HU209594B - Process for producing pyrrolo/2,3-d/pyrimidine derivatives and pharmaceutical preparations containing them - 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

Scope of the description: 18 pages (including 6 pages)

EN 209,594 B

The present invention relates to novel pyrrolo [2,3-d] pyrimidine derivatives, their pharmacologically acceptable salts and pharmaceutical compositions for the treatment of peripheral and central nervous system diseases in animals.

Japanese Patent Application Publication No. 23 394/1971 discloses aminopyrimidines of formula (A) wherein:

A is an alkylene group of up to 16 carbon atoms or an amino-substituted lower alkylene group or a C 2-5 acylamino group;

M is hydrogen, sodium, potassium, ammonium, magnesium, calcium, or an organic basic ammonium salt, n is the same as the M value, which has important therapeutic activity, especially as an anti-melancholic and psychoanaleptic agent in the field of psychosis.

According to Japanese Patent Application No. 22 044/1976, dichloro-lower carboxylic acid salts of 2-isopropylaminopyrimidine, such as 2-isopropylaminopyrimidine dichloroacetate, can be used as therapeutic agents for neurological disease.

According to Japanese Patent Application No. 100 477/1977, 2-isopropylaminopyrimidine phosphate can be used as a therapeutic agent of neurological diseases.

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

Japanese Patent Application Publication No. 122 768/1980 discloses hydroxyl derivatives of 2-isopropylaminopyrimidine of formula B, wherein A ⁴ , A ⁵ and A ⁶ are hydrogen or hydroxy, and at least one of them is a hydroxyl group that is a nerve regeneration and treatment of myodistrophy.

Japanese Patent Application No. 145,670/1980 discloses 2-isopropylaminohalo-pyrimidine derivatives of formula C wherein A ₄ ', A ₅ ' and A / are hydrogen or halogen and at least one of them represents a halogen atom. that can be used to treat various neurological diseases and myodistrophy.

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

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

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

U.S. Patent No. 26,880/1981 discloses the preparation of 2-isopropylaminopyrimidine by reacting bis (isopropyl guanidine) sulfate with 1,1,3,3-tetraethoxypropane.

Japanese Patent Application Publication No. 90 013/1981 discloses substituted pyrimidine derivatives useful as therapeutic agents for myodistrophy, myopathy, muscle stiffness and / or neuromuscular transmission, or for their acceptable therapeutic salts or metabolites. However, as an active ingredient, only the various salts, such as orthophosphate of 2-isopropylaminopyrimidine, are disclosed.

Japanese Patent Application 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 a herbicide in rice fields and on highland farms.

WO 87/04928 discloses certain 2-piperazinopyrimidine derivatives and pharmacologically acceptable salts thereof as therapeutic agents for neurological diseases.

The present invention relates to a process for the preparation of novel pyrrolo [2,3-d] pyrimidine derivatives and their pharmacologically acceptable salts, as well as to pharmaceutical compositions containing them for the treatment of neurological diseases and spinal nerves, and for the regeneration of nerve cells, peripheral nervous disorders, cerebral spinal injuries, \ t and can be used in diseases of the central nervous system that are different from psychosis and in which the abnormalities of the operating system or the abnormalities of the metabolic system of chemical transmitters can be considered as primary factors and for the treatment of cerebral diseases in which learning and memory are repaired and regenerated, and preparations have minor side effects such as liver damage.

The compounds of the present invention are represented by the formula (I) wherein R ²² is C 1 -C 7 alkyl;

A is a group of formula (t) or (u) wherein

R ²¹ is methylsulfonyl, triphenylmethyl, a or b, 0 or 1

R ²³ is halogen, hydrogen or C 1-7 alkyl;

R ²⁴ is hydrogen, C 1-7 alkyl, phenyl, 4-halophenyl or pyridyl, with the proviso that R ²³ and R ²⁴ cannot simultaneously represent hydrogen.

1-7 carbon atoms in R ²³ and R ²⁴ group is replaced are independently a linear or branched, preferably 1-4 carbon atoms. Examples include methyl, ethyl, η-propyl, isopropyl, η-butyl, sec-butyl and isobutyl.

Occurring ²⁴ wherein R is 4-halophenyl group such as 4-fluorophenyl, 4-chlorophenyl or 4-bromophenyl radical.

EN 209 594

In formula (I), R ²² is C 1-7 alkyl, represented by R ²³ . 1 can be 0 or 1.

Examples of compounds of formula I include the following compounds:

Compound 3100, Compound 3104, Hydrochloride of Compound 3100, Compound 3108, Compound 3112 Phthalene Sulfonate Compound 3108, Compound 3124 Compound (3124), Compound (3132), Compound (3136). the hydrochloride of the compound of formula (3132), the compound of formula (3140), the compound of formula (3144), the hydrochloride of formula (3140), the compound of formula (3148), the compound of formula (3152), the polyol sulfonate of the compound of formula (3148), Compound 3172, Compound 3176, Phthalene Sulfonate 3172, Compound 3180, Compound 3184, Phthalene Sulfonate 3180, Compound 3188 Compound 3192, ptolene sulfonate of compound 3188, Compound 3196, Compound 3200, hydrochloride of Formula 3196, Compound 3300, Compound 3400 (3404) Compound 3400, ptolol sulfonate, Compound 3408, Compound 3412, Hydrochloride 3408, Compound 3416, Compound 3420, (3416) a compound of Formula I: hydrochloride such acids.

