HU210001B - Process for producing 6-member heteroaromatic compounds comprising 1-3 nitrogen atoms and pharmaceutical compositions comprising same - Google Patents

Abstract

2-X-4-Y-5-Z-6-R1- Pyrimidine derivs. of formula (I) and their pharmaceutically acceptable salts are new: R1=H or lower alkyl; X=morpholino, 4-R2-piperazino, NR4R5 or a gp. of formula (a)-(e) R2=H, lower alkyl, phenyl, benzyl or alpha-(p-chlorophenyl)benzyl; x=1 or at least 2; each R3=lower alkyl, OH, phenyl opt. substd. by NO2, benzyl, benzoyloxy, benzoylamino, mono- or di-lower alkylamino, C(Ph)2OH, piperidino, lower hydroxyalkyl, PhSO2O, benzoyl opt. substd. by halo, lower alkylsulphonamido or lower alkoxycarbonyl; n=4-7; R4=H lower alkyl or benzyl; R5=lower alkyl, lower acyl, 2-fluroyl, benzyl, 4-piperidyl opt. substd. by benzoyl, phenethyl, a gp. of formula (g) or benzoyl opt. substd. by halo or NO2. Y= CH2R9, NR6R7 or a gp. of formula (g)-(j); R9=H, lower alkyl, lower alkoxy, lower alkylthio or di-lower alkylamino; R6=H, lower alkyl, phenyl, benzyl, lower alkoxy or 2-(N, N-dimethyl amino)ethyl; R7 = lower alkyl, lower acyl, cyclohexylcarbonyl, 2-fueoyl, lower alkoxycarbonyl, cinnamoyl, benzyl, benzylcarbonyl, tosyl, phenoxyacetyl, di-lower alkylcarbamoyl, 2-thienyl, COQ, CONHPh, COOPh, 4-lower alkyl piperazyl or benzoyl opt. substd. by halo, NO2, lower alkyl, lower alkoxy, NH2, benzoylamino or phenyl; provided that when R6=H, R7=benzoyl; Q=4-pyridyl, 1-piperidinyl or morpholino; R8=H, lower alkyl, phenyl or benzyl; m=4-7; Z=H halo, lower alkyl or lower alkoxycarbonyl; provided that (1) Y=CH2R9 only when Z=lower alkoxycarbonyl; (2) R4=H only when R5=lower alkyl, lower acyl, 2-furoyl, benzyl, phenethyl or benzoyl opt. substd. by halo or NO2 Y=CH2R9 and Z=lower alkoxycarbonyl or (3) Y=(g), (i) or (j) only when X=NR4R5 and R4= lower alkyl.

Description

The present invention relates to novel heteroaromatic compounds having 1-3 nitrogen atoms and their pharmacologically acceptable salts and to pharmaceutical compositions for treating diseases of the peripheral and central nervous system of animals.

Japanese Patent Application Laid-Open No. 23,394/1971 discloses aminopyrimidines of Formula A wherein

A is a C16 alkylene group or an amino substituted lower alkylene group or a C2-5 acylamino group,

M is hydrogen, sodium, potassium, ammonium, magnesium, calcium or an organic basic ammonium salt; n is equal to M, which has important therapeutic activity, particularly as anti-melancholic and psychoanaleptic agents in the field of psychosis.

Japanese Patent Application 22 044/1976 discloses dichloro-lower carboxylic acid salts of 2-isopropylaminopyrimidine, such as 2-isopropylaminopyrimidine dichloroacetate, as therapeutic agents for neurological disease.

Japanese Patent Application Publication No. 100477/1977 discloses that 2-isopropylaminopyrimidine phosphate may be used as a therapeutic agent for neurological disorders.

Japanese Patent Application Publication No. 393/1980 describes the preparation of 2-isopropylaminopyrimidine in high yield. One embodiment of this disclosure describes the preparation of 2-isopropylaminopyrimidine by 60%.

Japanese Patent Application Laid-Open No. 122,768/1980 discloses hydroxyl derivatives of 2-isopropylaminopyrimidine of formula B wherein

⁴ , A ⁵ and A ⁶ are hydrogen or hydroxy, and at least one of them is a hydroxyl group useful in the field of nerve regeneration and in the treatment of myodystrophy.

