JP2761048B2 - Nerve growth factor production promoter - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Purpose] (Industrial application field) The present invention relates to an excellent nerve growth factor (Nerve growth facto).
(hereinafter abbreviated as NGF)).

(Industrial application technology and conventional technology) NGF discovered by Levi-Montaleini et al. In 1954 is one of nutrients and growth factors necessary for the growth and maintenance of nerve tissue. In recent years, it has been known that the recovery of peripheral nerve damage is accelerated, and that it is effective for treating central dysfunction, particularly Alzheimer's dementia and cerebral ischemia pathological models. dimer 26,000), which poses problems in administration and safety.

It is known that catechol neurotransmitters such as adrenaline and noradrenaline and similar catechol compounds promote the production of NGF, but it is desirable that they have no action other than the intended physiological action, especially no nerve stimulating action.

(Problems to be Solved by the Invention) The inventors of the present invention have conducted intensive studies on the synthesis of a derivative having a nerve growth factor production / secretion promoting action and its pharmacological activity over many years, and as a result, have found that a known nerve growth factor production is obtained. The present inventors have found that a novel compound having a completely different structure from a secretagogue has an excellent nerve growth factor production / secretion promoting action, and completed the present invention.

〔Constitution〕

The novel nerve growth factor production enhancer of the present invention has the general formula [In the formula, X represents an oxygen atom, an imino group, a sulfur atom or a sulfonate group, and R ₁ represents a hydrogen atom or a protecting group of a hydroxyl group, an amino group or a mercapto group. R ₂ represents a cycloalkyl group, an aryl group, an aralkyl group or a heterocyclic group which may be condensed with an alkyl group or an aryl group;
Represents an integer of 1 to 2, and when m is 2, R _{2 at} the ortho position may form a condensed aromatic ring together with the ring carbon atom to which they are bonded. n represents an integer of 1 to 3, and when n is 2 or 3, X represents a combination of the same or different groups described above, but does not consist of only an oxygen atom. ] Or a pharmacologically acceptable salt thereof as an active ingredient.

As a preferable compound in the present invention, in the general formula (I), X represents an oxygen atom, an imino group, a sulfur atom or a sulfonate group.

R ₁ represents a hydrogen atom; a protecting group for forming a hydroxyl group, an amino group, or a mercapto group for a prodrug when administered to a living body; Fats such as alkylcarbonyl groups such as isovaleryl, octanoyl, lauroyl, palmitoyl, and stearoyl; halogenated alkylcarbonyl groups such as chloroacetyl, dichloroacetyl, trichloroacetyl, and trifluoroacetyl; and lower alkoxyalkylcarbonyl groups such as methoxyacetyl. Group acyl groups; arylcarbonyl groups such as benzoyl, α-naphthoyl and β-naphthoyl, 2-bromobenzoyl,
A halogenated arylcarbonyl group such as -chlorobenzoyl, a lower alkylated arylcarbonyl group such as 2,4,6-trimethylbenzoyl, 4-trioyl,
Lower alkoxylated arylcarbonyl groups such as anisoyl, nitrated arylcarbonyl groups such as 4-nitrobenzoyl and 2-nitrobenzoyl, lower alkoxycarbonylated arylcarbonyl groups such as 2- (methoxycarbonyl) benzoyl, 4-phenyl Aromatic acyl groups such as arylated arylcarbonyl groups such as benzoyl; tetrahydropyran-2-yl, 3-bromotetrahydropyran-2-yl, 4-methoxytetrahydropyran-4-yl, tetrahydrothiopyran-2-yl A tetrahydropyranyl or tetrahydrothiopyranyl group such as 4-methoxytetrahydrothiopyran-4-yl; tetrahydrofuranyl or tetrahydrothiol such as tetrahydrofuran-2-yl or tetrahydrothiofuran-2-yl Furanyl; methoxycarbonyl,
Lower alkoxycarbonyl groups such as ethoxycarbonyl, t-butoxycarbonyl and isobutoxycarbonyl, lower alkoxy substituted with halogen or tri-lower alkylsilyl groups such as 2,2,2-trichloroethoxycarbonyl and 2-trimethylsilylethoxycarbonyl An alkoxycarbonyl group such as a carbonyl group; an alkenyloxycarbonyl group such as vinyloxycarbonyl and allyloxycarbonyl;
The aryl ring is substituted with one or two lower alkoxy or nitro groups such as 4-methoxybenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl; Aralkyloxycarbonyl group; aliphatic acyloxyalkyloxycarbonyl group such as pivaloyloxymethyloxycarbonyl; esterified such as 3-carboxypropanoyl, 3-ethoxycarbonylpropanoyl and 4-carboxybutanoyl. A carboxy-substituted aliphatic acyl group;
An optionally alkylated carbamoyl group such as carbamoyl, dimethylcarbamoyl and diethylcarbamoyl; or an acylated amino acid such as glycyl, N-acetylglycyl, alanyl, N-acetylalanyl, arginyl and lysyl It represents an acyl group or the like.

