JPH08175992A - Agent for promoting production of nerve growth factor and condensed ring type oxazole compound - Google Patents

Abstract

PURPOSE: To obtain a nerve growth factor production promoting agent containing a specific condensed ring type oxazole compound as an active ingredient and useful for the therapy of spinal marrow damages, peripheral nerve damages, diabetic nerve disorders, amyotrophic lateral sclerosis, etc. CONSTITUTION: This agent contains a condensed ring type oxazole compound of formula (R<1> is H, 1-6C alkyl, aryl; the ring A is benzene, naphthalene or carbazole which may be substituted) as an active ingredient. The compound is obtained by condensing an o-quinone compound of formula III [R<4> is H, (substituted) 1-6C alkyl], derived from a carbazole compound of formula II (R<2> , R<3> are H, halogen, 1-6C alkyl), with an amino acid of formula: H2 NC(R<1> ) HCO2 H (R<1> is H, 1-6C alkyl) in the presence of a quaternary ammonium salt in an inactive solvent preferably at -78 to 200 deg.C. The agent is preferably administered at a dose of 1-1000mg/day for an adult in one to three portions a day.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to nerve growth factor (hereinafter referred to as N
A condensed ring type oxazole compound having a production promoting action (denoted by GF) and its use.

[0002]

2. Description of the Related Art NGF not only differentiates nerve cells in vitro to promote neurite generation and maintains nerve cells,
In animal experiments, it has been known that administration of NGF into the brain enhances memory or learning ability and also prevents death of neurons due to cerebral ischemia (J. Neurosc
i., 6, 2155 (1986). Brain Res., 293, 305 (1985), Sc
ience, 235, 214 (1986), Proc. Natl. USA, 63, 9231
(1986). Annals of Neurology, 120 275 (1986).).

In many cases of Alzheimer-type senile dementia, many cholinergic neurons of the Meinert nucleus, which is a nerve cell that controls memory and thought, die and are lost. Many researchers have revealed that NGF is essential for cell survival and differentiation (Pharmacia, 22, 147 (198).
6). Geriatric Psychiatry, 3, 751 (1986).). In fact, in a report by Lance-Olson et al. (Symposium for treatment of Alzheimer's disease in 1991), direct injection of NGF taken from a rat into the brain of a patient with Alzheimer's disease causing dementia improved the condition. Has been confirmed.
In the striatum of the brain of patients with Huntington's chorea, GA
It is known that the loss of cholinergic neurons is remarkable along with the loss of BA-acting neurons, and that NGF also acts on the endogenous cholinergic neurons of the striatum (Science,
234, 1341 (1986).).

Further, NGF acts as a trophic factor for not only central nerves but also peripheral sensory and sympathetic nervous systems, and is an essential factor for nerve regeneration. That is, it is considered that it can be used for the treatment of spinal cord injury, peripheral nerve injury, diabetic neuropathy, amyotrophic lateral sclerosis and the like.

However, NGF has a molecular weight of 10,000.
Since it is a protein exceeding 0 and NGF itself is used as a therapeutic agent, pharmacological and pharmacological restrictions are large. From these viewpoints, as a substantial and effective replacement therapy for NGF, it is important to search for a low molecular weight compound having the ability to induce the production and secretion of NGF in a specific tissue. Examples of the compound having such an action include catechol derivatives (Furukawa.Y., J. Bio).
l. Chem. , 1986, 261, 6039. JP-A-63-83020, JP-A-63-156751, JP-A-2-53767, JP-A-2-104568, JP-A-2-
149561, JP-A-3-99046, JP-A-3-83
921, JP-A-3-86853, JP-A-5-3264
6), quinones (JP-A-3-81218, JP-A-4-3)
30010) and the like are known, but the action is not sufficient, and a compound having higher activity is desired.

[0006]

An object of the present invention is to provide a substance having an action of promoting NGF production which is useful in vivo.

[0007]

Means for Solving the Problems As a result of earnest research, the inventors of the present invention have found that certain condensed ring type oxazole compounds have an excellent NGF production promoting action in vivo and completed the present invention.