Japanese Patent Application Laid-Open No. 145,670/1980 discloses 2-isopropylamino-halopyrimidine derivatives of the general formula (C) wherein A ₄ , A ₅ 'and Ag' are hydrogen or at least one of them; represents a halogen atom which can be used in the treatment of various neurological diseases and myodystrophy.

Japanese Patent Application Publication No. 145,671/1980 discloses the preparation of a hydroxyl derivative of 2-isopropylaminopyrimidine.

Japanese Patent Application Publication No. 151,571/1980 discloses 2-isopropylamino-5-halopyrimidines as important therapeutic agents for neurological diseases.

Japanese Patent Application Laid-Open No. 10,177/1981 discloses the preparation of 2-isopropylaminopyrimidine in essentially quantitative yield by aminating 2-methylsulfonylpyrimidine with isopropylamine.

Japanese Patent Application Laid-Open No. 26,880/1981 discloses the preparation of 2-isopropylaminopyrimidine by reacting bis (isopropylguanidine) sulfate with 1,1,3,3-tetraethoxypropane.

Japanese Patent Application Laid-Open No. 90,013/1981 discloses substituted pimidine derivatives, as well as therapeutically acceptable salts or metabolites thereof, for use as a therapeutic agent of myodystrophy, myopathy, muscle stiffness and / or neuromuscular transmission dysfunction. However, it discloses only different salts as active ingredient, such as orthophosphate of 2-isopropylaminopyrimidine.

Japanese Patent Application Publication No. 65 873/1986 discloses 2-piperazinopyrimidines of Formula D, wherein

R ¹ is hydrogen or aralkyl and Y is an organic group which can be used as herbicides in rice fields and on highland farms.

International PCT Application Publication No. WO 87/04928 discloses certain 2-piperazinopyrimidine derivatives and their pharmacologically acceptable salts as therapeutic agents for neurological diseases.

The present invention relates to novel heteroaromatic compounds having 1-3 nitrogen atoms and their pharmacologically acceptable salts, and to pharmaceutical compositions containing them, which are useful in the treatment of neurological diseases and spinal nerve collapse, in the regeneration of nerve cells, in peripheral nerve disorders, in brain injury. for use in diseases of the central nervous system, other than psychosis, in which abnormalities of the operating system or the metabolic system of chemical transmitters are considered as primary factors, and in the treatment of brain diseases in which learning and memory are improved or regenerated, and which has minor side effects, such as liver damage.

The compounds of the present invention are represented by formula (I) wherein A 1 is = CH- or -N =.

₂ is = CH-, -N =, or a)

₃ is = CH- or -N =,

R 1 is C 1-7 alkyl,

R ¹² is phenyl, 2-furyl optionally substituted with halo, singly or

2-thienyl, provided that when A! is -N =, then A ₂ is a group of formula (a), and A ₃ is -N =; if the! and A ₂ is = CH-, then A ₃ is = CH-; and when A ₂ is -N =, then A ₁ and A ₃ are = C 1 -C 7 alkyl groups at the place of R ¹¹ may be straight or branched, preferably C 1 -C 4, e.g.

EN 210 001 Β ethyl, ethyl, η-propyl, i-propyl, η-butyl, sec-butyl, 1-butyl and tert-butyl.

A 1 is = C- or = N-, and A ₂ is = CH or -N =, or a is a. ₃ is = CH or -N =. When A 1 and A ₂ are = CH-, then A ₃ is = CH-. When A ₂ is -N =, then A ₁ and A ₂ are = CH-.

Examples of compounds of formula (I) include, but are not limited to, the following:

Compound (2016), Compound (2020), ptoluenesulfonate of Compound (2016), Compound (2022), Compound (2022-1), Compound (2022), p-toluenesulfonate, Compound (2023) , A compound of formula (2023-1), a p-toluenesulfonate of a compound of formula (2023), a compound of a compound of formula (2024), a ptoluene sulfonate of a compound of formula (2024), a compound of a compound of formula (2036), compound 2032 is dipoluene sulfonate, compound 2040, compound 2044, ptoluenesulfonate compound 2040.

The preparation of compounds of formula (I) is described in detail in the Examples and generally in Scheme A.

