HU227116B1 - New propenecarboxylic acid amidoxime derivatives, process for their preparation and pharmaceutical compositions containing them - Google Patents

Abstract

Propenecarboxylic acid amidoxime derivatives (I) are new. Propenecarboxylic acid amidoxime derivatives of formula (I), their N-oxides, geometrical isomers, optical isomers, acid addition salts and/or quaternary derivatives are new. [Image] R : 1-20C alkyl, 5- or 6-membered heterocyclic group containing 1-2 N atom(s) or S atom, optionally fused with at least one benzene ring and/or heterocyclic group, or a phenyl optionally substituted with 1-3 substituents selected from T, (1-4C alkyl)amino, di-(1-4C alkyl)amino and/or (1-4C alkanoyl)amino; R' : H; or R+R' : 5-7C cycloalkyl optionally fused with benzene ring; R 4, R 5H, 1-5C alkyl, 1-5C alkanoyl or phenyl optionally substituted with 1-3 substituents selected from T; T : halo, 1-2C alkyl and/or 1-2C alkoxy; NR 4R 55- or 6-membered heterocyclic group optionally containing N, O and/or S atom and optionally fused with benzene ring (benzene ring and/or heterocyclic group optionally substituted with 1-2 T); R 1, R 2H; R 3H, OH, 1-5C alkoxy, halo, 1-5C alkylthio, 1-20C alkanoyloxy, 3-22C alkenyloxy containing at least one double bond, methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy; or R 1+R 2carbonyl or thiocarbonyl, the carbon atom of which is bound to the O atom adjacent to R 1and the N atom adjacent to R 2; or R 1+R 3valence bond between the O atom adjacent to R 1and the C atom adjacent to R 3; provided that: when R 1and R 2= H, R 3= H, OH or 1-5C alkoxy. An independent claim is also included for preparation of (I). ACTIVITY : Vasotropic; Immunosuppressive; Antidiabetic; Virucide; Hepatotropic; Antiinflammatory; Antiulcer; Gastrointestinal; Antipsoriatic; Dermatological; Antirheumatic; Antiarthritic; Nootropic; Neuroprotective; Antiparkinsonian. MECHANISM OF ACTION : Poly(adenosine diphosphate ribose) polymerase (PARP) inhibitor. In tests to determine PARP inhibition, 3-styryl-4-(3-pyrrolidino-2-hydroyxpropyl)-delta 2>-1,2,4-oxazolin-5-one hydrochloride displayed an I 0.5of 8 mg/l.

Description

R 'is hydrogen, or R and R'together are a C 5 -C 7 cycloalkyl group optionally fused to a benzene ring,

R ₄ and R ₅ each independently represent hydrogen, C 1 -C 5 alkyl, C 1 -C 5 alkanoyl or phenyl optionally substituted with 1 to 3 substituents wherein the substituent is halogen and / or C 1 -C 2 alkyl and / or C 1 -C 2 alkoxy, or R ₄ and R _{5 taken} together with the adjacent nitrogen atom form a one or six membered saturated or unsaturated heterocyclic group which may also contain a further nitrogen atom and / or oxygen and / or sulfur atom as heteroatom and may be fused to a benzene ring; the heterocyclic group or the benzene ring may carry one or two substituents wherein the substituent is C 1-2 alkyl and / or C 1-2 alkoxy and / or halogen,

CH-CH <

í ^K 5

The length of the description is 30 pages (including 4 pages)

HU 227 116 Β1

R-ι and R ₂ are hydrogen, and

R ₃ is hydrogen, hydroxy or C 1-5 alkoxy, or

R-ι and R ₂ together form a carbonyl or thiocarbonyl group, the carbon atom of which is bound to an oxygen atom adjacent to R 1 and a nitrogen atom adjacent to R ₂ , and

R ₃ is hydrogen, halogen, hydroxy, C 1 -C 5 alkoxy, C 1 -C 5 alkylthio, C 1 -C 20 alkanoyloxy, C 3 -C 22 alkenoyloxy having one or more double bonds a methylsulfonyloxy group, a benzenesulfonyloxy group or a toluenesulfonyloxy group, or

R ₂ is hydrogen, and

· R! and R ₃ together form a direct bond between the oxygen atom adjacent to R _r and the carbon atom adjacent to R ₃ .

The present invention relates to novel propenecarboxylic acid amidoxime derivatives, to processes for their preparation and to pharmaceutical compositions containing them as active ingredients. The novel compounds have valuable therapeutic effects and can be used in cellular deficiency conditions, diabetic complications, oxygen deficiency of the heart and brain, neurodegenerative diseases, in the treatment of autoimmune and / or viral diseases, and in the prevention of toxic effects.

More particularly, the invention relates to novel propenecarboxylic acid amidoxime derivatives of formula I wherein:

R is C 1 -C 20 alkyl, phenyl optionally substituted with 1 to 3 substituents - wherein the substituent is halogen and / or C 1 -C 2 alkyl and / or C 1 -C 2 alkoxy and / or amino; C 1 -C 4 alkylamino and / or di (C 1 -C 4 alkyl) amino and / or C 1 -C 4 alkanoylamino, and 5 or 6 membered, saturated or unsaturated a heterocyclic group containing one or more nitrogen or sulfur atoms as heteroatoms and optionally fused to one or more benzene rings and / or other heterocyclic groups, and

R 'is hydrogen or 40

R 'of the gap, taken together, is a C 5 -C 7 cycloalkyl group optionally fused to a benzene ring,

R ₄ and R ₅ are each independently hydrogen, C 1-5 alkyl, C 1-5 alkanoyl, or phenyl optionally substituted with 1-3 substituents wherein the substituent is halogen and / or C 1-2 alkyl. and / or C 1-2 alkoxy;

R ₄ and R ₅ together with the adjacent nitrogen atom form a 5 or 6 membered saturated or unsaturated heterocyclic group which may also contain a further nitrogen atom and / or oxygen atom and / or sulfur atom as heteroatom and may be fused to a benzene ring and the benzene ring may carry one or two substituents wherein the substituent is C 1-2 alkyl and / or C 1-2 alkoxy and / or halogen;

· R! and R ₂ is hydrogen; and

R ₃ is hydrogen, hydroxy or

C 1-5 alkoxy, or R ₁ and R ₂ together form a carbonyl or thiocarbonyl group whose carbon atom adjacent to the oxygen atom and R _r digested with the nitrogen atom adjacent to R _2, and

R ₃ is hydrogen, halogen, hydroxy, C 1 -C 5 alkoxy, C 1 -C 5 alkylthio, C 1 -C 20 alkanoyloxy, C 3 -C 22 alkenoyloxy having one or more double bonds a methylsulfonyloxy group, a benzenesulfonyloxy group or a toluenesulfonyloxy group, or

R ₂ is hydrogen, and

R-ι and R _{3 are taken} together to form a direct bond between the oxygen atom adjacent to R _r and the carbon atom adjacent to R ₃ , and pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof.

The preparation of some Δ ² -1,2,4-oxadiazolin-5-one derivatives without reference to biological activity is described in Chem. Bér., 103, 2330-2335 (1970). The preparation of 5,6-dihydro-4H-1,2,4-oxadiazine derivatives from these latter compounds is discussed in Chem. Bér., 108, 1911-1923 (1975), which also do not indicate the possible biological activity of the compounds.

Hungarian Patent No. 179,951 discloses 1,2,4-oxadiazolin-5-one derivatives having peripheral vasodilator, antianginal and antiarrhythmic activity. Hungarian Patent Application No. 180 708 discloses 1,2,4-oxadiazine derivatives having peripheral vasodilator and antihypertensive, antiarrhythmic, mild anti-inflammatory and diuretic action. However, the known 1,2,4-oxadiazolin-5-one and 1,2,4-oxadiazine derivatives do not contain an alkenyl group between their substituents, i.e. they are not derived from propenecarboxylic acid amidoxime.

It is an object of the present invention to provide novel compounds which have commercially effective effects.

It has now been found that this object is achieved by the novel propenecarboxylic acid amidoxime derivatives of formula (I).

As used herein and in the claims, C1-C20 alkyl is, for example, methyl, ethyl2

EN 227 116 1, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, n-pentyl, n-hexyl, n-heptyl. a n-decyl group, a dodecyl group, a hexadecyl group, an octadecyl group, and the like. means.

The halogen atom is preferably fluorine, chlorine or bromine, preferably chlorine or bromine.

C 1-2 alkyl is methyl or ethyl, while C 1-2 alkoxy is methoxy or ethoxy.

C 1-4 alkyl means methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl or isobutyl. In addition to the above, the C 1-5 alkyl group may also be n-pentyl.

Examples of (C1-C4) alkylamino include methylamino, ethylamino, isopropylamino and the like. Examples of di (C 1-4 alkyl) amino groups include dimethylamino, diethylamino, methylisopropylamino and the like.

C 1-4 alkanoyl is preferably formyl, acetyl, n-propionyl or n-butyryl. C 1-5 alkanoyl is, for example, n-pentanoyl.

A 5 or 6 membered saturated or unsaturated heterocyclic group containing one or two nitrogen atoms or one sulfur atom as a heteroatom, for example pyrrolyl, pyrazolyl, imidazolyl, thienyl, pyridyl, piperidyl, pyrimidinyl, piperazinyl and the like.

C 5 -C 7 cycloalkyl optionally fused to a benzene ring include, for example, cyclopentyl, cyclohexyl, cycloheptyl, indanyl or tetralin.

A 5 or 6 membered saturated or unsaturated heterocyclic group which may contain a nitrogen atom adjacent to the substituents R ₄ and R ₅ and a further nitrogen atom and / or oxygen and / or sulfur atom, for example a morpholino group in addition to the heterocyclic groups listed above.

C 1-20 alkanoyloxy groups include, for example, formyloxy, acetoxy, propionyloxy, butyryloxy, capryloxy, palmityloxy, stearyloxy and the like.

The C 3 -C 22 alkenoyloxy group will generally have 1 to 6 double bonds, and preferably linolenoyloxy group, linoloyloxy group, docosahexaenoyloxy group, eicosapentaenoyloxy group or arachidonoyloxy group.

The pharmaceutically acceptable acid addition salts of the propenecarboxylic acid amidoxime derivatives of formula I with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, etc., or with organic acids such as acetic acid, fumaric acid, lactic acid, succinic acid, succinic acid, succinic acid, with toluenesulfonic acid, etc. non-toxic salts thereof.

Quaternary derivatives of compounds of formula (I) are those compounds which have one or more nitrogen atoms quaternized, i.e., for example, an additional C 1 -C 4 alkyl group or phenyl (C 1 -C 4 alkyl) group attached to said nitrogen atom and the nitrogen atom is positive. becomes charged. Preferably, the nitrogen atom to which the R ₄ and R ₅ substituents are attached is one of the nitrogen atoms present in the compound of formula (I). When R in the formula I is a heterocyclic group containing nitrogen, such as pyridyl, its nitrogen may also be quaternized.

The N-oxides of the compounds of formula (I) are those compounds in which one or more nitrogen atoms are present in an oxidized form and one nitrogen atom is attached to said nitrogen atom. Among the nitrogen atoms present in the compound of formula (I), the nitrogen atom to which the R ₄ and R ₅ substituents are attached is suitably present as the N-oxide. When R is a heterocyclic group containing a nitrogen atom, such as a pyridyl group, the nitrogen atom thereof may also be present in the form of an N-oxide.

Due to the double bond present in the formula (I), the new propenecarboxylic acid amidoxime derivatives exhibit geometric isomerism and may therefore be present as cis or trans isomers or as mixtures thereof. The invention encompasses both pure geometric isomers and mixtures of isomers.

In addition to the geometric isomerism, some of the compounds of formula I have one or more chiral carbon atoms and may therefore exist as optical isomers. The invention also encompasses individual optical isomers and any mixtures thereof.

A preferred subgroup of the propenecarboxylic acid amidoxime derivatives of the present invention are the propenecarboxylic acid amidoxime derivatives of formula (Ia), wherein

R, and R ₂ is hydrogen,

R ₃ is hydrogen, hydroxy or

C 1-5 alkoxy,

R, R ', R ₄ and R ₅ are as defined above and further represent their geometric and / or optical isomers and / or their pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides.

According to the invention propenoate amidoximszármazékok Another preferred subgroup of the oxa-diazolin derivatives of the formula (Ib) wherein R ₁ and R ₂ together form a carbonyl or thiocarbonyl group having a carbon atom of the _Rf adjacent gel oxygen atom adjacent to the R ₂ with the nitrogen atom connects,

R ₃ is hydrogen, halogen, hydroxy, C 1 -C 5 alkoxy, C 1 -C 5 alkylthio, C 1 -C 20 alkanoyloxy, C 3 -C 22 alkenoyloxy having one or more double bonds , methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy,

HU 227 116 Β1

R, R ', R ₄ and R ₅ are as defined for formula (I), X is oxygen or sulfur, and their geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof. They are.

A further preferred subgroup of the propenecarboxylic acid amidoxime derivatives of the present invention are the oxadiazine derivatives of the general formula (le) wherein R ₂ is hydrogen,

R ι and R ₃ together form a bond adjacent to R _q -gyel oxygen atom and R ₃ malate between adjacent carbon atoms,

R, R ', R _4, and R ₅ are as defined in formula (I) and are geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof.

The propenecarboxylic acid amidoxime derivatives of formula (I) are prepared by:

a) for the preparation of a propoxycarboxylic acid amidoxime derivative of the general formula (Ia) wherein R _q and R ₂ are hydrogen, R ₃ is hydrogen or C 1-5 alkoxy, R, R ', R ₄ and R ₅ are a propene derivative of formula (II), wherein R, R ', R ₃ , R ₄ and R ₅ are as defined above, Y is halogen or a group of formula -SR ₆ wherein Rg is hydrogen or (C1-C4) -alkyl is reacted with hydroxylamine; obsession

b) for the preparation of a propoxycarboxylic acid amidoxime derivative of the general formula (Ia) wherein R _q and R ₂ are hydrogen, R ₃ is hydrogen, hydroxy or C 1-5 alkoxy, R, R ' R ₄ and R ₅ are as defined in formula (I), an oxa-diazolin derivative of formula (Ib) wherein R, R ', R _3, R ₄ and R ₅ are as defined above, X is oxygen or sulfur with an aqueous solution of an alkali metal hydroxide;

obsession

c) for the preparation of an oxadiazoline derivative of the Formula Ib wherein R ₃ is hydrogen or C 1 -C 5 alkoxy, X is O, R, R ', R ₄ and R ₅ is a Δ ² -1,2,4-oxadiazole derivative represented by formula (I): wherein R and R 'are as defined above, with an aminoalkyl halide of formula (IV) wherein Z is halogen; R ₃ , R ₄ and R ₅ are as defined above; obsession

d) for preparing an oxadiazoline derivative of the general Formula I wherein R ₃ is hydrogen, hydroxy or C 1 -C 5 alkoxy, X is O, R, R ', R ₄ and R ₅ is a Δ ² -1,2,4-oxadiazoline derivative represented by formula (I) wherein R and R 'are as defined above, a compound of formula (V) -dihalopropane, where Z and Zq are each independently halogen, and the resulting Δ ² -1,2,4-oxadiazolinylalkyl halide of formula VI wherein R, R 1, R 3 and Z is as defined above for reaction with an amine of formula VII wherein R ₄ and R ₅ are as defined above; obsession

e) for the preparation of an oxadiazoline derivative of the general formula (Ib), wherein R ₃ is a hydroxy group, X is an oxygen atom, R, R ', R ₄ and R ₅ are a compound of the general formula (I). as defined for formula, formula (III) of formula Δ ² -1,2, 4-oxa-diazolin derivative wherein R and R 'are as defined above, with epichlorohydrin, and the formed (VIII) formula ² Δ -1.2 Reacting 4-oxadiazolinylalkyl chloride, wherein R and R 'are as defined above, with an amine of formula VII wherein R ₄ and R ₅ are as defined above; obsession

f) for the preparation of an oxadiazoline derivative of the general formula (Ib), wherein R ₃ is a hydroxy group, X is an oxygen atom, R, R ', R ₄ and R ₅ are a compound of the general formula (I); a Δ ² -1,2,4-oxadiazolinylalkyl chloride of formula VIIIa, wherein R and R 'are as defined above, by reaction with an acid acceptor, and the resulting epoxide of formula IX, wherein R and R 'are reacted with an amine of formula (VII) above, wherein R ₄ and R ₅ are as defined above; obsession

g) for the preparation of an oxadiazoline derivative of the Formula Ib wherein R, R ', R ₄ and R ₅ are as defined for Formula I, R ₃ is hydrogen, hydroxy; or a C 1 -C 5 alkoxy group, X is oxygen or sulfur, a propoxycarboxylic acid amidoxime derivative of formula (Ia) wherein R, R ', R ₃ , R ₄ and R ₅ are as defined above with a carbonic acid derivative of formula (X) wherein X is as defined above, Z ₂ and Z ₃ are each independently halogen, C 1-4 alkoxy, or C 1-4 alkyl mercapto; obsession

h) for the preparation of an oxadiazine derivative of the Formula I wherein R, R ', R ₄ and R ₅ are as defined for Formula I, a compound of Formula Ib an oxadiazoline derivative wherein R, R ', R ₄ and R ₅ are as defined above, X is oxygen or sulfur, R ₃ is halogen, methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy; reacting a group with alkali metal hydroxide in the presence of water; obsession

i) for the preparation of an oxadiazine derivative of the general Formula I wherein R, R ', R ₄ and R ₅ are as defined for Formula I, for the preparation of a compound of Formula XI; a cyclic compound wherein R and R 'are as defined above, R ₇ is halogen, methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy with an amine of formula VII wherein R ₄ and R ₅ is as defined above; obsession

j) for the preparation of a quaternary derivative of a compound of formula I wherein X, R ',

