 2960, 2930, 2860, 1650, 1600,

1400 cm ^-1 . Yield: 40%.

Example 66

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-ejosan-6-yl-cystinyl] glycine

According to the general procedure described in Example 62, the title compound is obtained from 1.64 g of the corresponding N-trifluoroacetyl derivative (see Example 51).

IR (CH ₂ Cl ₂ ): 3300, 2930, 1860, 1730, 1670, 1600,

1400 cm ^- Yield: 96%.

Example 67

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-triclosen-6-ylcysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 960 mg of the corresponding N-trifluoroacetyl derivative (see Example 52).

IR (CH ₂ Cl ₂ ): 3250, 2940, 2870, 1680, 1600, 1400 cm ^-1 . Yield: 68%.

Example 67a

Sodium salt of N [S-4 (RS), 5 (3R) -4-hydroxy-6-cis-nonadecen-5-ylcysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the appropriate N-trifluoroacetyl derivative (see Example 54a).

M.p .: 180 'C. Yield: 78%.

Example 68

Sodium salt of N- [S-4 (RS), 5 (SR) -4-hydroxy-6-cis-ejosan-5-yl-cysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 55).

IR (CH ₂ Cl ₂ ): 3420, 2940, 2860, 1720, 1670, 1620,

1400 cm ^-1 . Yield: 82%.

Example 68a

S-4 (RS), 5 (SR) -4-coldloxy-6-eis-cis-cinn-5-yl-cystcinium potassium salt

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 65b).

153-156 ° C. Yield: 62%.

Example 68b

Potassium salt of N- [S-6 (RS), 7 (SR) -6-hydroxy-8-cis-ejosan-7-yl-cysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 56).

R f = 0.55 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 84%.

Example 68c

Sodium salt of S-6 (RS), 7 (SR) -6-hydroxy-8-cis-ejosan-7-yl-cysteine

By using the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 18b).

R f = 0.62 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 56%.

Example d

Sodium salt of N- [S-5 (RS), 6 (SR) -5-hydroxy-7-trans-9-cis-nonadecadien-6-yl-cysteinyl] glycine

It is obtained according to the general procedure described in Example 62

-2 141

196,364 of the title compound from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 57).

IR (KBr): 3350, 2960, 2920, 2875, 1650, 1605, 1540,

1400 cin ^-1 . Yield: 84%.

Example 68e

N- [S-5 (R), 6 (S) -5-yl-oxy-7-trans-9-cis-cis-cosadicn-6-yl-cis-lcinyl] -glycine, potassium salt

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 57b).

R f = 0.56 (isopropanol / CHCl ₃ / water).

Yield: 92%.

Example 68f

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7-trans-9-cis-eicosadien-6-yl-cystinyl] -glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 57c).

R f = 0.56 isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 69%.

Example 68g

Sodium salt of N- [S-4) RS), 5 (SR) -4-hydroxy-6-trans-8-cis-nonadecadien-5-yl-cysticyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 57d).

98-102 ° C. Yield: 77%.

Example 68lt ι

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7,9-trans-11-cis-eicosacrien-6-yl-cysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacelyl derivative (see Example 60b).

R f = 0.52 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 38%.

Example i

N- [S-5 (R), 6 (S) -5-Hydroxy-7,9-trans-11-cis-eicosacrien-6-yl-cysteinyl] -glycine potassium salt

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative and potassium carbonate (see Example 60a).

R f = 0.52 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 92%.

Example 68j

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7-cis-9-trans-11-cis-eicosacryl-6-yl-cystinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trihloroacetyl derivative (see Example 61).

R f = 0.50 (isopropanol / Cl 2 IC ₃ / water, 23: 12: 5). Yield: 44%.

68k pclda

Potassium salt of N- [S-5 (R), 6 (S) -5-hydroxy-7-cis-9-trans-11-cis-eicosacrien-6-yl-cysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetib derivative and potassium carbonate (see Example 61).

681 pclda

Potassium salt of N - (S-5 (S), 6 (R) -1,5-dihydroxy-7,9-trans-11-cis-eicosacrien-6-ylcystinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trilylacetyl derivative (see Example 61a).

R f = 0.44 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 67%.

68tn example

Potassium salt of N- [S-5 (R), 6 (S) -1,5-dihydroxy-7,9-trans-11-cis-eicosacryrien-6-yl-cysteinyl] glycine

Following the general procedure described in Example 62, the title compound is obtained from 200 mg of the corresponding N-trifluoroacetyl derivative (see Example 61b).

R f = 0.62 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 68%.

Subsequent simultaneous saponification of the N-tri-fluoroacetyl group and the terminal ester group

Example 69

Potassium salt of N- [S-5 (S), 6 (R) -1,5-dihydroxy-7,9-trans-11,14-cis-ejcatetraen-6-yl-cysteinyl] glycine

50 mg of the title compound N ^cis- trifluoroacetylmethyl ester (see Example 39, 5 (S), 6 (R) diastereomer) are dissolved in 4 ml of methanol and the solution is 170 mg of potassium carbonate in 10 ml. water. The reaction mixture was stirred under argon for 3 days and concentrated in vacuo at room temperature. The residue was taken up again in chloroform and evaporated in vacuo. Reverse phase chromatography on silica gel eluting with methanol / water (3: 1) gave the title compound.

-2 243

196,364

R f = 0.33 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 26%.

the example

Potassium salt of N- (S-5 (RS), 6 (SR) -1,5-dihydroxy-7-cis-octadecen-6-yl-cysteinyl] -glycine

Following the general procedure described in Example 69, the title compound is obtained from 30 mg of the corresponding N-trifluoroacetylmethyl ester (see Example 33).