The compounds of formula (I) may be prepared as shown in Schemes A and B.

Compounds of formula (VII) may be prepared as described in J. A. C. S., 71, 2731 (1949).

In Scheme A, the compound of formula (VII) is prepared in a solvent such as acetonitrile or dimethylformamide, or in the presence of a solvent, optionally in the presence of a base, at a temperature of 20-150 ° C, preferably 20-100 ° C, in a solvent such as acetonitrile or dimethylformamide. to give compounds of formula (VIII). An inorganic base such as sodium carbonate or potassium carbonate or an organic base such as triethylamine or pyridine may be used as the basic compound. The resulting compound of formula (VIII) is then hydrolyzed in the presence of an acid or a base to give compounds of formula (IX). The hydrolysis is carried out in a solvent such as water, methanol or ethanol at a temperature of 0-150 ° C, preferably 20-100 ° C.

Compound (IX) is reacted with a pyrimidine derivative of formula (II) followed by phosphorus oxychloride and (C1-C7) alkylamine.

The other part of the compounds of formula (I) can be prepared from the compounds of formula (X) as shown in Scheme B, i.e. a compound of formula (X) with a pyrimidine derivative of formula (II) followed by phosphorus oxychloride and (1) With C7-C7 alkyl).

Compounds of formula (X) may be prepared as described for Bleistein 20 (2), 73 (if 1 = 1) or Bleistein 20 (1), 35 (if 1 = 0).

We have found that compounds of formula (I) are useful in the treatment of neurological diseases.

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

The invention also relates to a process for the preparation of pharmacological compositions comprising as active ingredient a compound of formula (I) or a pharmacologically acceptable salt thereof. Pharmaceutically acceptable salts include acid addition salts and quaternary ammonium (or amine) salts.

The pharmacologically acceptable salts of the compounds of formula (I) are salts with acids which form pharmacologically acceptable nontoxic acid addition salts, for example, containing the following anions: chloride, bromide, sulfate, bisulfite, phosphate, hydrophosphate, acetate, maleate, fumarate, succinate, lactate, tartrate, benzoate, citrate, gluconate, methanesulfonate, p-toluenesulfonate, naphthalenesulfonate or hydrates thereof, and quaternary ammonium (or amine) salts, or hydrates thereof.

The pharmaceutical compositions of the present invention may be prepared, for example, in the form of tablets, capsules, powders, granules, pills, wafers, elixirs, emulsions, solutions, syrups, suspensions, aerosols, ointments aseptic injection, molten cataplasm, tape, soft or hard gelatin capsules, suppositories and aseptically packaged in powder form. Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, sorbitol, mannitol, corn starch, crystalline cellulose, gum arabic, calcium phosphate, polyvinylpyrrolidone, tragacanth resin, gelatin, syrup, methyl cellulose, carboxymethylcellulose, methyl hydroxybenzoic acid esters, propyl hydroxybenzoic acid esters, talc, magnesium stearates, inert polymers, water and mineral oils.

Solid and liquid compositions may contain the above fillers, binders, lubricants, wetting agents, disintegrating agents, emulsifiers, suspending agents, konzer3

EN 209 594 Β dispersants, flavorings and sweeteners. The compositions of the present invention may be formulated to deliver the active ingredient rapidly, continuously, or slowly when administered to a patient.

For oral administration, the compounds of formula (I) are mixed with a carrier or diluent into a tablet, capsule, and the like. For parenteral administration, the active ingredient is dissolved in 10% aqueous glucose solution, 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 in the vial or ampoule. Formulations for parenteral administration may also be formulated as a transdermal formulation, i.e. as an ointment or cataplasm. In this case, the molten cataplasm or ribbon is preferred.

The composition of the present invention may contain 0.1 to 2000 mg, preferably 0.51000 mg, of the active ingredient per dosage unit.

The compounds of formula I are effective over a wide range of doses. The daily dosage of the compound may vary, for example, from 0.03 mg / kg to 100 mg / kg. The actual dosage of the compound to be administered will be determined by the physician, for example, depending on the type of compound to be administered, the age of the subject to be treated, the body weight and general condition, and the route of administration.

Dosage can be 1-6 times daily, usually 1 to 4 times.

The compounds of formula (I) are useful for the treatment of disorders of the peripheral nervous system and the central nervous system. If desired, it can also be administered with at least one other active agent of similar potency. Examples of such agents include gangliosides, mecobalamin and isaxonine.

The following non-limiting examples illustrate the preparation of the compounds of formula (I) and their biological activity.

PREPARATION

Preparation of Diphenylmethylpiperazine

11.2 g (98 mmol) of 1-formylpiperazine are added to chloroform diphenyl methane (49 mmol) and the solution is stirred at room temperature for 48 hours, and the mixture is extracted with water and methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography.