HU 227 116 Β1

R _h R _2, R _3, R ₄ and R ₅ are as defined in formula (I), _R8 is C1-4 alkyl or phenyl (C1 -C4) alkyl, Y is a halogen atom or an R ₈ -SO ₄ , a Δ ² -1,2,4-oxadiazoline derivative of formula (III) wherein R and R 'are as defined above, with a quaternary alkyl halide of formula (XIII) wherein R _{3, R4, R5, R8} and Y are as defined above and Z is halogen, is reacted; obsession

k) for the preparation of the N-oxide of the formula I in the formula XIV wherein R, R ', R 1, R ₂ , R ₃ , R ₄ and R ₅ are as defined in formula I; A Δ ² -1,2,4-oxadiazole derivative of formula (III) wherein R and R 'are as defined above, with a compound of formula XV wherein R ₃ , R ₄ and R ₅ are as defined above, Z is halogen, reacting;

and, if desired, a compound of formula (Ib) wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined in formula (I), X is oxygen or sulfur by reacting R ₃ with halogen. containing a compound of formula Ib; or, if desired, a compound of formula (Ib) wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined for formula (I), X is oxygen or sulfur, with a C 1 -C 20 alkanecarboxylic acid halide or by reaction with a C ₃ -C ₂₂ alkenecarboxylic acid halide containing one or more double bonds to form a compound of formula Ib wherein R ₃ is C 1 -C 20 alkanoyloxy or C ₃ -C ₂₂ alkenoyloxy; or, if desired, a compound of formula Ib, wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined in formula I, X is oxygen or sulfur, a C 1 -C 5 alkyl group; reacting with a halide to form a compound of formula (Ib) wherein R ₃ is C 1-5 alkoxy; or, if desired, a compound of formula (Ib) wherein R ₃ is halogen, R, R ', R ₄ and R ₅ are as defined for formula (I), X is oxygen or sulfur, a C 1 -C 5 alkanol or reacting with an alkali metal salt of thioalkanol to form a compound of formula (Ib) wherein R ₃ is C 1-5 alkoxy or C 1-5 alkylthio. or, if desired, a resulting compound of formula (Ib) wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined in formula (I), X is oxygen or sulfur, with methylsulfonyl halide, benzenesulfonyl halide or reacting with toluenesulfonyl halide to form a compound of formula Ib wherein R ₃ is methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy; and optionally converting the resulting compound of formula (I) to a pharmaceutically acceptable acid addition salt thereof with an inorganic or organic acid and / or quaternizing one or more nitrogen atoms of the compound of formula (I) with an alkylating agent and / or A) is converted to N-oxide by oxidizing agent on one or more nitrogen atoms.

In the process (a) of the present invention, the reaction of the propene derivative (II) with the hydroxylamine in a solvent or mixture of solvents is carried out with a hydroxylamine base which can be liberated in situ by addition of a strong base of an acid addition salt of hydroxylamine. The resulting product of formula (Ia) is isolated in a manner known per se, for example by crystallization from the reaction mixture, evaporation of the reaction mixture, or isolation of the acid addition salt thereof.

When starting from a propene derivative of formula II wherein Y is a halogen, the solvent is an anhydrous organic solvent such as a halogenated hydrocarbon such as chloroform, dichloromethane, etc., a hydrocarbon such as benzene, toluene, etc., or other solvents customary for acylations, such as pyridine.

When starting from a propene derivative of the formula II in which Y is -SR ₆ , in addition to the solvent types listed above, for example, alkanols can be used as organic solvents.

The propene derivative of formula (II) wherein Y is a halogen atom, usually a chlorine atom, which is in fact an imidoyl halide, usually an imidoyl chloride, from the corresponding acid amide of formula (XVI) wherein R, R ', R ₄ and R ₅ are ), R ₃ is hydrogen or C 1-5 alkoxy, with a halogenating agent, preferably thionyl chloride, phosphorus trichloride, phosphorus pentachloride and the like. is prepared in a manner known in the art.

For example, the propene derivative of formula (II) wherein Y is a mercapto group may be prepared from the corresponding acid amide (XVI) with phosphorus pentasulfide in an organic solvent such as toluene, xylene or pyridine, as known in the art. A propylene derivative of formula II wherein Y is a mercapto group is used to alkylate a propylene derivative of formula II wherein Y is an alkylthio group.

In process (b) of the present invention, the oxadiazole ring is opened by aqueous alkaline hydrolysis using the procedure known in Chem. Bér., 103, 2330-2335 (1970). The alkali metal hydroxide is preferably potassium hydroxide or sodium hydroxide, to which an aqueous solvent, if desired an organic solvent, preferably an aliphatic alcohol such as methanol or ethanol, is added. Under the conditions of the process of the invention, the opening of the oxadiazole ring at the boiling point of the reaction mixture generally takes place in a short period of time and affords the compound of formula Ia in good yield which can be isolated in a manner known per se.

In the process c) according to the invention, the reaction is carried out in a reaction-inert organic solvent,

The reaction is carried out in the presence of an acid acceptor, usually at the boiling point of the reaction mixture. The inert organic solvent is, for example, an alkanol such as methanol or ethanol, a hydrocarbon such as benzene, toluene or xylene or a mixture thereof. Inorganic or organic bases may be used as acid binders. The reaction mixture can be worked up by conventional means, for example, after evaporation of the solvent, the product is crystallized or its acid addition salt is isolated.

The starting material A ² -1,2,4-oxadiazole derivatives of formula (III) can be prepared from the corresponding amidoximes with carbonic acid derivatives. Some representatives of amidoximes are known from Chem. Rews., 62, 155 (1962), and the new compounds can be prepared as described therein from the corresponding propenecarboxylic acid nitrile with hydroxylamine. The aminoalkyl halides of formula (IV) are largely known compounds and are either commercially available or can be readily prepared by reaction of 1,3-dihalopropane (V) with an amine (VII).

In process (d) of the present invention, both the alkylation, i.e. the reaction of the A ² -1,2,4-oxadiazoline derivative of formula (III) with the 1,3-dihalopropane of formula (V), and the amination, i.e. The reaction of the Δ ² -1,2,4-oxadiazolinylalkyl halide of formula (VI) with the amine of formula (VII) in an inert organic solvent, an acid scavenger, preferably an inorganic base such as sodium hydroxide or sodium carbonate in the presence of the reaction mixture, generally at the boiling point of the reaction mixture. The Δ ² -1,2,4-oxadiazolinylalkyl halide of formula (VI) formed during alkylation is crystallized or, after evaporation of the reaction mixture, used for the amination without crystallization. The resulting reaction product of formula (Ib) is isolated in a manner known per se by one of the methods described above. The inert organic solvent may be, for example, a hydrocarbon, a halogenated aliphatic or aromatic hydrocarbon such as chloroform, an alkanol such as methanol or ethanol, a ketone such as acetone, or a mixture of the solvent types listed.

In the process (e) of the present invention, the reaction of the A ² -1,2,4-oxadiazoline derivative of formula (III) with epichlorohydrin is carried out in an organic solvent inert or in the presence of a solvent, preferably in excess of . Inert organic solvents include, for example, hydrocarbons, ethers such as dioxane, tetrahydrofuran and the like. may. The catalysts used are bases such as sodium hydroxide, sodium carbonate and the like.

After completion of the reaction, the solvent was evaporated and the residue crystallized. The reaction of the resulting Δ ² -1,2,4-oxadiazolinylalkyl chloride (VIII) with the amine (VII) is carried out in a similar manner to the amination described in process d). The resulting reaction product of formula (Ib) is isolated in a manner known per se by one of the methods described above.

In the process (f) of the present invention, for example, an alkali metal carbonate such as sodium carbonate, potassium carbonate, etc., or an alkali metal hydroxide, such as sodium hydroxide, potassium hydroxide and the like, are used as acid scavengers. used. The reaction is carried out in an organic solvent inert to the reagents, preferably at the boiling point of the reaction mixture. Examples of inert organic solvents include hydrocarbon, acetone, ether such as tetrahydrofuran or dioxane, halogenated aliphatic or aromatic hydrocarbons, and the like. used. The reaction mixture is filtered, evaporated, the epoxide (IX) formed is crystallized, and then reacted with the amine (VII) as described for the amination in process (d), preferably in an alkanol. Alternatively, however, the epoxide of formula (IX) is not isolated but the amine of formula (VII) is added to the reaction mixture and the reaction mixture is further heated. The resulting reaction product of formula (Ib) is isolated in a manner known per se by one of the methods described above.

In the process g), the ring closure can be carried out with any carbonic or thiocarbonic acid derivative represented by the general formula (X) which is the oxygen atom of the hydroxy group at the portion of the propenecarboxylic acid amidoxime derivative (Ia) represented by the formula -C (= N-OH) -NH and to form a carbonyl or thiocarbonyl group between the nitrogen of the amino group. Suitable compounds for this purpose are the halides of carbonic acid and thiosenoic acid (e.g., phosgene and thiophosgene), halide esters (such as ethyl chloro-carbonic acid alkyl esters) or esters of carbonic acid and thiosenoic acid (e.g. such as dialkyl carbonates, mono-, di- and trithiocarbonates, xanthogenates, etc.). For the ring closure reaction, an inert organic solvent is used, although it may be employed without a solvent. The reaction mixture is cooled or heated, preferably at its boiling point. The resulting reaction product of formula (Ib) is isolated in a manner known per se by one of the methods described above.

In the case of the reaction using a carbonic acid derivative of the formula (X) wherein one or both of Z ₂ and Z ₃ are halogen, the inert organic solvent is preferably a hydrocarbon, a halogenated aliphatic or aromatic hydrocarbon or an ether. When Z ₂ and Z ₃ are both alkoxy or alkyl mercapto, the inert organic solvent may be, in addition to those listed above, alkanol.

In the process h) of the present invention, the oxadiazine ring is in fact transformed into an oxadiazine ring. For this, the method known from Chem. Bér., 108, 1911-1923 (1975) is used. The alkali metal hydroxide is preferably sodium hydroxide or potassium hydroxide. The reaction is carried out in a mixture of an organic solvent such as an alkanol and an aqueous alkali metal hydroxide solution at the boiling point of the reaction mixture. The resulting reaction product of formula (le) is itself

It is isolated in a manner known per se by one of the methods described above.

In the process (i) according to the invention, the reaction is carried out in an organic solvent inert to the reagents or in a mixture of several such solvents, in the presence or absence of an acid acceptor. The inert organic solvent is, for example, hydrocarbon, halogenated aliphatic or aromatic hydrocarbon, ether, alkanol, preferably butanol. The reaction may also be carried out in the absence of an organic solvent, in which case an excess of the amine of formula (VII) may serve as a solvent. The resulting reaction product of formula (le) is isolated in a manner known per se by one of the methods described above.

Processes j) and k) according to the invention are carried out in a manner similar to that described in process c). The quaternary alkyl halide of formula (XIII) may be prepared by quaternization of the corresponding aminoalkyl halide of formula (IV). The compound of formula (XV) may be prepared from the corresponding aminoalkyl halide of formula (IV) by oxidation.

R ₃ is appropriate diazolin oxa-hydroxy derivative (Ib) can be converted to the corresponding (Ib), a compound of formula R ₃ is halogen appropriate halogenating agent. Preferably the halogenating agent is thionyl chloride, phosphorus trichloride or phosphorus pentachloride and the reaction is carried out in conventional organic solvents or without the use of a solvent such as an excess of the halogenating agent. The reaction mixture is worked up after the halogenation reactions by conventional methods.

The oxadiazole derivative of formula (Ib) wherein R ₃ is hydroxy is an active acylating derivative of a C 1 -C 20 alkane carboxylic acid or a C 3 -C 22 alkenecarboxylic acid, such as a halide, anhydride, azide, etc., methylsulfonyl halide, halosulfonic acid or benzenesulfonyl may be reacted in an inert organic solvent, preferably an aromatic hydrocarbon or a halogenated aromatic or aliphatic hydrocarbon, with or without an acid acceptor. The resulting product of formula (Ib) may be isolated by conventional methods as described above.

Similarly, a compound of formula (Ib), wherein R ₃ is hydroxy, may be reacted with a C 1-5 alkyl halide. In this case, one or more nitrogen atoms of the compound may be quaternized simultaneously.

A compound of the formula and an alkanol or tioalkanol R ₃ is halogen, preferably chlorine atom, (Ib), the reaction of the alkali metal salt also can be carried out between the above-described reaction conditions.

If desired, the compound of formula (I) is converted into, or liberated from, a pharmaceutically acceptable acid addition salt. In the case of salt formation with an optically active organic acid such as camphoric acid, camphorsulfonic acid, tartaric acid or tartaric acid derivatives, it is possible to separate the stereoisomers of the compounds containing the chiral center. The resolution is then resolved in a manner known per se by crystallization of the acid addition salts with an optically active acid.

If desired, one or more nitrogen atoms of the propenecarboxylic acid amidoxime derivative of formula (I) are quaternized. To this end, for example, a compound of formula (I) is reacted with an alkylating agent of formula R ₈ -Y wherein R ₈ is C 1 -C 4 alkyl or phenyl (C 1 -C 4 alkyl), Y is halogen, and (XII) ), wherein R ₈ and Y are as defined above. The quaternization can also be carried out with a dialkyl sulfate of the formula (R ₈ ) ₂ SO ₄ , wherein R ₈ is as defined above. In this case, a quatemer derivative of formula (XII) is formed wherein Y is a group of formula R ₈ -SO ₄ . The quaternization is carried out with or without an inert organic solvent.

The reaction may also be carried out by quaternizing another nitrogen atom of the compound of formula I or by quaternizing another nitrogen atom. When R in the formula (I) is a heterocyclic group containing nitrogen, such as pyridyl, its nitrogen atom may be quaternized or its nitrogen atom quaternized.

The conversion of the compounds of formula (I) into N-oxide may be effected so that the nitrogen atom to which R ₄ and R ₅ are attached is conveniently oxidized to N-oxide. The oxidation is usually carried out with hydrogen peroxide, preferably in an alkanolic solution such as methanol, preferably at room temperature. Alternatively, or simultaneously, the nitrogen atom of the nitrogen-containing heterocyclic group present at the R site, such as pyridyl, may be converted to the N-oxide by an oxidizing agent. In the latter case, the oxidizing agent is preferably peroxy acid such as 3-chloroperbenzoic acid and the oxidation is carried out in an inert organic solvent, usually an aromatic hydrocarbon such as benzene or toluene, preferably at room temperature.

The pharmacological activity of the compounds of the invention is demonstrated by the following assays.

Investigation of PARP inhibition

Reactive oxidizing agents (ROS) (e.g., hydroxyl radicals, superoxide, peroxynitrite, hydrogen peroxide) are known to be produced continuously in living organisms (Richter, C., FEBS Lett., 241, 1-5 (1988)), and in small amounts. play a role in the regulation of important physiological processes (Beck, KF et al., J. Exp. BioL, 202, 645-53 (1999); McDonald, L.J. and Murad, F., Proc. Soc Exp Bioi. Med. 211: 1-6 (1996)] (such as vasodilation, platelet aggregation, white blood cell adhesion). The concentrations of reactive oxidants and nitric oxide are significantly increased in acute and chronic inflammation, resulting in autoimmune diseases.

HU 227 116 Β1 [Taraza, C., et al., Rom. J. Intem. Med., 35, 89-98 (1997)], postischemic heart injury, and ischemic brain conditions (Brain Pathology, 9, 119-131 (1999)). The source of the reactive oxidants is, in part, induction of inflammatory tissue adherent white blood cells and macrophages, and partly of normal tissue cells by the induction of inflammatory cytokines.

Reactive oxidants also damage DNA. DNA damage initiates a complex defense and repair process in the cell, an important element of which is the activation of the poly (adenosine diphosphate ribose) polymerase (PARP) enzyme. PARP is a nuclear-rich enzyme that is abundant in almost every cell and catalyses the transfer of the adenosine diphosphate ribose unit from nicotinic acid adenine dinucleotide (NAD) to protein and the construction of poly (adenosine diphosphate ribose) chains. The major substrates of the enzyme include itself [Gonzalez, R., et al., Mol. Cell. Biochem., 138, 33-37 (1994)], nuclear proteins, histones, topoisomerase I and II, transcription factors. The PARP enzyme activity is increased approximately 500-fold by DNA strand breakage (Menissier de Murcia, J. et al., J. Mol. Biol., 210, 229-233 (1989)]. Extreme DNA damage resulting in extreme PARP enzyme activation causes a critical decrease in NAD concentration. As a result, cellular synthesis of adenosine triphosphate (ATP) is reduced, while the use of ATP is increased as the cell attempts to restore NAD levels from adenosine diphosphate ribose and nicotinic acid amide. All this seriously damages the energy status of the cells and can lead to cell death. Inhibition of the PARP enzyme is therefore of great importance in the therapy of many diseases, such as autoimmune diseases [Szabó, C., et al., Trends Pharmacol. Sci., 19, 287-98 (1998)], ischemic heart and brain states, and neurodegenerative diseases. In fact, inhibition of PARP can eliminate NAD catabolism, thereby reducing the levels of nicotinic acid amide and adenosine diphosphate ribose in the cell and inhibiting the use of adenosine triphosphate for the synthesis of NAD; that is, by inhibiting the enzyme, we can avoid the above-mentioned cell damage and death.