Compounds of formula (II) used as starting materials in the reactions shown in Scheme A may be prepared from compounds of formula (XIII) according to J. Chem. Soc. 1965, 755-761. pages, as described in the literature.

The reaction shown in the scheme is preferably carried out at a temperature of from 20 to 150 ° C in a solvent such as toluene, dioxane, pyridine or water, optionally in the presence of a basic compound. As the basic compound, for example, an organic base (e.g. triethylamine, pyridine or 4-dimethylaminopyridine) or an inorganic base (such as sodium carbonate or potassium carbonate) can be used.

In our research, we have found that the compounds of formula I are useful in the treatment of neurological disorders.

The compounds of formula (I) may be administered in the form of pharmaceutical compositions by various routes, for example, orally, subcutaneously, intramuscularly, intravenously, intravenously, transdermally or rectally.

The present invention also relates to a process for the preparation of a pharmaceutical composition comprising as active ingredient a compound of formula (I) or a pharmacologically acceptable salt thereof. Pharmacologically acceptable salts include, for example, acid addition salts and quaternary ammonium (or amine) salts.

Pharmacologically acceptable salts of the compounds of formula I are salts with acids which form pharmacologically acceptable non-toxic acid addition salts which may contain, for example, the following anions: chloride, bromide, sulfate, bisulfite, phosphate, hydrophosphate, acetate, maleate, furmarate, succinate, lactate, tartrate, benzoate, citrate, gluconate, glucanate, methanesulfonate, p-toluenesulfonate, naphthyl sulfonate or hydrates thereof, and quaternary ammonium (or amine) salts or hydrates thereof.

The pharmaceutical compositions of the present invention may be formulated, for example, in the form of tablets, capsules, powders, granules, pills, wafers, capsules, elixirs, emulsions, solutions, syrups, suspensions, aerosols, ointments, aseptic injection, molten cataplasm, ribbon, soft gelatin capsules. and aseptically packaged in powder form. Examples of pharmacologically acceptable carriers are lactose, glucose, sucrose, sorbitol, mannitol, maize starch, crystalline cellulose, gum arabic, calcium phosphate, alginates, calcium silicate, microcrystalline cellulose, polyvinylate, polyvinylate, polyvinylate, cellulose, methyl hydroxybenzoic acid esters, propyl hydroxybenzoic acid esters, talc, magnesium stearates, inert polymers, water and mineral oils.

The solid and liquid compositions may contain the above fillers, as well as binders, lubricants, wetting agents, disintegrating agents, emulsifying agents, suspending agents, preserving agents, flavoring agents and sweetening agents. The compositions of the present invention may be formulated so that, when administered to a patient, the active ingredient is released rapidly, continuously, or slowly.

In the case of oral administration, the compounds of the formula I are mixed with a carrier or diluent to form tablets, capsules, and the like. For parenteral administration, the active ingredient is dissolved in 10% aqueous glucose, isotonic saline, sterile water, or the like, filled into a vial or ampoule for intravenous, infusion or injection or intramuscular injection. Preferably, the medium may comprise a local anesthetic, a preservative and a buffer. To improve stability, the composition may be lyophilized after filling into a vial or ampoule. Formulations for parenteral administration may also be presented as a transdermal formulation, i.e., as an ointment or cataplasm. In this case, the molten cataplasm or strip is preferred.

The composition according to the invention may contain from 0.1 to 2000 mg, preferably 0.51000 mg of active ingredient per unit dose.

The compounds of formula I are effective over a wide dosage range. For example, the daily amount of the compound may range from 0.03 mg / kg to 100 mg / kg. The compound will depend on the type of compound being administered, the age, weight and general condition of the subject being treated and the mode of administration.

HU 210 001 Β

Dosage can be 1-6 times a day, usually 1-4 times a day.

The compounds of formula I are useful in the treatment of disorders of the peripheral nervous system and the central nervous system. If desired, it may be co-administered with at least one other equally active agent. Examples of such agents include gangliosides, mecobalamin and isaxonin.

The preparation of compounds of formula I and their biological activity is illustrated by the following non-limiting examples.