Further, as an amino-protecting group, for example, N, N-dimethylaminomethylene, benzylidene, 4-methoxybenzylidene, 4-nitrobenzylidene, salicylidene,
-Substituted methylene groups forming Schiff bases such as -chlorosalicylidene, diphenylmethylene, (5-chloro-2-hydroxyphenyl) phenylmethylene.

When n is 2, di-acid acyl groups such as oxalyl, succinyl, maloyl, and glutaryl represented by R _{1 at} the ortho position can be also shown.

R ₂ is, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl,
A linear or branched alkyl group having 1 to 18 carbon atoms such as pentyl, hexyl, octyl, dodecyl and octadodecyl; a straight or branched alkyl group having 1 to 18 carbon atoms such as methoxymethyl, methoxyethyl and ethoxyethyl; Linear or branched alkoxyalkyl group; cyclopropyl,
An aryl group such as cyclobutyl, cyclohexyl, norbornyl, adamantyl and 2-indanyl, a cycloalkyl group having 3 to 10 carbon atoms which may be fused with an aryl group; an aryl group having 6 to 10 carbon atoms such as phenyl and naphthyl A lower alkoxy group such as methoxy, ethoxy, propoxy, isopropoxy; a hydroxyl group; fluorine; chlorine; bromine; a halogen atom such as iodine;
Trichloromethyl, difluoromethyl, dichloromethyl, dibromomethyl, fluoromethyl, 2,2,2-trichloroethyl, 2,2,2-trifluoroethyl, 2-bromoethyl, 2-chloroethyl, 2-fluoroethyl, 2,2
Halogenated lower alkyl groups such as dibromoethyl;
Ortho-substituted lower alkylenedioxy groups such as methylenedioxy, ethylenedioxy and propylenedioxy; nitro groups; amino groups; lower alkylamino groups such as dimethylamino and diethylamino; cyano groups; carboxy groups; Lower alkoxycarbonyl groups such as ethoxycarbonyl, propoxycarbonyl and isopropoxycarbonyl; carbamoyl groups; lower aliphatic acyl groups such as acetyl, propionyl and butyryl; aromatics such as benzoyl, p-chlorobenzoyl and p-anisoyl. Acyl group; benzyl, phenethyl,
An aralkyl group having 7 to 9 carbon atoms such as 3-phenylpropyl; or furyl, thienyl, benzo [b] thienyl, tetrahydrobenzo [b] thienyl, pyrrolyl,
Pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyranyl, pyridinyl,
One nitrogen atom, oxygen atom or sulfur atom such as pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, quinolyl, isoquinolyl, benzimidazolyl and purinyl
And represents an optionally condensed 5- to 7-membered heterocyclic group containing from 1 to 3.

When m is 2, it may represent a condensed aromatic ring such as indane, tetrahydronaphthalene, cycloheptabenzene or naphthalene formed together with the ortho-position R ₂ or the ring carbon atom to which they are bonded.