8.9 g (31.9 mmol) of the resulting formyl derivative are dissolved in 100 ml of ethanol and 6.6 g (64 mmol) of concentrated hydrochloric acid are added and the solution is refluxed for 1 hour. The solvent was then distilled off under reduced pressure and the residue was treated with potassium carbonate and water and extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and the solvent was distilled off under reduced pressure to give 6.8 g (55%) of the desired product. M.p .: 93-95 " C.

1 H-NMR (CDCl ₃ δ ppm): 2.33 (4H, m), 2.87 (4H, m)

4.19 (1H, s), 7.1-7.5 (10H, m).

Example 1

2- (4-Diphenylmethyl-piperazino) -5,6-dihydro-7-methyl-6-oxo- (7H) -pyrrolo [2,3-d] pyrimidine (Compound 3124).

1.9 g (7.5 mmol) of 1-diphenylmethylpiperazine and 1.7 g (7.5 mmol) of ethyl (2-methylthio-4-hydroxypyrimidin-5-yl) acetate are added 60 ml of n-amyl alcohol and the mixture was stirred at 170 ° C for 20 hours. The solvent was then evaporated under reduced pressure. To the residue was added 10 ml of phosphorus oxychloride and reacted for 2 hours at 100 ° C. After completion of the reaction, the mixture was gradually added to an aqueous solution of potassium carbonate and then extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and the solvent was evaporated under reduced pressure. The residue and a 40% methanolic solution of 15 ml of ethanol and methylamine were placed in a reactor and reacted at 140 ° C for 10 hours. The solvent was evaporated and the residue was purified by silica gel column chromatography to give 0.9 g (30% yield) of the title compound. Mp: 75-80 'C.

1 H-NMR (CDCl ₃ solution, δ ppm):

2.43 (4H, m), 3.13 (3H, s), 3.37 (2H, s) 3.80 (4H, m) 4.23 (1H, s), 7.08-7.52 (10H, m), 7.82 (1H, s).

Compounds prepared in the same manner and their physical data are summarized in Table 1 below.

Table 1

Compound number Extraction (%) Op. 1 H-NMR (CDCl ₃ solution, δ ppm) 3100 78 210-213 2.47 (4H, m), 3.17 (3H, s), 3.39 (2H, s), 3.50 (2H, s), 3.82 (4H, m), 7.28 (4H m) 7.88 (1H, s) 3108 56 82-86 0.74 (3H, d, J = 7Hz), 1.02 (3H, d, J = 7Hz), 2.38 (4H, m), 3.13 (3H, s), 3.35 (2H , s), 3.74 (4H, m), 4.10 (1H, m), 7.20 (5H, m), 7.80 (1H, s) 3132 38 80-85 2.42 (4H, m), 3] 15 (3H, s), 3.38 (2H, s), 3.81 (4H, m), 4.24 (1H, s), 6.8-7 , 5 (9H, m), 7.85 (1H, s) 3140 54 105-110 2.43 (4H, m), 3.15 (3H, s), 3.39 (2H, s), 3.82 (4H, m), 4.24 (1H, s), 7.32 (9H m) 7.88 (1H, s)

EN 209,594 B

Compound number Extraction (%) Op. 1 H-NMR (CDCl ₃ solution, δ ppm) 3148 35 - 2.29 (3H, s), 2.43 (4H, m), 3.14 (3H, s), 3.38 (2H, s), 3.80 (4H, m), 4.21 (1H , s), 6.9-7.5 (9H, m), 7.84 (1H, s) 3172 26 - 2.41 (4H, m), 3.14 (3H, s), 3.36 (2H, s), 3.80 (4H, m), 4.22 (1H, s), 7.28 (8H m) 7.84 (1H, s) 3180 12 91-94 240 (4H, m), 3.14 (3H, s), 3.37 (2H, s), 3.80 (4H, m), 4.23 (1H, s), 6.8-7.4 (8H, m), 7.83 (1H, s) 3188 44 170-175 2.50 (4H, m), 3.16 (3H, s), 3.40 (2H, s), 3.84 (4H, m), 4.44 (1H, s), 7.1-7 , 6 (8H, m), 7.84 (1H, s), 8.49 (1H, m) 3196 51 179-181 2.66 (4H, m), 3.12 (3H, s), 3.36 (2H, s), 3.76 (4H, m), 4.88 (1H, s), 7.28 (4H m) 7.70 (4H, m) 7.84 (1H, s) 3300 49 263-267 (decomp.) 2.36 (4H, m), 3.13 (3H, s), 3.36 (2H, s), 3.92 (4H, m), 7.1-7.6 (15H, m), 7 82 (1H, s) 3400 40 - 2.78 (3H, s), 3.18 (3H, s), 3.26 (4H, m), 3.40 (2H, s), 3.94 (4H, m), 7.87 (1H , s) 3408 68 - 1.1-2.0 (12H, m), 3.18 (3H, s), 3.32 (4H, m), 4.20 (2H, m), 7.84 (1H, s) 3416 61 - 1.2-2.4 (12H, m), 3.18 (3H, s), 3.3-3.8 (4H, m), 7.86 (1H, s)

Example 2

Preparation of 2- (4-Diphenylmethylpiperazino) -5,6-dihydro-7-methyl-6-oxo- (7H) -pyrrolo [2,3-d] pyrimidine hydrochloride (Compound 3128) 0.23 g (2 , 2M concentrated hydrochloric acid was added to a solution of 2- (4-diphenylmethylpiperazino) -5,6-dihydro-7-methyl-6-oxo- (7H) pyrrolo [2,3-d] pyrimidine in ethanol / methylene chloride. and the solution was stirred at room temperature for 1 hour. The solvent was then evaporated under reduced pressure. The residue was washed with ether to give the title compound (0.88 g, 90%). M.p .: 217-222 " C.