IR (CHCl ₃ ): 3400, 3140, 3050, 2925, 2870, 1675,

1595, 1410 cm ^-1 . Yield: 34%.

Example 69b

Potassium salt of N- [S-5 (RS), 6 (SR) -1,5-dihydroxy-7-cis-ejosan-6-yl-cysteinyl] glycine

Following the general procedure described in Example 69, the title compound is obtained from 30 mg of the appropriate N-trifluoroacetylmethyl ester (see Example 32).

IR (CHCl ₃ ): 3300, 2940, 2865, 1680, 1605, 1400 cm ^-1 . Yield: 41%.

Example 69c

Potassium salt of N- [S-5 (S), 6 (R) -1,5-dihydroxy-7-trans-9-cis-octadecadien-6-yl-cysteinyl] -glycine

Following the general procedure described in Example 69, the title compound is obtained from 30 mg of the appropriate N-trifluoroacetylmethyl ester (see Example 34).

IR (CHCl 3): 3285, 2930, 2855, 1675, 1600, 1390 cm ^-1 . Yield: 24%.

Example 69d

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7,9-trans-11-cis-hexadecatrien-6-yl cysteinyl] glycine

According to the general procedure described in Example 69, the title compound is obtained from 30 mg of the appropriate N-trifluoroalylmethyl ester (see Examples 24).

R f = 0.53 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 31%.

Example 69e

N - [S-5 (R), 6 (S) -5-Hydroxy-7,9-trans-11-cis-lexadecatrien-6-yl-cysteine] -glycine potassium salt

Following the general procedure described in Example 69, the title compound is obtained from 30 mg of the corresponding N-trifluoroacetylmethyl ester (see Example 24).

R f = 0.56 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 36%.

Example 69f

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7,11-cis-9-trans-hxadecalthien-6-ylcis (ethyl) glycine

Following the general procedure described in Example 69, the title compound is obtained from 30 mg of the appropriate N-trifluoroacetylmethyl ester (see Example 25).

R f = 0.55 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 31%.

Example 69g

The potassium salt of N- [S-5 (R), 6 (S) -5-hydroxy-7,11-cis-9-trans-hxadecatrien-6-yl-cysteinyl] glycine was obtained according to the general procedure of Example 69. 30 mg of the corresponding N-trifluoroacetylmethyl ester (see Example 25).

Yield: 23%. 129-130 ° C.

Example 70

N- [S-5 (R), 6 (S) -1,5-Dihydroxy-7,8-trans-11,14-cis-ejcatetraen-6-yl-cysteinyl] -glycine potassium salt

Following the general procedure described in Example 69, the title compound was obtained from 30 mg of the appropriate N-trifluoroacetylmethyl ester (see Example 40, [5 (R), 6 (S) diastereomer]).

R f = 0.37 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 55%.

Example 71

Potassium salt of N- [S-5 (S), 6 (R) -5-hydroxy-7,9-trans-11,14-cis-ejcatetetraen-6-yl-cysteinyl] glycine

Following the general procedure described in Example 69, the title compound was obtained from 33 mg of the corresponding N-trifluoroacetyl methyl ester (see Example 29, [BJ diastereomer).

UV (CH 3 OH): _λmax = 280 nm (ε = _48,600 ). Yield: 33%.

Example 72

Calcium salt of N- [S-5 (R), 6 (S) -5-hydroxy-7,9-trans-1, 1,14-cis-ejcatetetraen-6-ylcysteinyl] glycine

Following the general procedure described in Example 69, the title compound was obtained from 8 mg of the corresponding N-trifluoroacetyl methyl ester (see Example 29, [A] diastereomer).

UV _(CH3 OH): X = 280 nm _mnx (8 = 48600). Yield: 30%.

-2 345

Example 1 364% ¹ 73 I

Potassium salt of N- [S-5 (S), 6 (R) -1,5-dihydroxy-7,12,14-cis-9-trans -cis-trac-5-yl-cystinyl] -glycine (simultaneous cleavage of 3 protecting groups)

160 mg of N- [S-5 (S), 6 (R) -1-acetoxy-5-hydroxy-7,11,14-cis-9-trans-ejection tetraene-6-yl-N-trifluoroacetylcysteinyl] glycine methyl ester solution was added to a solution of 700 mg of potassium carbonate in 50 ml of water, stirred for 3 days under argon at room temperature and also concentrated in vacuo at room temperature. The residue was taken up in several portions in chloroform and the extract concentrated in vacuo. It was filtered through silica gel, dissolved in 1: 3 dichloromethane / methanol to give the desired title compound in solution.

IR (CII ₂ Cl ₂ ): 3400, 2940, 1690, 1600, 1440, 1220,

1190 cm ^-1 . Yield: 99%.

Example 74

Simultaneous Cleavage of N- [S-5 (R), 6 (S) -1,5-Dihydroxy-7,11,14,14-cis-9-trans-cis-cactenyl-cystinyl] glycine potassium salt (3 protecting groups) )

Similar to the previous example, 160 mg of N- [S-5 (R), 6 (S) -1-acetoxy-5-hydroxy-7,11,14-cis-9-trans-jectetetraen-6-yl] -N -trifluoroacetylcysteinyl] glycine methyl ester is hydrolyzed to the title compound.

IR (CH ₂ Cl ₂ ): 3420, 2940, 1685, 1615, 1420, 1220,

1190 cm- '.

Example 74a

Potassium salt of N- [S-5 (R), 6 (S) -1,5-dihydroxy-7-trans-9-cis-eicosaden-6-yl-cystinyl] -glycine (simultaneous cleavage of 3 vc groups)

In a manner similar to Example 73, 160 mg of N- [S-5 (R), 6 (S) -1-acetoxy-5-hydroxy-7-trans-9-cis-eicosadien-6-yl-N-trifluoroacetyl -cystinyl] -glycine inethyl ester (Example 23a) is hydrolyzed to the title compound.