In vitro determination of PARP inhibition by an isolated enzyme

Poly-ADP-ribose polymerase was isolated from rat liver by Shah. GM, Anal Biochem. 227, 1-13 (1995). PARP activity was determined in 130 µl reaction mixture consisting of 100 mM Tris-HCl buffer, pH 8.0, 10 mM MgCl ₂ , 10% glycerol, 1.5 mM DTT, 100 µg (32P), or (3H). ), NAD +, 10 pg activated DNA, 10 pg histone. After incubation for 10 min, the reaction was stopped with 8% perchloric acid and the protein was separated by centrifugation (10 min, 10,000 x g). The precipitate was washed three times with 8% perchloric acid and protein-bound radioactivity was measured on a Beckman scintillation counter. The results are shown in Table I.

/. spreadsheet

Compound PARP l _{0 5} mg / l Example 2 9 ± 2 Example 3 8 ± 1 Example 4 14 ± 2 Example 5 13 ± 2 Example 6 17 ± 3 Example 8 12 ± 2 Example 9 7 ± 1 Example 10 28 ± 4 Example 12 18 ± 3 Example 13 20 ± 3 Example 14 9 ± 2 Example 16 18 ± 3

Table I shows that some of the compounds of the invention tested are very good PARP inhibitors (10 ₅ <10 mg / L). Most of the compounds are good PARP inhibitors ( _{10 5} = 10-28 mg / L).

Effect of Compounds of Formula I on Ischemic Heart Rate and Reperfusion Arrhythmia

Myocardial cell damage, myocardial tissue destruction, is most often caused by an eating disorder. The main form of eating disorder is lack of oxygen. The resulting myocardial injury is myocardial ischemia, which can occur as a result of acute hypoxia / anoxia in the delimited area of the myocardium due to sudden coronary artery obstruction or cramping, or prolonged coronary heart disease. The ischemic phase of acute myocardial infarction is followed by an additional phase of blood flow, the so-called reperfusion phase. Reperfusion can be a fatal consequence of cardiac arrhythmias (ventricular tachycardia and fibrillation). It is also the first manifestation of reperfusion injury. Medication for the prevention of reperfusion arrhythmia may prevent the risk of life-threatening early post-infarction.

Our experiments were performed on male SPRD rats (body weight: 300-350 g) under anesthesia with 60 mg / kg pentobarbital [5-ethyl-5- (1-methylbutyl) -2,4,6 (1H, 3H, 5H) -pyrimidine tri] ] ip. administration. The animals' spontaneous breathing was maintained and assisted breathing through a tracheal cannula was performed using a ventilator (MTA Kutesz). Through the application of limb electrodes, ECG standard II. drainage was recorded. Through the right femoral artery cannula, blood pressure was measured using a blood pressure transducer (BPR-01, Experimetria, Hungary) and a pre-amplifier. The heart rate counter (HG-M Experimetria, Hungary) was triggered by a pulsating signal of blood pressure. For intravenous administration, a cannula was attached to the external jugular vein of the neck. After opening the chest and heart, surgical silk thread (4-0) was placed around the left anterior coronary artery branch. After a few minutes of stabilization a

The circular artery branch was closed for 5 minutes. The blockage was then lifted and a 10-minute reperfusion period followed. ECG was recorded during the initial normal state and during the periods of intervention indicated above. From the record, the duration (s) of ventricular tachycardia and ventricular fibrillation was determined. In addition, survival rates were recorded for the various groups of animals treated.

The results showed that the compounds of the invention are useful in the prevention of reperfusion arrhythmia. Thus, when the compound of Example 2 was administered, the life span of the animals was increased by 50% compared to the control.

Investigation of compounds of formula I in global cerebral ischemia

Following human ischemic insult, pyramidal cells in the hippocampal CA1 region are the most killed, while other cells in the region (CA2, CA3) are less sensitive [Crain, B.J., Westerkam, W.D., Hamison, A.H., Nadler, J.V .: Selective neuronal death after transient forebrain ischemia the Mongolian gerbil, a silver impregnation study. Neuroscience, 27, 402 (1988)]. Memory impairment has been reported by some authors to be associated with hippocampal cell death [Walker, A.E., Robins, M., and Weinfield, F.D .: The National Survey of Stroke NINCDS, NIH: Clinical Findings Stroke 12, Suppl. 1: 1-44 (1981)]. The mammalian central nervous system is less sensitive to ischemic damage. Due to the anatomical features of the Mongolian gerbil (Meriones unguiculatus), it is very suitable for the examination of cerebral ischemia, since 90% of this species lacks the pituitary anastomosis system (Circulus Willisi), so there is no connection between the carotid artery and vertebral artery. Therefore, extensive cerebral ischemia can be produced by suppressing the carotid arteries.

The purpose of the test is to determine whether compounds of formula I have a protective effect in global cerebral ischemia. Our experiments were performed on male Mongolian gerbils. The animals were anesthetized with a mixture of 2% halothane [2-bromo-2-chloro-1,1,1-trifluoroethane], 70% nitric oxide and 30% oxygen. Under anesthesia, we lowered the arterial carotid on both sides for 5 minutes. Nerve cells do not die immediately, so a four-day reperfusion period (late type of cell death) followed. On the fourth day after surgery, the cell damage in the CA1 pyramidal region was 80-90%.

To measure learning and memory abilities, and hypermotility, the animals were tested in a Y maze.

Cell death in the CA1 region was studied by histological section.

The animals were perfused with buffered formalin, brain removed and fixed in formalin. After staining of the brain slices, the size of CA1 dead areas was measured.

The following materials were used for the tests:

Reference substance GyKI-52466, 40 mg / kg ip. dose, 30 minutes after ischemia,

Nimodipine [1,4-dihydro-2,6-dimethyl-4- (3-nitrophenyl) 3,5-pyridinedicarboxylic acid (2-methoxyethyl) - (1-methylethyl) ester] (Reference substance ) 10 mg / kg ip. administered 5 to 30 minutes after ischemia,

The novel compound of Example 2, 25 mg / kg ip. administered 5 to 30 minutes after ischemia,

The novel compound of Example 9, 25 mg / kg ip. administered 5 to 30 minutes after ischemia,

The novel compound of Example 12, 25 mg / kg ip. dose 5 and 30 minutes after ischemia.

Study groups:

- ischemic control

- treated with reference material after ischemia

- treated with test substance after ischemia

- sham

The experiments showed that the compounds of the formula I have a protective effect in global cerebral ischemia.

Effect of compounds of formula I on autoimmune diseases

Autoimmune disease is defined as a disease in which the body elicits an immune response against one of its normal constituents [Ring, G.H., et al., Schem. Nephrol., 19, 25-33 (1999); Theofilopoulos, A. N., Ann. Ν. Y. Acad. Sci. 841: 225-235 (1998). Different autoimmune diseases differ from each other in terms of the antigen that initiates the process, but show a high degree of similarity in the cellular tissue destruction mechanism of the resulting autoimmune process [Szabó, C., et al., Proc. Natl. Acad. Sci. USA 95: 3867-3872 (1998). Autoimmune diseases include the following major diseases:

- hormonal disorders: insulin-dependent diabetes;

- liver disease: hepatitis;

skin disorders: bullous pemphigoid lupus, pemphigus vulgaris, psoriasis, scleroderma, vitiligo;

- haematopoietic disorders: sarcoidosis;

- joint diseases: rheumatoid arthritis;

vascular diseases: vasculitis, takayasu arteritis, polyarteritis nodosa, ankylosing spondylitis;

- intestinal diseases: ulcerative colitis;

- musculoskeletal disorders: multiple sclerosis, myasthenia gravis, chronic inflammatory demyelinating polyneuropathy.

Among the autoimmune diseases listed, prevention of streptozocin-induced type I diabetes mellitus in mice was investigated.

The β-cells of the Langerhans Islands of the pancreas produce and deliver insulin, the main regulator of the body's sugar metabolism. Selective damage or death of β-cells results in a reduction or elimination of insulin production, leading to the development of type I diabetes (that is, insulin-dependent diabetes). Β-cells are particularly sensitive to reactive oxy9

EN 227 116 Β1 Toxic effects of nitrogen oxides and nitrogen oxides. Studying nitric oxide-induced DNA damage has led to the discovery that overexpression of PARP and consumption of the NAD kit is responsible for β-cell death [Heller, B. et al., J. Biol. Chem., 1995, 270, 11176-180. By a similar mechanism, streptozotocin [2-deoxy-2- (3-methyl-3-nitrosoureido) -D-glucopyranose], which has been shown to be a model of type I diabetes in animal experiments, is damaging to insulin-producing β-cells [Yamamoto, H et al., Natura, 294, 284-286 (1981)]. Streptozotocin damages DNA by alkylation and formation of nitric oxide and consequently activates PARP as described above.

In our experiments, we investigated in mice whether a single dose of the hydroxy acid derivatives of the formula I prevents the blood glucose-raising effect of a single dose of streptozotocin. The experiments were performed on CD-1 male mice. We had three experimental groups with 8 animals in each group. The first group received 160 mg / kg streptozotocin (SZ) (Sigma) ip, the second group 160 mg / kg streptozotocin and 200 mg / kg compound (I), the third group was the control. Blood glucose levels were determined on the third day after treatment, and animals were sacrificed and serum samples were taken for insulin.

It has been found that the compound of the present invention significantly decreases the elevated blood glucose levels with the administration of streptozocin.

Effect against insulin resistance

Real folk disease in the II. type diabetes mellitus, which is not insulin dependent. It is essentially insulin-insensitive to peripheral tissues, particularly striated muscles and adipose tissue, which cannot, of course, be counteracted by hyperactivation of β-cells in the Langerhans islands of the pancreas. Insulin resistance, even without true diabetes, is a source of many cardiovascular regulatory disturbances, and is therefore considered to be an independent risk factor for coronary heart disease. Because of the central pathophysiological importance of insulin resistance, therapeutic approaches to enhance the sensitivity of peripheral tissues to insulin are of paramount importance. Currently, only one class of true insulin-sensitizing drug, thiazolidinediones, is used in clinical practice. However, their toxicity (mainly hepatotoxicity) significantly limits their use. These insulin-sensitizing drugs reduce the levels of glucose, triglycerides and insulin in the blood by increasing the sensitivity of target tissues (liver, striated muscles, adipose tissue) to insulin [Colca, JR and Morton, DR: Antihyperglycamic thiazolidines: analogues, in New Antidiabetic Drugs, edited by Bailey, CJ and Flatt, PR, Smith-Gordon, New York, 1990, p. 255-261],

We investigated the effects of the compounds of the invention on insulin sensitivity in healthy and hypercholesterolemic rabbits. New Zealand white male rabbits (3-3.5 kg) were used. One group of animals was fed normal rabbit food and the second group was fed 1.5% cholesterol-enriched diet for 8 weeks.

Both groups of animals were divided into four subgroups:

- untreated group,

- the group treated with the compound of the invention, 10 mg / kg iv. dose twice daily for 4 days,

- NOS inhibitor treated group: 5 mg / kg for 5 minutes iv. given 7-nitroindazole 5 minutes before the insulin infusion,

- a group co-treated with 7-nitroindazole and the test compound, as described above.

Test method used

During anesthesia, rabbits were cannulated into the bilateral cervical vein of the rabbit and one side of the carotid artery, and the ends of the cannula were removed from the wound at the cervical region. The cannulas were filled with heparin saline. The experiments were started after the animals were fully awake.

Tests on isolated veins

Rabbit aortic and cerebral arteries were made into 4 mm long vascular rings and mounted horizontally on two L-shaped glass hooks, one of which is connected to an isometric force meter. The experiments were performed in an oxygenated Krebs solution in an isolated organ bath. Initial resting tension was 20 and 10 mN in aortic and vascular preparations, respectively. The equilibration time was 60 minutes. Subsequently, noradrenaline was added to the preparations in increasing concentrations. After maximal contraction, norepinephrine was washed out of the organ bath and waited for resting tension to return. Vessel sensitivity to acetylcholine was examined by pre-contraction with noradrenaline EC ₅₀ concentration. Acetylcholine was also given in cumulatively increasing concentrations.

In another group of experiments, vascular sensitivity was also examined by electrical field stimulation in carotid arteries. The initial stress was 10 mN and the equilibration time was 1 hour. The contraction responses were elicited by two successive pulse sequences (100 paces, 20 V, 0.1 ms, 20 Hz). Electric field stimulation was repeated in the presence of 1 μΜ atropine and 4 μΜ guanethidine (the so-called “non-adrenergic, non-cholinergic” (NANC) solution). Studies were performed on intact endothelial and endothelial stripped blood vessels.

Determination of tissue cyclic GMP (guanyl monophosphate) content according to Szilvássy [Szilvássy, Z. et al., Coron art. Dis. 4: 443-452 (1993); Am. J. Physiol., 266, H2033-H2041 (1994)]: The muscle rings were immediately frozen with a pre-cooled Wollenberger gripper and pulverized in liquid nitrogen. The samples were then homogenized in 6% trichloroacetic acid (10 times the weight of the samples) in a mortar previously stored at -70 ° C. After thawing

The samples were processed at 4 ° C. After settling for 10 minutes in a Janetzki K-24 centrifuge using 15,000 g, the supernatant was extracted with 5 ml of water-saturated ether in a Wortex extractor for 5 minutes. The ether extract was separated and the extraction was repeated five times. The samples were then stripped under nitrogen and the cGMP content was determined with an Amershan RIA kit. Values were expressed as pmol / mg wet tissue weight.

Hyperinsulinemic euglycemic determination

In a hyperinsulinemic euglycemic glucose clamp assay, human insulin was infused at a constant rate (13 mU / kg min) over one venous catheter for 120 minutes. This insulin infusion provided 100 ± 5 pU / ml of plasma insulin immunoreactivity at dynamic equilibrium. Blood samples (0.3 ml) were taken at 10 minute intervals through the arterial cannula. Blood glucose concentrations were maintained at a constant (5.5 ± 0.5 mmol / L) level by variable rate glucose infusion through the other venous cannula. Once the glucose concentration has stabilized for at least 30 minutes, it is considered steady state and additional blood samples (0.5 mL volume) are taken at 10-minute intervals to determine plasma insulin levels. Insulin sensitivity was characterized by a fast steady state glucose infusion rate.

The above tests gave the following results:

1. The vasoconstrictor effect of cumulatively increased concentrations of acetylcholine (1 nM-10 μΜ) was not affected by the compound of the invention present at a concentration of 1 μΜ in normal animals.

2. Acetylcholine-induced vasoconstriction is significantly reduced by the compounds of the present invention.

3. As a result of space stimulation, tension in the carotid arteries increased in normal Krebs solution. An anesthetic response was observed in NANC solution for spatial stimulation. None of the responses was affected by the compound of the present invention.

4. After removal of the peritoneum (endothelial layer), vasoconstriction increased for vascular stimulation and NANC vasoconstriction decreased. The compound of the present invention reduced vasoconstriction induced by spatial stimulation and increased NANC vasoconstriction in vascular rings deprived of the endothelial layer.

5. In the carotid rings of hypercholesterolemic animals, vasoconstriction caused by field stimulation was increased, and the vasoconstrictor effect of NANC solution was reduced compared to responses of normal animals. The compound of the invention tested reduced vasoconstriction caused by electrical stimulation and increased NANC vasoconstriction in the arteries of hypercholesterolemic rabbits, whether intact or not.

6. Initial cyclic GMP levels in the vascular rings of hypercholesterolemic rabbits were significantly lower than in normal animals. This decrease was normalized by incubation with 1 μΜ of the compound of the invention. However, the test compound did not affect resting cyclic GMP levels in the vasculature of normal animals. CGMP levels were increased in vascular preparations from normal animals as a result of space stimulation. In hypercholesterolemic animals, cGMP levels were not elevated in the arteries. The compound of the present invention did not affect cGMP elevation in normal blood vessels, but significantly increased it in the arteries of hypercholesterolemic animals.

7. The cholesterol-rich diet significantly reduced insulin sensitivity in rabbits. The insulin sensitivity of hypercholesterolemic animals was almost completely normalized after four days of treatment with the compound of the invention. However, the compound of the invention tested had no effect on insulin sensitivity in normal animals.