1 H-NMR (CDCl ₃ , δ ppm):

3.18 (3H, s), 3.20 (4H, m), 3.52 (2H, s), 4.40 (4H,

m), 5.20 (1H, s) 7.2-7.9 (1H, m).

Compounds prepared in the same manner and their physical data are summarized in Table 2 below.

Table 2

Compound number Extraction (%) Op. 1 H-NMR (CDCl ₃ solution, δ ppm) 3104 82 > 300 (CDCl ₃ CD ₃ OD), 3.20 (3H, s), 3.30 (4H, m), 3.48 (2H, s), 4.32 (2H, s), 4.54 (4H m), 7.52 (4H, m), 7.93 (1H, s) 3136 80 238-243 (decomposes) (CDCl ₃ CD ₃ OD), 3.20 (3H, s), 3.24 (4H, m), 3.50 (2H, s), 3.8-4.6 (4H, m), 5 10 (1H, s), 7.08.0 (10H, m) 3144 93 238-245 (decomposes) 3.20 (3H, s), 3.29 (4H, m), 3.57 (2H, s), 4.58 (4H, m), 5.26 (1H, s), 7.40 (5H m, 7.90 (5H, m) 3200 73 244-245 (decomp.) 3.12 (3H, s), 3.14 (4H, m), 3.40 (2H, s), 4.56 (4H, m), 5.45 (1H, s), 7.17-8 , 27 (9H, m) 3412 70 250-252 (decomposes) 1.2-2.2 (12H, m), 3.26 (3H, s), 3.52 (4H, m), 4.50 (2H, m), 8.00 (1H, s) 3420 70 256-257 (decomposes) 1.2-2.5 (12H, m), 3.25 (3H, s), 3.3-4.2 (4H, m), 7.99 (1H, s)

EN 209 594

Example 3

2- {4- [a. (4-Methylphenyl) benzyl] piperazino} -5,6-dihydro-7-methyl-6-oxo- (7H) -pyrrolo [2,3-d] pyrimidinep- Production of toluenesulphonate (Compound No. 3152)

A solution of p-toluenesulfonic acid (0.13 g, 0.75 mmol) in ethyl acetate in methanol was added to 0.31 (0.75 mmol) of 2- {4- [α- (4-methylphenyl) benzyl] piperazine} A solution of -5,6-dihydro-7-methyl-6-oxo- (7H) -pyrrolo [2,3-d] pyrimidine in ethyl acetate in methanol and the mixture was stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure and the residue was washed with hexane to give 0.36 g (yield: 82%) of the title compound. Mp 150-155 ° C.

1 H-NMR (CDCl ₃ solution, ppm):

2.28 (3H, s), 2.34 (3H, s), 3.00 (4H, m), 3.11 (3H,

s), 3.39 (2H, s), 4.14 (4H, m), 4.73 (1H, s), 7.0-7.8 (13H, m), 7.93 (1H, s ).

Compounds prepared in the same manner and their physical data are summarized in Table 3.

Table 3

Compound number Extraction (%) Op. 1 H-NMR (CDCl ₃ solution, δ ppm) 3112 92 - 0.86 (3H, d, J = 7Hz), 1.16 (3H, d, J = 7Hz), 2.35 (3H, s), 3.12 (7H, m), 3.39 (2H, s), 4.20 (5H, m), 7.14 (2H, d, J = 7Hz), 7.35 (5H, m), 7.76 (2H, d, J = 7Hz), 7.87 (1H, s) 3176 90 145-150 2.34 (3H, s), 2.83 (4H, m), 3.13 (3H, s), 3.43 (2H, s), 4.06 (4H, m), 4.72 (1H , s), 7.0-7.5 (10H, m), 7.66 (2H, d, J = 7Hz), 8.03 (1H, s) 3184 91 124-130 2.34 (3H, s), 2.74 (4H, m), 3.14 (3H, s), 3.43 (2H, s), 4.04 (4H, m), 4.56 (1H , s), 6.8-7.8 (12H, m), 8.01 (1H, s) 3192 100 135-140 2.35 (3H, s), 3.0-3.4 (4H, m), 3.16 (3H, s), 3.44 (2H, s), 4.18 (4H, m), 5 , 34 (1H, s), 7.0-7.8 (8H, m), 7.11 (2H, d, J = 7Hz), 7.74 (2H, d, J = 7Hz), 7.96 (1H, s), 8.60 (1H, m) 3404 70 220-223 (decomposes) (CDCl ₃ CD ₃ OD), 2.38 (3H, s), 2.86 (3H, s), 3.27 (3H, s), 3.38 (4H, m), 3.60 (2H , s), 4.03 (4H, m), 7.16 (2H, d, J = 7Hz), 7.64 (2H, d, J = 7Hz), 8.02 (1H, s)

