JPH04330010A - Agent for promoting production or secretion of nerve growth factor - Google Patents

Abstract

PURPOSE:To obtain the subject agent useful for the treatment or prevention of progression of central regressive diseases by using an o-quinone derivative as an active component. CONSTITUTION:The objective agent contains, as an active component, a compound of formula [R<1> to R<4> are H, halogen, OH, alkyl, alkoxy, acyl or A-B (A is single bond or alkylene; B is carboxyl, alkyloxycarbonyl, aryloxycarbonyl, amino, alkylamino, alkyloxycarbonylamino, etc.); adjacent two groups of R<1> to R<4> may together form an aromatic ring], e.g. 4-methyl-1,2-benzoquinone. The compound passes through the blood-brain barrier and effectively exhibits the action on promoting production or secretion of nerve growth factor.

Description

[Detailed description of the invention]

0001

[Industrial Application Field] The present invention relates to the use of nerve growth factor (Nerve growth factor).
The present invention relates to an agent for promoting the production or secretion of ve Growth Factor (hereinafter abbreviated as NGF). Specifically, the compounds included in the present invention promote the production or secretion of NGF, thereby promoting the activation of nerve cells whose function has decreased due to degeneration, or the maintenance or enhancement of the function of remaining nerves. Specifically, it is useful as an agent for preventing or treating central nervous system degenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's chorea.

0002

BACKGROUND OF THE INVENTION NGF is known to promote the differentiation and growth of sensory nerve cells and sympathetic nerve cells, and is an essential factor for maintaining the function of nerve cells in the peripheral nervous system. However, it has recently been proven that NGF exists and functions in the brain, suggesting a link with aging, especially Alzheimer's disease [The Embo Journal (EMB
O. J. ) Volume 4, pp. 1389-1393, 198
5 years].

[0003] One of the important causes of Alzheimer's disease is
This includes degenerative loss of cholinergic neurons in the basal forebrain, which are important for memory and learning. Acetylcholinergic neurons send long projections to the neocortex and hippocampus and form a complex neural network, and NGF is deeply involved in promoting the differentiation or maintaining the function of these neurons.

For example, it has been reported that NGF has a promoting effect on choline acetyltransferase activity on septal cholinergic neurons in in vitro experiments [Developmental Brain Research (Developer)].
v. Brain Res. ), Volume 9, 45-5
2, 1983]. In addition, in animal experiments, it has been reported that administering NGF into the brains of model animals reversed memory impairment and maintained the survival of neurons [Behav.Brain Research]. Res.), Volume 17,
pp. 17-24, 1981, Nature
re), Vol. 329, pp. 65-18, 1987, The Journal of Neuroscience (J.N.
eurosci), vol. 6, p. 2155, 1986]
. In addition, there are reports that NGF content in the hippocampus is decreased in the brains of aged rats compared to young rats.
It has been suggested that NGF or NGF production or secretion promoters are effective agents for preventing or treating central nervous system degenerative diseases such as Alzheimer's disease.

Compounds included in the present invention, such as 4-methyl-1,2-benzoquinone, have been described in the literature, [Journal of Organic Chemistry (J. Org. Ch.
em. ), Vol. 36, pp. 1339-1341, 197
4], etc., the synthesis method is described. In addition, 3-methyl-1,2-benzoquinone is described in the literature [Journal of
The Chemical Society (C) (J. Chem.
Soc. (C)) 1401-1414, 1971], the synthesis method and reaction are described. However, there have been no reports regarding the production or secretion promoting effect of NGF related to the present invention.

[0006]

[Problems to be Solved by the Invention] NGF has a very large molecular weight and cannot pass through the blood-brain barrier, so there are limits to its use in the treatment of central nervous system degenerative diseases by direct administration. Therefore, the present invention provides effective NG that passes through the blood-brain barrier.
The purpose of the present invention is to develop a drug that has an F-producing or secretion-promoting effect, and can serve as a preventive or therapeutic agent for, for example, central nervous system degenerative diseases.

[0007]

[Means for Solving the Problems] As a result, the present inventors have discovered that o-quinone compounds have a strong NGF production or secretion promoting effect, and have completed the present invention.

That is, the present invention is based on the formula [1]

[Formula 3] [In the formula, R1, R2, R3, and R4 are each independently a hydrogen atom, a halogen atom, a hydroxy group, an alkyl group, an alkoxy group, an acyl group, or a formula [2]

embedded image (where A represents a single bond or an alkylene group, and B represents a carboxyl group, an alkyloxycarbonyl group, an aryloxycarbonyl group, a substituted aryloxycarbonyl group, an amino group, an alkylamino group, an alkyloxycarbonylamino group) group, aryloxycarbonylamino group, or substituted aryloxycarbonylamino group.)
or two adjacent groups selected from R1, R2, R3, and R4 together form an aromatic ring. The present invention relates to a nerve growth factor production or secretion promoter containing an o-quinone derivative represented by the following as an active ingredient.