That is, the present invention provides the following general formula (1)

[0009]

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(In the formula, R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or an aryl group, and the ring A represents an optionally substituted benzene ring, naphthalene ring or carbazole ring. The present invention provides a nerve growth factor production-promoting agent comprising a condensed ring-type oxazole compound represented by the formula (4) as an active ingredient.

The present invention also relates to the following general formula (2)

[0012]

[Chemical 4]

(In the formula, R ¹ represents a hydrogen atom or a C ₁ -C ₆ alkyl group, R ² and R ³ may be the same or different, and a hydrogen atom, a halogen atom or C ₁ -C ₆ In which R ⁴ represents a hydrogen atom or an optionally substituted C _{1 to} C ₆ alkyl group).

In the above general formula (1), ring A represents an optionally substituted benzene ring, naphthalene ring or carbazole ring, and the substituent thereof is a halogen atom or a C ₁ -C ₆ alkoxy group. A group or a C ₁ -C ₆ alkyl group may be mentioned. The nitrogen atom of the carbazole ring is a C _{1 to} C ₆ alkyl group, (CH ₂ ) _n CON.
(R ⁵ ) ₂ , (CH ₂ ) _n CO ₂ H, (CH ₂ ) _n OR ⁵ , (C
^{_{H 2) n OCOR 5, (}} CH 2) n N (R 5) 2, (n is 1 to
4, R ⁵ represents a hydrogen atom which may be the same or different, or a C ₁ -C ₆ alkyl group. ) Is represented. ) May be substituted.

The C ₁ -C ₆ alkyl group includes a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a s-butyl group, a t-butyl group, a pentyl group, an isopentyl group, a neopentyl group and a t-group. Examples thereof include a pentyl group, a hexyl group, a cyclopentyl group and a cyclohexyl group. As the substituent, CON (R ⁵ ) ₂ , C
^{_{O 2 R 5, OR 5,}} OCOR 5, N (R 5) 2 ( representing the R ⁵ each the same or different optionally hydrogen atom even though or C ₁ -C ₆ alkyl group.), And the like . Examples of the aryl group include a phenyl group, a naphthyl group, a biphenyl group and the like.

Further, examples of the halogen atom include atoms such as chlorine, bromine and fluorine. Furthermore, C _{1 to} C ₆
Examples of the alkoxy group include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, an isobutoxy group, a tert-butoxy group, a pentyloxy group, an isopentyloxy group and a hexyloxy group.

The compound represented by the general formula (1) according to the present invention is partially commercially available or the corresponding o-
It can be produced by reacting a quinone compound with an amino acid represented by the general formula: H ₂ NC (R ¹ ) HCO ₂ H.
The o-quinone compound used as a raw material can be manufactured by the method shown below.

[0018]

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(In the formula, R ² , R ³ and R ⁴ have the same meanings as described above, and X represents a halogen atom such as a chlorine atom, a bromine atom or an iodine atom.)

That is, the compound represented by the general formula (b) obtained by reacting the carbazole compound represented by the general formula (a) with a halogen compound (R ² X) in the presence of a base is used as a nitrating agent. The o-quinone compound (e) can be produced by treatment to give the nitrated form (c), then reduction and conversion to the amino form (d), followed by direct oxidation.

In the above scheme, some of the carbazole compounds used as raw materials are commercially available or are referred to in the literature (J. C.
hem, Soc., 530 (1945); Phytochem., 773 (1965); J.
Chem. Soc., 4831 (1965); Aust. J. Chem., 2053 (196
It can be produced by the method described in 8)).

In the method for producing a condensed ring type oxazole compound of the present invention, it is preferable to carry out the condensation reaction in the presence of a quaternary ammonium salt in order to efficiently produce the desired product. Four
Examples of the primary ammonium salt include cetyl trimethyl ammonium bromide, cetyl triethyl ammonium bromide, cetyl triethyl ammonium chloride and the like. As the solvent to be used, water or alcohols such as methanol, ethanol and propanol, halogenated hydrocarbons such as dichloromethane and dichloroethane, tetrahydrofuran,
Ethers such as diisopropyl ether, ethyl acetate,
In addition to esters such as butyl acetate, ketones such as acetone and ethyl methyl ketone, aprotic polar solvents such as dimethyl sulfoxide, N, N-dimethylformamide, and the like, any solvent that does not participate in the reaction can be used. The reaction temperature is preferably about -78 to 200 ° C.