Example 1B Tablets containing the active ingredient mg are prepared as follows:

agent tablet 10 mg Kukoricakeménytő 55 mg Crystalline cellulose 35 mg Polyvinyl pyrrolidone (10% aqueous solution) dat) 5 mg Carboxymethyl cellulose calcium 10 mg Magnesium stearate 4 mg talc 1 mg All: 120 mg The active ingredient, corn starch and crystalline

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

Silicon dioxide 7 mg

Magnesium stearate 1 mg

Total: 300 mg cellulose is passed through a 0.17 mm mesh sieve and mixed well. The mixed powder is granulated with a polyvinylpyrrolidone solution and passed through a 1.00 mm mesh sieve. The granules obtained were dried at 50-60 ° C and passed through a 1.00 mm mesh sieve. The carboxymethylcellulose calcium, magnesium stearate and talc, passed through a 0.17 mm mesh sieve, are added to the granulate. The resulting mixture was mixed homogeneously and converted to tablets of 120 mg on a tablet machine.

The components were passed through a 0.17 mm mesh sieve and thoroughly mixed. The resulting powder mixture is melted under pressure and converted to 300 mg tablets.

Example 3B

100 mg of active ingredient and tablets containing 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 were mixed, at 0.177 mm

Pass through a 20 mesh sieve and mix thoroughly. The resulting powder mixture is filled into capsules at 150 mg.

Example 4B: An injectable preparation comprising the active ingredient mg mg:

vial

Active ingredient 5 mg

Mannit 50 mg

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

Example 5B An injectable preparation of the following composition containing mg of active ingredient is filled into ampoules:

ampoule

Active ingredient 50 mg

Sodium chloride 18 mg

Total volume of injection distilled water: 2 ml

Example 6B

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

part of poly (ammonium acrylate) is dissolved in 60 parts by weight of water. 2 parts by weight of glycerol diglycidyl ether are dissolved in 10 parts of water by 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 aqueous glycerol diglycidyl ether and the active ingredient is added to the aqueous poly (ammonium acrylate) solution, and polyethylene glycol and water are added and mixed well. The resulting hydrogel solution is applied to a flexible plastic film in an amount sufficient to provide 0.5 mg of active ingredient per cm ² . Cover the surface with separating paper and cut into 35 cm ² pieces.

Example 7B A tablet containing the active ingredient mg is prepared by the following procedure.

An aqueous sol was prepared from 100 parts by weight of poly (sodium acrylate), 100 parts by weight of glycerol, 150 parts by weight of water, 0.2 parts by weight of triepoxypropyl isocyanurate, 100 parts by weight of ethanol, 25 parts by weight of isopropyl myristate, 25 parts by weight of propylene glycol and 15 parts by weight drug. The resulting stool is applied to a non-woven fabric surface of a non-woven fabric of non-woven rayon fabric and polyethylene film at a thickness of 100 μs to form an adhesive layer containing the active ingredient. The ratio of release components (isopropyl myristate and propylene glycol) in the layer is about 30% by weight. The adhesive layer is cured at 25 ° C for 24 hours and a removable film is applied to the surface of the adhesive layer. The resulting multilayer film is then cut into 35 cm ² pieces.

The biological activity of the compounds of formula (I) on neuronal cells was investigated in vitro. Mouse neuroblastoma neuro2a cell line (Dainippon Pharmaceutical Co., Ltd.) NS-20Y, etc. as test cells. cells were cultured for neuronal cell testing. Said nerve cells were grown in an incubator at 37 ° C in the presence of a 5% carbon dioxide gas and cultured for a specified period of time with the compounds of the invention. The results show that the compounds of the present invention exhibit growth-enhancing activity, neurite-forming activity, and effervescent activity on neurons, which activities are significantly higher than those of the control or the same as the control drug. isoxanine (Compound No. 28548/1984).

The biological activity of the compounds of the present invention on the rat PC-12 paraganglioma cell line was also investigated. When NGF is added to PC-12 cells, the neuritis burst. It has been found that when the compounds of the present invention are administered simultaneously, the binding of NGF to PC-12 cells and the uptake of NGF into the cells is enhanced and the neurite outgrowth is also increased.

When investigating the effect of the compounds of the present invention on binding of NGF to rabbit upper ganglion, the compounds have been found to enhance NGF binding.

Peripheral neural disorders were modeled with rats with obstructed nerve 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 foot muscle to normal mass.

In the central nervous system disorder, rat and mouse models were prepared and the pharmacological activity of the compounds of the present invention was investigated. Specifically, rat black dopamine cells were chemically deflated by injection of a very small amount of 6-hydroxy dopamine to reduce motion uncertainty. Two weeks later, embryonic brain dopamine cells were implanted on the damaged side of the rat brain wolf nunkleus and attempted to correct the motion disorder. Namely, from the day of implantation, the compounds of the present invention were administered intraperitoneally every two weeks, and the effect of the compounds of the present invention on improving motion uncertainty and growth of transplanted cells was investigated. It has been found that the compounds of the present invention promote the improvement of motion disorder.