Examples of the alkyl group include lower alkyl groups, and more specifically, methyl, ethyl, propyl, butyl, pentyl, hexyl, 1-methylethyl, 1-methylpropyl, 2-methylpropyl group, 1-ethylpropyl group, 2-ethylpropyl group, 1-methylbutyl group, 2-methylbutyl group, 3-methylbutyl group, 1-ethylbutyl group, 2-ethylbutyl group, 3-ethylbutyl group, 1- Examples include groups having 1 to 6 carbon atoms such as methylpentyl group, 2-methylpentyl group, 3-methylpentyl group, and 4-methylpentyl group.

[0010] Examples of the alkoxy group include lower alkoxy groups, and more specifically, methoxy, ethoxy, propoxy, and butoxy groups having 1 to 1 carbon atoms.
Four groups are mentioned.

[0011] Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

Examples of the acyl group include an alkanoyl group and an aroyl group. As an alkanoyl group,
Examples include lower alkanoyl groups, and more specifically groups having 1 to 6 carbon atoms such as formyl group, acetyl group, propanoyl group, butanoyl group, pentanoyl group, and hexanoyl group. Examples of the aroyl group include a group having 11 or less carbon atoms, and more specifically a benzoyl group.

Examples of the alkylene group include lower alkylene, more specifically methylene, ethylene, trimethylene, tetramethylene, 1-methylethylene, 2-methylethylene, 1,2-dimethylethylene, 1
Examples include groups having 1 to 4 carbon atoms, such as -ethylethylene, 2-ethylethylene, 1-methyltrimethylene, 2-methyltrimethylene, and 3-methyltrimethylene.

Examples of the alkyloxycarbonyl group include lower alkyloxycarbonyl groups, more specifically methyloxycarbonyl group, ethyloxycarbonyl group, propyloxycarbonyl group, butyloxycarbonyl group, and pentyloxycarbonyl group. , hexyloxycarbonyl group, 1-methylethyloxycarbonyl group, 1-methylpropyloxycarbonyl group,
2-methylpropyloxycarbonyl group, 1-ethylpropyloxycarbonyl group, 2-ethylpropyloxycarbonyl group, 1-methylbutyloxycarbonyl group,
2-methylbutyloxycarbonyl group, 3-methylbutyloxycarbonyl group, 1-ethylbutyloxycarbonyl group, 2-ethylbutyloxycarbonyl group, 3-ethylbutyloxycarbonyl group, 1-methylpentyloxycarbonyl group, 2- Examples include groups having 7 or less carbon atoms, such as a methylpentyloxycarbonyl group, a 3-methylpentyloxycarbonyl group, and a 4-methylpentyloxycarbonyl group.

Examples of the arylalkyloxycarbonyl group include a group having 11 or less carbon atoms, and more specifically a benzyloxycarbonyl group. As substituted arylalkyloxycarbonyl group,
Examples include arylalkyloxycarbonyl groups substituted with alkoxy groups, more specifically 3,5-
Examples include groups such as dimethoxybenzyloxycarbonyl group and p-methoxybenzyloxycarbonyl group.

Examples of the alkylamino group include an amino group substituted with an alkyl group, specifically an amino group substituted with a lower alkyl group, and more specifically, an amino group substituted with a lower alkyl group. Examples include amino groups substituted with ~6 alkyl groups.

Examples of the alkyloxycarbonylamino group include amino groups substituted with alkyloxycarbonyl groups, and more specifically amino groups substituted with lower alkyloxycarbonyl groups.

Examples of the aryloxycarbonylamino group include an amino group substituted with an aryloxycarbonyl group, and more specifically an amino group substituted with an aryloxycarbonyl group having 11 or less carbon atoms. It will be done. Examples of the substituted aryloxycarbonylamino group include an amino group substituted with a substituted aryloxycarbonyl group, and more specifically, an amino group substituted with an aryloxycarbonyl group substituted with an alkoxy group. It will be done.

Examples of the aromatic ring formed by two adjacent groups selected from R1, R2, R3 and R4 include a benzene ring. R1, R
Specific examples of o-quinone derivatives in which two adjacent groups selected from 2, R3, and R4 come together to form an aromatic ring include 1,2-naphthoquinone, 2,3
-Naphthoquinone and the like.

The nerve growth factor production or secretion promoting agent of the present invention can be administered orally or parenterally. That is, it can be administered orally in commonly used dosage forms, such as tablets, capsules, syrups, suspensions, etc., or it can be administered in the form of solutions, emulsions, suspensions, etc. by injection. It can be administered as a drug. It can also be administered rectally in the form of suppositories. Such dosage forms can be manufactured according to conventional methods by combining the active ingredient with conventional carriers, excipients, binders, stabilizers, etc. When used in injection form, buffers, solubilizing agents,
It is also possible to add isotonic agents and the like.