The NGF production promoter according to the present invention can be administered orally or parenterally. Examples of the dosage form include powders, granules, capsule tablets, pills, syrups, suspensions, injections and the like. These are conventional excipients (for example, starch, lactose, crystalline cellulose, magnesium aluminometasilicate, silicic acid anhydride, mannitol, etc.), binders (for example, hydroxy, depending on the patient's condition, age, and therapeutic purpose). Propylcellulose, polyvinylpyrrolidone etc.), lubricants (eg magnesium stearate, talc etc.), disintegrants (eg carboxymethylcellulose, etc.)
Carboxymethyl cellulose calcium, etc.), coating agents (eg hydroxyethyl cellulose), flavoring agents, solubilizers or solubilizing agents (eg distilled water for injection,
Physiological saline, propylene glycol, alcohol, fatty acid esters, etc., suspending agent (eg polysorbate 80)
Surfactants), pH adjusters (eg organic acids or metal salts thereof), adhesives (eg carboxyvinyl polymers, polysaccharides etc.), emulsifiers (eg surfactants),
It can be produced by using a stabilizer or the like and an ordinary production method (for example, a method defined in the 12th revised Japanese Pharmacopoeia). Furthermore, it is also possible to prepare a sustained-release preparation by a known technique. The dose of the compound according to the present invention as an NGF production promoter is 1 to 1000 when treating an adult.
mg, and it is preferable to administer this in 2 to 3 divided doses per day. This dosage can be adjusted according to the age, weight and condition of the patient.

The present invention will be described in more detail with reference to Reference Examples, Synthesis Examples and Test Examples. However, the present invention is not limited to them.

[0025]

【Example】

 Reference example 1

[0026]

[Chemical 6]

3-Amino-9-ethylcarbazole (0.16 g, 0.75 mmol) in acetone (75 m)
l) potassium nitrosodisulfonate (0.90
g, 3.36 mmol) of 1 / 6M dipotassium phosphate (19 ml) solution was added, and the mixture was stirred for 5 minutes. Brine (200 ml) was added to the reaction solution, extracted with ethyl acetate, washed with water, and dried over magnesium sulfate. The solvent was distilled off, and the residue was purified by column chromatography to give 9-ethylcarbazole-3,4-dione (0.06g). ¹ NMR (CDCl ₃ ): δ 1.51 (t, 1H),
4.28 (q, 2H), 6.26 (d, J = 10.2H
z, 1H), 7.34 (m, 3H), 7.35 (d, J
= 10.2 Hz, 1H), 8.18 (m, 1H). MS (m / e): 225 (M ⁺ ).

Synthesis Example 1

[0029]

[Chemical 7]

9-Ethylcarbazole-3,4-dione (0.145 g, 0.64 mmol) and glycine (1.
24 g, 16.5 mmol) of dichloromethane (40 m
l) In solution, cetyltrimethylammonium bromide (1.6
An aqueous solution (360 ml) prepared by dissolving 4 g, 4.5 mmol) in 1 / 6M disodium phosphate (75 ml) was added, and the mixture was stirred at room temperature for 24 hours. The reaction solution was extracted with ethyl acetate, washed with water, and dried over magnesium sulfate. The solvent was distilled off and the residue was purified by column chromatography to give 6-ethyl-1,3-oxazolo- [4,5-c].
Carbazole (0.021 g) was obtained. ¹ NMR (CDCl ₃ ): δ 1.48 (t, 3H),
4.47 (q, 2H), 7.35 (m, 1H), 7.4
5 (d, J = 8.7 Hz, 1H), 7.49 to 7.56
(M, 2H), 7.86 (d, J = 8.7Hz, 1
H), 8.20 (s, 1H), 8.33 (m, 1H). MS (m / e): 236 (M ⁺ ).