R f = 0.55 (isopropanol / CHCl ₃ / water, 23: 12: 5). Yield: 98%.

Example 75

N- [S-5 (RS), 6 (SR) -5-hydroxy-7-cis-heptadecen-6-yl-cysteine 1] -glycine

250 mg of the title compound sodium salt (see Example 62) are dissolved in 15 ml of dichloromethane and the solution is shaken vigorously with 15 ml of 5% aqueous acetic acid for 10 minutes at room temperature. The organic layer was separated, washed with water, dried over sodium sulfate and the solvent removed in vacuo. 220 mg of the title compound remain as an amorphous residue.

IR (CH ₂ C1 _2): 3300, 2960, 2870, 1680, 1620, 1410 cm ^- '.

2-74. The starting materials given in Examples 1 to 4 can be prepared as follows:

Λ Unsaturated aldehydes

ΛI 2-trans hepien

A solution of 44.8 ml of valeraldchide and 6.3 g of formylmethylene triphenylphosphorane (see S. Trippett and DM Walker, J. Chem. Soc. 1961, 1266) in 400 ml of tetrahydrofuran and 150 ml of chloroform were heated under argon for 24 hours. using a reflux condenser. The red solution was freed from the solvent in vacuo at room temperature and the residue was mixed with ether / hexane (1: 1). The solid residue was filtered and washed four times with 1: 1 ether / hexane. The filtrate was concentrated in vacuo and the residue was distilled in vacuo. The title compound is obtained as a colorless liquid.

(MP = 55-57 ° C / 22 mbar.)

In a similar manner, the following materials are obtained:

22 2-trans-hexenal (Fp = 33 ° C / 14 mbar.)

30.2 ml of butyraldehyde and 51 g of formylmethylene triphenylphosphorane.

A3 at 2-trans-octc (Fp = 65-69 ° C / 16 mbar.) G of capronaldehyde and 42.6 g of formylmethylene triphenylphosphorane.

7-Tetrahydropyranyloxy-2-trans-heptenal (m.p. = 106 ° C / 5 mbar) from 5-tetrahydropyranyloxy-pentanal.

[E. Corey, J., et al., J. Am. Chem. Soc. 92, 6635 (1970).]

26.1 g of formylmethylene triphenylphosphorane.

75 7-aceloxy-2-trans hepenal (oil)

3.9 g of 5-acetoxypentanal [H. Brown et al.,

Synthesis 1980, 151] and 8.2 g of formylmethylene

of crude product from triphenylphosphorane by chromatography on silica gel with hexane: ethyl acetate (3: 1).

B Epoxy aldehydes

BI 2 (RS), 3 (Sfi) -2,3-epoxy-hepania

Dissolve 9 g of 2-trans-heptanal (Al) in 200 ml of dichloromethane / methanol 1: 1, add 28 ml of 30% aqueous hydrogen peroxide and 800 mg of potassium carbonate and stir at room temperature for 6 hours. 100 ml of pH 8 phosphate buffer is added and the organic phase is separated. The aqueous phase is extracted three more times with 50 ml of dichloromethane. The combined organic phases are washed with 20 ml of phosphate buffer.

-2 447

196 364 washed and dried over magnesium sulfate. The solution was filtered through a small amount of Florisil and concentrated in vacuo. The title compound is obtained in the form of a colorless liquid.

IR (CIlCl 2): 2950, 2920, 2850, 1720, 1460, 850 cm ^-1 .

In a similar way we obtain the following materials:

B2 2 (RS), 3 (SR) -2,3-epoxyhexanal

16.2 g of 2-trans-hexenal (A2) as an oil.

(! CH ₂ C ₂₎ IR: 2980, 2950, 2890, 1740, 1475, 860 cm ^-1.

B3 2 (RS), 3 (SR) -2,3-octane

9.0 g of 2-trans-octenal (A3) as an oil.

IR (CH ₂ Cl ₂ ): 2970, 2950, 2870, 1740, 1475, 1025 cm ^-1 .

B4 2 (RS), 3 (SR) -2,3-epoxy-7-tetrahydropyranyloxyheptanal

8.1 g of 7-tetralyl-l-pyrrolinyloxy-2H / 7'-yl-leptene (A4) are obtained by chromatography on silica gel with hexane / ethyl acetate 2: 1 as an oil.

IR (CH ₂ Cl ₂ ): 2950, 2880, 1735, 1140, 1130, 1080, 1040 cm ^-1 .

B5 2 (RS), 3 (SR) -7-acetoxy-2,3-epoxy-heptanal

2.2 g of 7-acetoxy-2-ZranZ-heptenal (A5) are obtained as an oil by chromatography on silica gel with dichloromethane: ethyl acetate (93: 7).

IR (CH ₂ Cl ₂ ): 2960, 1740, 1375, 1240, 1050, 860 cm ^-1 .

C Once unsaturated epoxy aldehydes

Cl 4 (RS), 5 (RS) -4,5-epoxy-2-trans-nonenal

A solution of 7.4 g of 2 (RS), 3 (SR) -coxoxy-heptanal (B1) and 17.6 g of formylmethylene-triphenylphosphorane in 250 ml of tetrahydrofuran and 100 ml of chloroform was heated under reflux for 1.5 hours. heated. The cooled solution was removed in vacuo at room temperature and the residue was mixed with ether / hexane (4: 1). The slurry was filtered through a small amount of silica gel and washed with ether / hexane (4: 1). The filtrate was concentrated in vacuo and the residue was chromatographed on silica gel with 5: 1 hexane / ethyl acetate 1% triethylamine. The title compound is obtained as a colorless oil. IR (CH ₂ Cl ₂ ): 2970, 2940, 2870, 1690, 1650, 1115, 990 cm ^-1 .