Example 1

4- (N-Methyl-benzoylamino) -2- (4-phenyl-piperidino) -pyrimidine (Compound 164) (Reference Example)

A solution of benzoyl chloride (5.2 g, 0.037 mol) in tetrahydrofuran (50 ml) was added at room temperature over 30 minutes with 4-methylamino-2- (4-phenylpiperidino) pyrimidine (9.0 g, 0.034 mol) in tetrahydrofuran (90 ml). To a solution of triethylamine (5 ml) was added pyridine (1 ml) 2 hours after completion of the addition, the mixture was stirred for 2 days, extracted with dichloromethane, dried over anhydrous sodium sulfate and evaporated under reduced pressure. Purification by silica gel column chromatography gave 8.8 g (70%) of the title compound as an oil.

1 H-Nuclear magnetic resonance spectrum (deuterochloroform, δ ppm):

1.4-2.0 (4H, m), 2.5-3.0, (3H, m), 3.55 (3H, s), 4.62 (2H, broad d, J = 12.6 Hz), 6.14 (1H, d, J = 7.2 Hz), 7.1-7.6 (10H, m), 8.06 (1H, d, J = 7.2 Hz).

The physical data of the compounds of the invention prepared in the same manner are summarized in Table 1.

Table 1

Compound number Evade- pro- duction (%) Op. (° C) 1 H NMR (CDCl 3 solution, δ ppm) 2016 50 oil 1.4-2.0 (4H, m), 2.5-3.0 (3H, m), 3.53 (3H, s), 4.21 (2H, broad d, J = 12.6 Hz) ), 6.12 (1H, d, J = 7.2 Hz), 6.35 (1H, d, J = 7.2 Hz), 7.0-7.5 (11H, m) 2024 36 oil 1.4-2.0 (4H, m), 2.5-3.1 (3H, m), 3.54 (3H, s), 4.25 (2H, broad d, J = 12.6 Hz) ), 7.0-7.6 (10H, m), 7.41 (1H, s), 7.79 (1H, s) 2032 69 76-82 1.2-2.0 (8H, m), 2.4-3.9 (6H, m), 3.56 (3H, s), 4.50 (4H, m), 7.0-7, 52 (15H, m) 2040 28 oil 1.4-2.1 (4H, m), 2.5-3.2 (3H, m), 3.50 (3H, s), 4.38 (2H, broad d, J = 12.6 Hz) ), 6.32 (1H, dd, J = 3.6.2.0 Hz), 6.71 (1H, dd, J = 3.0.1.0 Hz), 7.24 (6H, m) , 7.58 (1H, s), 7.93 (1H, s)

Example 2

4- (N-Methyl-benzoylamino) -2- (4-phenyl-piperidino) -pyrimidine-p-toluenesulfonate (p-Toluenesulfonate of Compound 164) (Reference Example)

A solution of p-toluenesulfonic acid monohydrate (3.0 g, 0.022 mol) in ethyl acetate (300 ml) was added slowly at room temperature to 4- (N-methylbenzoylamino) -2- (4-phenylpiperidino) pyrimidine (6.0 g). 100 ml of ethyl acetate. Upon administration, a suspension is formed. After the addition, the suspension was stirred for 10 minutes. The resulting solid was collected by filtration, washed with ethyl acetate and diethyl ether and dried to give 6.8 g (83%) of the title product. Mp: 180-182 ° C.

1 H-NMR (deuterochloroform, δ ppm):

1.4-2.1 (4H, m), 2.35 (3H, s), 2.6-3.3 (3H, m), 3.56 (3H, s), 4.55 (2H, broad d, J = 12.6 Hz), 6.60 (1H, d, J = 7.2 Hz), 7.0-7.9 (14H, m), 8.36 (1H, d, J = 7.2 Hz).

The physical data of the compounds of the present invention prepared in the same manner are summarized in Table 2.