Test Example 1. NGF for rat submandibular gland
Production promoting activity Wistar male rats (8-10 weeks old, 200-250
The test compound was intraperitoneally administered to g) once or twice (once / day), and the submandibular gland was extracted 24 hours after the final administration. After weighing the submandibular gland, 20 times the amount of phosphate buffer (NaCl 8 g
/ L, KCl 0.2 g / L, Na ₂ HPO ₄ 1.15 g /
L, KH ₂ PO ₄ 0.2 g / L) and ultrasonic crusher (BR
Homogenized with CELL DISRUPTOR 185, manufactured by ANSON,
Then, centrifugation (10,000 × G, 30 minutes) was performed to obtain a supernatant. The NGF content of this supernatant was measured by the enzyme immunoassay method shown below.

[Method of measuring NGF] 96 made of polystyrene
Hole plate (MS-3496F made by Sumitomo Bakelite Co., Ltd.)
Anti-mouse beta NGF polyclonal antibody (produced by the inventors according to a conventional method using beta NGF prepared from mouse submandibular gland as an antigen, [S. Furukawa, I. Kamo, Y.
Furukawa, S. Akazawa, E. Satoyoshi, K. Itoh and
K. Hayashi, J. Neurochem., 40, 734-744 (1983)])
50 microliters of the solution (pH 8.3) was dispensed into each well and left at 37 ° C. for 4 hours. After removing the antibody not adsorbed on the microplate, each well was washed three times with a washing solution. Standard beta NGF (Toyobo) solution or
40 microliters of the above-mentioned centrifugal supernatant was dispensed into each well and allowed to stand at 4 ° C. for 18 hours, after which the standard solution or sample solution was removed. Further, each hole was washed 3 times. Beta-galactosidase-labeled anti-beta NGF monoclonal antibody (Boehringer Mannheim) solution (40 mU
50 mL / mL, pH 7.6) was dispensed into each well and left at 37 ° C. for 4 hours, then the enzyme-labeled antibody was removed and each well was washed 3 times in the same manner as above. Four
-Methylumbelliferyl-beta-D-galactoside (manufactured by Sigma) solution (20 microgram / mL, pH
7.6) into each well by 100 microliters,
After reacting for 1.5 hours at room temperature, 0.2 microliters of glycine-sodium hydroxide buffer (pH 10.3) was dispensed into each well by 100 microliters to stop the reaction.
-The fluorescence intensity of methyl umbelliferone was measured with a plate reader, and the amount of NGF was calculated from the standard curve.

The results were expressed as the magnification of the submandibular gland NGF content when the test compound was administered to the submandibular gland NGF content when no drug was administered. The results are shown in Table 1.

[0034]

[Table 1] Table 1 ─────────────────────────────────── Compound dose × number of doses NGF production promotion Magnification ─────────────────────────────────── Benzooxasol 1 mg / kg × 2 times 5.2 2-Phenylbenzazosol 1 mg / kg × 2 times 3.0 2,5-Dimethylbenzazoxazole 0.2 mg / kg × 2 times 2.0 2-methylnaphthoxasole 1 mg / kg × 1 time 3.4 Compound of Synthesis Example 1 0.15 mg / kg × 2 times 6.0 ── ──────────────────────────────────

All the compounds shown in Table 1 showed the activity of increasing the NGF content of rat submandibular gland at a low dose.

[0036]

The fused oxazole compound according to the present invention has an excellent nerve growth factor production promoting action and is useful as a nerve growth factor production promoting agent. Moreover, some fused ring type oxazole compounds are novel compounds.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical indication C07D 263/60 (C07D 498/04 209: 88 263: 32) (72) Inventor St. Kondo Kanagawa 5-16-4 Chuorinkan, Yamato-shi

Claims (2)

[Claims] 1. A compound represented by the general formula (1): (In the formula, R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group, or an aryl group, and the ring A may be substituted;
It represents a benzene ring, a naphthalene ring, or a carbazole ring. ) A nerve growth factor production promoter comprising a fused oxazole compound represented by the formula (4) as an active ingredient. 2. A compound of the general formula (2) (In the formula, R ¹ represents a hydrogen atom or a C ₁ -C ₆ alkyl group, and R ² and R ³ may be the same or different,
Represents a hydrogen atom, a halogen atom or a C ₁ -C ₆ alkyl group, and R ⁴ represents a hydrogen atom or an optionally substituted C ₁
It represents an alkyl group having -C _6. ) A condensed ring type oxazole compound represented by