In a similar manner, the following materials were obtained: Cla 4 - (RS), 5 (RS) -4,5-epoxy-2-transxzoctenal

2 (RS), 3 (SR) -2,3-epoxyhexanal (B2), as an oil.

C2 4 (RS), 5 (RS) -4,5-epoxy-9-trihydropyranyloxy-2-trans-nonenal g 2 (RS), 3 (SR) -2,3-cpoxy-7-tetrahydropyranyloxy from heptanal (B4) as an oil. IR (CH ₂ Cl ₂ ): 2950, 2880, 1700, 1125, 1040 cm ^-1 .

C2a4 (RS), 5 (RS) -9-actooxy-4,5-epoxy-2-trans-nonenal

2 (RS), 3 (SR) -7-acetoxy-2,3-epoxy-heptanal (B5), as an oil.

D Ke'-unsaturated epoxy aldehydes

D16 (RS), 7 (RS) -6,7-epoxy-2,4-indecadiene:

2-trans-4-trans isomer (Dia) and 2-trans-4-cis isomer (Dlb)

A solution of 1.75 g of 2 (RS), 3 (SR) -2,3-epoxy-heptanal (B1) in 40 ml of dichloromethane under stirring under argon-thiospheres was treated with 4.6 g of 4-triphenylphosphorinylidene-2-trans-butc [ MJ Bcrcnguer ct al., Tetrahedron Lett. 1975, 495] was added dropwise over 1 hour at room temperature. After stirring for another hour, the solvent was evaporated in vacuo at room temperature. The residue was stirred with 4: 1 ether / hexane, filtered through a small amount of silica gel and washed with 4: 1 ether / hexane. After evaporation of the solvent in vacuo, the residue is chromatographed on silica gel with hexane / ethyl acetate (4: 1, 1% triethylamine). It is obtained in the form of a light yellow oil, ca. the same amounts of the 2-trans-4-cis (D1b) and 2-trans-4-trarts (Dia) isomers of the title compound, both having the same spectral maxima: IR (CH ₂ Cl ₂ ): 2950, 2920, 1680, 1640 , 1110, 990 cm ^-1 .

In a similar way we obtain the following materials:

D2 6 (RS), 7) RS) -6,7-epoxy-11-tetrahydropyranyloxy-2,4-trans-undecadienal

1.4 g of 2 (RS), 3 (SR) -2,3-epoxy-7-tetrahydropyranyloxy-heptanal (B4) are obtained as an isomeric mixture which is then added in dichloromethane solution until a permanent color is obtained. The solution is stirred at room temperature for 1 hour. After filtration on silica gel, the solvent was distilled off and the title compound was isolated as an oil.

IR (CH ₂ Cl _a ): 2960, 2880, 1690, 1650, 1580, 1120, 1040 cm ^-1 .

D3 6 (RS), 7 (RS) -1-acetoxy-6,7-epoxy-2,4-undecadienal

2-trans-4-trans isomer (D3a) and 2-trans-4-cis-isomer (D3b). A mixture of both isomers

-2 549

196,364

0.54 g of 2 (RS), 3 (SR) -7-actoxy-2,3-cpoxy-heptane (B5) is obtained in a ratio of about 1: 2 (D3a) :( D3b) and separated by chromatography.

Ep Epoxy-olefins of formula (II): E5 (RS), 6 (RS) -5,6-epoxy-7-cis-heptadecene

9.4 g of n-decyltriphenylphosphonium bromide (CT Eyles and S. Trippctt, J. Chem. Soc. [C] 1966, 67) are dissolved in 50 ml of tetrahydrofuran, cooled to -30 ° C and added dropwise. added

12.2 ml of 1.6 M n-butyllithium, dissolved in hexane, with stirring under argon, maintained at -25 to -30 ° C. The red solution was allowed to warm to room temperature and was stirred at this temperature for 10 minutes, cooled to -78 ° C and added dropwise over 2 minutes in 2 ml of 2 (RS), 3 (SR) -2,3-epoxy in 10 ml of tetrahydrofuran. -heptanate (B1). The solution was allowed to warm to room temperature and stirred for 1 hour. The solvent was evaporated in vacuo at 40 ° C and the residue was dissolved in dichloromethane. To the solution was added Kieselgel (as much as all the solvents were absorbed), then slurried with ether and filtered. Wash four times with ether / hexane (1: 1) and evaporate the filtrate in vacuo. The residue was purified by chromatography on silica gel with hexane / ethyl acetate (30: 1) containing 1% triethylamine. The title compound is obtained as a colorless oil.

IR (CH ₂ Cl ₂ ): 2980, 2940, 2870, 1475, 875 cm ^-1 .

In a similar way we obtain the following materials:

E2 5 (RS), 6 (RS) -5,6-epoxy-7-cis-undecene

1.3 g of 2 (RS), 3 (SR) -2,3-epoxy-heptane (B1) and 5.5 g of n-butyl triphenylphosphonium bromide (R. Mechoulam and F. Sondheimer, J. Am. Chetn. Soc. 80, 4386 (1958)].

IR (CH ₂ Cl ₂ ): 2970, 2940, 2880, 1740, 875 cm ^-1 .

E3 5 (RS), 6 (RS) -5,6-epoxy-7-cis-idecene

1.2 g of 2 (RS), 3 (SR) -2,3-cpoxy-heptane (B1) and 5 g of n-hexyltriphenylphosphonium bromide. [C. Hauser F. et al., J. Org. Chem., 28, 372 (1963)].