Mercury poisoning has resulted in nerve damage in rats and mice and the activity of the compounds of this invention has been investigated. The compounds of the present invention have been found to promote chemically in7

It also helps to improve the recovery and improvement of learning and memory.

The compounds of the present invention are therefore suitable for the treatment of various neurological disorders in mammals, such as peripheral and central nervous system disorders, as well as for the improvement of learning and memory.

Such neurological disorders include various types of nerve disorders, including, for example, various nervous disorders associated with motion, sensory or objective bending deceleration, as well as alcohol or drug-induced, diabetic and metabolic or idiopathic peripheral nerve disorders, including accidental, inflammatory or immunological nerve root damage. For example, facial nerve paralysis, sciatic paralysis, spinal cord atrophy, muscle nutrition disorder, musculoskeletal weakness, multiple sclerosis, muscular sclerosis lateral, acute multi-centered encephalitis, Guillan-Barre syndrome, post-vaccination brain-spinal inflammation, SMON disease, dementia, Alzheimer's syndrome , post-cranial injury, cerebral ischemia, cerebral infarction or stroke, brain and spinal injury, rheumatism. These examples are not limiting.

In the toxicity studies, the compounds of the present invention have only been shown to be poorly toxic and have poor side effects and are therefore suitable and safe as active ingredients of pharmaceutical compositions.

Experimental Example 1

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

10% FCS modified by Dulbecco

Eagle medium (DMEM, 100 units / ml penicillin G sodium and 100 micrograms / ml streptomycin sulfate) was injected with 48 wells with neuro 2a mouse cells at a log depth of 1000 cells / well and per well Cultured on 0.25 ml medium in 5% CO 2 gas at 37 ° C. A 4% aqueous glutaraldehyde solution (0.25 mL) was then added to each well and the cultured liquid was allowed to stand for 2 hours at room temperature. It was then washed with water and stained with 0.05% aqueous methylene blue. Cells containing incremental neurites, i.e., cells containing at least one neurite having a length of at least twice the larger diameter of the cell, were counted under a microscope, and the ratio of such cells was calculated for all cells. A recess was observed in 5 or more fields of vision (at least 2% of the total surface of the recess) continuously to the right and to the left of the signal placed in the center of the recess, and more than 200 cells were counted. All compounds were tested at 6 different concentrations and 3 concentrations were performed at each concentration. The results are expressed as a standard deviation of 30 and are summarized in Table 4.

Table 4 Effect on neuro-2a

Number of experiments Compound Number of cells with neuritis of at least twice the diameter of the cell relative to total cells (%) (concentration of compound) 35 3104 2.8 ± 0.8 (0.03 mmol / l), 4.0 ± 2.2 (0.1 mmol / l), 7.2 ± 2.8 (0.3 mmol / l), 6, 1 ± 1.6 (1 mmol / l) isaxonine 16.8 ± 3.4 (3 mmol / l) control 2.6 + 3.4 (3 mmol / 1) 36 3144 20.8 ± 4.7 (0.03 mmol / l), 34.1 ± 3.8 (0.1 mmol / l), 44.5 + 9.7 (0.3 mmol / l), 32, 2 ± 1.6 (1 mmol / l) isaxonine 30.8 + 2.9 (10 mmol / 1) control 3.2 ± 1.6 37 3200 3.6 ± 0.5 (0.03 mmol / l) isaxonine 23.3 ± 2.9 (10 mmol / l) control 2.3 ± 0.6 38 3300 6.1 ± 1.2 (0.3 mmol / l), 7.8 ± 1.0 (1 mmol / l) 3200 2.7 + 0.7 (0.01 mmol / l), 2.5 ± 2.3 (0.03 mmol / l) isaxonine 19.4 + 3.1 (3 mmol / 1) control 3.2 ± 1.2 39 3136 5.8 ± 0.6 (0.01 mmol / l), 13.8 + 4.8 (0.03 mmol / l), 20.9 ± 5.3 (0.1 mmol / l), 7, 1 + 3.0 (0.3 mmol / 1) isaxonine 22.6 ± 0.5 (10 mmol / l) control 1.8 ± 1.4