Table 2

Compound number Evade- pro- duction (%) Op. (° C) 1 H-NMR (CDCl ₃ solution, δ ppm) 2020 100 54-60 1.7-2.2 (4H, m), 2.34 (3H, s), 2.5-3.3 (3H, m), 3.4-3.7 (2H, m), 3, 55 (3H, s), 6.9-7.9 (17H, m), 8.35 (1H, m) 2028 100 60-70 2.4-3.1 (4H, m), 2.35 (3H, s), 2.5 - 3.2 (3H, m), 3.50 (3H, s), 4.24 (2H, broad d, J = 12.6 Hz), 7.0-7.5 (12 H, m), 7.75 (2 H, m), 8.04 (2 H, m) 2036 100 54-58 1.4-2.1 (8H, m), 2.32 (6H, m), 2.2-3.3 (6H, m), 3.55 (3H, s), 4.50 (4H, m), 6.95-7.8 (23H, m), 9.1 (2H, m) 2044 100 62-72 1.4-2.2 (4H, m), 2.35 (3H, s), 2.5-3.4 (3H, m), 3.59 (3H, s), 4.36 (2H, br d, J = 12.6 Hz), 6.48 (1H, dd, J = 3.6, 2.0 Hz), 7.0-7.6 (9H, m), 7.76 (2H, d, J = 7 , 2 Hz), 7.97 (1H, s), 8.15 1H, s), 10.92 (1H, m) 2022-1 46 118- 121 1.75-2.90 (4H, m), 2.35 (3H, s), 2.95-3.40 (3H, m), 3.55 (3H, s), 3.68 (2H, broad d, J = 12.6 Hz), 6.9-8.0 (16H, m), 8.70 (1H, broad s) 2023-1 100 57-64 1.8-3.0 (4H, m), 2.38 (3H, s), 3.1-4.0 (5H, m), 3.59 (3H, s), 6.70-8, 0 (15H, m)

Example IB Tablets containing mg of active ingredient are prepared as follows:

HU 210 001 Β

tablet agent 10 mg Corn starch \ 55 mg Crystalline cellulose 35 mg Polyvinylpyrrolidone (10% aqueous ol dat) 5 mg Carboxymethyl cellulose calcium 10 mg Magnesium stearate 4 mg talc 1 mg Altogether: 120 mg

The active ingredient, corn starch and crystalline cellulose were passed through a sieve with a mesh size of 0.177 mm and mixed well. The mixed powder is granulated with polyvinylpyrrolidone solution and passed through a 1.00 mm sieve. The resulting granules were dried at 50-60 ° C and re-passed through a 0.177 mm mesh screen. Carboxymethylcellulose calcium, magnesium stearate and talc, passed through a sieve having a mesh size of 0.177 mm, are added to the granulate. The resulting mixture was homogeneously mixed and converted into 120 mg tablets on a tabletting machine.

Example 2B

Tablets containing 200 mg of active ingredient are prepared as follows:

tablet

Active ingredient: 200 mg

Corn starch 50 mg

Crystalline cellulose 42 mg

Silica 7 mg

Magnesium stearate 1 mg

Total: 300 mg

The components were passed through a 0.177 mm mesh sieve and thoroughly mixed. The resulting powder mixture was melt-pressurized and converted to tablets of 300 mg.

Example 3B

Tablets containing 100 mg of active ingredient and the following components are prepared:

capsule

Active ingredient: 100 mg

Corn starch 40 mg

Lactose 5 mg

Magnesium stearate 5 mg

Total: 150 mg

The components are mixed, passed through a sieve having a mesh size of 0.177 mm and mixed thoroughly. The resulting powder mixture is filled into capsules at 150 mg.

Example 4B An injectable formulation of mg of active ingredient is prepared as follows:

vial

Active ingredient 5 mg

Mannit 50 mg

Before use, the components are dissolved in 1 ml distilled water for injection and added.

Example 5B: An injectable formulation of mg of active ingredient is filled into ampoules of:

ampoule

Active ingredient 50 mg

Sodium chloride 18 mg

Required amount of distilled water for injection Total: 2 ml

Example 6B

A patch containing 17.5 mg of active ingredient is prepared as follows.

parts by weight of poly (ammonium acrylate) are dissolved in 60 parts by weight of water. 2 parts by weight of glycerol diglycidyl ether are dissolved in 10 parts by weight of water with heating. In addition, 10 parts by weight of polyethylene glycol (degree of polymerization 400), 10 parts by weight of water and 1 part by weight of the active ingredient are mixed until dissolved. While stirring, a solution of the aqueous glycerol diglycidyl ether and the active ingredient is added to the aqueous poly (ammonium acrylate) solution, then polyethylene glycol and water are added and mixed well. The resulting hydrogel solution is applied to a flexible plastic film in an amount such that the active ingredient is 0.5 mg per cm ² . Cover the surface with detachable paper and cut it into 35 cm ² pieces.