IR (CH ₂ Cl ₂ ): 2970, 2940, 2870, 1475, 875 cm -1.

E4 5 (RS), 6 (RS) -5,6-epoxy-7-cis-pentadecene

1.25 g of 2 (RS), 3 (RS) -2,3-epoxy-heptanal (B1) and 5.6 g of n-octyl triphenylphosphonium bromide [C. T. Eyles and S. Trippett, J. Chem. Soc. (C) 7966, 67].

IR (CH ₂ Cl ₂ ): 2970, 2930, 2870, 1475, 875 cm -1.

E5 5 (RS), 6 (SR) -5,6-epoxy-7-cis-eicosene g from 2 (RS), 3 (SR) -2,3-epoxy-heptanal (B1) and 12.3 g of n- from tridecyl triphenylphosphonium bromide26 [J. Ciigg et al., J. Chem. Soc. 7966, 1872].

IR (CH ₂ Cl ₂ ): 2970, 2940, 2860, 1475, 875 cm -1. E6 from 5 (RS), 6 (RS) -5,6-epoxy-7-cis-iricosene from 2 (RS), 3 (SR) -2,3-epoxy-heptane (B1) and 5.5 g of n- from licxadecyltriphenylphosphonium bromide [13]. Jcrcliel and J. Kimmig, Chem. Bcr. 83, 277 (1950).

IR (CH ₂ Cl ₂ ): 2970, 2935, 2870, 1470, 875 cm -1. E7 from 4 (RS), 5 (RS) -4,5-epoxy-6-cis-telradecene g from 2 (RS), 3 (SR) -2,3-epoxy-hexane (B2) and 10 g of n-octyl- prepared from triphenylphosphonium bromide.

IR (CH ₂ Cl ₂ ): 2990, 2960, 2880, 1480, 910 cm -1.

E8 4 (RS), 5 (RS) -4,5-epoxy-6-cis-nonadecene g from 2 (RS), 3 (SR) -2,3-cpoxy-hexanal (B2) and

3.6 g of n-tridecyl triphenylphosphonium bromide are prepared.

IR (CH ₂ Cl ₂ ): 2980, 2940, 2870, 1475, 905 cm -1.

E9 4 (RS), 5 (RS) -4,5-epoxy-6-cis-e / cosene

0.58 g of 2 (RS), 3 (SR) -2,3-epoxyhexanal (B2) and 3.8 g of n-tetradecyl triphenylphosphonium bromide [E. J. Reist and P. H. Christie, J. Org. Chem. 35, 3521 (1970)].

IR (CH ₂ Cl ₂ ): 2940, 2870, 1740, 905 cm -1.

E10 from 6 (RS), 7 (RS) -6,7-cpoxy-8-cis-eicosene g from 2 (RS), 3 (SR) -2,3-epoxyoctane (B3) and

4.1 g of n-dodecyl triphenylphosphonium bromide [D. Jerchel and J. Kimmig, Chem. 83, 277 (1950)].

IR (CH ₂ Cl ₂ ): 2940, 2870, 1465, 875 cm -1.

Ely 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyloxy-7-cisoctadecene g 2 (RS), 3 (SR) -2,3-epoxy-7-tetrahydropyranyloxy -heptane (B4) and 6.6 g of n-undcyl-triphenylphosphonium bromide (prepared in a manner similar to n-dcyl-tryphenylphosphonium brotnide).

IR (CH ₂ Cl ₂ ): 2930, 2860, 1465, 1040 cm ^-1 .

Ella 5 (RS), 6 (RS) -I-acctoxy-5,6-epoxy-7-cis-octadecene

It is prepared under the same conditions as in the previous example, but from 2 (RS), 3 (SR) -7-acetoxy-2,3-epoxy-heptanal (B5) instead of THP derivative B4: oil.

Ε125 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyloxy-7-cis-eicosene g 2 (RS), 3 (SR) -2,3-epoxy-7 tetrahydropyraryl-2651

196,364

oxypeptane (B4) and 6.6 g of n-tridecyltrienylphosphonium bromide.

IR (CH ₂ Cl ₂ ): 2930, 2860, 1465, 1040 cm -1.

El2a 5 (RS), 6 (RS) -1-Aceloxy-5,6-cpoxy-7-cis-cis

Prepared under the same conditions as described in the previous example, but from 2 (RS), 3 (SR) -7-acetoxy-2,3-lithptanal (135): ι B4 instead of THP derivative: oil.

EI3 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-nottadecadiene g from 4 (RS), 5 (RS) -4,5-epoxy-2-trans-nonenal (Cl ) and prepared from 3.8 g of n-dcyl-triphenylphosphonium bromide.

E13a from 5 (RS), 6 (RS) -5,6-epoxy-7-trans-9-cis-eicosadiene g from 4 (RS), 5 (RS) -4,5-epoxy-7-trans-nonenal (Cl ) and prepared from 3.9 g of n-undecyl triphenylphosphonium bromide: oil.

N 3b 4 (RS), 5 (RS) -4,5-epoxy-6-trans-8-cis-nonadecadiene Ig 4 (RS), 5 (RS) -4,5-epoxy-7-trans-octenal (Cla. ) and prepared from 3.9 g of ii-undecyltriphenylphosphonium bromide: oil.

E14-5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydropyranyloxy-7-tris-9-cis-octadecadiene

0.8 g of 4 (RS), 5 (RS) -4,5-epoxy-9- (tetrahydropyranyloxy-2-nzzz-nonenal) (C2) and 2.2 g of n-nonyl triphenylphosphonium bromide ( ... Ohloff, G. et al, Helv.Chim.Acta 60, 1161 (1977)] according to IR (CI 1 ₂ CI _2). 2940,2870,1585,1460,1040 cm⁻¹.