8. 7-Nitroindazole, a neuronal nitric oxide synthase inhibitor, also developed insulin resistance in normal animals. The tested compound of the invention did not affect the insulin resistance induced by 7-nitroindazole. 7-Nitroindazole inhibited the insulin resistance-relieving effect of the compound of the invention in hypercholesterolemic rabbits.

findings

The compound of the present invention potentiated the hypoglycemic effect of insulin on hypercholesterolemic insulin resistance in alert rabbits. The results show that this insulin sensitizing effect is closely related to nitrergic mechanisms, the importance of which has recently been recognized in the regulation of insulin sensitivity (Shankar, R.R. et al., Diabetes 49: 684-687 (2000)). Accordingly, neurohormonal regulation of peripheral insulin sensitivity in the liver is conceived as follows [Lautt, W. W., Can. J. Physiol., 77, 553-562 (1999)]:

- increase in insulin levels after a meal,

- this stimulus triggers a parasympathetic reflex in the liver,

- release of acetylcholine, which activates muscarinic receptors.

- Muscarinic excitement leads to the release of nitric oxide.

- (In a saturated organism), nitric oxide causes the release of a hepatic insulin sensitizing substance (HISS). It has an insulin synergistic or insulin-like effect.

- HISS released increases glucose uptake in skeletal muscles. The HISS mechanism is sensitive to inhibition of nitric oxide synthesis and can be activated by exogenous nitric oxide donor. It is likely that the HISS mechanism binds to the sensory (sensory) fibers that run into the liver. The increase in postprandial plasma insulin levels activates the nitrergic subpopulation of sensory fibers that pass into the liver, causing the release of additional sensory neurotransmitters from the surrounding nerve fibers, which, when released into the circulation, increase the insulin sensitivity of the tissues.

HU 227 116 Β1

Obesity and insulin resistance have recently become known, and very common in patients with obesity. The exact molecular biological basis of the epidemiological link between the onset of type 2 diabetes. Fat cells secrete a peptide hormone called resistin, which reduces insulin sensitivity in important target tissues (Steppan, C.M. et al., Naturre, 409, 307-312 (2001)). Of the known drugs, only thiazolidinedione-structured compounds (so-called insulin sensitizers) are capable of inhibiting the secretion of resistance by activating the peroxisome proliferation receptor.

The compounds of formula I, through their action on insulin sensitivity, are capable of eliminating insulin resistance through a nitrergic mechanism and sensory neurotransmitters. The restoration of insulin sensitivity is of pathological importance in diseases as important as those in Part II. diabetes, high blood pressure, coronary artery disease, obesity and some endocrine disorders.

Use of Compounds of the Invention to Prevent Toxic Effects 1. Effect of Compounds on Endotoxin-Induced Lethality in Mice

Septic shock is one of the most common deaths in intensive care units. Infections caused by Gram-negative bacteria lead to hypotension, malfunctioning of several organs, and eventually to collapse of the body. Injection of the bacterial membrane component lipopolysaccharide (LPS) into experimental animals produces a shock-like condition and eventually results in death. LPS activates the NF-κB / Rel transcriptional family, which regulates the production of several transporters involved in the pathomechanism of shock (TNF-α, interleukins, NO synthase) [Oliver, FJ, Murcia, JM, Nacci, C, Decker, P ., Andriantsitohaina, R., Müller, S., Rubia, G., Stodet, J.C., Murcia, G.: Resistance to endotoxic shock as a consequence of defective NF-κΒ activation in poly (ADP / ribose) polymerase-1 deficient. mice, EMBO J., 18 (16), 4446-4454. (1999)]. The PARP-1 gene is functionally linked to NF-κB, thus, in the absence of PARP, NF-κΒ-dependent transcription does not take place, thus the release of inflammatory mediators is also downregulated in endotoxin shock. The purpose of the assay is to determine whether inhibition of PARP-1 by compounds of Formula I prevents endotoxin lethality.

C57BL / 6 mice (Charles River Breeding LTD) were used for the experiments. The dose and type of LPS used in the assay are the same as those described above by F. J. Oliver in Lipopolysaccharide Escherichia coli 0111: B4 (Sigma) (LPS).

3-Aminobenzamide (Sigma) was also used in the studies. 24-hour survival was monitored at least twice. Compounds of formula (I) were administered orally 1 and 6 hours after LPS treatment.

It has been found that the compounds of formula (I) tested significantly reduce endotoxin-induced lethality.

2. Effect of the compounds on acetaminophen (paracetamol) -induced hepatotoxicity

Various non-steroidal antirheumatic drugs are known [Peters et al., Clin. Inves., 71, 875-881 (1993)] and analgesics have significant hepatotoxicity [Kroger, H., et al., Gen. Pharmac., 27, 167-170 (1996)]. Paracetamol induces liver and kidney failure at high doses [Meredeth, T.J., et al., Arch. Inter. Med., 141, 397-400 (1981)]. In recent years, it has become evident that poly (ADP-ribose) polymerase inhibitors prevent paracetamol-induced liver damage [Kroger, H., et al., Gene. Pharmac., 27, 167-170 (1996)].

It is known in the literature that paracetamol is an inducer of cytochrome P-450. The effect of paracetamol on the cytochrome P-450 system generates reactive quinone imines which bind to the sulfhydryl group of proteins and cause rapid depletion of intracellular glutathione (Jollow, D. J. et al., Pharmacol., 12, 251-271 (1974)). Thus, inactivated proteins lead to the death of liver cells and liver necrosis. Intracellular glutathione is one of the most important antioxidants and the strongest eliminator of reactive oxidants. Weakening of the glutathione-dependent antioxidant defense system leads to the intracellular growth of free oxygen radicals (Miesel, R. et al., Inflamatin, 17, 283-294 (1993)). Free oxygen radicals are potent inducers of PARP, which affect the post-translocation of proteins. Enhanced PARP activation depletes cellular NAD stores and may induce apoptosis (Hoschino, J. et al., J. Steroid Mol. Biok., 1993, 44, 113-119. Therefore, nicotinic acid amide, a selective inhibitor of the PARP enzyme, suppresses the hepatic release of GOT and GPT enzymes, which have been detected in mice with paracetamol-induced hepatitis [Kroger, H. and Ehrlich, W., L-Tryptophan: Current Prospects in Medicine and Drug , edited by Kochen W. and Steinhart, H. Verlag Berlin, 1994],

We tested whether paracetamol-induced liver injury could be prevented with the novel compounds of formula (I). In the experiment, hepatic injury is characterized by an increase in GOT and GTP enzymes induced by paracetamol. A protective effect is observed when GOT and GTP enzymes are less potent when the test compound is administered. The experiments were performed on male NMRI mice weighing 30-40 g. The animals were pre-treated orally with the compounds of formula (I) for 7 days. On day 8, the mice were fasted for 12 hours, followed by administration of paracetamol (500 mg / kg po) and a given dose of a compound of formula (I). The animals were bled after 16 hours and serum levels of GOT and GPT were measured. The results were analyzed using the Mann-Whitney nonparametric test. The mean and deviation (S.D) for the results were reported, where p Z0.05 was considered significant.

We found that once administered orally, paracetamol increased serum GOT and GPT activity.

EN 227 116 Β1 in male NMRI mice compared to saline-treated control animals. At the same time, the compounds of formula (I), after seven days of oral pre-treatment, significantly reduced the activity of the GOT and GPT enzymes. Thus, at a dose of 50 mg / kg, the compound of Example 12 already provided very favorable liver protection.

3. Effects of compounds on paraquat toxicity in vitro and in vivo

Paraquat [1,1'-dimethyl-4,4'-bipyridinium], a compound formerly used as a pesticide, exerts its toxic effect by the formation of a superoxide radical. Oxidoreductase enzymes using NADH and NAD (P) H as electron donors (NAD (P) H is a reduced form of β-nicotinamide adenine dinucleotide phosphate) are involved in the formation of superoxide radicals. The p66 protein plays an important role in mediating the cellular response to paraquat-induced oxidative stress. [E. Migligaccio et al., Natura, 402, 309-313]. Mechanisms that promote superoxide inactivation (e.g., elevation of superoxide dismutase) effectively reduce paraquat toxicity. Superoxide radicals play an important role in the pathomechanism of many diseases (ischemiareoxygenation, infarction, inflammatory diseases). Paraquat administration provides a simple model of the superoxide load in these diseases.

Effect on paraquat toxicity in vitro

Hepa-1 hepatoma cells were cultured in RPMI-1640 supplemented with 10% calf serum, RPMI-1640 supplemented with 10% calf serum and 5% equine serum at 37 ° C in air containing 5% carbon dioxide. In 100 μΙ medium, 5 × 10 ³ cells were plated in 96 well (Costar) culture plate wells. Some cultures were untreated and served as controls. Cells were treated partly with increasing concentrations of paraquat, partly with the same concentration of paraquat and 3, 10 and 30 pg / ml of test substances. After an additional 3 days of culture, the growth of the cultures was determined by SRB staining. Higher paraquat concentrations used resulted in cell death, while lower concentrations resulted in partial growth inhibition. The effect of the test compound was determined by reducing paraquat toxicity.

Effect on paraquat toxicity in vivo

Paraquat has significant toxicity in mice. An intraperitoneal dose of 70 mg / kg results in death of the animals within 2 days. Mechanisms that reduce superoxide formation, neutralization, and oxidative stress can also reduce paraquat toxicity in vivo.

Groups of CFLP mice weighing 20-22 g were treated with 50 and 70 mg / kg paraquat ip. Some of the groups were also treated with test substances for 6 hours before and after the administration of paraquat. Test substances were administered at a dose of 100 mg / kg po. We added. The survival of the mice was determined in the morning and evening. The efficacy of the test substance was determined by increasing the survival of the mice.

It has been found that the compounds of formula I are effective in reducing the toxicity of paraquat.

Effect of compounds of formula I on neurodegenerative diseases

As mentioned above, during damage to DNA caused by reactive oxidants, PARP is activated, whereupon the cell loses NAD, resulting in cell death. Excessive PARP activation is not only observed during ischemia-induced neuronal death, such as cerebral ischemia, but has also been shown to play a role in other neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease or amyotrophic lateral sclerosis [Love et al., Neuropol. Appl. Neurobiol. 25: 98-108 (1999); Ellásson et al., Nat. Med. 10: 1089-1095 (1997)].

Effect on experimental amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal, progressive, neurodegenerative disease. ALS is the most common cause of adult motor neuron disease in industrialized countries. Motoneuron damage in the cortex, brainstem, and spinal cord causes skeletal muscle atrophy, paralysis and death [Rowland, L.P. in Neurodegenerative diseases, p. 507-521 (1994)]. In some cases of ALS, the disease is inherited. Inherited cases are partly caused by a missense mutation of the Cu / Zn superoxide dismutase-1 (SOD-1) gene [Deng, R.H. et al. Science, 261, 1047 (1993)]. SOD-1, an abundant cytoplasmic enzyme in nerve tissue, plays an important role in preventing cellular damage caused by oxygen radicals. The mutated enzyme retains almost normal levels of activity. In vitro studies have shown that the mutation results in a gain-of-function change, increasing the free radical production of the enzyme.

Mice carrying the transgenic mutant SOD-1 gene develop symptoms similar to ALS. Several variants of the mutant human gene (G93A, V148G) have been used to generate transgenic mice, which induce increased expression of the pathogenic gene in animals. These animals have been used in screening for anti-ALS drugs [Gurney, Μ. E. J. Neurol. Sci., 152 Suppl 1.67-73 (1997)].

Studies on hereditary ALS

The assays were performed on transgenic mice expressing the mutant SOD-1 gene (G93A). The animals were purchased from Jackson Laboratory (USA). The animals were treated with the compounds of formula (I) before the onset of symptoms at 4 weeks of age. The test compounds were administered at three dose levels, daily by po. animals were administered until the end of the experiment. The disease is prog13

Weekly monitoring of motor function (extension reflex, loaded grid, rotarod test), animal survival, and histological and biochemical examination of the motoneuron at the end of the experiment (120 days) were used to monitor the progression.

Compounds of Formula I have been found to have a moderate delay in the onset of reflex, coordination, and muscle loss in transgenic ALS animals. The effect was dose-dependent. Treatment also delayed the onset of paralysis and end-stage disease. The histological findings support the observed clinical effect. Degeneration and death of motoneurons and subtantia nigra neurons were less extensive in the treated group than in the control group. Based on the results, it is expected that the compounds of formula (I) will have a beneficial therapeutic effect in ALS.

Studies on autoimmune ALS

In sporadic ALS, no alteration was found in the SOD-1 gene. This suggests that other root causes lead to the same cycle.

The vast majority of patients with sporadic ALS have antibodies against calcium channels. This observation supports the notion that the autoimmune process against motoneurons and calcium channels plays a primary causal role in the development of sporadic ALS cases. Engelhardt et al. (1995), by immunization with motoneuron, followed by the use of immune serum alone, induced disease with specific lesions of ALS (Engelhardt, J. et al., Synapse, 20, 185-199). This model was used to investigate the efficacy of the compounds of formula (I) against sporadic ALS.

Hartley-bred guinea-pigs were immunized with anterior horn homogenate of bovine spinal cord. Immunization was performed with a suspension of complete Freund's adjuvant (CFA) containing motoneurons. The treatment was given by 10 subcutaneous or intradermal injections of 0.1 ml. After one month, the injections were repeated, but incomplete Freund's adjuvant was used to prepare the suspension. Two weeks after the second immunization, severe weakness developed in animals, particularly in the hind limbs, within 1 to 3 days. Their weight gain stopped and their mobility decreased. The animals did not change for the next two weeks. Two weeks after immunization, the animals were bled. Serum collected by centrifugation of the collected blood was used to induce ALS in mice.

Groups of 5-5 albino male mice (CFLP subtype, 25-30 g body weight) were tested. An amount of 1 ml of the above serum was injected into the abdominal cavity of the animals to damage the motoneuron. One group of animals received serum only, the other groups of animals were treated with the compound of the formula I tested at a dose of 100 mg / kg intraperitoneally in addition to serum administration. An additional group of animals received only the compound of the formula (I) under test without injecting serum.

The serum-only animals slowed their movement, their hind legs became difficult to use and then paralyzed. Animals treated with serum and test compound (I) showed no symptoms of motor deficits, similar to animals treated with test compound (I). All this indicates that the compounds of formula I prevent the development of immunosuppressive motor disorders.

Effect on experimental Parkinson's disease model

Parkinson's disease (PB) is a common disabling neurodegenerative disorder characterized by tremor, disability, stiffness and imbalance. Behavioral disorders are caused by a decrease in dopamine levels in the brain resulting from the death of dopaminergic neurons in the substantia nigra pars compacta. Examination of the effect of selective neurotoxicity on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has significantly contributed to understanding the pathomechanism of Parkinson's disease. MPTP causes parkinsonian symptoms in humans and animals [Dexter, A., et al. Ann. Neurol., 35, 38-44 (1994)]. MPTP treatment also results in the loss of dopaminergic cells of the substantia nigra pars compacta. Lewy body-like, eosinophilic inclusions appear in the damaged neurons and at the same time mitochondrial I complex activity is reduced in these cells. All these lesions are typically the consequence of oxidative stress [Shapira, A., Adv. Neurol. 69: 161-165 (1996)]. The biologically active metabolite of MPTP is MPP (1-methyl-4-phenylpyridine). MPP directly inhibits mitochondrial I complex and results in increased formation of the superoxide anion. The data suggest that oxidative stress plays a central role in the development of both MPTP-induced and naturally occurring Parkinson's disease. The enzyme (PARP) is activated by oxidative stress and appears to be actively involved in the pathomechanism of Parkinson's disease. The sensitivity of PARP knockout mice to the induction of parkinsonism by MPTP is greatly reduced [Mandir, A. et al. Proc. Natl. Acad. Sci. USA 96: 5774-5779 (1999). These observations suggest that PARP inhibition may result in a therapeutic effect in Parkinson's disease.

The C57BI mice used for our study were obtained from Charles River Hungary. Mice weighing 20 g were injected with MPTP at 20 mg / kg ip. was administered four times at 2 hour intervals. The compounds to be tested were po. was administered 30 minutes prior to MPTP injections. Control animals received only vehicle at the same schedule. Seven days after MPTP treatment, the animals were sacrificed and the brain was rapidly removed. The striatum was immediately excised on an ice-cold Petri dish. The tissues were frozen immediately and stored at -80 ° C until use.

HU 227 116 Β1

The tissue pieces were homogenized by sonication in 50 volumes of 0.1 M perchloric acid. After centrifugation (14000xg for 10 min, 4 ° C), 20 μΙ of the supernatant was injected onto a reverse phase catecholamine column (ESA, Bedford) and the dopamine content was determined.

Two hours after the last MPTP treatment, the ventrolateral midbrain and striatum were excised and homogenized in sucrose / DTT buffer and centrifuged (14,000 x g for 5 min). The pellet was resuspended in buffer. After determination of the protein concentration, an equal amount of protein was run on SDS / PAGE gel. Protein was transferred from the gel to a nitrocellulose membrane and subjected to immuno-staining specific for poly (ADP-ribose) polymer. Specific binding was visualized by chemiluminescence.

Studies have shown that MPTP treatment resulted in a drastic reduction (80%) in striatal dopamine content. The test compounds of formula I partially inhibited MPTP-induced dopamine loss (20-40%). MPTP treatment resulted in the formation of a poly (ADP-ribose) polymer adduct in the striatum. Treatment with compounds of formula I partially inhibited this process (20-70%). Therefore, the compounds of formula (I) are expected to have a therapeutic effect in the treatment of Parkinson's disease.