Example 7B A patch containing mg of active ingredient was prepared by the following procedure.

An aqueous sol is prepared from the following components: 100 parts polysodium acrylate, 100 parts glycerol, 150 parts water, 0.2 parts triepoxypropyl isocyanurate, 100 parts ethanol, 25 parts isopropyl myristate, 25 parts propylene glycol and 15 parts by weight of active ingredient. The resulting sol is applied to a non-woven fabric surface of a composite film of a nonwoven rayon fabric and a polyethylene film at a thickness of 100 µm to form an adhesive layer containing the active ingredient. The proportion of release components (isopropyl myristate and propylene glycol) in the layer is about 30% by weight. The adhesive layer was cured at 25 ° C for 24 hours and a removable film was placed on the surface of the adhesive layer. The formed multilayer film is then cut into 35 cm ³ pieces.

The biological activity of the compounds of formula (I) on neuronal cells was tested in vitro. As test cells, the mouse neuroblastoma neuro2a cell line (Dainippon Pharmaceutical Co., Ltd.) NS20Y, and the like. cells cultured for the examination of neuronal cells were used. Said nerve cells were grown at 37 ° C in the presence of 5% carbon dioxide gas in an incubator and cultured with the compounds of the present invention for a defined period of time. The results show that the compounds of the present invention exhibit growth-promoting activity, neurite-forming activity, and clot-promoting activity on nerve cells, which are significantly higher than control or identical,

EN 210 001 Β or higher than the activity of the control compound isoxanin (Japanese Patent Application Publication No. 28 548/1984).

The biological activity of the compounds of the present invention on the rat PC-12 paraganglioma cell line was also tested. When NGF is added to PC-12 cells, neurites proliferate. It has been found that when the compounds of the present invention are administered concurrently, the binding of NGF to PC-12 cells and the uptake of NGF into cells is enhanced and also increased by neurite proliferation.

When the effect of the compounds of the present invention on the binding of NGF to rabbit cervical ganglion was investigated, it was found that the compounds enhanced NGF binding.

Rats with crushed sciatic nerve were modeled for peripheral nerve disorders, and the effect of the compounds of the present invention was investigated. It has been shown that the compounds of the present invention promote the restoration of the distance between the two fingers and the restoration of the soleus muscle to normal weight.

Rat and mouse models of CNS disorders were prepared and tested for pharmacological activity of the compounds of the present invention. Namely, very small amounts of rat brain black dopamine cells

By injection of 6-hydroxydopamine it was chemically degraded to induce motility insecurity. Two weeks later, embryonic brain dopamine cells were implanted on the injured side of rat rat tail nucleus and attempts were made to repair locomotor disorder. Specifically, starting from the day of implantation, the compounds of the invention were administered intraperitoneally every day for two weeks, and the compounds of the invention were administered intraperitoneally every two weeks, and the effects of the compounds of the invention on improving motility and for the growth of transplanted cells. It has now been found that the compounds of the present invention promote the improvement of motor dysfunction.

Mercury poisoning has been used to induce nerve damage in rats and mice and to investigate the activity of the compounds of this invention. It has been found that the compounds of the present invention promote the healing and restoration of chemically-induced disorder and promote the restoration and improvement of learning and memory.

The compounds of the present invention are thus useful in the treatment of various neurological disorders in mammals, such as peripheral and central nervous disorders, and in the improvement of learning and memory.

Such nerve disorders include various types of nerve disorders, including, for example, various nerve disorders associated with motor, sensory or objective bending retardation, and alcohol- or drug-induced, diabetic and metabolic or idiopathic peripheral nerve disorders, including accidental, inflammatory, or immunological. More specifically, for example, facial nerve palsy, sciatic nerve palsy, spinal muscular atrophy, muscle eating disorder, weight loss muscle, multiple sclerosis, muscular wasting sclerosis lateral, acute polycentric encephalitis, Alzheimer's disease, Guillan-Barre's syndrome, syndrome, post-cranial trauma, cerebral ischemia, cerebral infarction or cerebral haemorrhage, brain and spinal cord injury, rheumatism. These examples are not limiting.