[0021] The dose and frequency of administration vary depending on symptoms, age, body weight, administration form, etc., but when administered orally, the dose is usually 10 to 500 mg per day for adults, and when administered parenterally, the dose is usually 10 to 500 mg per day. ~100 mg can be administered once or in divided doses.

Manufacturing method

[Formula 5] (In the formula, R1, R2, R3, and R4 have the same meanings as above.) In an inert solvent, a catechol compound [3]
is oxidized using an oxidizing agent to obtain o-quinone derivative [1]. Examples of oxidizing agents include silver salts such as silver carbonate, silver oxide, and silver nitrate, cerium (IV) sulfate, and cerium nitrate (
IV) Cerium salts such as ammonium, iron(III) chloride
, iron salts such as potassium ferricyanide, chloranil, o-
Chloranil, 2,3-dichloro-5,6-dicyano-1
, 4-benzoquinone (DDQ), and other organic oxidizing agents. Examples of inert solvents include methanol, ethanol,
Alcohols such as 2-propanol, esters such as ethyl acetate, ketones such as acetone, ethers such as tetrahydrofuran, dioxane and diethyl ether, halogenated hydrocarbons such as dichloromethane and chloroform,
Examples include aromatics such as benzene and toluene. The reaction temperature can be conveniently selected from -70°C to the boiling point of the solvent.

[0023]

[Examples] Next, the present invention will be specifically explained with reference to experiments and examples.

Example 1 <NGF production or secretion promoting effect on mouse LM cells> Method of Furukawa et al. [Y. Furukawa et al., Journal of Biological Chemistry (J.
Biol. Chem. ) 261, 6039-60
47, 1986] with some modifications. That is,
199 medium supplemented with 0.5% Bactopeptone (Flow
LM cells were precultured in a 24-well culture plate (manufactured by Falcon, culture area per culture hole: 2.1 cm2) at approximately 1.6 x 105 cells.
Cells/culture hole are seeded and cultured at 37° C. for 3 days to reach complete confluence (approximately 5×10 5 cells/culture hole). The medium was supplemented with 0.5% bovine serum albumin (fifth fraction,
199 medium (0.5 ml/culture hole) supplemented with Armor (manufactured by Armor). The test compound is contained in the culture medium at a predetermined concentration, and the NGF concentration in the culture medium is measured 24 hours later.

[0025] The NGF concentration was measured using the enzyme immunoassay method of Furukawa et al. [S. Furukawa et al.: Journal of
Neurochemistry (J. Neurochem.)
40, 734-744, 1983) with some modifications. That is, a 96-hole plate (Falc
30 ng/well of an anti-mouse NGF monoclonal antibody (manufactured by Boehringer Mannheim) was added to the plate to adsorb the antibody to the bottom of the plate. After 1 hour, unadsorbed antibodies were washed and removed, and 20 μl/well of standard NGF (Sigma) solution (1 x 10-12 to 3 x 10-6 g/ml) or the culture medium to be measured was added, and incubated at 4°C. Leave it overnight. Unreacted NGF is washed and removed, and β-galactosidase-labeled anti-mouse NGF monoclonal antibody (manufactured by Boehringer Mannheim) is added at 6.6×10 −3 U/hole. After 3 hours, unreacted antibodies were washed and removed, and 33.3 μg
20 μl/ml of 4-methylumbelliferyl-β-galactoside was added to each well and reacted at 37° C. for 10 minutes. The reaction is stopped by adding 100 μl/hole of 0.1M glycine-NaOH (pH 10.3) aqueous solution, and the fluorescence of the 21-fold diluted solution is measured at 450 nm (excitation light 360 nm). A standard curve is drawn using the measured values of the standard NGF solution, and the NGF concentration in the culture medium is calculated from it. Thereafter, the amount of NGF per amount of cell protein was determined.

[0026] The results were expressed as a ratio of the amount of NGF in the culture medium of a control cultured in a medium not containing the test compound. The detection limit of this enzyme immunoassay is 10 pg/ml, and the control NGF concentration is usually 1.5 to 2.0 ng.
/0.5 ml/culture hole. Values are shown as the average of 5 trials using the same cell preparation.

The test results are shown in FIG.