Investigation of cytoprotective effect

Many medications can cause nerve damage as a side effect when used permanently or frequently. Among the long list of compounds with such side effects (chloramphenicol, dapsone, disulfiram, dichloroacetate, ethionamide, glutethimide, sodium aurothiomalate, hydralazine, isoniazid, metronidazole, nitrofurantoin, nitric oxide, cisplatin, pyridoxine, vincristine), neuropathy caused by isoniazid, pyridoxine and vincristine, and cisplatin; there are also drugs such as chloramphenicol, where this side effect can be stopped by stopping treatment. However, premature discontinuation of the medication jeopardizes the success of the therapy and threatens the relapse of the disease. The risk of therapeutic change due to such side effects is particularly high for anticancer chemotherapeutic agents. Thus, so-called chemoprotective or cytoprotective agents which limit only the adverse side effects of the vital drug without reducing the therapeutic potency are of great importance.

The most important side effect of cisplatin treatment given to cancer patients for chemotherapy is peripheral nerve damage (peripheral neuropathy). The occurrence of an adverse reaction may prevent complete cisplatin treatment, limit the success of chemotherapy and further impair the quality of life of the patients. The presence and extent of nerve damage can be determined clinically and experimentally by measuring nerve conduction velocity. The neurotoxic side effect of cisplatin [cis-diamine-dichloroplatinum] mainly affects large peripheral nerves with a myelin sheath and manifests itself in sensory nerve damage (sensory neuropathy). However, there have been recent reports of vegetative nerve damage from cisplatin treatment and, in very rare cases, motor neuropathy has been reported in treated cancer patients. Cisplatin often causes sensory dysfunction as a result of direct damage to the dorsal horn nerve ganglia (ganglia) and the large sensory nerves. In rats, chronic treatment with cisplatin causes sensory neuropathy, which is manifested by a decrease in the sensory conduction velocity of the mixed nerve in the ischiadicus. The compounds of formula (I) have been hypothesized to have cell-protective (cytoprotective) activity and to prevent the development of a serotoxic side effect of antineoplastic agents, based on the factual biochemical attack point (s) described above, predominantly to prevent free radical damage. Therefore, in rat studies, cisplatin is administered subcutaneously (1 and 2 mg / kg) ip. administered for 10 weeks and investigated the development of peripheral (sensory and motor) neuropathy and how different doses of the compounds affect the impairment of nerve function (conduction velocity).

In order to detect cisplatin-induced motor and sensory nerve damage in rats, nerve conduction velocity was measured using the modified Miyoshi method in the tail of rats. The modification consisted in measuring the conduction velocity at room temperature instead of 37 ° C. The pacing velocity of the sensory and motor nerves was measured before cisplatin treatment (as a control) and at weeks 5 and 10. For measurement, the animals were superficially anesthetized with ether and two pairs of needle electrodes were applied to the tail nerve, 50 mm apart. At supramaximal stimuli, afferent (sensory) and efferent (motor) nerve action potentials were recorded. The pacing rate was determined offline by averaging 10 action potentials using the following formula:

NCV = v / l [m / s], where v = distance between electrode pairs (stimulus and recorder), l = latency of onset of action potential (ms),

NCV = conduction velocity (m / s).

In the study, we found that 1 and 2 mg / kg 10 weeks ip. cisplatin treatment at week 10 significantly reduced the weight of the treated animals compared to controls. This weight loss was also observed in animals treated with compounds of formula (I). There was no difference in the overall behavior of the treated and untreated animals compared to the combined treatment (cisplatin and the new compound). The pacing velocity of the sensory and motor nerves did not differ in the control group at the three measurement points. In cisplatin-treated animals, there was a clear and significant reduction in both 5 and 10 weeks of treatment with 1 mg / kg cisplatin. After treatment with 2 mg / kg cisplatin, a more pronounced decrease in the conduction velocity was observed.

HU 227 116 Β1

Neuropathy has also developed in motor nerves. During 10 weeks of cisplatin treatment, the motor conduction velocity was significantly reduced in the 1 and 2 mg / kg cisplatin groups. The decrease was dose-dependent. The decrease in sensory nerve conduction velocity when treated with 1 mg / kg cisplatin and compounds of Formula I was significantly less than in the 1 mg / kg cisplatin group alone, thus improving nerve function with greater the stronger the damage was. In the groups receiving different doses of cisplatin 2 mg / kg and the compounds of formula I at week 5, the decrease in sensory nerve conduction velocity did not differ from that seen in the group receiving only 2 mg / kg cisplatin. However, at week 10, the rate of excitation of animals receiving cisplatin 2 mg / kg alone was significantly further decreased, whereas in animals treated with cisplatin and the new compounds tested, the reduction was dose-dependent compared to cisplatin 2 mg / kg alone. Decreases in efferent nerve conduction velocity were less pronounced by week 10, particularly in the cisplatin-treated groups.

In summary, the deterioration of the sensory and motor nerve conduction velocity induced by cisplatin treatment was reduced by concomitant administration of the compounds of formula (I) and prevented the progressive damage (from week 5 to week 10). This protective effect was sometimes dose-dependent. Thus, the neuroprotective effect of the compounds of formula I can be demonstrated in both types of sensory and motor nerves.

Biological activity of carnitine palmitoyltransferase (CPTI)

CPTI is the key enzyme at the junction of fatty acid metabolism. Coenzyme A (CoA) esters of free fatty acids (FFA) have two possible pathways: 1. triglyceride synthesis by reaction with glycerol, or 2. combustion, the first step of which is the formation of acylcarnitine by the CPTI enzyme [see McGarry, JD. , Woeltje, KF., Kuwajima, M. and Foster, D., Diabetes, 5, 271-284 (1989); McGarry, JD. and Foster, D., Ann. Port. Biochem., 49, 395-420 (1980)]. CPTI is localized on the outer side (or outer membrane) of the inner membrane of the mitochondria and catalyzes the following reaction:

FFA-CoA + L-kamitin-> FFA-carnitine + CoA

Inhibition of the burning of fatty acids results in an increase in the breakdown and burning of glucose, which is two particularly beneficial interventions with a very high pathological condition in morbidity and mortality, and in myocardial ischemia and diabetes. In myocardial ischaemia and subsequent reoxygenation, increased fatty acid oxidation is detrimental to the extra oxygen demand and the membrane damaging effect of the resulting acyl carnitines [Busselen, P., Sercu, D. and Verdonck, F., J. Mol. Cell. Cardiol., 20, 905-916 (1988); Ford, DA „Han, X., Horner, CC. and Gross, R.W., Biochemistry, 35, 7903-7909 (1996);

Reeves, KA., Dewar, G.H., Rad-Niknam, M. and Woodward, B., J. Pharm. Pharmacol., 1995, 48, 245-248. Based on numerous experimental data, it is now accepted that glucose uptake is beneficial at the expense of fatty acid oxidation, both in restoring myocardial mechanical function and in metabolic parameters (enzyme release, lipid peroxidation) [Lopaschuk, GD., Spafford, MA, Davies, NJ]. . and Wall SR., Circ. 66: 546-553 (1990); Kennedy, JA. Unger, SA. and Horowitz, JD. Biochem. Pharmacol. 52: 273-280 (1996)]. Influencing of this myocardial substrate selection, i.e., the choice between glucose and fatty acid, toward glucose and thus improving myocardial energy is also achieved by CPTI inhibitors [Lopaschuk, GD., Wall, SR., Olley, PM. and Davies, NJ., Circ. Res. 63: 1036-1043 (1988); Carregal, M. "Varela, A., Dalaimon, V., Sacks, S. and Savino, EA., Arch. Phys. Biochem., 103, 45-49 (1995); Lopaschuk, GD., Spafford, M., Circ. 65: 378-387. (1989); Pauly, DF., Kirk, KA. and McMillin, JB., Circ. Res. 68: 1085-1094 (1991).

Our studies show that the enzyme catalyzing the rate-determining reaction of fatty acid oxidation can be inhibited at a submillimolar concentration with the compounds of formula (I). Our studies also indicate that the compounds tested influence the substrate selection of the heart and other tissues and also alter the substrate selection by postischemic injury.

Biological role of oxygen-sensitive genes regulated primarily by bHLH transcription factors

Preventing the harmful effects of hypoxia requires a series of coherent defenses at both the cellular and body level. The HIF-1 / ARNT transcriptional complex plays a central but not exclusive role in the regulation of the gene expression change induced by hypoxia. Oxygen-sensitized, genes with a coordinated expression include erythropoietin (Wang, G.L., et al., PNAS 92, 5510 (1995)) and erythropoietin (Goldberg, M.A. and Schneider, T.J., J. Biol.). Chem., 269, 4355 (1994)] enhancing VEGF, more glycolytic enzymes such as GAPDH, LDH (lactate dehydrogenase) [Rolfs. A. et al., J. Bioi. Chem., 272, 20055 (1997)] and Glut-1 glucose transporter.

The compounds of formula I are believed to be capable of influencing the activation of genes involved in alarm (hypoxia-sensitive) states by binding to ARNT and / or HIF-1 transcription factors. It is believed that compounds express heat shock proteins, which play an important role in alarm situations, via this signal transduction pathway.

The synthesis of heat shock proteins (HSPs) is initiated by a variety of cellular stress factors, helping to prevent cellular life-threatening events and restore potential damage [Car16

HU 227,116,11 diovascular Rev. 578, (1993), J. Clin. Invest; Neurosci. Lett 163: 135-137 (1993)].

Agents that adjust the alert response to hypoxia, reoxygenation, and depleted adaptation can reduce tissue damage caused by hypoxia (e.g., infarction, arteriosclerosis, diabetes).

Specification of HSP-70

HSP-70 activity was assayed by introducing a reporter gene into a DNA hybrid. The promoter region of the HSP-70 gene encoding the heat shock protein is linked to a reporter gene by a well-measurable enzyme activity gene sequence. The luciferase enzyme was used as a reporter gene, the activity of which can be well measured by measuring luminescence. If the promoter of the luciferase enzyme gene is replaced by the promoter of the HSP-70 gene, the change in luciferase enzyme activity, i.e., the frequency of transcription of the gene, will correlate with the frequency of transcription of the HSP-70 gene under given conditions. In this way, the effect of a substance or process on HSP-70 gene expression can be studied by measuring luciferase enzyme activity. The effect of the test substances on HSP-70 expression was studied in such an experimental system.

To carry out the measurements, we constructed a circular double-stranded DNA molecule, a plasmid, which contains a nearly six hundred base region of the promoter of the mouse HSP-70 gene from its transcriptional start site to a 5 'end that contains several activating protein binding sites for HSP-70 gene transcription. contain. The DNA coding for the luciferase enzyme from Photymus pyralis was inserted into this promoter region. The entire heterologous gene construct was incorporated into a pBR-based plasmid selectable for neomycin. This plasmid HSP-70-luc was introduced into L929 mouse fibroblast cells. The measurements were as follows:

L929 cells containing the HSP-70-luc plasmid were grown in DMEM (Dulbecco's Modified Eagle's Medium) 5% FCS (Fetal Calf Serum). Cells were then plated per 24 ml COSTAR plates with a cell number of 104 in each sample. The test substances were prepared in PBS (Phosphate Saline Buffer) in a 10 ^-2 M stock solution and, after adherence (about 3-4 hours), 10 μΙ were pipetted into cells. The cells are incubated at 37 ° C for 30 minutes in a CO ₂ thermostat and the medium is replaced with fresh medium of the same composition except that the test substance is no longer present. The cells were allowed to regenerate for one hour at 37 ° C and then washed once with PBS. After removal of PBS, 40 μΙ of 1x lysis buffer was added to the cells and the samples were placed on ice for 30 minutes. The samples are then pipetted into Eppendorf tubes and centrifuged at 14,000 rpm for 20 minutes. ΜΙ of the supernatants were added to 25 μΙ of luciferase buffer and the samples were weighed in a luminometer for 25 seconds.

Luciferase meter buffer composition:

20.00 mM Tricin [N- (2-hydroxy-1,1-bis (hydroxymethyl) ethyl] glycine], pH 7.8

1.07 mM (MgCO ₃ ) ₄ Mg (OH) ₂ .5H ₂ O

2.67 mM MgSO ₄

0.10 mM EDTA [ethylenediaminetetraacetic acid]

3.33 mM DTT

270 μΜ Lithium salt of Coenzyme A

470 μΜ luciferin

530 μΜ ATP

5 * l flavor buffer composition:

125 mM Tris-H ₃ PO ₄ pH 7.8 mM CDTA (trans-1,2-diaminocyclohexane-N, N, Ν ', N'-tetraacetic acid) mM DTT

50% glycerol

5% Triton X-100

Investigation of hypoxia-sensitive genes

In cell culture (Hepa and HepG2 cells), we investigated the effect of compounds of formula I on xenobiotic and hypoxia (1% O ₂ ) -induced gene expression changes at mRNA and protein levels. Compounds of formula (I) have been found to increase methyl cholanthrene-induced HSP-70 expression in Hepa cells. The novel compounds enhance the expression of oxygen-sensitive genes such as VEGF, GAPDH, and LDH on Hepa and HepG2 cells by hypoxia.

The compounds of formula I enhance the expression of several oxygen-sensitive genes under hypoxia. This suggests that the compounds interfere with the central, general pathway of regulation of oxygen-sensitive genes.

Hypoxia, compounds of formula (I) that promote adaptation to stress induced by hypoxia reoxygenation, are useful in preventing the deleterious effects of hypoxia, hypoxia reoxygenation. The compounds are expected to have therapeutic effects in cardiovascular disorders, vasoconstriction, arteriosclerosis, vasospasm, infarction, embolism, vascular occlusion, hypotension, shock, burn, frostbite tissue damage. The compounds of the invention may also be effective in hypoxic states secondary to degenerative processes, metabolic disorders (Alzheimer's disease, diabetes).

Effect on LDH enzyme levels exposed to hypoxia

HepG2 cells

HepG2 cells were cultured in DMEM medium supplemented with 10% FCS in air containing 5% CO ₂ at 37 ° C. 10 ⁵ cells were plated in 24-well Costar culture plate wells in 1 ml of culture medium. The next day, the cultures were treated with the test compounds at a concentration of 30 pg / ml and the cells were exposed to hypoxia (1% O ₂ , 5% CO _{2 in} nitrogen) for 24 hours. Some of the control cultures were treated with water as solvent and the other cultured without hypoxia. Hypoxic treatment is protected17

The 116 Β1 gene was aspirated from the cells and washed twice with cold PBS. The cells were then lysed in 0.05 M phosphate buffer containing 0.05% Triton Χ-100. After centrifugation (2 ', 20,000 xg), the supernatant LDH activity was determined from NADH consumption using sodium pyruvate substrate.

Hypoxia treatment resulted in a 3-fold increase in cellular LDH levels. The compounds of formula I, in addition to hypoxia treatment, additively increased the cellular LDH level.

Antiviral effect

The retroviral genome consists of a single-stranded RNA molecule that replicates via a double-stranded DNA intermediate. A crucial step in the virus's life cycle is the insertion of double-stranded DNA into the host cell genome during a transposition-like event.

The enzyme responsible for generating the initial DNA copy of RNA is the reverse transcriptase. The enzyme copies linear double-stranded DNA from RNA in the cytoplasm of the infected cell. The linear DNA is then introduced into the nucleus. One or more copies of it are integrated into the host cell genome. The enzyme called integrase is responsible for this integration. The integrated proviral DNA then generates viral RNAs using the host cell enzymes, which appear on the one hand as mRNAs and, on the other hand, as genomes upon insertion into virions.

The smooth functioning of reverse transcriptase is vital for retroviruses. Therefore, inhibition of the growth of these viruses, including HIV, can be effectively achieved by inhibiting this enzyme.

Some of the currently marketed, sometimes effective, drugs for the treatment of HIV infection work by inhibiting reverse transcriptase. The most popular and most effective anti-HIV drugs today are the so-called anti-HIV drugs. One or two components of combination drugs are reverse transcriptase inhibitors. Reverse transcriptase inhibitors are divided into two broad classes. One of them is the so-called nucleoside analogue substance, the best known of which is azidothymidine, AZT for short. These inhibit the activity of the enzyme when bound to the nucleotide binding site of the enzyme. The other group is the so-called non-nucleoside analogue material, which is not bound to the substrate binding site of the enzyme but to another point on the protein, but there is a specific and relatively stable interaction with certain amino acid residues that deform the active center. thereby losing much of its specific activity.

The reverse transcriptase inhibitory activity of the compounds of the invention was assayed as follows. These substances are classified as non-nucleoside analogue reverse transcriptase inhibitors. The experiments were performed with Moloney murine reverse transcriptase, which is a good model for the HIV reverse transcriptase enzyme. The experimental set-up was as follows:

The activity measurement is based on the assay for the incorporation of (3H) dTTP into cDNA starting from the poly (rA) oligo (dT) 12-18 template primer. The reaction volume was 20 µl. The composition of the reaction mixture:

μΙ 10 * buffer pg / ml template primer μΜ dTTP pCi (3H) dTTP Test substance: 1 * dissolved in buffer

The reaction was initiated by the addition of 5U reverse transcriptase.