EN 209,594 B

40 3128 14.2 ± 1.9 (0.1 mmol / l), 11.9 ± 4.5 (0.3 mmol / l) 3112 6.3 ± 0.4 (0.03 mmol / l), 6.2 ± 3.4 (0.1 mmol / l) 3152 11.4 ± 3.1 (0.03 mmol / l), 6.8 ± 4.2 (0.1 mmol / l) 3176 7.3 ± 3.3 (0.03 mmol / l), 23.1 ± 4.8 (0.1 mmol / l), 21.2 ± 4.9 (0.3 mmol / l), 14, 6 ± 5.0 (1 mmol / l) 3184 3.7 + 1.1 (0.003 mmol / l), 13.9 ± 2.3 (0.1 mmol / l) 18.4 ± 1.9 (0.3 mmol / l), 17.0 ± 2 , 1 (1mmol / 1) 3192 9.7 ± 1.1 (0.1 mmol / l), 3.7 + 3.2 (0.3 mmol / l) isaxonine 19.4 + 3.1 (10 mmol / 1) control 2.2 ± l, 0 41 3404 3.3 ± 1.3 (0.03 ±, 3.4 ± 1.4 80.3 mmol / l) 3412 2.9 + 0.9 (0.03 mmol / l), 20.6 ± 8.2 (0.1 mmol / l) 3420 4.2 ± 1.9 (0.03 mmol / l), 8.7 ± 2.3 (0.1 mmol / l) isaxonine 19.4 + 2.4 (10 mmol / 1) control l, 7 ± 0.9

Experimental Example 2

Therapeutic effect in rats with a crushed nerve

The therapeutic effect of the compound of the present invention was investigated in rats with a crushed nerve disorder as a peripheral nerve disorder model, in which they investigated (1) a change in the action of the rear paw with a crushed sciatic nerve as an index of the process of degeneration and regeneration of peripheral nerves.

In the experiment, seven Wistar male rats of six weeks of age were used per group. The nerve nerve is similar to the method of Yamatsu et al. [Kiomi Yamatsu, Takenori Kaneko, Akifumi Kitahara and Isao Ohkawa, Journal of Japanese Pharmacological Society, 72, 259-268 (1976)] and Hasegawa et al., Kazuo Hasegawa, Naoji Mikuni and Yutaka Sakai , Journal of Japanese Pharmacological Society, 74, 721-734 (1978)]. More specifically, the anesthetic on the left side of the thigh was detected in anesthesia (40 mg / kg, i. P.) With pentobarbital, and the occluded nerve was pressed at a special location with a vascular tube for 30 seconds. After squeezing or squeezing, the site was immediately stitched. Vincristine was then administered intraperitoneally at a dose of 100 mg / kg once a week, which inhibits peripheral nerve regeneration in a known manner.

The compounds of the present invention were administered intraperitoneally once daily from day of disintegration to day 30. The control group received only 0.9% saline.

1) Functional change in the back paw with a crushed nerve

The jerking tension, which is a transient tension with the contraction of the dominant muscles, and which occurs as the electrical stimulation or the like of the nerve nerves, is considered as a parameter of functional change in the nerves and muscles, as is the case with the distance between the finger tips to be described.

Thus, 30 days later, treatment with chloral hydrate (400 mg / kg, i. p.) was measured by sensing the rat tension by Kern et al. Neurosci. Methods, 19, 259 (1987)]. More specifically, the hair was shaved from the hind paws of rats and embedded with Cardiocream (product of K. K. K.). Then, electrodes are connected to the back paw skin with crocodile clip. The cathode is attached to the bottom of the butt, and the anode is 1 cm rearward from the cathode and 1 cm from the back. The rat was fastened to its back and the hind paw to be measured was fixed vertically. A finger of about 20 cm in length has one end of the silk pattern, tied to the third finger of the hind paw to be measured, and the other end to a tension converter. The isotonic contractions of the third digitus flexus muscle were recorded with a polygraph. Electric stimulation was performed at 100 V for 1 msec continuously with 2 Hz rectangular waves. The static tension was 15-30 g and 10 stimuli were repeated three times at 15 second intervals. The contraction force was expressed as tension (g). The percent recovery (%, left / right) of the contraction force of the two paws with the crushed seat nerve was calculated. The results are summarized in Table 5 below.

Table 5 Tension tension

Compound Dose (Mg / kg) Cases number Jerking left / right (%) Day 18 Day 23 Physiological saline - 7 44.2 ± 17.6 49.8 ± 14.8 3144 30 8 54.5 ± 17.1 57.9 + 15.5

The test compounds apparently improved the regeneration of the jerky tension, which is an electrophysiological index and improved symptoms compared to the control group.

The distance between the fingers was measured because it is a good index for functionally showing the degeneration and regeneration of the nerves, and its change can be measured in time.

Hasegawa Method [Hasegawa, K., Experien9

Similarly, the distance between the first and fifth fingers in the hind paw was measured by a similar method.

The distance between the measured distance and the distance between the fingers of a normal hind paw was calculated and expressed as a percentage. The distance between the fingers of the hind paw with the crushed nerves was less than 50% of the value of a normal hind paw immediately after crushing. The regeneration of the distance between the fingers began after 12-16 days, and increased regeneration was observed in the treated group relative to the control group.

From day 17 to last measurement day (day 26).

An example is shown in Table 6.

Table 6

Therapeutic effect in rats with crushed nerve

Compound Dose (mg / kg, p. 1) Regeneration between fingers Day 18 Day 23 physiological saline 1 ml / kg 67.6 ± 16.1 76.5 20.2 + 3144 30 72.7 ± 14.0 83.8 ± 12.2

The compounds of the present invention are therefore useful as therapeutic agents for the treatment of disorders of the peripheral nerves.