In toxicity studies, the compounds of the present invention have been shown to be only slightly toxic and have minor side effects, thus being suitable and safe as active ingredients in pharmaceutical formulations.

Experimental Example 1

The effect of the compounds of the invention on neuroblastoma cells was investigated by the following method.

Dulbecco's modified Eagle's medium (DMEM with 100 units / ml penicillin G sodium and 100 micrograms / ml streptomycin sulphate) containing 10% FCS was inoculated with neuro 2a mouse cells in a logarithmic growth phase so that the cell number should be 1000 / well and grown in 0.25 ml media per well in 5% CO2 gas at 37 ° C. An equal volume of 4% aqueous glutaraldehyde solution (0.25 mL) was then added to each well and the culture fluid was allowed to stand at room temperature for 2 hours to fix the cells. The cells were then washed with water and stained with 0.05% aqueous methylene blue. Cells containing growth neurites, i.e. cells containing at least one neurite at least twice the length of the larger cell, were counted under a microscope, and the proportion of such cells relative to all cells was calculated. One well was observed in 5 or more fields of view (at least 2% of the total surface of the well) continuously to the right and left of the signal in the center of the well, and more than 200 cells were counted. Each compound was tested at 6 different concentrations and 3 replicates were performed at each concentration. Results are expressed as mean ± standard deviation and are summarized in Table 3.

Mouse neuroblastoma NS-20Y cells were similarly cultured in a bowl coated with polyiornitine and tested for activity. The results obtained after 24 and 48 hours from the start of culture are summarized in Table 4.

HU 210 001 Β

Table 3 Effect on neuro-2a

Number of experiments Vegyü- being Number of cells with neurites of at least twice the cell diameter (%) (concentration of compound) 33 2020 4.6 ± 1.1 (0.03 mmol / L), 33.8 ± 9.6 (0.1 mmol / L), 30.5 ± 9.5 (0.3 mmol), 15.1+ 5.0 (1 mmol / l) 2028 4.0 ± 0.8 (0.03 mmol / L), 12.5 ± 1.2 (0.1 mmol / L) 2036 3.9 + 0.8 (0.03 mmol / L), 5.4 + 1.1 (0.1 mmol / L) 2044 3.7 + 0.7 (0.01 mmol / L), 10.3 + 4.7 (0.1 mmol / L) izaxo- nin 19.5 ± 3.6 (10 mmol / L) control 2.7 ± 0.6 44 2022-1 23.1 + 4.8 (0.1 mmol / L), 18.1 ± 2.8 (0.3 mmol / L), 19.8 + 2.1 (1 mmol / L) 2023-1 8.3 + 2.1 (0.1 mmol / L), 7.0 ± 0.5 (0.3 mmol / L) izaxo- nin 20.1 + 3.0 (10 mmol / L) control 3.2 ± 0.9

Table 4

Activity against NS-20Y cells

Vegyü- being Number of cells in which neurites were expressed, total cells (compound concentration) 24 hours 48 hours 2022-1 12/50 (0.2 mmol / L) 5/50 (0.4 mmol / L) 2/50 (0.1 mmol / L) control 0/50 0/50 2023-1 2/50 (0.1 mmol / L) 2/50 (0.2 mmol / L) control 0.50 0/50

Experimental Example 2

Improving Learning and Memory in Mice with Mercury-Induced Nervous Disorders 7-week-old male BalbC mice were trained in a T-shaped maze three times a week to run from a starting point to a safe area. Methyl mercuric chloride (MMC) was then administered orally at a dose of 6 mg / kg / day for 6 days. Mice in the control group received saline at a dose of 0.1 ml / 10 g / day. Starting the day after MMC administration, the compounds of the invention were administered intraperitoneally for 10 days. At day 6 (i.e., day 12 from the start of the experiment), after drug administration, the T-shaped labyrinth was restarted and the running behavior of the mice was observed. Mice that were able to experiment in the T-shaped maze on days 10 and 11 (i.e., days 21 and 22 from the start of the experiment) were counted and denominated. Mice that ran at least eight times out of 10 attempts within 5 seconds to a secure location were counted and counted. The decrease in the number of test animals was a consequence of MMC poisoning. The time (seconds) needed for the animals to run to a safe place was measured and the mean ± standard error (SE) was calculated.