Composition of 10 * reverse transcriptase buffer:

500 mM Tris-HCl (pH 8.3) mM MgCl ₂ 300 mM KCl

100 mM DTT

The reaction mixture was incubated for 40 minutes at 37 ° C. Subsequently, 15 μΙ of the reaction mixture was transferred to a Whatman DE81 filter paper disk, washed with 5% disodium hydrogen phosphate buffer, water, then 96% ethanol. After drying, the disks were placed in a 5 ml scintillation cocktail (OptiPhase HiSafe 3, Wallac) and the radioactivity of the samples was measured in a Packard Tri-Carb 2200 CE scintillation counter.

In the experiments, AZT, a nucleoside analogue RT inhibitor, or a non-nucleoside-based inhibitor known as Nevirapin, which binds to the so-called benzodiazepine binding site of the enzyme, was used as a positive control (known as an inhibitor).

From the experimental results, the following conclusions can be drawn: In the presence of the compounds of the invention, the reverse transcriptase enzyme of the Moloney murine leukemia virus can be inhibited. Inhibition concentrations ranged from 0.2 to 2.0 micrograms per ml. Based on the dose-dependent reverse transcriptase inhibition, the inhibitory effect of the new compounds is greater than that of the comparator Nevirapine but less than that of the nucleoside analogue AZT. Because the test enzyme is a model for a similar HIV enzyme, the results obtained herein can be evaluated as anti-HIV effects.

Recent data indicate that PARP is required for integration of the retroviral genome into a host cell, and inhibition of PARP prevents integration of the retroviral genome into host cell DNA. Therefore, non-toxic PARP inhibitors may inhibit virulent retroviruses and stop the spread of retroviruses such as HIV and non-type B hepatitis.

From the above experimental results, it can be concluded that the compounds of the present invention can be used as multi-targeting antiviral agents because of their reverse transcriptase inhibiting activity and PARP inhibitory activity.

Accordingly, the novel propenecarboxylic acid amidoxime derivatives can be used as active ingredients in pharmaceutical formulations. Therefore, the present invention also provides a pharmaceutical composition comprising as active ingredient a propenecarboxylic acid amide oxime derivative of formula (I) in addition to the usual carriers in pharmaceutical compositions.

HU 227 116 Β1

The pharmaceutical composition according to the invention generally contains from 0.1 to 95% by weight of the active ingredient, preferably

It is present in an amount of from 1 to 50% by weight, preferably from 5 to 30% by weight, and is suitable for treating diseases based on oxygen deficiency, energy deprivation, PARP inhibition, in particular autoimmune or neurodegenerative diseases and / or viral diseases, and for preventing toxic effects.

The pharmaceutical composition may be a solid or liquid composition suitable for oral, parenteral, rectal or transdermal administration or topical administration.

Solid pharmaceutical compositions for oral administration may be in the form of powders, capsules, tablets, film-coated tablets, microcapsules, etc., and may contain as excipients excipients such as gelatin, sorbitol, polyvinylpyrrolidone, etc .; fillers such as lactose, glucose, starch, calcium phosphate, etc .; tabletting aids such as magnesium stearate, talc, polyethylene glycol, silicon dioxide, etc .; wetting agents such as sodium lauryl sulfate and the like.

Liquid pharmaceutical compositions for oral administration include solutions, suspensions or emulsions which contain, for example, suspending agents such as gelatin, carboxymethylcellulose, etc .; an emulsifier such as sorbitan monooleate; solvents such as water, oils, glycerol, propylene glycol, ethanol; preservatives such as methyl or propyl p-hydroxybenzoic acid and the like. They contain.

Pharmaceutical compositions for parenteral administration generally comprise a sterile solution of the active ingredient.

The above exemplified and other dosage forms are known per se, see, for example, Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Co., Easton, USA (1990).

The pharmaceutical composition generally contains a unit dose. A typical daily dose for an adult is from 0.1 to 1000 mg of the compound of formula I or a pharmaceutically acceptable acid addition salt and / or quaternary derivative and / or N-oxide thereof per kg of body weight. The daily dose may be administered in one or more divided doses. The actual dose will depend on many factors and will be determined by your doctor.

The pharmaceutical composition is prepared by mixing an active compound consisting of a compound of formula (I) or a pharmaceutically acceptable acid addition salt and / or a quaternary derivative and / or N-oxide thereof with one or more excipients, and then converting the resulting mixture into a pharmaceutical composition. Applicable methods are known in the art, such as the Remington's Pharmaceutical Sciences manual mentioned above.

The present invention also encompasses a pharmaceutical use wherein a patient suffering from an oxygen deficiency, energy deficiency, PARP inhibition disorder, in particular an autoimmune or neurodegenerative disorder and / or a viral disease or a toxic effect, is a propenecarboxylic acid amidoxime derivative of formula (I). and a non-toxic amount of its geometric and / or optical isomer and / or pharmaceutically acceptable acid addition salt and / or quaternary derivative and / or N-oxide thereof.

In addition, the present invention relates to the use of novel propenecarboxylic acid amidoxime derivatives of formula (I), and their geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof in an oxygen deficient, for the manufacture of a medicament for the treatment or prevention of toxic effects of diseases based on inhibition, in particular autoimmune or neurodegenerative diseases and / or viral diseases.

The invention is illustrated in detail by the following examples.

Example 1

3-Styryl-4- (3-piperidinopropyl) -Δ ² -1,2,4-oxadiazolin-5-one hydrochloride

0.94 g (0.005 mol) of 3-styrid-Δ ² -1,2,4-oxadiazolin-5-one is dissolved in 6 ml of acetone, and 1.19 g (0.006 mol) of 1-chloro-3-piperidino- is added. propane hydrochloride, 0.76 g (0.0055 mol) of anhydrous potassium carbonate, 1 ml of methanol and 0.05 g of potassium iodide. The reaction mixture is refluxed for 20 hours, the inorganic salts are filtered off and the solution is concentrated in vacuo. The residual oily crude product was dissolved in isopropanol, acidified by the addition of hydrochloric acid isopropanol, allowed to stand overnight in a refrigerator, and the precipitated crystals were filtered. 1.05 g of the title compound is obtained. Mp: 203-205 ° C.

IR (KBr) v = 2550-2650 (NH ⁺ ), 1768 (CO), 1639 (C = N) cm ^-1 .

¹ H-NMR (DMSO-d ₆ ) δ = 1.3-1.85 (6H, m, piperidine-3,4,5-CH ₂ ), 2.11 (2H, m, propyl-CH ₂ ), 2, 82 (2H, m, piperidine-CH ₂ ), 3.09 (2H, m, -CH ₂ -N), 3.36 (2H, m, piperidine-CH ₂ ), 3.87 (2H, m, - O-CH ₂ ), 7.12 (1H, d, Ar-CH = CH-), 7.4-7.5 (3H, m, Ar-H), 7.60 (1H, d, Ar-CH). = CH), 7.8 (2H, m, ArH), 10.3 (1H, br, ⁺ NH).

Example 2

3-Styryl-4- (3-piperidino-2-hydroxypropyl) -A ² -1,2,4-oxadiazolin-5-one hydrochloride

THE)

3-styryl-4, 2.25 g (0.008 mole) of (3-chloro-2-hydroxypropyl) ^{-N-2-oxa-1,2,4} diazolin-5-one in 10 ml of ethanol was added to the solution Piperidine (0.68 g, 0.008 mol) was heated to 50 ° C and a solution of sodium hydroxide (0.32 g, 0.008 mol) in water (2 ml) was added dropwise with stirring. The reaction mixture was stirred at 50-55 ° C for an additional 2 hours, the precipitated crystalline material was filtered off, dried, then dissolved in ethanol with heating, and the solution was acidified by the addition of hydrochloric acid isopropanol. After standing overnight in the refrigerator, the precipitated crystals are filtered off and dried. 1.09 g of the title compound are obtained. Mp: 234-235 ° C.

EN 227 116 Β1 (3-Styryl-4- (3-chloro-2-hydroxypropyl) -A ² -1,2,4-oxadiazolin-5-one used as starting material 3-styryl-A ² -1 , 2,4-oxadiazolin-5-one and epichlorohydrin were prepared according to the method described in the literature (Chem. Bér., 108, 1911 (1975)).

¹ H-NMR (DMSO-d ₆ ) δ = 1.3-1.9 (6H, m, piperidine-3,4,5CH ₂ ), 2.9-3.6 (6H, m, 3CH ₂ ), 3.8 (2H, m, propyl-1-CH _2), 4.35 (1H, m, CH-OH), 6.3 (1H, br, OH), 7.19 (1H, d, Ar-CH = CH), 7.37-7.47 (3H, m, Ar-H), 7.57 (1H, d, Ar-CH = CH-), 7.84 (2H, m, Ar-H), 10. , 25 (1H, br, ⁺ NH).

B)

0.65 g (0.0027 mole) of 3-styryl-4- (2,3-epoxypropyl) -A ^2, 1,2,4-oxadiazolin-5-one was dissolved in 2 ml of methanol and 0.24 g. g of piperidine (0.0028 mol) was added. The reaction mixture, which is heated, is allowed to stand for one hour. The precipitated crystals are filtered off and dissolved in isopropanol. The solution was acidified by addition of hydrochloric acid isopropanol. 0.38 g of the title product crystallizes, which is identical to the compound obtained in A) above. Mp 234-235 ° C.

[3-Styryl-4- (2,3-epoxypropyl] -A ² -1,2,4-oxadiazolin-5-one used as starting material was prepared as follows:

1.5 g (0.0053 mole) of 3-styryl-4- (3-chloro-2-hydroxypropyl) -Δ ² -1,2,4-oxa-diazolin-5-one in 5 ml of acetone was To the solution was added 0.73 g of anhydrous potassium carbonate, the reaction mixture was refluxed for 16 hours, filtered and concentrated in vacuo. 1.41 g of 3-styryl-4- (2,3-epoxypropyl) -A ² -1,2,4-oxadiazolin-5-one are obtained in the form of an oil.]

C.)

3-Styryl-η ² -1,2,4-oxadiazolin-5-one (0.3 g, 0.0016 mol) was dissolved in acetone (3 ml), 3-piperidino-2- (0.41 g, 0.0019 mol). hydroxy-1-chloropropane hydrochloride, 0.48 g of anhydrous potassium carbonate, 1 ml of methanol and 0.05 g of potassium iodide are added. The reaction mixture was refluxed for 40 hours, filtered and the solvent evaporated in vacuo. The residue was dissolved in 5% aqueous hydrochloric acid, filtered and made basic with 10% aqueous sodium hydroxide. The precipitated product was extracted with chloroform, the solution was dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo. The residue was dissolved in isopropanol and acidified with hydrochloric acid isopropanol. 0.18 g of product is crystallized which is the same as the title compound prepared according to A) above. Mp 234-235 ° C.

Example 3

3-Styryl-4- (3-pyrrolidino-2-hydroxypropyl) -A ² -1,2,4-oxadiazolin-5-one hydrochloride

8.4 g (0.03 mol) of 3-styryl-4- (3-chloro-2-hydroxypropyl) Δ ² -1,2,4-oxadiazolin-5-one are dissolved in 30 ml of ethanol, Pyrrolidine (2.55 g, 0.036 mol) was added dropwise at 60 ° C with stirring

A solution of 1.2 g (0.03 mol) of sodium hydroxide in 8 ml of water. The reaction mixture was stirred for an additional hour at 60 ° C and the ethanol was evaporated in vacuo. The residue was acidified with concentrated aqueous hydrochloric acid, the solution clarified with charcoal, filtered and basified with 2N aqueous sodium hydroxide. The oily precipitate was extracted with chloroform, and the solution was dried over anhydrous sodium sulfate, filtered and evaporated. The residue is dissolved in isopropanol and the solution is acidified by the addition of hydrochloric acid isopropanol. The crystals are filtered off and dried. 1.8 g of the title compound are thus obtained. Mp 188-189 ° C.

¹ H-NMR (DMSO-d ₆ ) δ = 1.8-2.0 (4H, m, pyrrolidine 3,4CH ₂ ), 3.06-3.55 (6H, m, 3CH ₂ ), 3.82 (2H, d, propyl 1 -CH ₂ ), 4.21 (1H, m, -CH-OH), 6.25 (1H, d, -OH), 7.14 (1H, d, Ar-CH = CH). -), 7.4-7.5 (3H, m, Ar-H), 7.58 (1H, d, Ar-CH = CH-), 7.82 (2H, m, ArH), 10.3 (1H, br, ⁺ NH).

Example 4

3-Styryl-4- (3-hexamethylenimino-2-hydroxypropyl) -Δ ² -1,2,4-oxadiazolin-5-one hydrochloride

2.8 g (0.01 mol) of 3-styryl-4- (3-chloro-2-hydroxypropyl) Δ ² -1,2,4-oxadiazolin-5-one 1.19 g (0.012 mol) ) with hexamethyleneimine as described in Example 3. The hydrochloric acid salt is separated from the isopropanol solution by the addition of hydrochloric acid isopropanol. 1.0 g of the title compound are obtained. Mp 202-203 ° C.

¹ H-NMR (CDCl ₃ + DMSO-d ₆ ) δ = 1.6-2.0 (8H, m, hexamethylenimine-3,4,5,6-CH ₂ ), 3.1-3.6 (6H, m, _3-CH2), 3.8 (2H, m, _{propyl-1-CH2),} 4.35 (1H, m, -CH-OH), 6.21 (1H, d, OH); 7.11 (1H, d, Ar-CH = CH-), 7.4 (3H, m, Ar-H), 7.57 (1H, d, Ar-CH = CH-), 7.77 (2H). , m, Ar-H), 10.0 (1H, br, ⁺ NH).

Example 5

3-Styryl-4- (3-morpholino-2-hydroxypropyl) - \ ^2--1,2,4oxa diazolin-5-one hydrochloride

4.2 g (0.015 mol) of 3-styryl-4- (3-chloro-2-hydroxypropyl) Δ ² -1,2,4-oxadiazolin-5-one with 1.6 g (0.018 mol) of morpholine is reacted as described in Example 3. The hydrochloric acid salt is separated from ethanol by the addition of hydrochloric acid isopropanol. 0.67 g of the title compound is obtained. Mp: 232-234 ° C.

¹ H-NMR (DMSO-d ₆ ) δ = 3.1-3.55 (6H, m, morpholine-3.5CH ₂ , propyl-3-CH ₂ ), 3.8-4.0 (6H, m , morpholine-2,6CH ₂ , propyl-1-CH ₂ ), 4.37 (1H, m, -CH-OH), 6.3 (1H, br, OH), 7.12 (1H, d, Ar -CH = CH-), 7.4 (3H, m, ArH), 7.6 (1H, d, Ar-CH = CH-), 7.8 (2H, m, ArH),

10.6 (1H, br, ⁺ NH).

Example 6

3-Styryl-4- [3- (tert-butylamino) -2-hydroxypropyl] - \ ^2-oxa-1,2,4 diazolin-5-one hydrochloride

6.6 g (0.024 mol) of 3-styryl-4- (3-chloro-2-hydroxypropyl) - ² -1,2,4-oxadiazolin-5-one 2.63 g (0.036 mol) of tert-butyl amine as described in Example 3.

The hydrochloric acid salt is separated from the isopropanol solution by the addition of hydrochloric acid isopropanol. 1.8 g of the title compound are thus obtained. Mp 244-246 'C.

¹ H NMR (DMSO-d 6) δ = 1.3 (9H, s, tert-butyl), 2.9-3.15 (2H, m, propyl-3-CH ₂ ), 3, 85 (2H, m, _{propyl-1-CH2),} 4.15 (1H, m, CH-OH), 6.08 (1H, d, OH), 7.12 (1H, d, Ar-CH = CH-), 7.40 (3H, m, Ar-H), 7.55 (1H, d, Ar-C7 = CH), 7.8 (2H, m, Ar-H), 8.55 (1H, br, NH), 8.85 (1H, br, NH).

Example 7

3-Styryl-4- [3- (4-methyl-1-piperazinyl) -2-hydroxypropyl] \ ^{2-oxa-1,2,4-5-one} diazolin dihidmkloríd

8.4 g (0.03 mol) of 3-styryl-4- (3-chloro-2-hydroxypropyl) Δ ² -1,2,4-oxadiazolin-5-one 3.6 g (0.036 mol) ) With N-methylpiperazine as described in Example 3. The hydrochloric acid salt is separated from the isopropanol solution by the addition of hydrochloric acid isopropanol. 2.08 g of the title compound are obtained. M.p .: 206-208'C.

¹ H-NMR (DMSO-d 6) δ = 2.77 (3H, s, CH ₃ ), 3.0-3.1 (2H, m, propyl-3-CH ₂ ), 3.6 (8H, m , piperazine-CH ₂ ), 3.8 (2H, m, propyl-1-CH ₂ ), 4.23 (1H, m, -CH-OH), 6.2 (1H, br, OH), 7, 03 (1H, d, Ar-CH = CH-), 7.4 (3H, m, Ar-H), 7.58 (1H, d, Ar-CH = CH-), 7.77 (2H, m). , ArH), 11.8 (2H, br, 2x ⁺ NH).