E14a 5 (RS), 6 (RS) -1-Acetoxy-5,6-epoxy-7-trans-9-cis-octadecadicn

Prepared under the same conditions as in the previous example, except for 4 (RS), 5 (RS) -acetoxy-4,5-epoxy-2-trans-noncne (C2a) instead of the THP derivative (C2). : oil.

El5 5 (RS), 6 (RS) -5,6-cpoxy-1-tetralide -mpyranyloxy-7-trans-9-cis-eicosadiene

0.8 g of 4 (RS), 5 (RS) -4,5-epoxy-9-tetrahydropyranyl ooxide from 2-O-nonenal (C2) and 2.4 g of n-undcyl-triphenylphosphonium bromide.

IR (CH ₂ Cl ₂ ): 2940, 2870, 1585, 1450, 1040 cm -1.

E15a 5 (RS 1, 6 (RS) -] - Acetoxy-5,6-cpoxy-7-trans-9-cis-eicosadiene

Prepared under the same conditions as in the previous example, except for 4 (RS), 5 (RS) -9-acetoxy-4,5-epoxy-2-trans-nonenal. (C2a) instead of THP derivative (C2): oil.

E16 5 (RS), 6 (RS) -5,6-epoxy-7,9-trans-11-cis-hexadecairicn

0.95 g of 6 (RS), 7 (RS) -6,7-epoxy-2,4-lance-undckadicol (Dia) and 2.8 g of n-pentylyl triphenylphosphonine bioinide [L. Jaenickc et al., Liebigs Am. Chem. J973, 1252].

IR (C ₁₂ H ₁₂ Cl ₂ ): 2980, 2940, 2880, 1470, 1000, 870 cm -1.

E175 (RS), 6 (RS) -5,6-epoxy-7,1H-cis-9-trans-Texexadecal.

0.78 g of 6 (RS), 7 (RS) -6,7-epoxy-2-trans-4-cis-undecadienal (D1b) and 2.3 g of n-pentyl triphenylphosphonium bromide are obtained.

IR (C 11 H ₁₃ Cl ₂ ): 2975, 2940, 2880, 1470, 1000, 870 cm -1.

f

E17a 5 (RS), 6 (RS) -5,6-epoxy-7,9-trans-11-cis-octadecatriene

Prepared under the same conditions as in the example above, starting from leptyltriphenylphosphonium bromide instead of phenyl derivatives, an oil is obtained.

E18 δ (RS), δ (RS) -5,6-epoxy-7,9-thians-11-cis-eiccatriene

0.65 g of 5 (RS), 7 (RS) -6,7-cpoxy-2,4-trans-undecadiene (Dia) and 4.65 g of n-nonyl triphenylphosphonium bromide are prepared.

Ε 19 5 (RS), 6 (RS) -5,6-epoxy-7,11-cis-9-trans-ejectorate

0.65 g of 6 (RS), 7 (RS) -6,7-epoxy-2-trans-4-cis-undecadienal (D1b) and 4.65 g of n-nonyltriphenylphosphonium bromide are prepared.

IR (CH ₂ Cl ₂ ):

E20 5 (RS), 6 (RS) -5,6-epoxy-1-tetrahydrapyranyl-axi-7,9-trans-11-cis-ejkozaticin

0.66 g of 6 (RS), 7 (RS) -6,7-epoxy-22-telhydroxypyranyloxy-2,4-trans-undecadienal (D2) and

2.8 g of n-nonyl trifluorophosphonyl bromide are obtained.

E20a 5 (RS), 6 (RS) -1-Aceloxy-5,6-cpoxy-7,9-trans-11-cis-cis-cytosine

Prepared under the same conditions as in the previous example, only 6 (RS), 7 (RS) - 11 -acctoxy-6,7-epoxy-2,4-trans-undecadined (D3a) was used to prepare the TI IP derivative. Instead of (D2): oil is obtained.

E21 5 (R.S), 6 (RS) -5-Epoxy-7,9-trans-11,14-cis-eicosacrene

125 mg of 6 (RS), 7 (RS) -6,7-cpoxy-2,4-trans-undecadienal (Dia) and 435 mg of 3-cis-nonenyl-triphenylphosphonium iodide [E. J. Corey et al.,

-2 753

196 3t1

J. Am. Chem. Soc. IR (CH ₂ Cl ₂ ): 2910, 2840, 1450, 990 cm -1.

E22 5 (RS), fi (RS) -5,6-cpoxy-1-tetrahydropyranyloxy-7,9-trans-11,14-cis-ejcatetraene

0.5 g of 6 (RS), 7 (RS) -6,7-epoxy-11-telrahydropyranyloxy-2,4-trans-undckadic (D2) and

2.1 g of 3-cis-noncycl-triflemylphosphonyl iodine are prepared.

E23 5 (RS), 6 (RS) -1-Acetoxy-5,6-epoxy-7,11,14-cis-9-trans-ejectacetraene

0.3 g of 6 (RS), 7 (RS) -11-acetoxy-6,7-cpoxy-2-trans-4-cis-imidecadienal (D3b) and 0.8 g of 3-cis-nonenyl-triphenyl- prepared from phosphonium iodide.