The above aryl group, aralkyl group and heterocyclic group are
The ring may have 1 to 4 substituents selected from the following, for example, methyl, ethyl,
Lower alkyl groups such as propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, hexyl, lower alkoxy groups such as methoxy, ethoxy, propoxy, isopropoxy, hydroxyl group, fluorine, chlorine, bromine, iodine Such as halogen atom, trifluoromethyl, trichloromethyl, difluoromethyl, dichloromethyl, dibromomethyl, fluoromethyl, 2,2,2-trichloroethyl, 2,2,2-trifluoroethyl, 2-bromoethyl, 2-bromoethyl Chloroethyl, 2-fluoroethyl,
A halogenated lower alkyl group such as 2,2-dibromoethyl, an ortho-substituted lower alkylenedioxy group such as methylenedioxy, ethylenedioxy and propylenedioxy, a nitro group, an amino group, dimethylamino and diethylamino; Such di-lower alkylamino group, cyano group,
Carboxy group, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, lower alkoxycarbonyl group such as isopropoxycarbonyl, carbomoyl group, acetyl, propionyl, lower aliphatic acyl group such as butyryl, benzoyl, p-chlorobenzoyl, p
-An aromatic acyl group such as anisoyl.

The compound (I) of the present invention can be a pharmacologically acceptable non-toxic salt, and such a salt is preferably an alkali metal such as a sodium salt, a potassium salt or a calcium salt or an alkaline earth salt. Salts of a class of metals; hydrofluoric acid salts,
Inorganic acid salts such as hydrohalides such as hydrochloride, hydrobromide and hydroiodide, nitrates, perchlorates, sulfates and phosphates; methanesulfonate, trifluoromethanesulfonic acid Salts, lower alkyl sulfonates such as ethane sulfonate, benzene sulfonates, aryl sulfonates such as p-toluene sulfonate, acetate, fumarate, succinate, citrate, tartaric acid Salts, organic acid salts such as oxalate and maleate, and amino acid salts such as glutamate and aspartate can be mentioned.

When the compound of the present invention contains an asymmetric carbon in R ₁ or R ₂ , stereoisomers exist, and each of them or a mixture thereof is included in the present invention.

Particularly preferred compounds represented by the above general formula (I) are those wherein X is an oxygen atom, an imino group or a sulfur atom, and R ₁ is a hydrogen atom; acetyl, benzoyl, pivaloyloxymethyloxycarbonyl, 3-carboxypropane Noil,
Examples of the compound include a protecting group that forms a prodrug such as glycyl, R ₂ represents an alkyl, cycloalkyl, or phenyl group, and n represents 2 or 3.

Compound (I) of the present invention can be obtained according to a known production method as described in Production Examples described later.

〔effect〕

(Measurement of the effect) Furukawa et al. Found that the fibroblast cell line LM cells derived from mouse connective tissue produced and secreted relatively large amounts of NGF, and that catecholamines promoted the production and secretion of NGF. (J. Biol. Chem., 261 , 6039-6047, 1
986). Therefore, NGF production of the test compound according to this report,
The presence or absence of a secretion promoting effect was examined.

For culture of LM cells, 199 medium containing 0.5% peptone was used. The LM cells were placed in a 24-well culture plate at about 5 × each well.
10 ⁴ plants were sown and cultured in a CO incubator (37 ° C., 5% CO ₂ ) until they reached confluence. After removing the culture,
Cells were washed once with 199 medium containing 0.5% bovine serum albumin (Fraction V. Sigma). The test compound was contained at a specified concentration in 199 medium containing 0.5% bovine serum albumin, and 0.5 ml
LM cells were treated. After culturing the LM cells in a CO ₂ incubator for 24 hours, the culture solution is collected and NG in the culture solution is recovered.
F was quantified.

NGF was measured by oxygen immunoassay (Korsching and Thoenen, Proc. Nat.
l. Acad. Sci. USA, 80 , 3513-3516, 1983).
An anti-mouse βNGF antibody (B
oehringer Mannheim) solution (0.3 μg / ml, pH 9.6)
Each 5 μl was dispensed and left at room temperature for 1 hour. After removing the antibody, each well was washed three times with a washing solution. 50 μl of a standard βNGF (Wako Pure Chemical) solution or a sample solution was dispensed into each well, and left at room temperature for 6 to 8 hours. After removing the standard βNGF or the sample solution and washing three times in each hole, the β-Galactosidas
eStandard anti-βNGF monoclonal antibody (Boehringer Mannhei
m) Dispense 50 μl of solution (100 mU / ml, pH 7.0) into each hole
For 15-18 hours. After removing the enzyme-labeled antibody and washing three times, Chlorophenolred-β-D-galactopy was used.
rano-side (Boehringer Mannheim) solution (1 mg / ml, pH7.
3) was dispensed at 100 μl per well. After an appropriate color development was obtained (after 2-3 hours at room temperature), the absorbance at 570 nm was measured. The amount of NGF was calculated from the standard curve, and the result was a relative value (%) to the amount of NGF secreted from the production and secretion of test compound-untreated cells.
Indicated by

Numerical values (% Control) are shown as average values of three experiments (3 wells) of a control (no compound added).