Example 8

3-Styryl-4- [3- (1,2,3,4-tetrahydro-2-isoquinolyl) -2-hydroxypropyl] -A ² -1,2,4-oxadiazolin-5-one hydrochloride

2.8 g (0.01 mol) of 3-styryl-4- (3-chloro-2-hydroxypropyl) Δ ² -1,2,4-oxadiazolin-5-one 1.6 g (0.012 mol) ) With 1,2,3,4-tetrahydroisoquinoline as described in Example 3. The hydrochloric acid salt is separated from the isopropanol solution by the addition of hydrochloric acid isopropanol. 0.83 g of the title compound is obtained. M.p. 208-210 ° C.

¹ H-NMR (DMSO-d 6) δ = 3.0-3.6 (8H, m, isoquinoline-CH ₂ , propyl-3-CH ₂ ), 3.84 (2H, m, propyl-1-CH _2). ), 4.4 (1H, m, -CH-OH), 6.3 (1H, br, OH), 7.13 (1H, d, Ar-CH-CH-), 7.2-7.5 (7H, m, ArH), 7.60 (1H, d, Ar-CH = CH-), 7.8 (2H, m, Ar-H), 10.6 (1H, br, ⁺ NH).

Example 9

3-Styryl-4- [3- (2,6-dimethylanilino) -2-hydroxypropyl] -Δ ² -1,2,4-oxadiazolin-5-one hydrochloride

8.4 g (0.03 mole) of 3-styryl-4- (3-chloro-2-hydroxypropyl) - ² -1,2,4-oxadiazolin-5-one 4.36 g (0.036 mole) ) With 2,6-dimethylaniline instead of ethanol in 50 ml of methanol solution as described in Example 3. The hydrochloric acid salt is separated from the isopropanol solution with hydrochloric acid isopropanol and then recrystallized from a 1: 1 mixture of isopropanol and ethanol. 1.62 g of the title compound are obtained. Mp 182-184 ° C.

¹ H-NMR (DMSO-d ₆ ) δ = 2.44 (6H, s, CH ₃ ), 3.16-3.53 (2H, m, propyl-3-CH ₂ ), 3.86 (2H, m, _{propyl-1-CH2),} 4.21 (1H, m, CH-OH), 6.0 (1H, br, OH), 7.10 (1H, d, Ar-CH = CH-); 7.14 (3H, m, ArH), 7.4-7.5 (3H, m, ArH), 7.59 (1H, d, Ar-CH = CH-), 7.78 (2H, m, ArH), 9.0 (2H, br, ⁺ NH ₂ ).

Example 10

3- (3,4-Dimethoxy-styryl) -4- (3-piperidino-2-hydroxypropyl) -Δ ² -1,2,4-oxadiazolin-5-one hydrochloride

3.4 g (0.01 mol) of 3- (3,4-dimethoxystyryl) -4- (3-chloro-2-hydroxypropyl) -Δ ² -1,2,4-oxadiazolin-5-one 1.02 g (0.012 mol) of piperidine were reacted as in Example 3. The hydrochloric acid salt is separated from the ethanolic solution with hydrochloric acid isopropanol. 1.8 g of the title compound are thus obtained. Mp 187-188 ° C.

¹ H-NMR (CDCl ₃ + DMSO-d ₆ ) δ = 1.4-2.0 (6H, m, piperidine-3,4,5-CH ₂ ), 3.18-3.31 (6H, m , piperidine-2,6CH ₂ , propyl-3-CH ₂ ), 3.85-3.92 (2H, m, propyl-1CH ₂ ), 3.89 (3H, s, -OCH ₃ ), 3.96 (3H, s, _-OCH3);

4.45 (1H, m, CH-OH), 6.27 (1H, br, OH), 6.93 (1H, m, ArH), 6.98 (1H, d, Ar-CH = CH-). , 7.21 (1H, m, ArH), 7.42 (1H, m, ArH), 7.48 (1H, d, Ar-CH = CH-),

9.8 (1H, br, ⁺ NH).

3- (3,4-Dimethoxystyryl) -4- (3-chloro-2-hydroxypropyl) -A ² -1,2,4-oxadiazol-5-one used as starting material was prepared from 3- (3,4- Dimethoxy-styI) -Δ ² -1,2,4-oxadiazolin-5-one was prepared using epichlorohydrin according to the method known in the literature [Chem. Bér., 108, 1911 (1975)].

Example 11

3-Styryl-4- [3- (1-methyl-4-piperazinyl) -2-hydroxypropyl] -Δ ² -1,2,4-oxadiazolin-5-one dihydrochloride

3- (3,4-Dimethoxy-styryl) -4- (3-chloro-2-hydroxy-propyl) -A ² -1,2,4-oxadiazolin-5-one (2.0 g, 0.0059 mol) 0.71 g (0.0071 mol) of N-methylpiperazine was reacted as in Example 3. The hydrochloric acid salt is separated from the isopropanol solution with hydrochloric acid isopropanol. 0.8 g of the title compound is obtained. M.p. 192-193 ° C.

¹ H-NMR (DMSO-d 6) δ = 2.8 (3H, s, N-CH ₃ ), 3.2-3.8 (10H, m, piperazine-CH ₂ , propyl-3-CH ₂ ), 3.80 (3H, s, methoxy), 3.83 (2H, m, propyl-1-CH ₂ ), 3.86 (3H, s, OCH ₃ ), 4.31 (1H, m, -CH-). OH), 6.3 (1H, br, OH), 7.0 (1H, m, ArH), 7.03 (1H, d, Ar-CH = CH-), 7.30 (1H, m, ArH) ), 7.50 (1H, m, ArH), 7.51 (1H, d, Ar-CH = CH-), 11.8 (2H, br, 2x ⁺ NH).

Example 12

N- (3-piperidino-2-hydroxy-propyl) cinnamic acid amidoxime

1.91 g (0.006 mol) of 3-styryl-4- (3-piperidino-2-hydroxypropyl) -Δ ² -1,2,4-oxadiazolin-5-one (prepared as in Example 2). Ethanol (10 mL) and 10% aqueous sodium hydroxide solution (10 mL) were added and the reaction mixture was refluxed for 2 hours. The ethanol was evaporated in vacuo and the residue was adjusted to pH 8 with aqueous hydrochloric acid. The precipitated semisolid product was decanted and the residue was dissolved in methanol. After dilution with water, 0.79 g of the title compound crystallizes out of solution. 114-115 ° C.

¹ H-NMR (DMSO-d 6) δ = 1.7-1.9 (6H, m, piperidine-3,4,5CH ₂ ), 2.1-2.3 (6H, m, piperidine-2.6) -CH ₂ , propyl-3CH ₂ ), 3.2-3.33 (2H, m, propyl-1-CH ₂ ), 3.83 (1H, m,

CH-OH), 5.6 (1H, br, OH), 6.55 (1H, d,

Ar-CH = CH-, 7.15 (1H, d, Ar-CH = CH-), 7.8-7.32 (3H, m, ArH), 7.44 (2H, m, ArH).

HU 227 116 Β1

Example 13

N- (3-Morpholino-2-hydroxy-propyl) cinnamic acid maleate amidoximdihidrogén

3-styryl-4- (3-morpholino-2-hydroxypropyl) -Δ ² -1,2,4-oxa-diazolin-5-one (2.76 g, 0.0075 mole) (as in Example 5 10 ml of ethanol and 10 ml of 10% aqueous sodium hydroxide solution are added and the reaction mixture is refluxed for 2 hours. The ethanol was evaporated in vacuo and the residue was adjusted to pH 8 with aqueous hydrochloric acid. The precipitated oily product is extracted with dichloromethane, the solution is dried over anhydrous sodium sulfate and the solvent is evaporated. The maleic salt is separated from the acetone solution by adding maleic acid. This way

1.1 g of the title compound are obtained. Mp 128-130 ° C. ¹ H-NMR (CDCl ₃ + DMSO-d ₆ ) δ = 2.8-3.2 (6H, morpholine3.5-CH ₂ , propyl-3-CH ₂ -), 3.3-3.8 (6H , m, morfolin2.6- _{CH2, propyl-1-CH2),} 4.10 (1H, m, CH-OH), 6.05 (4H, s, maleic acid-CH), 6.75 (1H, d, Ar-CH = CH-),

7.30 (1H, d, Ar-CH = CH-), 7.3 (3H, m, ArH), 7.50 (2H, m, ArH).

Example 14

N- [3- (1-Methyl-4-piperazinyl) -2-hydroxypropyl] cinnamic acid amidoxime trihydrogen maleate 1,17 g (0.0025 mol) of 3-styryl-4- [3- (4- methyl-1-piperazinyl) -2-hydroxypropyl] -A ² -1,2,4-oxadiazolin-5-one (prepared as in Example 7) was reacted with sodium hydroxide as in Example 13. . The maleic salt is separated from the acetone solution by adding maleic acid. 0.63 g of the title compound is obtained. M.p. 142-143 ° C.

¹ H-NMR (DMSO-d ₆ ) δ = 2.7 (3H, s, CH ₃ ), 2.5-3.2 (1 OH, m, piperazine-CH ₂ , propyl-3-CH ₂ ), 3.31-3.42 (2H, m, _{propyl-1-CH2),} 3.81 (1H, m, CH-OH), 6.14 (6H, s, maleic acid-CH), 6.90 ( 1H, d, Ar-CH = CH-), 7.28 (1H, d, Ar-CH = CH), 7.4 (3H, m, ArH), 7.65 (2H, m, ArH).

Example 15

N- (3-Morpholino-2-hydroxypropyl) -3,4-dimethoxycinnamic acid amidoxime

3.91 g (0.01 mole) of 3- (3,4-dimethoxystyryl) -4- (3-morpholino-2-hydroxypropyl) -Δ ² -1,2,4-oxadiazolin-5 was reacted with sodium hydroxide as described in Example 13.

1.2 g of the title compound are obtained in the form of an oil.

¹ H-NMR (DMSO-d ₆ ) δ = 3.18-3.35 (6H, morpholine-3.5CH ₂ , propyl-3-CH ₂ ), 3.60 (2H, m, propyl-1-CH). ₂ ),

3.82 (3H, s, OCH ₃ ), 3.86 (3H, s, OCH ₃ ), 3.92 (4H, m, morpholine-2,6-CH ₂ ), 4.34 (1H, m, -CH-OH), 6.3 (1H, br, -CH-OH), 6.98 (1H, d, Ar-CH = CH-), 7.02 (1H, m, Ar-3H), 7 , 24 (1H, m, Ar-2H), 7.35 (1H, d, Ar-CH = CH-), 7.40 (1H, m, Ar-6H).

Example 16

3-Styryl-6- (piperidinomethyl) -4H-5,6-diazine-1,2,4oxa

1.65 g (0.0043 mole) of 3-styryl-4- (3-piperidino-2-chloropropyne) -Δ ² -1,2,4-oxa-diazolin-5-one hydrochloride in 33 ml of ethanol and 33 ml of A 2N aqueous sodium hydroxide solution was added and the reaction mixture was heated under reflux for 30 minutes with stirring. The reaction mixture was concentrated in vacuo and the residue suspended in water (10 mL). The crystallized material is filtered off, washed with water and dried. 1.06 g of the title compound are obtained. M.p. 147-149 ° C.

¹ H-NMR (DMSO-d 6) δ = 1.3-1.7 (6H, m, piperidine-3,4,5CH ₂ ), 2,3-2,7 (6H, m, piperidine-2,6 -CH ₂ , 6-CH ₂ ),

3.25 to 3.6 (2H, m, _{5-oxa-diazine-CH2),} 3.95 (1H, m, 6-oxa-diazine-CH), 5.0 (1H, br-oxa- diazine -4-ΝΗ), 6.5 (1H, d, Ar-CH = CH-), 6.9 (1H, d, Ar-CH = CH-), 7.2-7.5 (5H, m, Ar-H).

The starting material used is 3-styryl-4- (3-piperidino-2-chloropropyl) -A ² -1,2,4-oxadiazolin-5-one hydrochloride 3-styryl-4- (3-piperidino) -2-Hydroxypropyl) -A ² 1,2,4-oxadiazolin-5-one hydrochloride (prepared as described in Example 2) was prepared with thionyl chloride according to a method known in the art [Chem. Bér., 108, 1911 (1975)].

Example 17

3-Styryl-6-morpholino-methyl-4H-5,6-dihydro-1,2,4-oxadiazine dihydrogen maleate

To 1.5 g (0.0039 mol) of 3-styryl-4- (3-morpholino-2-chloropropyl) -A ² -1,2,4-oxadiazolin-5-one hydrochloride 9 ml ethanol and 9 ml A 10% aqueous solution of sodium hydroxide was added and the reaction mixture was refluxed for half an hour. The ethanol was evaporated in vacuo and the residue acidified with 5% aqueous hydrochloric acid. The solution was clarified with activated carbon, filtered, and basified with 10% aqueous sodium hydroxide. The oily precipitate is extracted with chloroform, the organic solvent solution is dried over anhydrous sodium sulfate, filtered and the solvent is evaporated. The residue was dissolved in ethyl acetate and a solution of maleic acid (0.66 g) in ethyl acetate was added. 1.0 g of the expected product are obtained. Mp 137 ° C. ¹ H-NMR (DMSO-d 6) δ = 3.1-3.44 (8H, m, 5-CH ₂ , 6-CH ₂ , morpholine-3 and -5-CH ₂ ), 3.83 (4H , m, morpholine-2 and -6-CH ₂ ), 4.08 (1H, m, 6-CH), 6.18 (4H, s, maleic CH), 6.43 (1H, d, Ar -CH = CH-), 7.25 (1H, br, 4H), 7.33-7.51 (5H, m, 5 Ar-H), 13-14 (br, maleic-OH).

(3-Styryl-4- (3-morpholino-2-chloropropyl) -A ² -1,2,4-oxadiazolin-5-one hydrochloride used as starting material was prepared from the 3-styryl-4 ( 3-morpholino-2-hydroxypropyl) -A ² -1,2,4-oxadiazolin-5-one hydrochloride was prepared by heating with thionyl chloride and evaporating the reaction mixture.)

Example 18

3-Styryl-6- (1,2,3,4-tetrahydro-2-isoquinolyl) methyl-4H5,6-dihydro-1,2,4-oxadiazine dihydrogen maleate

1.1 g (0.0025 mol) of 3-styryl-4- [3- (1,2,3,4-tetrahydro-2-isoquinolyl) -2-chloropropyl-A ² -1,2,4-oxa- To the diazolin-5-one hydrochloride was added ethanol (8 ml) and 10% sodium hydroxide (8 ml), and the reaction mixture was refluxed for half an hour. The ethanol was evaporated in vacuo and the residue was dissolved in 5% aqueous hydrochloric acid. The solution was clarified with charcoal, filtered, 10% aqueous sodium 22

It is made alkaline with a solution of 116 µl of sodium hydroxide. After extraction with ethyl acetate, the combined ethyl acetate solution was dried over anhydrous sodium sulfate, filtered and evaporated. The residue was dissolved in ethyl acetate and 0.36 g of maleic acid was added. 0.55 g of the expected product is obtained. 151-153 ° C.

¹ H-NMR (DMSO-d ₆ ) δ = 3.10 (2H, m, isoquinoline-4-CH ₂ ),

3.15-3.50 (2H, m, _{5-oxa-diazine-CH2),} 3.28-3.39 (2H, m, _{6-oxa-diazine-CH2} -N =), 3.44 -3.6 (2H, m, _{isoquinoline-3-CH2),} 4.2 (1H, m, 6-oxa-diazine-CH), 4.4 (2H, s, _{isoquinoline-1-CH2);} 6.13 (4H, s, maleic-CH), 6.44 (1H, d, Ar-CH = CH-), 7.15 (1H, d, Ar-CH = CH-), 7.2-7 , 33 (4H, m, isoquinoline Ar-H), 7.33-7.52 (5H, m, phenyl Ar-H).

Example 19

3- (3,4-Dimethoxystyryl) -4- [3- (tert-butylamino) -2-hydroxy-propyl] - / \ ^2-oxa-1,2,4 diazolin-5-one hydrochloride

8.54 g (0.025 mol) of 3- (3,4-dimethoxystyryl) -4- (3-chloro-2-hydroxypropyl) -Δ ² -1,2,4-oxadiazolin-5-one in 40 ml dissolved in acetone, and 3.46 g (0.025 mol) of anhydrous potassium carbonate are added. The reaction mixture was refluxed for 6 hours, cooled, the inorganic salts filtered off and the solvent evaporated in vacuo. The residue was crystallized by trituration with methanol (25 mL). 6.5 g of 3- (3,4-dimethoxystyryl) -4- (2,3-epoxy-propyl) -Δ ² -1,2,4-oxa-diazolin-5-one. 113-115 ° C.