Examples of pharmaceutical compositions and suitable finished pharmaceutical compositions

As used herein, the term "active ingredient" is intended to be understood as meaning the compounds of formula (I) of the present invention, in particular those of claims 1-75. are described in examples such as:

S- (5 (RS), 6 (SR) -5-hydroxy-7-cis-pentadecen-6-yl-cysteine methyl ester);

N- {S- [5 (RS), 6 (SR) -5-Hydroxy-7-cis-heptadecen-6-yl] -N-trifluoroacetyl-cysteinyl} -glycine methyl ester; N- {S- [5 (RS), 6 (SR) -5-Hydroxy-7-trans-9-cis-nonadck: dicn-6-yl] -N-trinoroacetylcystinyl] glycine sodium salt;

N- {S- [5 (RS), 6 (SR) -5-Hydroxy-7,9-trans-11-cis-eccosacrien-6-yl] -N-trifluoroacetylcysteinyl} glycine -methyl ester;

N- (S- [5 (RS), 6 (SR) -5-Hydroxy-7,9-trans-1,14-cis-ejcatetraen-6-yl] -cysteinyl} glycine (also in optically active form) ; and

N- {S- (5 (RS), 6 (SR) -1,5-dihydroxy-7,9-trans-11,14-cis-ejcatetraen-6-yl) -cysteinyl; -glycine potassium salt (optically active form).

Example A)

A solid acrosole inhalation suspension containing 0.1% by weight of the active ingredient.

got included. composition Sidy% Active ingredient, micronized 0.1 Sorbitan trioleate 0.5 The active ingredient (trichlorotrifluoroethane) 4.4 Active substance B (dichlorodifluoromethane and 15.0 1,2-dichloro-tetrafluoroethane) 80.0

manufacture:

The active ingredient is suspended in trichlorotrifluoroethane with the addition of sorbitan trioleate, using a standard homogenizer, to remove moisture, and the suspension is filled into an acrosol container with a metering valve; it is sealed by filling it with active ingredient B under pressure.

Example B)

Suitable for inhalation, approx. With 2% aqueous active ingredients in the form of our sodium or potassium salts. composition

Active ingredient (K or Na salt) 2000 mg

Disodium salt of ethylenediaminetetraacelic acid 10 mg

Benzalkonium chloride 10 mg

V-z, give freshly distilled 100 ml

manufacture:

Approx. It is dissolved in 60 ml of freshly distilled water and the stabilizer (disodium salt of ethylenediaminetetraacetic acid) and the preservative (benzalkonium chloride) are added. After all components have dissolved, the resulting beaker is made up to 100 ml, filled into a pressure bottle and sealed. The active ingredient may be added in gaseous form under pressure or in liquid form as required.

Pharmacological test methods

Guinea pig bronchoconstriction test (in vivo aerosol)

Male guinea pigs (400-700 g) are anesthetized intrapcritonally with 1.4 g / kg urethane and a polyethylene cannula is introduced into the jugular vein. Another polyethylene cannula is introduced into the trachea. With the help of a cannula inserted into the supply line - a Statham transducer! connected - record the pressure in the supply line. The animal is placed in four airtight plexiglas enclosures connected to a No. 000 Fleisch tube and an MP 45-1 Validyne transducer. This arrangement measures the flow.

After surgical preparation of the experimental animal, a period of time is allowed for pulmonary function to stabilize. The test compound is finally administered according to the following protocol. The test animals are treated for one minute with a 1% aerosol solution (w / v) or with distilled water (for control) of the test compound. For each test compound administered by inhalation, a Monaghanull loudspeaker device (Model 670) having a particle size of 1 to 8 microns and a majority of 3 microns is used.

Aqueous solutions are always freshly prepared and introduced into the chamber of the spray vessel by means of an on-stream medication vessel. The spray mist produced is administered to the experimental animals over a 65 ml glass chamber connected to the trachea by a cannula. After the treatment time, a second Moiiaghan ultrasound device (Model 670) and a similar LTD over ₄ glass chambers (0.3 / cg / ml) are added over 2 minutes. The decrease at 3 minutes after LTD _{4 is} read and compared

-2 855

196,364 is the mean value obtained from the three animals with the mean value obtained from the three control animals and the percent inhibition is calculated using the following formula:

,,,,, (100 - picpiliriluiilelJesKiiúíiiy) l {K>

(100 - control! Power)

If different drug concentrations are tested, the percent inhibition at each concentration is recorded and the log is plotted against the abscissa. concentration as a percentage of inhibition on ordinates. IC ₅₀ is then determined by linear regression analysis.

In vitro assay for inhibition of A ₂ phospholipase in human leukocytes

Neutrophilic polymorphic human leukocytes were isolated and deep-frozen starting from "Buffy coals". The phospholipase A ₂ is extracted by homogenization of cell suspension in ice-cold 2N NaCl in 0.36 N H ₂ SO ₄ with and, after centrifugation at 10 x g VCL resulting supernatant was dialyzed to pH 4.5 with sodium acetate buffer against .

To determine enzyme activity, the enzyme (10-30 pg protein) in 0.1 M Tris / HCl buffer, pH 7, with addition of 1 mM CaCl ₂ and substrate ^{, was} radiolabeled with ⁴ C oleic acid in Eserhia coli. (2 μπϊόΙ) phospholipids are incubated at - 37 ° C for 1 hour. The reaction is stopped by the addition of Dola reagent (isopropanol / heptane / 1N H ₂ SO ₄ 40: 10: 1, v / v) and the ¹⁴ C-oleic acid selectively liberated by the A ₂ phospholipase is extracted. Similarly, the co-extracted substrate is completely removed by filtration of the extract on a silica gel column. The ¹⁴ C-oleic acid in the eluate is determined radiochemically.

To determine the inhibitory effect of test substances on A ₂ phospholipase, in solution, in water, dimethylsulfoxide (final concentration in the mixture up to 5% v / v) or ethanol (final concentration in the mixture up to 2,5% v / v) v) added dissolved to the incubation system. The potency of the test substance is characterized by the IC ₅₀ data, i.e. the concentration that exerts a 50% inhibitory effect on the control activity. The IC ₅₀ is determined graphically by applying the percentage inhibitory effect to the ordinate against the log concentration (μπϊόΙ) on the abscissa.