The compound (I) of the present invention has an excellent nerve growth factor production / secretion promoting action and has low toxicity, so that it has dementia,
It is useful as a therapeutic agent for cerebral ischemic injury and various nerve injuries.

Administration forms of the compound of the present invention include, for example, tablets,
Oral administration by capsules, granules, powders, syrups, and the like, and parenteral administration by injections, suppositories, and the like can be mentioned. These preparations are manufactured by a known method using additives such as excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents. The amount used varies depending on symptoms, age and the like, but 1 to 1000 mg per day can be usually administered to an adult once or several times a day. Formulation example (capsule) 4-Methylphenylenediamine 25 mg (Test Example 8 compound) Lactose 153.6 mg Corn starch 100 mg Magnesium stearate 1.4 mg Total 280 mg Mix powders of the above formulation and pass through 60 mesh sieve After that, 280 mg of this powder was placed in a No. 3 gelatin capsule to prepare a capsule.

Production Example 1 (a) 4-Ethyl-2-nitro-acetanilide 32 g of 4-ethyl-acetanilide are dissolved in 35 ml of acetic anhydride and 80 ml of acetic acid.

At 25 ° C. to 35 ° C., 23 ml of fuming nitric acid (d = 1.5) are added in 30 minutes. The reaction solution was poured into 800 ml of ice water, and the resulting precipitate was collected by filtration and recrystallized from ether-n-hexane to obtain 30 g of the title compound.

(B) Preparation of 15 g of 4-ethyl-2-amino-acetanilide Example 1 (a) A compound was dissolved in 100 ml of methanol, and 1 ml of acetic acid and 1.5 g of 10% Pd-carbon were added thereto. I do. The contact was filtered, the solvent was distilled off, and the residue was purified by silica gel column chromatography and recrystallized from ethyl acetate-n-hexane to obtain 10 g of the title compound.

Melting point: 95 to 97 ° C. [Production Example 2] A compound of Production Example 2 (2 g of 4-ethyl-o-phenylenediamine) is refluxed in 20 ml of concentrated hydrochloric acid for 2 hours. The solvent is distilled off, dissolved in 3 ml of water and activated carbon is added. Addition of ethanol to the filtrate gave 1.0 g of the title compound.

Melting point> 220 ° C. [Production Example 3] (a) 2-Nitro-4-propylphenol When 30 g of 4-propylphenol was nitrated in the same manner as in Production Example 1 (a), 20 g of the title target compound was obtained as an oil. Was done.

(B) Preparation Example 3 (a) of 15 g of 2-amino-4-propylphenol The compound was reduced in the same manner as in Preparation Example 1 (b). The product was recrystallized from methylene chloride-n-hexane to give 10 g of the title compound.

 140-141 ° C

Continuation of front page (56) References JP-A-1-151514 (JP, A) JP-A-1-238524 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 31 / 05 A61K 31/00 CA (STN) MEDLINE (STN)

Claims (1)

(57) [Claims] (1) General formula [In the formula, X represents an oxygen atom, an imino group, a sulfur atom or a sulfonate group, and R ₁ represents a hydrogen atom or a protecting group of a hydroxyl group, an amino group or a mercapto group. R ₂ represents a cycloalkyl group, an aryl group, an aralkyl group or a heterocyclic group which may be condensed with an alkyl group or an aryl group;
Represents an integer of 1 to 2, and when m is 2, R _{2 at} the ortho position may form a condensed aromatic ring together with the ring carbon atom to which they are bonded. n represents an integer of 1 to 3, and when n is 2 or 3, X represents a combination of the same or different groups described above, but does not consist of only an oxygen atom. ] A nerve growth factor production and secretion enhancer comprising a substituted benzene derivative represented by the following formula or a pharmacologically acceptable salt thereof as an active ingredient.