3.04 g (0.01 mol) of 3- (3,4-dimethoxy-styryl) -4- (2,3-epoxy-propyl) -Δ ² -1,2,4-oxadiazolin-5-one Methanol (15 mL) and tert-butylamine (0.73 g, 0.01 mol) were added and the reaction mixture was refluxed for 4 hours and the solvent evaporated in vacuo. The residue was dissolved in 10 ml of a 5% aqueous hydrochloric acid solution and the solution was decanted from the insoluble residue. The aqueous solution was made basic with 10% aqueous sodium hydroxide solution, the oily precipitate was dissolved in dichloromethane, the solution was dried over anhydrous sodium sulfate, filtered and the solvent was evaporated in vacuo. 1.7 g of an oily product are obtained, the hydrochloric acid salt of which is separated from the absolute ethanolic solution by the addition of hydrochloric acid isopropanol. 1.0 g of the title compound are obtained. Mp: 225-227 ° C.

¹ H-NMR (DMSO-d ₆ ) δ = 1.3 (9H, s, 3 * CH ₃ ), 2.93-3.17 (2H, m, propyl-3-CH ₂ ), 3.80 ( 3H, s, -OCH ₃ ), 3.85 (3H, s, -OCH ₃ ), 3.9 (2H, d, propyl-1-CH ₂ ), 4.16 (1H, m, -CH-OH). ), 6.12 (1H, d, -OH), 7.03 (1H, d, Ar-CH = CH-), 7.51 (1H, d, Ar-CH = CH-), 7.00 ( 1H, m, ArH), 7.29 (1H, m, ArH), 7.49 (1H, m, ArH), 8.58 (1H, br, NH), 8.86 (1H, br, NH) ).

Example 20

3- (3,4-Dimethoxystyryl) -4- (3-morpholino-2-hydroxypropyl) - / \ ^2-oxa-1,2,4 diazolin-5-one hydrochloride

3.04 g (0.01 mol) of 3- (3,4-dimethoxy-styryl) -4- (2,3-epoxy-propyl) -Δ ² -1,2,4-oxadiazolin-5-one (prepared as in Example 19) was treated with 0.96 g (0.011 mol) of morpholine in the same manner as in Example 19. 0.8 g of the title compound is obtained. Mp 228-230 ° C.

¹ H-NMR (DMSO-d 6) δ = 3.2-3.34 (6H, m, morpholine-3.5CH ₂ , propyl-3-CH ₂ ), 3.82 (3H, s, OCH ₃ ), 3.86 (2H, d, propyl-1-CH ₂ ), 3.88 (3H, s, OCH ₃ ), 3.90 (4H, m, morpholine-2,6-CH ₂ ), 4.43 ( 1H, m, -CH-OH), 6.4 (1H, br, OH), 7.0 (1H, d, Ar-CH), 7.03 (1H, d, Ar-CH = CH), 7.29 (1H, m, ArH), 7.48 (1H, m, ArH), 7.50 (1H, d, Ar-CH = CH-), 10.7 (1H, br, + NH).

Example 21

N- [3- (2,6-Dimethyl-phenylamino) -2-hydroxypropyl] -fahéjsavamidoxim

Following the procedure of Example 12, except that 1.5 g (0.004 mole) of 3-styryl-4- [3- (2,6-dimethyl-phenylamino) -2-hydroxypropyl] -N ² -1,2,4-oxadiazolin-5-one (prepared according to Example 9). 0.55 g of the title compound is obtained. Mp: 143-144 ° C.

¹ H-NMR (DMSO-d 6) δ = 2.20 (6H, s, 2 * CH ₃ ), 2.82-3.03 (2H, m, propyl-3-CH ₂ ), 3.15-3. 27 (2H, m, propyl-1-CH _2), 3.64 (1H, m, -CH-OH), 3.80 (1H, m, NH-aniline), 5.15 (1H, br, -CH -OH), 5.58 (1H, br, NH-amidoxime), 6.68 (1H, d, Ar-CH = CH-), 6.69 (1H, m, ArH), 6.90 (2H, m, ArH), 7.01 (1H, d, Ar-CH = CH-), 7.25-7.55 (5H, m, ArH), 9.57 (1H, S, = N-OH).

Example 22

N- (3-pyrrolidino-2-hydroxy-propyl) cinnamic acid amidoxime

Example 12 was repeated except that 1.23 g (0.0035 mol) of 3-styryl-4- (3-pyrrolidino-2-hydroxypropyl) -A ² -1,2,4 starting from oxadiazolin-5-one hydrochloride (prepared as described in Example 3). 0.65 g of the title compound is obtained. 139-141 ° C (from 96% ethanol).

¹ H-NMR (CDCl ₃ + DMSO-d ₆ ) δ = 1.75 (4H, m, pyrrolidine- _3,4 -CH ₂ ), 2.50-2.64 (6H, m, pyrrolidine-2,5-CH ₂ , propyl-3-CH ₂ ), 3.2-3.35 (2H, m, propyl-1-CH ₂ ), 3.75 (1H, m, CH-OH), 5.6 (1H, t , NH), 6.58 (1H, d, Ar-CH = CH-), 7.08 (1H, d, Ar-CH = CH-), 7.25-7.45 (5H, m, Ar- H), 9.0 (1H, br, = N-OH).

Claims (9)

PATENT CLAIMS A propoxycarboxylic acid amidoxime derivative of formula (I) wherein: R is C 1-20 alkyl, phenyl optionally substituted with 1 to 3 substituents wherein the substituent is halogen and / or C 1-2 alkyl and / or C 1-2 alkoxy and / or amino and / or (1-4). C 1 -C 4 alkylamino and / or di (C 1 -C 4 alkyl) amino and / or C 1 -C 4 alkanoyl-amino and a 5 or 6 membered saturated or unsaturated heterocyclic group containing one or two nitrogen atoms or one sulfur atom as a heteroatom and optionally fused to one or more benzene rings and / or other heterocyclic groups, and R 'is hydrogen, or HU 227 116 Β1 R 'of the gap, taken together, is a C 5 -C 7 cycloalkyl group optionally fused to a benzene ring, R ₄ and R ₅ each independently represent hydrogen, C 1 -C 5 alkyl, C 1 -C 5 alkanoyl or phenyl optionally substituted with 1 to 3 substituents wherein the substituent is halogen and / or C 1 -C 2 alkyl and / or C 1-2 alkoxy, or R ₄ and R _{5 taken} together with the adjacent nitrogen atom form a 5 or 6 membered saturated or unsaturated heterocyclic group which may also contain a further nitrogen atom and / or oxygen and / or sulfur atom as a heteroatom and may be fused to a benzene ring and the benzene ring may carry one or two substituents wherein the substituent is C 1-2 alkyl and / or C 1-2 alkoxy and / or halogen, · R! and R ₂ is hydrogen; and R ₃ is hydrogen, hydroxy or C 1-5 alkoxy, or R ₁ and R ₂ together form a carbonyl or thiocarbonyl group whose carbon atom adjacent to the oxygen atom and R _r digested with the nitrogen atom adjacent to R _2, and R ₃ is hydrogen, halogen, hydroxy, C 1 -C 5 alkoxy, C 1 -C 5 alkylthio, C 1 -C 20 alkanoyloxy, C 3 -C 22 alkenoyloxy having one or more double bonds a methylsulfonyloxy group, a benzenesulfonyloxy group or a toluenesulfonyloxy group, or R ₂ is hydrogen, and R ₁ and R ₃ together form a direct bond between the oxygen atom adjacent to R ₄ and the carbon atom adjacent to R ₃ , and also its geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof. . The propenecarboxylic acid amidoxime derivative of the formula (Ia) according to claim 1, wherein R ₁ and R ₂ is hydrogen, R ₃ is hydrogen, hydroxy or C 1-5 alkoxy, R, R ', R ₄ and R ₅ are as defined in claim 1, and also geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof. The oxadiazole derivative of the Formula Ib according to claim 1, wherein Ri and _R2 taken together form a carbonyl or thiocarbonyl group whose carbon atom adjacent to the oxygen atom and R _r digested with the nitrogen atom adjacent to R _2, R ₃ is hydrogen, halogen, hydroxy, C 1 -C 5 alkoxy, C 1 -C 5 alkylthio, C 1 -C 20 alkanoyloxy, C 3 -C 22 alkenoyloxy having one or more double bonds , methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy, R, R ', R ₄ and R ₅ are as defined in claim 1, X is oxygen or sulfur, and its geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof. An oxadiazine derivative of the general formula (Ie) according to claim 1, wherein R ₂ is hydrogen, R ₁ and R ₃ together form a direct bond between the oxygen atom adjacent to R ₁ and the carbon atom adjacent to R ₃ , R, R ', R ₄ and R ₅ are as defined in claim 1, and also geometric and / or optical isomers and / or pharmaceutically acceptable acid addition salts and / or quaternary derivatives and / or N-oxides thereof. A process for preparing the propoxycarboxylic acid amide oxime derivative of formula (I) wherein R, R ', R 1, R ₂ , R ₃ , R ₄ and R ₅ are as defined in claim 1, and pharmaceutically acceptable acid addition salts, quaternary derivatives and N- oxides, characterized in that: (a) for the preparation of propenecarboxylic acid amidoxime derivatives of the general formula (Ia), wherein R _{1 is;} R ₂ and R ₃ are hydrogen, R, R ', R ₄ and R ₅ are propylene derivatives of the general formula (II) as defined herein, wherein R, R', R ₃ , R ₄ and R ₅ are as defined above, Y is halogen or a group of the formula -SR ₆ wherein R ₆ is hydrogen or C 1-4 alkyl, is reacted with hydroxylamine; obsession b) for the preparation of a propoxycarboxylic acid amidoxime derivative of the general formula (Ia), wherein R ₁ and R ₂ are hydrogen, R ₃ is hydrogen or hydroxy, R, R ', R ₄ and R ₅ is an oxadiazole derivative of the formula Ib as defined herein wherein R, R ', R ₃ , R ₄ and R ₅ are as defined above, X is oxygen or sulfur, an aqueous solution of an alkali metal hydroxide; obsession c) forming a narrower group of the compounds of formula (I) oxa-diazolin derivative of formula (Ib) are compounds wherein R ₃ is hydrogen, X is oxygen, R, R ', _R4 and _R5 are as defined above, a Δ ² -1,2,4-oxadiazole derivative of formula (III) wherein R and R 'are as defined above, with an aminoalkyl halide of formula (IV) wherein Z is halogen, R ₃ , R ₄ and _R5 are as hereinbefore defined; obsession d) for the preparation of an oxadiazoline derivative of the Formula Ib, wherein R ₃ is hydrogen or hydroxy, X is O, R, R ', R ₄ and R ₅ are within the scope of the present invention; defined above, (III) of general formula a ^2-oxa-1,2,4 diazolin derivative wherein R and R 'are as defined above, 1.324 (V) of the formula The reaction is carried out with 1-1 dihalopropane, where Z and -L 1 are independently halogen, and the resulting Δ ² -1,2,4-oxadiazolinylalkyl halide of formula VI where R, R ', R ₃ and Z are as defined above, with an amine of formula VII wherein R ₄ and R ₅ are as defined above; obsession e) for the preparation of a narcotic oxadiazoline derivative of the Formula Ib wherein R ₃ is hydroxy, X is O, R, R ', R ₄ and R ₅ are as defined in the foregoing, reacting a Δ ² -1,2,4-oxadiazoline derivative of formula (III) wherein R and R 'are as defined above with epichlorohydrin and then forming the Δ ² -1,2,4-oxa-derivative of formula (VIII) diazolinil alkyl chloride, wherein R and R 'are as defined above, with an amine (VII) wherein R ₄ and R ₅ are as defined above, is reacted; obsession f) for the preparation of a narcotic oxadiazoline derivative of the Formula Ib wherein R ₃ is hydroxy, X is O, R, R ', R ₄ and R ₅ are as defined in the foregoing, an Δ ² 1,2,4-oxadiazolinylalkyl chloride of formula VIII wherein R and R 'are as defined above, is reacted with an acid acceptor and the resulting epoxide of formula IX wherein R and R' are reacting the above amine of formula VII wherein R ₄ and R ₅ are as defined above; obsession g) for the preparation of an oxadiazoline derivative of the Formula Ib wherein R, R ', R ₄ and R ₅ are as defined above, R ₃ is hydrogen or hydroxy, X is a compound of Formula I; oxygen atom or sulfur atom, a (Ia) propenoic acid amidoxime derivative of the formula wherein R, R ', R _3, R ₄ and R ₅ are as defined above, (X) of carbonic acid of the formula wherein X is as defined above, Z ₂ and Z ₃ is, independently, halogen, C 1-4 alkoxy, or C 1-4 alkyl mercapto; obsession h) for the preparation of an oxadiazine derivative of the general Formula I wherein R, R 1, R ₄ and R ₅ are as defined herein, an oxadiazole of the Formula Ib a compound wherein R, R ', R ₄ and R ₅ are as defined above, X is oxygen or sulfur, R ₃ is halogen, methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy with alkali metal hydroxide in the presence of water; obsession i) for the preparation of an oxadiazine derivative of the general Formula I wherein R, R ', R ₄ and R ₅ are as defined herein, a cyclic compound of the Formula XI; wherein R and R 'are as defined above, R ₇ is halogen, methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy, with an amine of formula VII wherein R ₄ and R ₅ are above, reacting; obsession j) for the preparation of a quaternary derivative of a compound of formula (I) wherein R, R ', R _q , R ₂ , R ₃ , R ₄ and R ₅ are as defined for formula (I), R ₈ is C 1 -C 4 alkyl or phenyl (C 1 -C 4 alkyl), Y is halogen or a group R ₈ -SO ₄ , a Δ ² -1,2,4-oxadiazole of formula III a derivative wherein R and R 'are as defined above, with a quaternary alkyl halide of formula XIII wherein R ₃ , R ₄ , R ₅ , R ₈ and Y are as defined above, Z is halogen; obsession k) for the preparation of the N-oxide of the formula I in the formula XIV wherein R, R ', R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are as defined for formula (I); A Δ ² -1,2,4-oxadiazole derivative of formula III wherein R and R 'are as defined above, with a compound of formula XV wherein R ₃ , R ₄ and R ₅ are as defined above, Z is halogen , reacting; and, if desired, a compound of the formula (Ib) wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined herein, X is oxygen or sulfur, reacted with a halogenating agent to replace R ₃ ) to a compound of the formula; or, if desired, a compound of formula Ib, wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined herein, X is oxygen or sulfur, with C 1-20 alkanecarboxylic acid halide or 3- Reaction with a C 22 alkane carboxylic acid halide containing one or more double bonds to form a compound of formula Ib wherein R ₃ is C 1-20 alkanoyloxy or C 3-22 alkenoyloxy; or, if desired, a compound of formula Ib, wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined in formula I, X is oxygen or sulfur, a C 1 -C 5 alkyl group; reacting with a halide to form a compound of formula (Ib) wherein R ₃ is C 1-5 alkoxy; or, if desired, a compound of formula (Ib) wherein R ₃ is halogen, R, R ', R ₄ and R ₅ are as defined for formula (I), X is oxygen or sulfur, a C 1 -C 5 alkanol or reacting with an alkali metal salt of thioalkanol to form a compound of formula (Ib) wherein R ₃ is C 1-5 alkoxy or C 1-5 alkylthio. or, if desired, a compound of formula Ib, wherein R ₃ is hydroxy, R, R ', R ₄ and R ₅ are as defined herein, X is oxygen or sulfur, with methylsulfonyl halide, benzenesulfonyl, reacting with a halide or toluenesulfonyl halide to form a compound of formula Ib wherein R ₃ is methylsulfonyloxy, benzenesulfonyloxy or toluenesulfonyloxy; and optionally converting the resulting compound of formula (I) to a pharmaceutically acceptable acid addition salt thereof with an inorganic or organic acid and / or quaternizing one or more nitrogen atoms of the compound of formula (I) with an alkylating agent and / or ) general The compound of formula Β1 is converted to N-oxide by oxidizing agent on one or more nitrogen atoms. Pharmaceutical composition, characterized in that the active ingredient is a propoxycarboxylic acid amidoxime derivative of the formula (I), in which R, R ', R _{1 (} R ₂ , R ₃ , R 4 and R ₅₎ are as defined in claim 1 or and / or an optical isomer (s) and / or a pharmaceutically acceptable acid addition salt and / or a quaternary derivative and / or N-oxide thereof in addition to one or more conventional carriers. 7. The therapeutic use of the propenecarboxylic acid amidoxime derivatives of the formula I, characterized in that a patient suffering from an oxygen deficient, energy deficient, PARP inhibitory disease, in particular an autoimmune or neurodegenerative disease and / or a viral disease, is affected or toxic. The propenecarboxylic acid amidoxime derivative of formula (I) wherein R, R ', R 1, R ₂ , R ₃ , R 4 and R ₅ are as defined in claim 1 or a geometric and / or optical isomer (s) thereof and / or a pharmaceutically acceptable acid addition salt 5 and / or a non-toxic amount of a quaternary derivative and / or its N-oxide. The propenecarboxylic acid amidoxime derivative of formula (I) wherein R, R ', R _h, R ₂ , R ₃ , R ₄ and R ₅ are as defined in claim 1 or And / or its optical isomer (s) and / or pharmaceutically acceptable acid addition salt and / or quaternary derivative and / or N-oxide thereof for the treatment of diseases based on oxygen deficiency, energy deficiency, PARP inhibition, in particular autoimmune or neurodegenerative diseases and / or viral diseases. for the preparation of a pharmaceutical composition for preventing toxic effects.