Under the experimental conditions described, mepacrine inhibits A ₂ phospholipase with a 1600 gm IC ₅₀ .

In vitro assay for inhibition of phospholipase C in human platelets

Human platelets are obtained from "Buffy coats" by fractional centrifugation and finally freezing. Phospholipase C is released by sonication of the cell suspension and, after ultracentrifugation (150,000 x g for 1 hour), is soluble in the supernatant.

To determine enzyme activity, the enzyme (20-100 pg protein) in 0.025 M Tris / maleate ptiffer pH 6 with 0.2 mM CaCl ₂ and 0.02 mM radiolabeled substrate - phoslatidyl [ ¹⁴ C] -isolite Incubate at 37 ° C for 5 minutes. The reaction was quenched by extraction with CHCl ₃ / CH ₃ OH 2: 1 (v / v). Thus, the unused substrate is extracted into the organic phase while the ¹⁴ C-isonite phosphate reaction product remains in the aqueous phase and an aliquot can be measured radiometrically.

To determine the inhibitory effect of the test substance on phospholipase C, they are added in solution in water, dimethylsulfoxide (final concentration in the mixture up to 5% v / v) or ethanol (final concentration in the mixture 2.5% v / v). for the incubation system. L the degree of saturation of the test substances is characterized by the IC ₅₀ , that is, the concentration which exerts a 50% inhibitory effect on the control activity. The IC ₅₀ is plotted by plotting the percentage inhibitory effect on the ordinate against the log concentration (μιηύΙ) on the abscissa.

Under the experimental conditions described, mepacrine inhibits phospholipase C by a value of 20 μιηόϊο5 lC ₅₀ .

Claims (7)

Claims Process 1 is a process for preparing a compound of formula (I): wherein: R 'is a C 1-3 alkyl group or a C 1-3 hydroxyalkyl group whose hydroxyl group may be esterified with a 1-4 carbon monoxide alkanecarboxylic acid or protected with a tetrahydropyranyl group, R ² is a C 3 -C 15 saturated aliphatic group or C 8 -C 12 unsaturated aliphatic group, R ³ is hydroxy, C 1-4 alkoxy or -NH-CH ₂ -CO-R ³ (-Rb) substituted amino, nitol R ³ , hydroxy or C 1-4 alkoxy cs -X- is a single bond, methylene or an optionally N-acylated primary amino methylene group of the formula CH-NH-R 4 (-X <r-) in which N is hydrogen or an acyl group of a polilineactive (C 2 -C 5) -alkanecarboxylic acid with the oxygen atom of the hydroxy group relative to the sulfur atom in the trans configuration is - as well as alkali or aliphatic ammonium salt compounds have the salt-forming properties, characterized in that (II) traiisz-epoxidol formula - wherein R ¹ cs R ² have the same meanings as defined above and wherein an optional hydroxyl group may be in protected form - a mercaptoalkanecarboxylic acid derivative of formula (III) al - wherein R ³ and -X- have the same meaning as previously defined and in which the optionally present amino group may be in protected form, and if necessary or desired, the hydroxyl and / or amino group (s) is protected 29 -2 957 196 364 groups or protecting groups thereof Ichasftjuk and / or cj. ' esterifying the compound in its ester form to the free acid or salt and / or converting the resulting free compound having salt-forming properties to the salt and / or liberating the compound from the corresponding salt. _{(E. M4} "jj. A process for the preparation of compounds of formula (I) according to claim 1 wherein R ¹ is C 1 -C 3 alkyl, R ² , R ³ and -X- are as defined in claim 1, characterized in that A compound of Formula II wherein R 1 and R ² are as defined above, with a compound of Formula III wherein R ³ and -X- are as defined above (E .: June 28, 1984). The process for the preparation of compounds of formula I according to claim 1, wherein R 'is ethyl or β-hydroxyethyl which may be esterified with a C 1-4 alkanoic acid, R ² , R ³ and -X - as defined in claim 1, characterized in that a compound of formula (II) wherein R 'and R ² are as defined above, a compound of formula (III) in which R ³ and -X- are as defined above - with. _(E . _; June 28 _, 1984) A scintillational process for the preparation of compounds of general formula (I) wherein R 'is ethyl, (S-hydroxyethyl or (O-acetoxyethyl), R ² , R ³ and -X- are as defined in claim 1, characterized in that (II) compound in which R ¹ and R ^z are as defined above, (III) as defined above - are reacted - in which R ³ and X- are as defined above. (E .: June 28, 1984) The process for the preparation of ten compounds of formula I according to claim 1, wherein R ² is C 5 -C 15 alkyl or the corresponding one, two or three double bonds, C 8 -C 12, R ¹ , R ³ and -X- are as defined in claim 1, wherein a compound of formula (II) wherein R 'and R ² are as defined above is a compound of formula (III) with R ³ and -X- as defined above. _(E. ] _{984. 06 2} sec.) The process for the preparation of compounds of formula (I) according to claim 1, wherein R ² is a non-branched C 5 -C 15 alkyl group, R 1, R ² and -X- are as defined in claim 1, characterized in that a compound of formula (II) in which R ¹ and R ² is as defined above, with a compound of formula (III) in which R ³ and -X- meaning above - we react. _{£ · _{19g4 θ6 2g} 5g} A process for the preparation of a compound of formula (I) according to claim 1, wherein R ² is a non-branched C 1 -C 12 alkenyl group, R ', R ³ and -X- are as defined in claim 1, characterized in that a compound of formula (II) wherein R' and R ² are as defined above with a compound of formula (III) in which R ³ and - X - is as defined above.