Experimental Example 3

Effect on improving the motion uncertainty due to damage to brain cells in rats by transplanting embryonic brain cells Left nigal dopaminergic neurons of the brain of the 4-week-old female Wistar rats weighing 100 g were injected with very small amounts of 6-hydroxy dopamine. The rats spontaneously rotated in the opposite direction to the injured side for a few days, but no abnormal movement was observed. Following the addition of 5 mg / kg methamphetamine to rats with damaged nigral dopaminergic cells i. p., turned to the injured side.

Two weeks after the drug was destroyed, the trunk of the crustal body containing dopamine cells (i.e. the substantia nigrate and the abdominal tagment) was cut from the brain in a 14-17 day old rat embryo, finely shredded and treated with trypsin. The extracted tissue was then incubated at 37 ° C for 30 minutes and the tissue was suspended. A 5 µΐ suspension was introduced into the wounded nucleos of the injured site (a total of 10 µΐ, approximately 10 ⁵ cells) at two sites.

Compound 168 of the present invention was 156 mg / kg i. p. doses were administered for 4 days after transplantation and for 10 days at 50 mg / kg i. p. dose for 10 days. day.

Compound 3144 was administered at 135 mg / kg and then at 45 mg / kg in the same manner.

The rotational movement induced by methamphetamine was investigated 2 weeks before, 1 week, 2 (or 3), 4.6 and 8 weeks after transplantation and drug administration. The number of rotations after the methamphetamine dosing was counted in the first minute at 10 minute intervals, and all rotations of the 6 calculations were averaged to give an average rotation number. The results are shown in Table 7.

The results show that the test compounds are useful as therapeutic agents for improving central nervous system disorders.

Table 7

Compound -1 week 3 weeks 4 weeks 6 weeks 8 weeks 3144 14.1 + 4.7 7.7 ± 4.7 4.0 ± 5.6 0.2 ± 2.2 1.1 ± 4.1 Physiological saline 16.7 + 9.1 11.2 + 9.6 5.3 ± 8.3 2.8 ± 5.4 2.2 ± 6.0

Experimental Example 4

Improving and regenerating the learning and memory of mice with mercury-induced neural disorder 7-week-old male BalbC mice were taught 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 orally administered at 6 mg / kg / day for 6 days. Mice of the control group received saline at a dose of 0.1 ml / 10 g / day. From the day after MMC administration, the compounds of the invention were administered intraperitoneally for 10 days. After administration of the drug on day 6 (i.e., 12 days after the start of the experiment), the learning of the T-shaped labyrinth was restarted and the running behavior of the mice was observed. Mice that were able to experiment with the T-shaped maze on day 10 and day 11 (i.e. day 21 and day 22 from the start of the experiment) were counted. We counted mice that ran to a safe place at least eight times out of 10 attempts within 5 seconds and took them to the counter. The decrease in the number of test animals was a consequence of MMC poisoning. The time (seconds) that the animals needed to run to a safe place and the mean ± standard error (SE) was measured.

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

Experimental Example 5

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

As experimental animals, male mice of 5 weeks ddy strain were used, 4-6 animals per group. The compounds were administered intraperitoneally and the toxicity was administered

24 hours after dosing. The results are shown in Table 8.

Table 8 Acute mouse toxicity

Compound LD ₅₀ (mg / kg, ip) 3144.3176, 3184 and 3420 500-1000

Compounds of Formula I of the present invention promote neuronal proliferation and formation, neurite proliferation, nerve regeneration, and regeneration of motion functions in rats and mice with neurodegenerative disorders, and are suitable for disorders of the nervous system such as peripheral nerves and central nerves. and to cure mental illness. They are suitable for the healing and regeneration of neurological diseases of neurons and cells with sensory and sensory function and autonomous function.

It has been found that the compounds of the present invention have the same or better biological activity than isaxonine as shown in Experimental Example 1 and Table 4. The compounds of the present invention are therefore highly active and safe, with low toxicity.

Claims (2)

PATENT CLAIMS A process for the preparation of a compound of formula (I) or a pharmacologically acceptable salt thereof, wherein: R ²² is C 1 -C 7 alkyl, A is a group of formula (t) or (u) wherein 1 is 0 or 1, R ²¹ is methylsulfonyl, triphenylmethyl or a group of formula (b) wherein R ²³ is halogen or hydrogen or C 17 alkyl; R ²⁴ is hydrogen, (C 1 -C 7) -alkyl, phenyl, 4-halophenyl or 2-pyridyl, provided that R ²³ and R ²⁴ are not hydrogen at the same time, provided that one (IX) or reacting a compound of formula (X) with a pyrimidine derivative of formula (II) followed by phosphorus oxychloride and a (C 1 -C 7) alkylamine and, if desired, the resulting compound of formula (I) into a pharmacologically acceptable salt converted. A process for the preparation of a pharmaceutical composition comprising the compound of the formula (I) obtained by the process of claim 1, wherein R ²² and A are the pharmaceutically acceptable carriers, diluents and pharmaceutically acceptable salts thereof as defined in claim 1. / or mixed with other excipients to form a pharmaceutical composition.