The results show that the compounds of the present invention improve the learning and memory of mice and their regeneration.

Experimental Example 3

The acute toxicity of the compounds of the invention was investigated by the following method.

Male 5-week-old ddy mice, 4-6 animals per group, were used as experimental animals. The compounds were administered intraperitoneally and toxicity was evaluated 24 hours after dosing. The results are shown in Table 5.

Table 5 Acute toxicity in mice

Compound LD ₅₀ (mg / kg, ip) 2020 500-1000 2028 500-1000 2044 500-1000

The compounds of formula (I) produced by the process of the present invention promote proliferation and formation of neurons, neurite proliferation, nerve regeneration and regeneration of motor function in rats and mice, which are useful in the treatment of neurodegenerative disorders, e.g. and cure a mental illness. They are suitable for the treatment and regeneration of neurological diseases of neurons and cells with sensory and perceptual functions and autonomous functions.

The compounds of the present invention have been found to have the same or better biological activity as isaxonine and mecobalamin as shown in Experimental Examples 1 and 2 and in Examples 3-4. Table. Thus, the compounds of the present invention can be used as high activity and safe active ingredients with low toxicity.

Claims (4)

PATENT CLAIMS A process for the preparation of compounds of the formula I and their pharmacologically acceptable salts, wherein A 1 is = CH- or -N = ₂ is = CH-, -N =, or a) ₃ is = CH- or -N =, R ¹ 'is C 1-7 alkyl, ^R12 is optionally substituted by halogen egysze7 EN 210 001 és substituted phenyl, 2-furyl or 2-thienyl, provided that when A 1 is -N =, then A ₂ is a group of formula (A) and A ₃ is -N =, if A ₁ and A ₂ are = CH-, then A ₃ A = CH-, and when A ₂ is -N =, A) and A ₃ are = CHaz in that a compound of formula II is reacted with a compound of formula R ₁₂ -COC 1 - wherein R _{1b is} R ₁₂ , A _b A ₂ and A ₃ are as stated above - in the case, and the desired compound of formula (I) into a pharmaceutically acceptable salt. (Priority: 12/22/1989) 2. A process for the preparation of a pharmaceutical composition comprising the compound of formula (I) or a pharmacologically acceptable salt thereof prepared by the process of claim 1 in admixture with a carrier, diluent and / or other excipients customary in the pharmaceutical formulation. (Priority: December 22, 1989) A process for the preparation of compounds of formula I and their pharmacologically acceptable salts, wherein Ai is -CH = or -N =, ₂ is -CH =, -N = or a) ₃ is -CH = or -N =, R ¹ 'is C 1-7 alkyl, R ¹² is phenyl or 2-furyl optionally substituted by halogen, provided that when A 1 is -N =, then A ₂ is a group of formula (A), and A ₃ is -N = if A ₁ is -N =. and A ₂ is -CH =, then A ₃ is -CH =, and if A ₂ is -N =, then A ₁ is -CH =. and A ₃ is -CH =, wherein a compound of formula II is reacted with a compound of formula R ¹² -COC 1 - wherein R ¹¹ , R ¹² , A _b A ₂ and A ₃ are as defined in the scope, and optionally converting the resulting compound of formula (I) into a pharmacologically acceptable salt. (Priority: February 23, 1989) A process for the preparation of compounds of formula I and their pharmacologically acceptable salts, wherein A 1 is -CH = or -N =, ₂ is -CH =, -N = or a) ₃ is -CH = or -N =, R ¹ 'is C 1-7 alkyl, R ¹² is phenyl or 2-furyl, provided that when A 1 is -N =, then A ₂ is a group of formula (a), and A ₃ is -N =, if A, and A ₂ is -CH =. then a ₃ is -CH =, and when a ₂ is -N =, then Ai and a ₃ is -CH =, wherein (II) is reacted with a compound of formula ^I2 R -COC1 compound of formula - wherein R, R ¹² , A 1, A ₂ and A ₃ are within the scope of the invention and, if desired, converting the resulting compound of formula I into a pharmacologically acceptable salt. (Priority: December 29, 1988)