Reference Example 1 Fetizone reagent (hereinafter referred to as Fetizon) used in the reaction
Reagent) is published in Journal of Organic Chemistry (J. Org. Chem.) Volume 36,
It can be produced according to the method described in 1974, pp. 1339-1341. Add a solution of 34 g (200 mmol) of silver nitrate in 200 ml of distilled water to 30 g of Celite (washed thoroughly with methanol containing 10% concentrated hydrochloric acid, then washed with distilled water until neutral and dried).
Stir. Add 300ml of distilled water into the mixture while stirring.
30 g of sodium carbonate decahydrate (105 mm
Slowly add the solution containing ol). Once complete addition is complete, stirring is continued for an additional 10 minutes, and the resulting yellow solid is filtered off and dried. 1 g of reagent contains approximately 1 mmol of silver carbonate.

Reference example 2

[Chemical formula 6] Add 200 ml of dichloromethane to 40 g (about 40 mmol) of Fetizon reagent and cool it from 0°C to 5°C.
4-methyl-1,2-benzenediol 1g (8.01
mmol) dissolved in 10 ml of dichloromethane is added and stirred vigorously. The insoluble matter was kept at the same temperature for 3 hours, filtered out, and the filtrate was concentrated under reduced pressure.
-0.98 g of benzoquinone (melting point 61-62°C) was obtained. 1H-NMR spectrum (CDCl3) δ: ppm
2.18 (3H, d, J=1.65 Hz), 6.2
5 (1H, s), 6.36 (1H, d, J = 1.23
Hz), 6.89 (1H, dd, J=2.31 Hz
, J=10.23 Hz)

Reference example 3

[Chemical formula 7] Add 200 ml of dichloromethane to 40 g (about 40 mmol) of Fetizon reagent, cool from 0°C to 5°C,
1 g of 3-methyl-1,2-benzenediol (8.01
mmol) dissolved in 10 ml of dichloromethane is added and stirred vigorously. Incubate at the same temperature for 4 hours, filter out insoluble matter, concentrate the filtrate under reduced pressure, and 3-methyl-1,2
-0.85 g of benzoquinone (melting point 111-113°C) was obtained. 1H-NMR spectrum (CDCl3) δ: ppm
2.00 (3H, d, J=0.99 Hz), 6.2
9 (1H, dd, J=0.66 Hz, J=10.8
9 Hz), 6.79 (1H, m), 6.98 (1H
, dd, J=6.27 Hz, J=9.90 Hz)

Reference example 4

[Chemical formula 8] Add 200 ml of dichloromethane to 33 g (about 33 mmol) of Fetizon reagent and cool it from 0°C to 5°C.
4-propyl-1,2-benzenediol 1g (6.5
Add a solution of 7 mmol) dissolved in 10 ml of dichloromethane and stir vigorously. Keep warm at the same temperature for 3 hours,
Insoluble materials were filtered off, the filtrate was concentrated under reduced pressure, and 4-propyl-1
, 0.89 g (oil) of 2-benzoquinone was obtained. 1H-NMR spectrum (CDCl3) δ: ppm
1.02 (3H, t, J=7.26 Hz), 1.6
3 (2H, sex, J=7.58 Hz), 2.
38 (2H, t, J = 7.58 Hz), 6.23 (1
H, t, J = 0.66 Hz), 6.37 (1H, d
, J=9.57 Hz), 6.91 (1H, dd, J
= 2.31 Hz, J = 10.22 Hz)

[0032]

[Brief explanation of drawings]

[Figure 1] Figure 1 shows 4-methyl-1,2-
Benzoquinone, 3-methyl-1,2-benzoquinone, 4
- When each test product was changed in a concentration-dependent manner using propyl-1,2-benzoquinone, mice L-M
FIG. 3 is a diagram in which the rate of increase in the content of NGF produced by cells was measured and the changes were expressed in a graph. Each line in the figure represents the results when using the following test products. 1 -○- 3-methyl-1,2-benzoquinone 2 -●- 4-methyl-
1,2-benzoquinone 3 -■- 3
-propyl-1,2-benzoquinone

Claims (2)

[Claims] Claim 1: General formula [1] [Formula 1] [In the formula, R1, R2, R3, and R4 are each independently a hydrogen atom, a halogen atom, a hydroxy group, an alkyl group, an alkoxy group, an acyl group, or a formula [ 2] [Formula 2] (wherein A represents a single bond or an alkylene group, and B represents a carboxyl group, an alkyloxycarbonyl group, an aryloxycarbonyl group, a substituted aryloxycarbonyl group, an amino group, an alkylamino group, an alkyloxy Represents a carbonylamino group, aryloxycarbonylamino group, or substituted aryloxycarbonylamino group.)
or two adjacent groups selected from R1, R2, R3, and R4 together form an aromatic ring. A nerve growth factor production or secretion promoter containing an o-quinone derivative represented by the following as an active ingredient. 2. The nerve growth factor production or secretion promoter according to claim 1, which is an agent for preventing or treating the progression of central nervous system degenerative diseases.